U.S. patent application number 16/446868 was filed with the patent office on 2020-02-06 for compounds.
This patent application is currently assigned to CTxT Pty Limited. The applicant listed for this patent is CTxT Pty Limited. Invention is credited to Ylva Elisabet Bergman Bozikis, Michelle Ang Camerino, Richard Charles Foitzik, Catherine Fae Hemley, Helen Rachel Lagiakos, Benjamin Joseph Morrow, Paul Anthony Stupple, Scott Raymond Walker.
Application Number | 20200039945 16/446868 |
Document ID | / |
Family ID | 63042663 |
Filed Date | 2020-02-06 |
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United States Patent
Application |
20200039945 |
Kind Code |
A1 |
Lagiakos; Helen Rachel ; et
al. |
February 6, 2020 |
Compounds
Abstract
A compound of formula (I), or a pharmaceutical salt thereof:
##STR00001##
Inventors: |
Lagiakos; Helen Rachel;
(Parkville, AU) ; Morrow; Benjamin Joseph;
(Parkville, AU) ; Foitzik; Richard Charles;
(Parkville, AU) ; Hemley; Catherine Fae;
(Parkville, AU) ; Camerino; Michelle Ang;
(Parkville, AU) ; Stupple; Paul Anthony;
(Parkville, AU) ; Bozikis; Ylva Elisabet Bergman;
(Parkville, AU) ; Walker; Scott Raymond;
(Parkville, AU) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CTxT Pty Limited |
Melbourne |
|
AU |
|
|
Assignee: |
CTxT Pty Limited
Melbourne
AU
|
Family ID: |
63042663 |
Appl. No.: |
16/446868 |
Filed: |
June 20, 2019 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 413/10 20130101;
C07D 417/12 20130101; A61P 35/00 20180101; C07D 413/12 20130101;
C07D 261/20 20130101; C07D 413/04 20130101 |
International
Class: |
C07D 261/20 20060101
C07D261/20; C07D 413/12 20060101 C07D413/12; C07D 417/12 20060101
C07D417/12; C07D 413/10 20060101 C07D413/10 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 20, 2018 |
GB |
1810092.5 |
Claims
1. A compound of formula (I), or a pharmaceutically acceptable salt
thereof: ##STR00383## wherein: R.sup.1, R.sup.2, R.sup.3 and
R.sup.4 are independently selected from the group consisting of:
(i) H; (ii) C.sub.1-3 alkyl, optionally substituted by: hydroxy,
C.sub.1-2 alkoxy, optionally substituted by one or more fluoro
groups, NH.sub.2, phenyl, C.sub.5-6 heteroaryl, C.sub.1-4 alkyl
carbamoyl, acylamido, or one or more fluoro groups; (iii) C.sub.1-3
alkoxy, optionally substituted by C.sub.3-6 cycloalkyl or by one or
more fluoro groups; (iv) C.sub.3-6 cycloalkyl; (v) halo; (vi)
COR.sup.C, where R.sup.C is selected from NR.sup.N1R.sup.N2, where
R.sup.N1 and R.sup.N2 are independently selected from H and methyl;
(vii) cyano, NH.sub.2, or NO.sub.2; and (viii) phenyl or C.sub.5-6
heteroaryl, optionally substituted by methyl, cyano, hydroxy or
methoxy; Ar is selected from the group consisting of: (a)
5-ethyl-2-methoxyphenyl; (b) 5-CF.sub.3-2-methoxyphenyl; and (c)
2,6-dimethoxyphenyl.
2. The compound or salt according to claim 1, wherein at least one
of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is C.sub.1-3 alkyl,
optionally substituted by: hydroxy, C.sub.1-2 alkoxy, optionally
substituted by one or more fluoro groups, NH.sub.2, phenyl,
C.sub.5-6 heteroaryl, C.sub.1-4 alkyl carbamoyl, acylamido, or one
or more fluoro groups.
3. The compound or salt according to claim 1, wherein at least one
of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is C.sub.1-3 alkoxy,
optionally substituted by C.sub.3-6 cycloalkyl or one of more
fluoro groups.
4. The compound or salt according to claim 1, wherein at least one
of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is C.sub.3-6
cycloalkyl.
5. The compound or salt according to claim 1, wherein at least one
of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is halo.
6. The compound or salt according to claim 1, wherein at least one
of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is COR.sup.C, where
R.sup.C is selected from NR.sup.N1R.sup.N2, where R.sup.N1 and
R.sup.N2 are independently selected from H and methyl.
7. The compound or salt according to claim 1, wherein at least one
of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is cyano, NH.sub.2 or
NO.sub.2.
8. The compound or salt according to claim 1, wherein at least one
of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is phenyl or C.sub.5-6
heteroaryl, optionally substituted by methyl, cyano, hydroxy or
methoxy.
9. The compound or salt according to claim 1, wherein: (a) R.sup.4
is methoxy, R.sup.2 is CH.sub.2OCH.sub.3 or
CH.sub.2OCH.sub.2CH.sub.3, and R.sup.1 and R.sup.3 are H; (b)
R.sup.4 is methoxy, R.sup.2 is phenyl, optionally substituted by
methyl or methoxy, and R.sup.1 and R.sup.3 are H; (c) R.sup.4 is
methoxy, R.sup.2 is C.sub.5-6 heteroaryl, optionally substituted by
methyl; (d) R.sup.4 is methoxy and R.sup.1, R.sup.2 and R.sup.3 are
H; (e) R.sup.4 is chloro, R.sup.2 is C.sub.1-3 alkyl or bromo, and
R.sup.1 and R.sup.3 are H; (f) R.sup.4 is chloro and R.sup.1,
R.sup.2 and R.sup.4 are H; or (g) R.sup.3 is C.sub.1-3 alkyl and
R.sup.1, R.sup.2 and R.sup.4 are H.
10. The compound or salt according to claim 1, wherein Ar is
2,6-dimethoxyphenyl.
11. A method of treating cancer comprising administering to a
patient in need of treatment a compound of formula (I), or a
pharmaceutically acceptable salt thereof: ##STR00384## wherein:
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are independently selected
from the group consisting of: (i) H; (ii) C.sub.1-3 alkyl,
optionally substituted by: hydroxy, C.sub.1-2 alkoxy, optionally
substituted by one or more fluoro groups, NH.sub.2, phenyl,
C.sub.5-6 heteroaryl, C.sub.1-4 alkyl carbamoyl, acylamido, or one
or more fluoro groups; (iii) C.sub.1-3 alkoxy, optionally
substituted by C.sub.3-6 cycloalkyl or by one or more fluoro
groups; (iv) C.sub.3-6 cycloalkyl; (v) halo; (vi) COR.sup.C, where
R.sup.C is selected from NR.sup.N1R.sup.N2, where R.sup.N1 and
R.sup.N2 are independently selected from H and methyl; (vii) cyano,
NH.sub.2, or NO.sub.2; and (viii) phenyl or C.sub.5-6 heteroaryl,
optionally substituted by methyl, cyano, hydroxy or methoxy; Ar is
selected from the group consisting of: (a) 5-ethyl-2-methoxyphenyl;
(b) 5-CF.sub.3-2-methoxyphenyl; and (c) 2,6-dimethoxyphenyl.
12. The method according to claim 11, wherein at least one of
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is C.sub.1-3 alkyl,
optionally substituted by: hydroxy, C.sub.1-2 alkoxy, optionally
substituted by one or more fluoro groups, NH.sub.2, phenyl,
C.sub.5-6 heteroaryl, C.sub.1-4 alkyl carbamoyl, acylamido, or one
or more fluoro groups.
13. The method according to claim 11, wherein at least one of
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is C.sub.1-3 alkoxy,
optionally substituted by C.sub.3-6 cycloalkyl or one of more
fluoro groups.
14. The method according to claim 11, wherein at least one of
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is C.sub.3-6 cycloalkyl.
15. The method according to claim 11, wherein at least one of
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is halo.
16. The method according to claim 11, wherein at least one of
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is COR.sup.C, where R.sup.C
is selected from NR.sup.N1R.sup.N2, where R.sup.N1 and R.sup.N2 are
independently selected from H and methyl.
17. The method according to claim 11, wherein at least one of
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is cyano, NH.sub.2 or
NO.sub.2.
18. The method according to claim 11, wherein at least one of
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is phenyl or C.sub.5-6
heteroaryl, optionally substituted by methyl, cyano, hydroxy or
methoxy.
19. The method according to claim 11, wherein: (a) R.sup.4 is
methoxy, R.sup.2 is CH.sub.2OCH.sub.3 or CH.sub.2OCH.sub.2CH.sub.3,
and R.sup.1 and R.sup.3 are H; (b) R.sup.4 is methoxy, R.sup.2 is
phenyl, optionally substituted by methyl or methoxy, and R.sup.1
and R.sup.3 are H; (c) R.sup.4 is methoxy, R.sup.2 is C.sub.5-6
heteroaryl, optionally substituted by methyl; (d) R.sup.4 is
methoxy and R.sup.1, R.sup.2 and R.sup.3 are H; (e) R.sup.4 is
chloro, R.sup.2 is C.sub.1-3 alkyl or bromo, and R.sup.1 and
R.sup.3 are H; (f) R.sup.4 is chloro and R.sup.1, R.sup.2 and
R.sup.4 are H; or (g) R.sup.3 is C.sub.1-3 alkyl and R.sup.1,
R.sup.2 and R.sup.4 are H.
20. The method according to claim 11, wherein Ar is
2,6-dimethoxyphenyl.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application claims the benefit of GB Patent Application
No. 1810092.5 filed Jul. 20, 2018, which is incorporated by
reference herein in its entirety.
[0002] This application incorporates by reference the contents of a
16.2 kb text file created on Oct. 16, 2019 and named
"16446868sequencelisting.txt" which is the sequence listing for
this application.
FIELD OF THE INVENTION
[0003] The present invention relates to compounds which act as
Lysine Acetyl Transferase (KAT) inhibitors of the MYST family.
BACKGROUND TO THE INVENTION
[0004] The MYST family is the largest family of KATs and is named
after the founding members in yeast and mammals: MOZ, Ybf2/Sas3,
Sas2 and TIP60 (Dekker 2014). MYST proteins mediate many biological
functions including gene regulation, DNA repair, cell-cycle
regulation and development (Avvakumov 2007; Voss 2009). The KAT
proteins of the MYST family play key roles in post-translational
modification of histones and thus have a profound effect on
chromatin structure in the eukaryotic nucleus (Avvakumov 2007). The
family currently comprises five mammalian KATs: TIP60 (KAT5;
HTATIP; MIM 601409), MOZ (KAT6A; MIM 601408; MYST3), MORF (KAT6b;
QKF; MYST4), HBO (KAT8; HBO1; MYST2) and MOF (KAT8; MYST1) (Voss
2009). These five members of the MYST family are present in humans
and malfunction of MYST proteins is known to be associated with
cancer (Avvakumov 2007). The most frequently used names for members
of the MYST family are:
TABLE-US-00001 Common MYST Systematic name name name MOF MYST1 KAT8
HBO MYST2 KAT7 MOZ MYST3 KAT6A MORF MYST4 KAT6B TIP60 KAT5
[0005] MYST Functional Domains
[0006] MYST proteins function in multisubunit protein complexes
including adaptors such as ING proteins that mediate DNA binding
(Avvakumov 2007). For instance, TIP60 is affiliated to the NuA4
multiprotein complex (which embraces more than 16 members) (Zhang
2017). However, there have also been some reports of a
helix-turn-helix DNA-binding motif within the structure of the MOZ
protein itself (Holbert 2007), which suggests the capacity to bind
directly to DNA.
[0007] The acetyltransferase activity of MYST proteins is effected
by the MYST domain (the catalytic domain). The MYST domain contains
an acetyl-coenzyme A binding motif, which is structurally conserved
with other HATs, and an unusual C.sub.2HC-type zinc finger (Voss
2009). The highly conserved MYST domain, including the acetyl-CoA
binding motif and zinc finger, is considered to be the defining
feature of this family of enzymes (Avvakumov 2007).
[0008] Role of MYST Proteins
[0009] Acetylation of histone residues is generally associated with
transcriptional activation. However, in some instances,
transcriptional repression has also been attributed to MYST
proteins (Voss 2009). The individual members of the MYST family are
known to participate in a broad range of important biochemical
interactions:
[0010] HBO1 positively regulates initiation of DNA replication
(Avvakumov 2007; Aggarwal 2004; Doyon 2006; Iizuka 2006) via
acetylation of histone substrates, which presumably leads to a more
accessible chromatin conformation (Avvakumov 2007, Iizuka 2006).
HBO1 is also known to play a role in the pathogenesis of breast
cancer by promoting an enrichment of cancer stem-like cells (Duong
2013) and by destabilising the estrogen receptor .alpha.
(ER.alpha.) through ubiquinitiation, which proceeds via the
histone-acetylating activity of HBO1 (Iizuka 2013). HBO1 has also
been implicated in Acute myeloid leukaemia (AML) (Shi 2015).
[0011] TIP60 (KAT5) is the most studied member of the MYST family.
TIP60 plays an important role not only in the regulation of
transcription but also in the process of DNA damage repair,
particularly in DNA double-strand breaks (DSB) (Gil 2017). TIP60
can acetylate p53, ATM and c-Myc. TIP60 and MOF specifically
acetylate lysine 120 (K120) of p53 upon DNA damage (Avvakumov
2007). TIP60 has also been implicated in being important for
regulatory T-cell (Treg) biology. FOXP3 is the master regulator in
the development and function of Tregs and it has been shown that
acetylation of FOXP3 by TIP60 is essential for FOXP3 activity (Li
2007, Xiao 2014). Underscoring this, conditional TIP60 deletion in
mice leads to a scurfy-like fatal autoimmune disease, mimicking a
phenotype seen in FOXP3 knock out mice (Xiao 2014). In cancer, Treg
cells can facilitate tumour progression by suppressing adaptive
immunity against the tumour.
[0012] MOF ("males absent on the first") was originally identified
as one of the components of the dosage compensation in Drosophila,
and was classified as a member of the MYST family based on
functional studies and sequence analysis (Su 2016). The human
ortholog exhibits significant similarity to drosophila MOF;
containing an acetyl-CoA-binding site, a chromodomain (which binds
histones) and a C.sub.2HC-type zinc finger (Su 2016). MOF is a key
enzyme for acetylating histone H4K16, and MOF-containing complexes
are implicated in various essential cell functions with links to
cancer (Su 2016). Besides the global reduction of histone
acetylation, depletion of MOF in mammalian cells can result in
abnormal gene transcription, particularly causing abnormal
expression of certain tumor suppressor genes or oncogenes,
suggesting a critical role of MOF in tumorigenesis (Su 2016). For
example, KAT activity of MOF has been shown to be required to
sustain MLL-AF9 leukemia and may be important for multiple AML
subtypes (Valerio 2017).
[0013] KAT6B (Querkopf) was first identified in a mutation screen
for genes regulating the balance between proliferation and
differentiation during embryonic development (Thomas 2000). Mice
homozygous for the KAT6B mutant allele have severe defects in
cerebral cortex development resulting from a severe reduction in
both proliferation and differentiation of specifically the cortical
progenitor population during embryonic development. KAT6B is
required for the maintenance of the adult neural stem cell
population and is part of a system regulating differentiation of
stem cells into neurons (Merson 2006). KAT6B is also mutated in
rare forms of leukaemia (Vizmanos 2003).
[0014] The MOZ locus ranks as the 12th most commonly amplified
region across all cancer types (Zack 2013). MOZ is within the
8p11-p12 amplicon, which is seen at frequencies around 10-15% in
various cancers, especially breast and ovarian (Turner-Ivey 2014).
MOZ was first identified as a fusion partner of the CREB-binding
protein (CBP) during examination of a specific chromosomal
translocation in acute myeloid leukaemia (AML) (Avvakumov 2007;
Borrow 1996). MOZ KAT activity is necessary for promoting the
expression of MEIS1 and HOXa9, proteins that are typically seen
overexpressed in some lymphomas and leukaemias. Increased survival
of MOZ.sup.+/- heterozygote mice in the E.mu.-Myc transgenic model
of B-cell lymphoma is seen, where loss of a single MOZ allele leads
to a biologically relevant reduction in Meis1 and Hoxa9 levels in
pre-B-cells (Sheikh 2015).
[0015] Inhibitors of some MYSTs are known. For example, the
following Anacardic acid derivative is reported (Ghizzoni 2012) as
inhibiting TIP60 (IC.sub.50=74 .mu.M) and MOF (IC.sub.50=47
.mu.M):
##STR00002##
[0016] Other known inhibitors include (Zhang 2017):
##STR00003##
[0017] In light of the established role of KATs in general, and
MYSTs in particular, in diseases such as cancer, a need exists for
new inhibitors of these proteins.
DISCLOSURE OF THE INVENTION
[0018] The present invention provides compounds which inhibit the
activity of one or more KATs of the MYST family, i.e., TIP60,
KAT6B, MOZ, HBO1 and MOF.
[0019] A first aspect of the present invention provides a compound
of formula (I), or a pharmaceutically acceptable salt thereof, for
use in a method of therapy:
##STR00004##
wherein: R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are independently
selected from: [0020] (i) H; [0021] (ii) C.sub.1-3 alkyl,
optionally substituted by: [0022] hydroxy, [0023] C.sub.1-2 alkoxy,
optionally substituted by one or more fluoro groups, [0024]
NH.sub.2, [0025] phenyl, [0026] C.sub.5-6 heteroaryl, [0027]
C.sub.1-4 alkyl carbamoyl, [0028] acylamido, or [0029] one or more
fluoro groups; [0030] (iii) C.sub.1-3 alkoxy, optionally
substituted by C.sub.3-6 cycloalkyl or by one or more fluoro
groups; [0031] (iv) C.sub.3-6 cycloalkyl; [0032] (v) halo; [0033]
(vi) COR.sup.C, where R.sup.C is selected from NR.sup.N1R.sup.N2,
where R.sup.N1 and R.sup.N2 are independently selected from H and
methyl; [0034] (vii) cyano, NH.sub.2, or NO.sub.2; and [0035]
(viii) phenyl or C.sub.5-8 heteroaryl, optionally substituted by
methyl, cyano, hydroxy or methoxy; Ar is a phenyl, napthyl, or
C.sub.5-10 heteroaryl group, which groups are optionally
substituted by one or more groups selected from: [0036] (i)
C.sub.1-4 alkyl, optionally substituted by hydroxy, C.sub.1-2
alkoxy, NH.sub.2, C.sub.1-4alkyl carbamoyl, or by one or more
fluoro groups; [0037] (ii) C.sub.3-6 cycloalkyl; [0038] (iii)
hydroxy; cyano; NR.sup.N3R.sup.N4, where R.sup.N3 and R.sup.N4 are
independently selected from H and methyl; or acylamido; [0039] (iv)
halo; [0040] (v) C.sub.1-3 alkoxy, optionally substituted by
hydroxy, C(O)NH.sub.2, C.sub.3-6 cycloalkyl, phenyl, C.sub.5-8
heteroaryl, or by one or more fluoro groups; [0041] (vi) phenoxy,
optionally substituted by fluoro; [0042] (vii) phenyl, or C.sub.5-8
heteroaryl [0043] (viii) SF.sub.5 or SO.sub.2CH.sub.3; [0044] (ix)
--(CH.sub.2).sub.n--Y--, where Y is O or CH.sub.2, and n is 2 or 3;
or [0045] (x) C.sub.1-4 alkyl ester.
[0046] A first aspect also provides a pharmaceutical composition
comprising a compound of formula (I), or a pharmaceutically
acceptable salt thereof, as defined and a pharmaceutically
acceptable excipient.
[0047] A second aspect of the present invention provides a method
of treatment of cancer, comprising administering to a patient in
need of treatment, a compound, or a pharmaceutically acceptable
salt thereof, as defined in the first aspect of the invention or a
pharmaceutical composition of the first aspect of the invention.
The second aspect of the present invention also provides the use of
a compound, or a pharmaceutically acceptable salt thereof, as
defined in the first aspect of the invention in the manufacture of
a medicament for treating cancer, and a compound, or a
pharmaceutically acceptable salt thereof, as defined in the first
aspect of the invention or pharmaceutical composition thereof for
use in the treatment of cancer.
[0048] As described below, the compound as defined in the first
aspect may be administered simultaneously or sequentially with
radiotherapy and/or chemotherapy in the treatment of cancer.
[0049] A third aspect of the invention provides a compound of
formula (I), or a pharmaceutically acceptable salt thereof:
##STR00005##
wherein: R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are independently
selected from: [0050] (i) H; [0051] (ii) C.sub.1-3 alkyl,
optionally substituted by: [0052] hydroxy, [0053] C.sub.1-2 alkoxy,
optionally substituted by one or more fluoro groups, [0054]
NH.sub.2, [0055] phenyl, [0056] C.sub.5-6 heteroaryl, [0057]
C.sub.1-4 alkyl carbamoyl, [0058] acylamido, or [0059] one or more
fluoro groups; [0060] (iii) C.sub.1-3 alkoxy, optionally
substituted by C.sub.3-6 cycloalkyl or by one or more fluoro
groups; [0061] (iv) C.sub.3-6 cycloalkyl; [0062] (v) halo; [0063]
(vi) COR.sup.C, where R.sup.C is selected from NR.sup.N1R.sup.N2,
where R.sup.N1 and R.sup.N2 are independently selected from H and
methyl; [0064] (vii) cyano, NH.sub.2, or NO.sub.2; and [0065]
(viii) phenyl or C.sub.5-8 heteroaryl, optionally substituted by
methyl, cyano, hydroxy or methoxy; Ar is a phenyl, napthyl, or
C.sub.5-10 heteroaryl group, which groups are optionally
substituted by one or more groups selected from: [0066] (i)
C.sub.1-4 alkyl, optionally substituted by hydroxy, C.sub.1-2
alkoxy, NH.sub.2, C.sub.1-4alkyl carbamoyl, or by one or more
fluoro groups; [0067] (ii) C.sub.3-6 cycloalkyl; [0068] (iii)
hydroxy; cyano; NR.sup.N3R.sup.N4, where R.sup.N3 and R.sup.N4 are
independently selected from H and methyl; or acylamido; [0069] (iv)
halo; [0070] (v) C.sub.1-3 alkoxy, optionally substituted by
hydroxy, C(O)NH.sub.2, C.sub.3-6 cycloalkyl, phenyl, C.sub.m
heteroaryl, or by one or more fluoro groups; [0071] (vi) phenoxy,
optionally substituted by fluoro; [0072] (vii) phenyl or C.sub.5-8
heteroaryl; [0073] (viii) SF.sub.5 or SO.sub.2CH.sub.3; [0074] (ix)
--(CH.sub.2).sub.n--Y--, where Y is O or CH.sub.2, and n is 2 or 3;
or [0075] (x) C.sub.1-4 alkyl ester; with the proviso that: (a) at
least one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is not H, and
optionally that R.sup.3 is not CF.sub.3; or
(b) R.sup.4 is OMe; or
[0076] (c) R.sup.4 is Cl, and either R.sup.1, R.sup.2 and R.sup.3
are H, or R.sup.2 is C.sub.1-3 alkyl or bromo, and R.sup.1 and
R.sup.3 are H; or (d) R.sup.3 is C.sub.1-3 alkyl and R.sup.1,
R.sup.2 and R.sup.4 are H.
[0077] A fourth aspect of the present invention provides the
synthesis of compounds as defined in the first or third aspects of
the invention, as described below.
Definitions
[0078] Unless otherwise specified, the term "substituted" as used
herein, pertains to a parent group which bears one or more
substituents. The term "substituent" is used herein in the
conventional sense and refers to a chemical moiety which is
covalently attached to, or if appropriate, fused to, a parent
group. The phrase "optionally substituted" as used herein, pertains
to a parent group which may be unsubstituted or which may be
substituted.
[0079] C.sub.5-12 heteroaryl: The term "C.sub.5-12 heteroaryl" as
used herein, pertains to a monovalent moiety obtained by removing a
hydrogen atom from an aromatic structure having from 5 to 12 rings
atoms, of which from 1 to 3 are ring heteroatoms. The term
`aromatic structure` is used to denote a single ring or fused ring
systems having aromatic properties, and the term `ring heteroatom`
refers to a nitrogen, oxygen or sulphur atom.
[0080] In this context, the prefixes (e.g. C.sub.5-12, C.sub.5-6,
etc.) denote the number of atoms making up the aromatic structure,
or range of number of atoms making up the aromatic structure,
whether carbon atoms or heteroatoms.
[0081] Examples of C.sub.5-12 heteroaryl structures include, but
are not limited to, those derived from:
N.sub.1: pyrrole (azole) (C.sub.5), pyridine (azine) (C.sub.6);
pyridone (C.sub.6); indole (C.sub.9); quinoline (C.sub.10);
S.sub.1: furan (oxole) (C.sub.5); S.sub.1: thiophene (thiole)
(C.sub.5); N.sub.1O.sub.1: oxazole (C.sub.5), isoxazole (C.sub.5),
isoxazine (C.sub.6); N.sub.2O.sub.1: oxadiazole (furazan)
(C.sub.5); N.sub.1S.sub.1: thiazole (C.sub.5), isothiazole
(C.sub.5); N.sub.2S.sub.1: thiadiazole (C.sub.5) N.sub.2: imidazole
(1,3-diazole) (C.sub.5), pyrazole (1,2-diazole) (C.sub.5),
pyridazine (1,2-diazine) (C.sub.6), pyrimidine (1,3-diazine)
(C.sub.6) (e.g., cytosine, thymine, uracil), pyrazine (1,4-diazine)
(C.sub.6); benzimidazole (C.sub.9) N.sub.3: triazole (C.sub.5),
triazine (C.sub.6).
[0082] Halo: The term "halo" as used herein, refers to a group
selected from fluoro, chloro, bromo and iodo.
[0083] Cyano: The term "cyano" as used herein, refers to a group
--C.ident.N.
[0084] Hydroxy: the term "hydroxyl" as used herein, refers to a
group --OH.
[0085] Phenyl: the term "phenyl" as used herein, pertains to a
monovalent moiety obtained by removing a hydrogen atom from a
single aromatic ring structure having 6 carbon ring atoms
(--C.sub.6H.sub.5).
[0086] Phenoxy: the term "phenoxy" as used herein, pertains to a
monovalent moiety obtained by removing a hydrogen atom from the
oxygen atom of phenol (--O--C.sub.6H.sub.5).
[0087] C.sub.1-4 alkyl: The term "C.sub.1-4 alkyl" as used herein,
pertains to a monovalent moiety obtained by removing a hydrogen
atom from a carbon atom of a saturated hydrocarbon compound having
from 1 to 4 carbon atoms.
[0088] Examples of saturated alkyl groups include, but are not
limited to, methyl (C.sub.1), ethyl (C.sub.2), propyl (C.sub.3),
and butyl (C.sub.4).
[0089] Examples of saturated linear alkyl groups include, but are
not limited to, methyl (C.sub.1), ethyl (C.sub.2), n-propyl
(C.sub.3), and n-butyl (C.sub.4).
[0090] Examples of saturated branched alkyl groups include
iso-propyl (C.sub.3), iso-butyl (C.sub.4), sec-butyl (C.sub.4) and
tert-butyl (C.sub.4).
[0091] C.sub.3-6 cycloalkyl: The term "C.sub.3-6 cycloalkyl" as
used herein, pertains to a monovalent moiety obtained by removing a
hydrogen atom from a carbon atom of a saturated cyclic hydrocarbon
compound having from 3 to 6 carbon atoms. Examples of C.sub.3-6
cycloalkyl groups include, but are not limited to, cyclopropyl
(C.sub.3), cyclobutyl (C.sub.4), cyclopentyl (C.sub.5) and
cyclonexyl (C.sub.6).
[0092] C.sub.1-4 alkoxy: The term "C.sub.1-4 alkoxy" as used
herein, pertains to a monovalent moiety obtained by removing a
hydrogen atom from an oxygen atom of a saturated alcohol compound
having from 1 to 4 carbon atoms. It can be represented as
--O--C.sub.1-4 alkyl. Examples of C.sub.1-4 alkoxy groups include,
but are not limited to, methoxy (C.sub.1), ethoxy (C.sub.2),
propyloxy (C.sub.3), and butyloxy (C.sub.4).
[0093] C.sub.1-4 alkyl carbamoyl: --NHC(.dbd.O)OR wherein R is a
C.sub.1-4 alkyl group as defined above. Examples of C.sub.1-4 alkyl
carbamoyl include, but are not limited to,
--N(H)C(.dbd.O)OCH.sub.3, --N(H)C(.dbd.O)OCH.sub.2CH.sub.3, and
--N(H)C(.dbd.O)OC(CH.sub.3).sub.3.
[0094] Acylamido: --NR(C.dbd.O)R' wherein R and R' are
independently selected from H and C.sub.1-4 alkyl as defined above.
R and R' may also be --(CH.sub.2).sub.n--, where n is 3 or 4.
Examples of an acylamido group include, but are not limited to,
--N(H)C(.dbd.O)CF.sub.3, N(H)C(.dbd.O)Me, and:
##STR00006##
[0095] C.sub.1-4 alkyl ester: The term "C.sub.1-4 alkyl ester" as
used herein, pertains to a monovalent moiety obtained by removing a
hydrogen atom from an oxygen atom of a saturated carboxylic acid
compound having from 1 to 5 carbon atoms. It can be represented as
--O--C(O)--C.sub.1-4 alkyl. Examples of C.sub.1-4 alkyl ester
groups include, but are not limited to, acetoxy
(--O--C(O)--CH.sub.3), propanoyloxy (--O--C(O)--CH.sub.2CH.sub.3),
butanoyloxy (--O--C(O)--CH.sub.2CH.sub.2CH.sub.3) and pentanoyloxy
(--O--C(O)--CH.sub.2CH.sub.2CH.sub.2CH.sub.3).
[0096] Includes Other Forms
[0097] Unless otherwise specified, included in the above are the
well known ionic, salt, solvate, and protected forms of these
substituents. For example, a reference to carboxylic acid (--COOH)
also includes the anionic (carboxylate) form (--COO.sup.-), a salt
or solvate thereof, as well as conventional protected forms.
Similarly, a reference to an amino group includes the protonated
form (--N.sup.+HR.sup.1R.sup.2), a salt or solvate of the amino
group, for example, a hydrochloride salt, as well as conventional
protected forms of an amino group. Similarly, a reference to a
hydroxyl group also includes the anionic form (--O.sup.-), a salt
or solvate thereof, as well as conventional protected forms.
[0098] Salts
[0099] It may be convenient or desirable to prepare, purify, and/or
handle a corresponding salt of the active compound, for example, a
pharmaceutically-acceptable salt. Examples of pharmaceutically
acceptable salts are discussed in Berge 1977. For example, if the
compound is anionic, or has a functional group which may be anionic
(e.g. --COOH may be --COO.sup.-), then a salt may be formed with a
suitable cation. Examples of suitable inorganic cations include,
but are not limited to, alkali metal ions such as Na.sup.+ and
K.sup.+, alkaline earth cations such as Ca.sup.2+ and Mg.sup.2+,
and other cations such as Al.sup.+3. Examples of suitable organic
cations include, but are not limited to, ammonium ion (i.e.
NH.sub.4+) and substituted ammonium ions (e.g. NH.sub.3R.sup.+,
NH.sub.2R.sub.2.sup.+, NHR.sub.3.sup.+, NR.sub.4.sup.+). Examples
of some suitable substituted ammonium ions are those derived from:
ethylamine, diethylamine, dicyclohexylamine, triethylamine,
butylamine, ethylenediamine, ethanolamine, diethanolamine,
piperazine, benzylamine, phenylbenzylamine, choline, meglumine, and
tromethamine, as well as amino acids, such as lysine and arginine.
An example of a common quaternary ammonium ion is
N(CH.sub.3).sub.4.sup.+.
[0100] If the compound is cationic, or has a functional group which
may be cationic (e.g. --NH.sub.2 may be --NH.sub.3.sup.+), then a
salt may be formed with a suitable anion. Examples of suitable
inorganic anions include, but are not limited to, those derived
from the following inorganic acids: hydrochloric, hydrobromic,
hydroiodic, sulfuric, sulfurous, nitric, nitrous, phosphoric, and
phosphorous.
[0101] Examples of suitable organic anions include, but are not
limited to, those derived from the following organic acids:
2-acetyoxybenzoic, acetic, ascorbic, aspartic, benzoic,
camphorsulfonic, cinnamic, citric, edetic, ethanedisulfonic,
ethanesulfonic, fumaric, glucheptonic, gluconic, glutamic,
glycolic, hydroxymaleic, hydroxynaphthalene carboxylic, isethionic,
lactic, lactobionic, lauric, maleic, malic, methanesulfonic, mucic,
oleic, oxalic, palmitic, pamoic, pantothenic, phenylacetic,
phenylsulfonic, propionic, pyruvic, salicylic, stearic, succinic,
sulfanilic, tartaric, toluenesulfonic, trifluoroacetic acid and
valeric. Examples of suitable polymeric organic anions include, but
are not limited to, those derived from the following polymeric
acids: tannic acid, carboxymethyl cellulose.
[0102] Solvates
[0103] It may be convenient or desirable to prepare, purify, and/or
handle a corresponding solvate of the active compound. The term
"solvate" is used herein in the conventional sense to refer to a
complex of solute (e.g. active compound, salt of active compound)
and solvent. If the solvent is water, the solvate may be
conveniently referred to as a hydrate, for example, a mono-hydrate,
a di-hydrate, a tri-hydrate, etc.
[0104] Isomers
[0105] Certain compounds of the invention may exist in one or more
particular geometric, optical, enantiomeric, diasteriomeric,
epimeric, atropic, stereoisomeric, tautomeric, conformational, or
anomeric forms, including but not limited to, cis- and trans-forms;
E- and Z-forms; c-, t-, and r-forms; endo- and exo-forms; R-, S-,
and meso-forms; D- and L-forms; d- and I-forms; (+) and (-) forms;
keto-, enol-, and enolate-forms; syn- and anti-forms; synclinal-
and anticlinal-forms; .alpha.- and .beta.-forms; axial and
equatorial forms; boat-, chair-, twist-, envelope-, and
halfchair-forms; and combinations thereof, hereinafter collectively
referred to as "isomers" (or "isomeric forms").
[0106] The term "chiral" refers to molecules which have the
property of non-superimposability of the mirror image partner,
while the term "achiral" refers to molecules which are
superimposable on their mirror image partner.
[0107] The term "stereoisomers" refers to compounds which have
identical chemical constitution, but differ with regard to the
arrangement of the atoms or groups in space.
[0108] "Diastereomer" refers to a stereoisomer with two or more
centers of chirality and whose molecules are not mirror images of
one another. Diastereomers have different physical properties, e.g.
melting points, boiling points, spectral properties, and
reactivities. Mixtures of diastereomers may separate under high
resolution analytical procedures such as electrophoresis and
chromatography.
[0109] "Enantiomers" refer to two stereoisomers of a compound which
are non-superimposable mirror images of one another.
[0110] Stereochemical definitions and conventions used herein
generally follow S. P. Parker, Ed., McGraw-Hill Dictionary of
Chemical Terms (1984) McGraw-Hill Book Company, New York; and
Eliel, E. and Wlen, S., "Stereochemistry of Organic Compounds",
John Wiley & Sons, Inc., New York, 1994. The compounds of the
invention may contain asymmetric or chiral centers, and therefore
exist in different stereoisomeric forms. It is intended that all
stereoisomeric forms of the compounds of the invention, including
but not limited to, diastereomers, enantiomers and atropisomers, as
well as mixtures thereof such as racemic mixtures, form part of the
present invention. Many organic compounds exist in optically active
forms, i.e., they have the ability to rotate the plane of
plane-polarized light. In describing an optically active compound,
the prefixes D and L, or R and S, are used to denote the absolute
configuration of the molecule about its chiral center(s). The
prefixes d and I or (+) and (-) are employed to designate the sign
of rotation of plane-polarized light by the compound, with (-) or I
meaning that the compound is levorotatory. A compound prefixed with
(+) or d is dextrorotatory. For a given chemical structure, these
stereoisomers are identical except that they are mirror images of
one another. A specific stereoisomer may also be referred to as an
enantiomer, and a mixture of such isomers is often called an
enantiomeric mixture. A 50:50 mixture of enantiomers is referred to
as a racemic mixture or a racemate, which may occur where there has
been no stereoselection or stereospecificity in a chemical reaction
or process. The terms "racemic mixture" and "racemate" refer to an
equimolar mixture of two enantiomeric species, devoid of optical
activity.
[0111] In the present invention, the carbon atom to which R.sup.1
and Cy are bound may be a stereochemical centre, i.e. when R.sup.1
is not H and R.sup.1 and Cy are different. The compounds of the
present invention may be a racemic mixture, or may be in
enantiomeric excess or substantially enantiomerically pure.
[0112] Note that, except as discussed below for tautomeric forms,
specifically excluded from the term "isomers", as used herein, are
structural (or constitutional) isomers (i.e. isomers which differ
in the connections between atoms rather than merely by the position
of atoms in space). For example, a reference to a methoxy group,
--OCH.sub.3, is not to be construed as a reference to its
structural isomer, a hydroxymethyl group, --CH.sub.2OH. Similarly,
a reference to ortho-chlorophenyl is not to be construed as a
reference to its structural isomer, meta-chlorophenyl. However, a
reference to a class of structures may well include structurally
isomeric forms falling within that class (e.g. C.sub.1-7 alkyl
includes n-propyl and iso-propyl; butyl includes n-, iso-, sec-,
and tert-butyl; methoxyphenyl includes ortho-, meta-, and
para-methoxyphenyl).
[0113] The above exclusion does not pertain to tautomeric forms,
for example, keto-, enol-, and enolate-forms, as in, for example,
the following tautomeric pairs: keto/enol (illustrated below),
imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime,
thioketone/enethiol, N-nitroso/hyroxyazo, and nitro/aci-nitro.
##STR00007##
[0114] The term "tautomer" or "tautomeric form" refers to
structural isomers of different energies which are interconvertible
via a low energy barrier. For example, proton tautomers (also known
as prototropic tautomers) include interconversions via migration of
a proton, such as keto-enol and imine-enamine isomerizations.
Valence tautomers include interconversions by reorganization of
some of the bonding electrons.
[0115] Note that specifically included in the term "isomer" are
compounds with one or more isotopic substitutions. For example, H
may be in any isotopic form, including .sup.1H, .sup.2H (D), and
.sup.3H (T); C may be in any isotopic form, including .sup.12C,
.sup.13C, and .sup.14C; O may be in any isotopic form, including
.sup.16O and .sup.18O; and the like.
[0116] Examples of isotopes that can be incorporated into compounds
of the invention include isotopes of hydrogen, carbon, nitrogen,
oxygen, phosphorous, fluorine, and chlorine, such as, but not
limited to .sup.2H (deuterium, D), .sup.3H (tritium), .sup.11C,
.sup.13C, .sup.14C, .sup.15N, .sup.18F, .sup.31P, .sup.32P,
.sup.35S, .sup.36Cl, and .sup.125I. Various isotopically labeled
compounds of the present invention, for example those into which
radioactive isotopes such as 3H, 13C, and 14C are incorporated.
Such isotopically labelled compounds may be useful in metabolic
studies, reaction kinetic studies, detection or imaging techniques,
such as positron emission tomography (PET) or single-photon
emission computed tomography (SPECT) including drug or substrate
tissue distribution assays, or in radioactive treatment of
patients. Deuterium labelled or substituted therapeutic compounds
of the invention may have improved DMPK (drug metabolism and
pharmacokinetics) properties, relating to distribution, metabolism,
and excretion (ADME). Substitution with heavier isotopes such as
deuterium may afford certain therapeutic advantages resulting from
greater metabolic stability, for example increased in vivo
half-life or reduced dosage requirements. An 18F labeled compound
may be useful for PET or SPECT studies. Isotopically labeled
compounds of this invention and prodrugs thereof can generally be
prepared by carrying out the procedures disclosed in the schemes or
in the examples and preparations described below by substituting a
readily available isotopically labeled reagent for a
non-isotopically labeled reagent. Further, substitution with
heavier isotopes, particularly deuterium (i.e., 2H or D) may afford
certain therapeutic advantages resulting from greater metabolic
stability, for example increased in vivo half-life or reduced
dosage requirements or an improvement in therapeutic index. It is
understood that deuterium in this context is regarded as a
substituent. The concentration of such a heavier isotope,
specifically deuterium, may be defined by an isotopic enrichment
factor. In the compounds of this invention any atom not
specifically designated as a particular isotope is meant to
represent any stable isotope of that atom.
[0117] Unless otherwise specified, a reference to a particular
compound includes all such isomeric forms, including (wholly or
partially) racemic and other mixtures thereof. Methods for the
preparation (e.g. asymmetric synthesis) and separation (e.g.
fractional crystallisation and chromatographic means) of such
isomeric forms are either known in the art or are readily obtained
by adapting the methods taught herein, or known methods, in a known
manner.
[0118] Inhibition
[0119] The compounds of the present invention inhibit the activity
of one or more KATs of the MYST family, i.e., TIP60, KAT6B, MOZ,
HBO1 and MOF.
[0120] The inhibitory activity of the compounds of the invention is
likely to vary between the KATs of the MYST family.
[0121] The compounds of the present invention may selectively
inhibit the activity of one or more KATs of the MYST family over
other KATs of the MYST family, i.e. the inhibitory activity of the
compound may be higher for one or more of the KATs of the MYST
family over one or more of the other KATs of the MYST family.
[0122] Compounds of the present invention may (selectively) inhbit
the activity of a single KAT of the MYST family. Thus, compounds of
the present invention may inhibit the activity of TIP60, MORF, MOZ,
HBO1 or MOF.
[0123] Compounds of the present invention may inhibit the activity
of two KATs of the MYST family, for example MOZ and MORF.
[0124] Compounds of the present invention may inhibit the activity
of three KATs of the MYST family, for example MOZ, MORF and
HBO1.
[0125] Compounds of the present invention may inhibit the activity
of four KATs of the MYST family, for example MOZ, MORF, HBO1 and
TIP60.
[0126] Compounds of the present invention may inhibit the activity
of all five KATs of the MYST family, thus the compounds may inhibit
the activity of TIP60, MORF, MOZ, HBO1 and MOF.
[0127] Compounds of the present invention may, in particular,
inhibit the activity of MOZ and/or KAT6B and/or HBO1 and/or
TIP60.
[0128] Therapeutic Indications
[0129] Compounds disclosed herein may provide a therapeutic benefit
in a number of disorders, in particular, in the treatment or
prevention of cancers.
[0130] Cancer
[0131] Inhibitors of post-translational lysine acetylation mediated
by KATs of the MYST family are considered to be promising
anti-neoplastic agents and therefore may be useful therapeutic
agents, e.g. for use in the treatment of cancer. Such agents may
also be useful as therapeutic agents for the treatment of cancers
which exhibit overexpression of MYST proteins.
[0132] A "cancer" may be any form of cancer. In particular, a
cancer can comprise any one or more of the following: leukemia,
acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML),
chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML),
non-Hodgkin's lymphoma, Hodgkin's disease, prostate cancer, lung
cancer, melanoma, breast cancer, colon and rectal cancer, colon
cancer, squamous cell carcinoma and gastric cancer.
[0133] Alternatively, the cancer may comprise adrenocortical
cancer, anal cancer, bladder cancer, blood cancer, bone cancer,
brain tumor, cancer of the female genital system, cancer of the
male genital system, central nervous system lymphoma, cervical
cancer, childhood rhabdomyosarcoma, childhood sarcoma, endometrial
cancer, endometrial sarcoma, esophageal cancer, eye cancer,
gallbladder cancer, gastrointestinal tract cancer, hairy cell
leukemia, head and neck cancer, hepatocellular cancer,
hypopharyngeal cancer, Kaposi's sarcoma, kidney cancer, laryngeal
cancer, liver cancer, malignant fibrous histiocytoma, malignant
thymoma, mesothelioma, multiple myeloma, myeloma, nasal cavity and
paranasal sinus cancer, nasopharyngeal cancer, nervous system
cancer, neuroblastoma, oral cavity cancer, oropharyngeal cancer,
osteosarcoma, ovarian cancer, pancreatic cancer, parathyroid
cancer, penile cancer, pharyngeal cancer, pituitary tumor, plasma
cell neoplasm, primary CNS lymphoma, rectal cancer, respiratory
system, retinoblastoma, salivary gland cancer, skin cancer, small
intestine cancer, soft tissue sarcoma, stomach cancer, stomach
cancer, testicular cancer, thyroid cancer, urinary system cancer,
uterine sarcoma, vaginal cancer, vascular system, Waldenstrom's
macroglobulinemia and/or Wilms' tumor.
[0134] Cancers may be of a particular type. Examples of types of
cancer include lymphoma, melanoma, carcinoma (e.g. adenocarcinoma,
hepatocellular carcinoma, medullary carcinoma, papillary carcinoma,
squamous cell carcinoma), astrocytoma, glioma, medulloblastoma,
myeloma, meningioma, neuroblastoma, sarcoma (e.g. angiosarcoma,
chrondrosarcoma, osteosarcoma).
[0135] The cancer may be a MYST overexpressing cancer. The cancer
may overexpress MYST protein relative to non-cancerous tissue. In
some cases, the cancer overproduces MYST mRNA relative to
non-cancerous tissue. The overexpressed MYST protein or MYST mRNA
may be any one KATs of the MYST family, i.e. any one of TIP60,
KAT6B, MOZ, HBO1 and MOF. In some embodiments, the cancer may
overexpress more than one KATs of the MYST family, e.g. two or more
selected from the group consisting of TIP60, KAT6B, MOZ, HBO1 and
MOF. The cancer may be a cancer that evades immune recognition,
e.g. via tumor-associated Treg cells.
[0136] Alternatively or additionally, the cancer may be a
bromodomain overexpressing cancer: The cancer cell may overexpress
one or more bromodomain-containing proteins (herein referred to as
"bromodomain proteins") relative to non-cancerous tissue. It may
overproduce one or more bromodomain mRNA as compared to
non-cancerous tissue. In some cases, the level of bromodomain
protein and/or mRNA in the cell is at a level approximately
equivalent to that of a non-cancerous cell. The cancer may
overexpress one or more bromodomain proteins selected from the
group consisting of; a bromodomain protein (namely BRD2, BRD3,
BRD4, BRD7, BRD8, BRD9 and BRDT), TAF1/TAF1L, TFIID, SMARC2 (also
called BRM) and SMARC4 (also called BRG1). For example, some colon
cancers overexpress BRD8. Some acute myeloid leukemia cells
overexpress BRD4.
[0137] Treq Cells as a Cancer Target
[0138] Treg cells are immunosuppressive cells, which act to prevent
autoimmunity in the healthy mammalian immune system. However, some
cancers act to upregulate Treg activity to evade the host immune
system. Infiltration of Tregs in many tumour types correlates with
poor patient prognoses and Treg cell depletion in tumour models
demonstrates increased anti-tumour immune responses (Melero 2015).
Tumour-associated Treg suppression of the host immune system has
been reported in lung (Joshi 2015), (Tso 2012), breast (Gobert
2009; Yan 2011), prostate (Miller 2006) & pancreatic (Wang X
2016) cancers. FOXP3 is considered to be the master regulator of
Treg differentiation, development and function of Treg cells.
[0139] Several studies have demonstrated that acetylation of FOXP3
plays a critical role in the stability of the FOXP3 protein and in
regulating its ability to access DNA; and FOXP3 acetylation is
mediated by KATs (Dhuban 2017). Decreases in TI P60-mediated FOXP3
acetylation has been shown to attenuate Treg development,
suggesting a further mechanism by which the inhibition of the
acetylating activity of MYST proteins could be used to intervene in
diseases such as cancer.
[0140] Combination Therapies
[0141] The agents described herein may be useful in combination
with other anti-cancer therapies. They may act synergistically with
chemo- or radiotherapy, and/or with targeted therapies, including
but not limited to FGFR1 inhibitors and therapies targeting nuclear
hormone receptors. For example, the agents described herein may be
useful in combination with bromodomain targeted drugs including BET
inhibitors. BET inhibitors reversibly bind the bromodomains of the
BET proteins BRD2, BRD3, BRD4 and BRDT.
[0142] Inhibition of KAT proteins of the MYST family, to reduce the
extent of lysine acetylation of histones (and other nuclear
proteins described herein) will likely sensitize tumour cells to
chemo- and radiotherapy by attenuating the process of DNA damage
repair, e.g. the repair of DNA double-strand breaks (DSB), thus
increasing the frequency of chemo- and radiotherapy induced cancer
cell death. Therefore, it is likely that inhibition of KAT proteins
of the MYST family would synergize well with low dose chemo- or
radiotherapy.
[0143] Thus, in some cases, a MYST protein antagonist disclosed
herein may be administered in conjunction with a radiotherapeutic
or chemotherapeutic regime. It may be administered simultaneously
or sequentially with radio and/or chemotherapy. Suitable
chemotherapeutic agents and radiotherapy protocols will be readily
appreciable to the skilled person. In particular, the compound
described herein may be combined with low dose chemo or radio
therapy. Appropriate dosages for "low dose" chemo or radio therapy
will be readily appreciable to the skilled practitioner.
[0144] In particular, where the compounds of the present
application are used to abrogate Treg suppression, these may be
combined with with immune checkpoint inhibitors (Melero 2015, Wang
L 2016). Furthermore, where compounds of the present invention
which abrogate Treg suppression may be used in combination with
radiotherapy, to reduce the depletion of Treg function in tumours
(Persa 2015, Jeong 2016)
[0145] Methods of Treatment
[0146] The compounds of the present invention may be used in a
method of therapy. Also provided is a method of treatment,
comprising administering to a subject in need of treatment a
therapeutically-effective amount of a compound of the invention.
The term "therapeutically effective amount" is an amount sufficient
to show benefit to a patient. Such benefit may be at least
amelioration of at least one symptom. The actual amount
administered, and rate and time-course of administration, will
depend on the nature and severity of what is being treated.
Prescription of treatment, e.g. decisions on dosage, is within the
responsibility of general practitioners and other medical
doctors.
[0147] As described above, the anti-cancer treatment defined herein
may be applied as a sole therapy or may involve, in addition to the
compound of the invention, conventional surgery or radiotherapy or
chemotherapy. Such chemotherapy may include one or more of the
following categories of anti-tumour agents:--
(i) other antiproliferative/antineoplastic drugs and combinations
thereof, as used in medical oncology, such as alkylating agents
(for example cisplatin, oxaliplatin, carboplatin, cyclophosphamide,
nitrogen mustard, melphalan, chlorambucil, busulphan, temozolamide
and nitrosoureas); antimetabolites (for example gemcitabine and
antifolates such as fluoropyrimidines like 5 fluorouracil and
tegafur, raltitrexed, methotrexate, cytosine arabinoside, and
hydroxyurea); antitumour antibiotics (for example anthracyclines
like adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin,
idarubicin, mitomycin-C, dactinomycin and mithramycin); antimitotic
agents (for example vinca alkaloids like vincristine, vinblastine,
vindesine and vinorelbine and taxoids like taxol and docetaxel
(Taxotere) and polokinase inhibitors); and topoisomerase inhibitors
(for example epipodophyllotoxins like etoposide and teniposide,
amsacrine, topotecan and camptothecin); (ii) cytostatic agents such
as antioestrogens (for example tamoxifen, fulvestrant, toremifene,
raloxifene, droloxifene and iodoxyfene), antiandrogens (for example
bicalutamide, flutamide, nilutamide and cyproterone acetate), LHRH
antagonists or LHRH agonists (for example goserelin, leuprorelin
and buserelin), progestogens (for example megestrol acetate),
aromatase inhibitors (for example as anastrozole, letrozole,
vorazole and exemestane) and inhibitors of 5*-reductase such as
finasteride; (iii) anti-invasion agents (for example c-Src kinase
family inhibitors like
4-(6-chloro-2,3-methylenedioxyanilino)-7-[2-(4-methylpiperazin-1-yl)ethox-
y]-5-tetrahydropyran-4-yloxyquinazoline (AZD0530; International
Patent Application WO 01/94341),
N-(2-chloro-6-methylphenyl)-2-{6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-met-
hylpyrimidin-4-ylamino}thiazole-5-carboxamide (dasatinib,
BMS-354825; J. Med. Chem., 2004, 47, 6658-6661 and and
4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxy-7-(3-(4-methylpiperazin-
-1-yl)propoxy)quinoline-3-carbonitrile (bosutinib, SKI-606; Cancer
research (2003), 63(2), 375-81), and metalloproteinase inhibitors
like marimastat, inhibitors of urokinase plasminogen activator
receptor function or antibodies to Heparanase); (iv) inhibitors of
growth factor function: for example such inhibitors include growth
factor antibodies and growth factor receptor antibodies (for
example the anti erbB2 antibody trastuzumab [HerceptinT], the
anti-EGFR antibody panitumumab, the anti erbB1 antibody cetuximab
[Erbitux, C225] and any growth factor or growth factor receptor
antibodies disclosed by Stern 2005; such inhibitors also include
tyrosine kinase inhibitors, for example inhibitors of the epidermal
growth factor family (for example EGFR family tyrosine kinase
inhibitors such as
N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-
-amine (gefitinib, ZD1839),
N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine
(erlotinib, OSI 774) and
6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)-quinazol-
in-4-amine (CI 1033), erbB2 tyrosine kinase inhibitors such as
lapatinib, inhibitors of the hepatocyte growth factor family,
inhibitors of the platelet-derived growth factor family such as
imatinib, inhibitors of serine/threonine kinases (for example
Ras/Raf signalling inhibitors such as farnesyl transferase
inhibitors, for example sorafenib (BAY 43-9006)), inhibitors of
cell signalling through MEK and/or AKT kinases, inhibitors of the
hepatocyte growth factor family, c-kit inhibitors, abl kinase
inhibitors, IGF receptor (insulin-like growth factor) kinase
inhibitors; aurora kinase inhibitors (for example AZD1152,
PH739358, VX-680, MLN8054, R763, MP235, MP529, VX-528 AND AX39459)
and cyclin dependent kinase inhibitors such as CDK2 and/or CDK4
inhibitors; (v) antiangiogenic and antilymphangiogenic agents such
as those which inhibit the effects of vascular endothelial growth
factor, [for example the anti vascular endothelial cell growth
factor A (VEGFA) antibody bevacizumab (AvastinT), the anti vascular
endothelial cell growth factor A (VEGFA) antibody ranibizumab, the
anti-VEGF aptamer pegaptanib, the anti vascular endothelial growth
factor receptor 3 (VEGFR3) antibody IMC-3C5, the anti vascular
endothelial cell growth factor C (VEGFC) antibody VGX-100, the anti
vascular endothelial cell growth factor D (VEGFD) antibody VGX-200,
the soluble form of the vascular endothelial growth factor receptor
3 (VEGFR3) VGX-300 and VEGF receptor tyrosine kinase inhibitors
such as
4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)qu-
inazoline (vandetanib; ZD6474; Example 2 within WO 01/32651),
4-(4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-1-ylpropoxy)-
quinazoline (cediranib; AZD2171; Example 240 within WO 00/47212),
vatalanib (PTK787; WO 98/35985), pazopanib (GW786034), axitinib
(AG013736), sorafenib and sunitinib (SU11248; WO 01/60814),
compounds such as those disclosed in International Patent
Applications WO97/22596, WO 97/30035, WO 97/32856 and WO 98/13354
and compounds that work by other mechanisms (for example linomide,
inhibitors of integrin avb3 function and angiostatin)]; (vi)
vascular damaging agents such as Combretastatin A4 and compounds
disclosed in International Patent Applications WO 99/02166, WO
00/40529, WO 00/41669, WO 01/92224, WO 02/04434 and WO 02/08213;
(vii) antisense therapies, for example those which are directed to
the targets listed above, such as ISIS 2503, an anti-ras antisense;
(viii) gene therapy approaches, including for example approaches to
replace aberrant genes such as aberrant p53 or aberrant BRCA1 or
BRCA2, GDEPT (gene directed enzyme pro drug therapy) approaches
such as those using cytosine deaminase, thymidine kinase or a
bacterial nitroreductase enzyme and approaches to increase patient
tolerance to chemotherapy or radiotherapy such as multi drug
resistance gene therapy; and (ix) immunotherapy approaches,
including for example ex vivo and in vivo approaches to increase
the immunogenicity of patient tumour cells, such as transfection
with cytokines such as interleukin 2, interleukin 4 or granulocyte
macrophage colony stimulating factor, approaches to decrease T cell
anergy, approaches using transfected immune cells such as cytokine
transfected dendritic cells, approaches using cytokine transfected
tumour cell lines and approaches using anti idiotypic
antibodies
[0148] Administration
[0149] The active compound or pharmaceutical composition comprising
the active compound may be administered to a subject by any
convenient route of administration, whether
systemically/peripherally or at the site of desired action,
including but not limited to, oral (e.g. by ingestion); topical
(including e.g. transdermal, intranasal, ocular, buccal, and
sublingual); pulmonary (e.g. by inhalation or insufflation therapy
using, e.g. an aerosol, e.g. through mouth or nose); rectal;
vaginal; parenteral, for example, by injection, including
subcutaneous, intradermal, intramuscular, intravenous,
intraarterial, intracardiac, intrathecal, intraspinal,
intracapsular, subcapsular, intraorbital, intraperitoneal,
intratracheal, subcuticular, intraarticular, subarachnoid,
intravitreal and intrasternal; by implant of a depot, for example,
subcutaneously, intravitreal or intramuscularly. The subject may be
a eukaryote, an animal, a vertebrate animal, a mammal, a rodent
(e.g. a guinea pig, a hamster, a rat, a mouse), murine (e.g. a
mouse), canine (e.g. a dog), feline (e.g. a cat), equine (e.g. a
horse), a primate, simian (e.g. a monkey or ape), a monkey (e.g.
marmoset, baboon), an ape (e.g. gorilla, chimpanzee, orang-utan,
gibbon), or a human.
[0150] Formulations
[0151] While it is possible for the active compound to be
administered alone, it is preferable to present it as a
pharmaceutical composition (e.g. formulation) comprising at least
one active compound, as defined above, together with one or more
pharmaceutically acceptable carriers, adjuvants, excipients,
diluents, fillers, buffers, stabilisers, preservatives, lubricants,
or other materials well known to those skilled in the art and
optionally other therapeutic or prophylactic agents.
[0152] Thus, the present invention further provides pharmaceutical
compositions, as defined above, and methods of making a
pharmaceutical composition comprising admixing at least one active
compound, as defined above, together with one or more
pharmaceutically acceptable carriers, excipients, buffers,
adjuvants, stabilisers, or other materials, as described
herein.
[0153] The term "pharmaceutically acceptable" as used herein
pertains to compounds, materials, compositions, and/or dosage forms
which are, within the scope of sound medical judgement, suitable
for use in contact with the tissues of a subject (e.g. human)
without excessive toxicity, irritation, allergic response, or other
problem or complication, commensurate with a reasonable
benefit/risk ratio. Each carrier, excipient, etc. must also be
"acceptable" in the sense of being compatible with the other
ingredients of the formulation.
[0154] Suitable carriers, excipients, etc. can be found in standard
pharmaceutical texts, for example, Remington's Pharmaceutical
Sciences, 18th edition, Mack Publishing Company, Easton, Pa.,
1990.
[0155] The formulations may conveniently be presented in unit
dosage form and may be prepared by any methods well known in the
art of pharmacy. Such methods include the step of bringing into
association the active compound with the carrier which constitutes
one or more accessory ingredients. In general, the formulations are
prepared by uniformly and intimately bringing into association the
active compound with liquid carriers or finely divided solid
carriers or both, and then if necessary shaping the product.
[0156] Formulations may be in the form of liquids, solutions,
suspensions, emulsions, elixirs, syrups, tablets, losenges,
granules, powders, capsules, cachets, pills, ampoules,
suppositories, pessaries, ointments, gels, pastes, creams, sprays,
mists, foams, lotions, oils, boluses, electuaries, or aerosols.
[0157] Formulations suitable for oral administration (e.g. by
ingestion) may be presented as discrete units such as capsules,
cachets or tablets, each containing a predetermined amount of the
active compound; as a powder or granules; as a solution or
suspension in an aqueous or non-aqueous liquid; or as an
oil-in-water liquid emulsion or a water-in-oil liquid emulsion; as
a bolus; as an electuary; or as a paste.
[0158] A tablet may be made by conventional means, e.g.,
compression or moulding, optionally with one or more accessory
ingredients. Compressed tablets may be prepared by compressing in a
suitable machine the active compound in a free-flowing form such as
a powder or granules, optionally mixed with one or more binders
(e.g. povidone, gelatin, acacia, sorbitol, tragacanth,
hydroxypropylmethyl cellulose); fillers or diluents (e.g. lactose,
microcrystalline cellulose, calcium hydrogen phosphate); lubricants
(e.g. magnesium stearate, talc, silica); disintegrants (e.g. sodium
starch glycolate, cross-linked povidone, cross-linked sodium
carboxymethyl cellulose); surface-active or dispersing or wetting
agents (e.g. sodium lauryl sulfate); and preservatives (e.g. methyl
p-hydroxybenzoate, propyl p-hydroxybenzoate, sorbic acid). Moulded
tablets may be made by moulding in a suitable machine a mixture of
the powdered compound moistened with an inert liquid diluent. The
tablets may optionally be coated or scored and may be formulated so
as to provide slow or controlled release of the active compound
therein using, for example, hydroxypropylmethyl cellulose in
varying proportions to provide the desired release profile. Tablets
may optionally be provided with an enteric coating, to provide
release in parts of the gut other than the stomach.
[0159] Formulations suitable for topical administration (e.g.
transdermal, intranasal, ocular, buccal, and sublingual) may be
formulated as an ointment, cream, suspension, lotion, powder,
solution, past, gel, spray, aerosol, or oil. Alternatively, a
formulation may comprise a patch or a dressing such as a bandage or
adhesive plaster impregnated with active compounds and optionally
one or more excipients or diluents.
[0160] Formulations suitable for topical administration in the
mouth include losenges comprising the active compound in a
flavoured basis, usually sucrose and acacia or tragacanth;
pastilles comprising the active compound in an inert basis such as
gelatin and glycerin, or sucrose and acacia; and mouthwashes
comprising the active compound in a suitable liquid carrier.
[0161] Formulations suitable for topical administration to the eye
also include eye drops wherein the active compound is dissolved or
suspended in a suitable carrier, especially an aqueous solvent for
the active compound.
[0162] Formulations suitable for nasal administration, wherein the
carrier is a solid, include a coarse powder having a particle size,
for example, in the range of about 20 to about 500 microns which is
administered in the manner in which snuff is taken, i.e. by rapid
inhalation through the nasal passage from a container of the powder
held close up to the nose. Suitable formulations wherein the
carrier is a liquid for administration as, for example, nasal
spray, nasal drops, or by aerosol administration by nebuliser,
include aqueous or oily solutions of the active compound.
[0163] Formulations suitable for administration by inhalation
include those presented as an aerosol spray from a pressurised
pack, with the use of a suitable propellant, such as
dichlorodifluoromethane, trichlorofluoromethane,
dichoro-tetrafluoroethane, carbon dioxide, or other suitable
gases.
[0164] Formulations suitable for topical administration via the
skin include ointments, creams, and emulsions. When formulated in
an ointment, the active compound may optionally be employed with
either a paraffinic or a water-miscible ointment base.
Alternatively, the active compounds may be formulated in a cream
with an oil-in-water cream base. If desired, the aqueous phase of
the cream base may include, for example, at least about 30% w/w of
a polyhydric alcohol, i.e., an alcohol having two or more hydroxyl
groups such as propylene glycol, butane-1,3-diol, mannitol,
sorbitol, glycerol and polyethylene glycol and mixtures thereof.
The topical formulations may desirably include a compound which
enhances absorption or penetration of the active compound through
the skin or other affected areas. Examples of such dermal
penetration enhancers include dimethylsulfoxide and related
analogues.
[0165] When formulated as a topical emulsion, the oily phase may
optionally comprise merely an emulsifier (otherwise known as an
emulgent), or it may comprises a mixture of at least one emulsifier
with a fat or an oil or with both a fat and an oil. Preferably, a
hydrophilic emulsifier is included together with a lipophilic
emulsifier which acts as a stabiliser. It is also preferred to
include both an oil and a fat. Together, the emulsifier(s) with or
without stabiliser(s) make up the so-called emulsifying wax, and
the wax together with the oil and/or fat make up the so-called
emulsifying ointment base which forms the oily dispersed phase of
the cream formulations.
[0166] Suitable emulgents and emulsion stabilisers include Tween
60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl
monostearate and sodium lauryl sulphate. The choice of suitable
oils or fats for the formulation is based on achieving the desired
cosmetic properties, since the solubility of the active compound in
most oils likely to be used in pharmaceutical emulsion formulations
may be very low. Thus the cream should preferably be a non-greasy,
non-staining and washable product with suitable consistency to
avoid leakage from tubes or other containers. Straight or branched
chain, mono- or dibasic alkyl esters such as di-isoadipate,
isocetyl stearate, propylene glycol diester of coconut fatty acids,
isopropyl myristate, decyl oleate, isopropyl palmitate, butyl
stearate, 2-ethylhexyl palmitate or a blend of branched chain
esters known as Crodamol CAP may be used, the last three being
preferred esters. These may be used alone or in combination
depending on the properties required.
[0167] Alternatively, high melting point lipids such as white soft
paraffin and/or liquid paraffin or other mineral oils can be
used.
[0168] Formulations suitable for rectal administration may be
presented as a suppository with a suitable base comprising, for
example, cocoa butter or a salicylate.
[0169] Formulations suitable for vaginal administration may be
presented as pessaries, tampons, creams, gels, pastes, foams or
spray formulations containing in addition to the active compound,
such carriers as are known in the art to be appropriate.
[0170] Formulations suitable for parenteral administration (e.g. by
injection, including cutaneous, subcutaneous, intramuscular,
intravenous and intradermal), include aqueous and non-aqueous
isotonic, pyrogen-free, sterile injection solutions which may
contain anti-oxidants, buffers, preservatives, stabilisers,
bacteriostats, and solutes which render the formulation isotonic
with the blood of the intended recipient; and aqueous and
non-aqueous sterile suspensions which may include suspending agents
and thickening agents, and liposomes or other microparticulate
systems which are designed to target the compound to blood
components or one or more organs. Examples of suitable isotonic
vehicles for use in such formulations include Sodium Chloride
Injection, Ringer's Solution, or Lactated Ringer's Injection.
Typically, the concentration of the active compound in the solution
is from about 1 ng/mL to about 10 .mu.g/mL, for example from about
10 ng/ml to about 1 .mu.g/mL. The formulations may be presented in
unit-dose or multi-dose sealed containers, for example, ampoules
and vials, and may be stored in a freeze-dried (lyophilised)
condition requiring only the addition of the sterile liquid
carrier, for example water for injections, immediately prior to
use. Extemporaneous injection solutions and suspensions may be
prepared from sterile powders, granules, and tablets. Formulations
may be in the form of liposomes or other microparticulate systems
which are designed to target the active compound to blood
components or one or more organs.
[0171] Dosage
[0172] It will be appreciated by one of skill in the art that
appropriate dosages of the compound, and compositions comprising
the compound, can vary from patient to patient. Determining the
optimal dosage will generally involve the balancing of the level of
therapeutic benefit against any risk or deleterious side effects.
The selected dosage level will depend on a variety of factors
including, but not limited to, the activity of the particular
compound, the route of administration, the time of administration,
the rate of excretion of the compound, the duration of the
treatment, other drugs, compounds, and/or materials used in
combination, the severity of the condition, and the species, sex,
age, weight, condition, general health, and prior medical history
of the patient. The amount of compound and route of administration
will ultimately be at the discretion of the physician,
veterinarian, or clinician, although generally the dosage will be
selected to achieve local concentrations at the site of action
which achieve the desired effect without causing substantial
harmful or deleterious side-effects.
[0173] Administration can be effected in one dose, continuously or
intermittently (e.g., in divided doses at appropriate intervals)
throughout the course of treatment. Methods of determining the most
effective means and dosage of administration are well known to
those of skill in the art and will vary with the formulation used
for therapy, the purpose of the therapy, the target cell(s) being
treated, and the subject being treated. Single or multiple
administrations can be carried out with the dose level and pattern
being selected by the treating physician, veterinarian, or
clinician.
[0174] In general, a suitable dose of the active compound is in the
range of about 100 ng to about 25 mg (more typically about 1 .mu.g
to about 10 mg) per kilogram body weight of the subject per day.
Where the active compound is a salt, an ester, an amide, a prodrug,
or the like, the amount administered is calculated on the basis of
the parent compound and so the actual weight to be used is
increased proportionately.
[0175] In one embodiment, the active compound is administered to a
human patient according to the following dosage regime: about 100
mg, 3 times daily.
[0176] In one embodiment, the active compound is administered to a
human patient according to the following dosage regime: about 150
mg, 2 times daily.
[0177] In one embodiment, the active compound is administered to a
human patient according to the following dosage regime: about 200
mg, 2 times daily.
[0178] However in one embodiment, the active compound is
administered to a human patient according to the following dosage
regime: about 50 or about 75 mg, 3 or 4 times daily.
[0179] In one embodiment, the active compound is administered to a
human patient according to the following dosage regime: about 100
or about 125 mg, 2 times daily.
[0180] Treatment
[0181] The term "treatment," as used herein in the context of
treating a condition, pertains generally to treatment and therapy,
whether of a human or an animal (e.g., in veterinary applications),
in which some desired therapeutic effect is achieved, for example,
the inhibition of the progress of the condition, and includes a
reduction in the rate of progress, a halt in the rate of progress,
regression of the condition, amelioration of the condition, and
cure of the condition. Treatment as a prophylactic measure (i.e.,
prophylaxis, prevention) is also included.
[0182] The term "therapeutically-effective amount," as used herein,
pertains to that amount of an active compound, or a material,
composition or dosage from comprising an active compound, which is
effective for producing some desired therapeutic effect,
commensurate with a reasonable benefit/risk ratio, when
administered in accordance with a desired treatment regimen.
[0183] Similarly, the term "prophylactically-effective amount," as
used herein, pertains to that amount of an active compound, or a
material, composition or dosage from comprising an active compound,
which is effective for producing some desired prophylactic effect,
commensurate with a reasonable benefit/risk ratio, when
administered in accordance with a desired treatment regimen.
[0184] The Subject/Patient
[0185] The subject/patient may be an animal, mammal, a placental
mammal, a marsupial (e.g., kangaroo, wombat), a monotreme (e.g.,
duckbilled platypus), a rodent (e.g., a guinea pig, a hamster, a
rat, a mouse), murine (e.g., a mouse), a lagomorph (e.g., a
rabbit), avian (e.g., a bird), canine (e.g., a dog), feline (e.g.,
a cat), equine (e.g., a horse), porcine (e.g., a pig), ovine (e.g.,
a sheep), bovine (e.g., a cow), a primate, simian (e.g., a monkey
or ape), a monkey (e.g., marmoset, baboon), an ape (e.g., gorilla,
chimpanzee, orangutang, gibbon), or a human.
[0186] Furthermore, the subject/patient may be any of its forms of
development, for example, a foetus. In one preferred embodiment,
the subject/patient is a human.
[0187] General Synthesis Methods
[0188] The compounds of the invention can be prepared by employing
the following general methods and using procedures described in
detail for the examples. The reaction conditions referred to are
illustrative and non-limiting, for example one skilled in the art
may use a diverse range of synthetic methods to synthesise the
desired compounds such as but not limited to methods described in
literature (for example but not limited to March's Advanced Organic
Chemistry: Reactions, Mechanisms, and Structure, 7th Edition or
Larock's Comprehensive Organic Transformations: Comprehensive
Organic Transformations: A Guide to Functional Group
Preparations).
[0189] Compounds of formula (I), as described above, can be
prepared by synthetic strategies outlined below, wherein the
definitions above apply.
[0190] General Synthesis 1
[0191] Scheme 1A illustrates the formation of a sulfonamide bond to
form compounds with the structure I by coupling a relevant sulfonyl
chloride compound of structure G2 with a primary or secondary amine
such as benzisoxazole amine G3.
##STR00008##
[0192] Methods to form such sulfonamides will be apparent to those
skilled in the art, but include for example the use of a suitable
base such as but not limited to pyridine, LiHMDS, n-BuLi or NaH and
the use of activated forms of the sulfonic acid such as the
corresponding sulfonyl halide. Formation of sulfonyl chlorides of
structure G2 from the corresponding acids of structure G1 can be
achieved by for example use of thionyl chloride or cyanuric
chloride.
[0193] Alternatively, the activated form of a sulfonic acid such as
but not limited to a pentafluorophenyl sulfonate ester or
trichlorophenyl sulfonate ester with the structure G5 can be
coupled with the relevant primary or secondary amine, such as
benzisoxazole amine G3 (Scheme 1B).
##STR00009##
[0194] Formation of the sulfonate ester in G5 from the
corresponding sulfonyl chloride G2 and relevant phenol (R.sup.5 may
be for example pentaflurorphenyl or trichlorophenyl) can be
achieved using a suitable base such as but not limited to pyridine
or triethylamine. Methods to form the sulfonamides in I will be
apparent to those skilled in the art, but include for example the
use of a suitable base such as but not limited to LiHMDS.
[0195] General Synthesis 2
[0196] Scheme 2A illustrates the formation of a sulfonyl chloride
such as G2, as a substituent which is part of Ar.
##STR00010##
[0197] This can be achieved by reacting a relevant aryl compound
with the structure G6 with for example but not limited to
chlorosulfonic acid. Alternatively, the aryl compound G6 may be
sequentially treated with a base, such as but not limited to
n-BuLi, and sulphur dioxide to furnish the lithium arylsulfinate
which is further oxidised by for example sulfuryl chloride to give
the desired sulfonyl chloride in G2. The product G2 may be isolated
by methods know to those skilled in the art or may be formed in
situ and used immediately in subsequent step.
[0198] In addition, the sulfonyl chloride in G2 may be formed from
an aryl thiol in structure G8 illustrated in Scheme 2B.
##STR00011##
[0199] Methods to form G2 include for example the use of a suitable
oxidant such as but not limited to hydrogen peroxide and potassium
nitrate in the presence of a chloride source such as but not
limited to chlorotrimethylsilane or thionyl chloride. A thiol of
structure G8 may be synthesised from a compound of structure G7
where (X) may be a halogen by methods known to those skilled in the
art, including but not limited to nucleophilic displacement in the
presence or absence of a transition metal. Alternatively,
sulfonation of an aryl compound such as G6 may give the
corresponding sulfonic acid of structure G1. This can be achieved
by any suitable reagent known to those skilled in the art, for
example sulphur trioxide or sulfuric acid.
##STR00012##
[0200] The sulfonic acid G1 may be converted to the sulfonyl
chloride G2 by methods outlined in General Synthesis 1, Scheme
1A.
[0201] General Synthesis 3
[0202] Scheme 3A illustrates the formation of a benzisoxazole amine
such as G3 from an aryl nitrile with an ortho substituent X, such
as structure G9. The group (X) may be, but is not limited to, a
halogen such as a chloro or a fluoro group and is chosen to be
suitable for the reaction employed.
##STR00013##
[0203] For example, the starting material G9 may be reacted with an
oxime such as but not limited to acetone oxime or with for example
acetohydroxamic acid, in the presence of a suitable base such as
but not limited to potassium tert-butoxide, to form the
benzisoxazole amine G3.
[0204] General Synthesis 4
##STR00014##
[0205] Scheme 4A illustrates the addition of an R.sup.7 group to a
compound of structure G10 (where R.sup.6 represents H or a suitable
protecting group including but not limited to 2,4-dimethoxybenzyl
(DMB); methods for the removal of said protecting groups will be
known to those skilled in the art (for example Greene's Protective
Groups in Organic Synthesis, 4th Edition)), as a substituent which
is part of Ar. This can be achieved using any suitable coupling
reaction known to the person skilled in the art, for example Suzuki
coupling. The groups R.sup.7B.sup.1 and X are chosen to be suitable
for the coupling reaction employed. For example, in the case of a
Suzuki coupling reaction, (X) may be a halogen, triflate or other
suitable group and B.sup.1 represents a suitable boron compound
including but not limited to a boronic acid or boronate ester.
[0206] Examples of B.sup.1 that can be used in the Suzuki coupling
include, but are not limited to, those shown below.
##STR00015##
[0207] The types of R.sup.7B.sup.1 compounds that can be used in
the Suzuki coupling include, but are not limited to, those shown
below.
##STR00016##
[0208] In addition to scheme 4A, the position of the (X) and
(B.sup.1) can be reversed as shown below in scheme 4B, to give the
same final compound G11. Similarly to Scheme 2A, the groups denoted
by R.sup.7X and B.sup.1 are chosen to be suitable for the coupling
reaction employed. For example, in the case of a Suzuki coupling
reaction (X) may be a halogen, triflate or other suitable group and
B.sup.1 represents a suitable boron compound including, but not
limited to, a boronic acid or boronate ester.
##STR00017##
[0209] A variety of coupling reactions may be used to introduce the
R.sup.7 group other than Suzuki coupling, such as for example
transition metal catalysed coupling reactions or for example tin
(Stille type reaction) and zinc (Negishi type reaction) compounds.
In addition, a Chan-Lam type coupling may be used when the group
(X) is for example, but not limited to, a phenol (O--H) or primary
or secondary amine (R'R''N--H).
[0210] The transitions described in Scheme 4A and 4B may also be
carried out with substituent R.sup.1, R.sup.2, R.sup.3 or R.sup.4
on the benzisoxazole moiety in structure G13, represented by Scheme
4C below.
##STR00018##
[0211] In addition, substituent R.sup.7 may be introduced prior to
sulfonamide and benzisoxazole formation on the nitrile precursor G9
(when R.sup.1 or R.sup.2 or R.sup.3 or R.sup.4.dbd.X or B.sup.1) in
General Synthesis 3, Scheme 3D.
[0212] General Synthesis 5
[0213] Scheme 5A illustrates the addition of a nitrogen linked
R.sup.8 group, as a substituent which is part of Ar or on the
benzisoxazole moiety to give a compound of structure G16. This can
be achieved using any suitable coupling reaction known to the
person skilled in the art, for example, by SnAr displacement or
Buchwald coupling. The group denoted by (X) may be, but not limited
to, a halogen and is chosen to be suitable for the coupling
reaction employed.
##STR00019##
[0214] Alternatively, to synthesise ether linked compounds, a
similar strategy can be employed as shown in Scheme 5B. This can be
achieved using any suitable coupling reaction known to a person
skilled in the art, for example, by a SnAr or Ullman-type coupling
to give compounds with structure G17.
##STR00020##
[0215] Both the above couplings may also be reversed, such that the
group added is R.sup.8--X.
[0216] In addition, substituent OR.sup.8 or NHR.sup.8 may be
introduced prior to sulfonamide and benzisoxazole formation on the
nitrile precursor G9 (when R.sup.1 or R.sup.2 or R.sup.3 or
R.sup.4.dbd.X) in General Synthesis 3, Scheme 3D.
[0217] General Synthesis 6
##STR00021##
[0218] Scheme 6A illustrates the addition of an amine
(HNR.sup.10R.sup.11)) to form the corresponding amide, as a
substituent which is a part of Ar or on the benzisoxazole moiety to
give a compound of structure G19. This can be achieved by coupling
a relevant carboxylic acid with a primary amine or a secondary
amine NHR.sup.10R.sup.11. Methods to form such amides will be
apparent to those skilled in the art, but include for example the
use of reagents such as HATU, HBTU, T3P and EDCl/HOBt, and the use
of activated forms of the carboxylic acid such as the corresponding
acyl halide, mixed anhydride or N-hydroxysuccinimide ester. Amide
G19 may also be synthesised directly from the ester compound (when
R.sup.12=Alk, such as but not limited to methyl or ethyl).
Formation of the carboxylic acid (when R.sup.12.dbd.H) from the
corresponding ester can be achieved by for example hydrolysis with
a base such as an alkali metal hydroxide or an acid for example
aqueous hydrochloric acid.
[0219] From the amide compound G19, further transformations may be
carried out, such as but not limited to, reduction of the amide to
form the amine or dehydration of the amide to form the nitrile.
Methods to perform such transformation will be known to those
skilled in the art.
[0220] In addition, an amide CONR.sup.10R.sup.11 may be introduced
prior to sulfonamide and benzisoxazole formation on the nitrile
precursor G9 (when R.sup.1 or R.sup.2 or R.sup.3 or
R.sup.4.dbd.CO.sub.2R.sup.12, R.sup.12=Alk or H) in General
Synthesis 3, Scheme 3D.
[0221] General Synthesis 7
[0222] Conversion of (X) in structure G20 in Scheme 7A into an
ester in structure G18 (R.sup.12=Alkyl, such as methyl or ethyl)
will be apparent to those skilled in the art, but include for
example a carbonylation reaction which can be achieved by the use
of carbon monoxide in the presence of an transition metal catalyst
such as but not limited to PdCl.sub.2dppf.DCM; and an alcoholic
solvent such as but not limited to methanol or ethanol. Formation
of the carboxylic acid in structure G18 (R.dbd.H) can be achieved
by for example hydrolysis with a base such as an alkali metal
hydroxide or an acid for example aqueous hydrochloric acid.
##STR00022##
[0223] The ester or acid G18 in Scheme 7A may be reduced to the
hydroxyl compound such as structure G21. Methods for such
transformation will be known to those skilled in the art but
include for example the use of reducing agents such as lithium
aluminium hydride (for the ester and carboxylic acid) and borane
(for the carboxylic acid).
[0224] From the hydroxyl compound G21, further transformations may
be carried out, such as but not limited to, Mitsunobu or
nucleophilc substitution reactions. Methods to perform such
transformation will be known to those skilled in the art.
[0225] In addition, an ester group may be introduced prior to
sulfonamide and benzisoxazole formation on the nitrile precursor G9
(when R.sup.1 or R.sup.2 or R.sup.3 or R.sup.4.dbd.X) in General
Synthesis 3, Scheme 3D.
[0226] General Synthesis 8
[0227] Scheme 8A illustrates the reduction of a nitro group in
structure G22 to form the corresponding amine in structure G23, as
a substituent which is part of the Ar or on the benzisoxazole
moiety.
##STR00023##
[0228] Reduction of the nitro group to the primary amine G23 will
be apparent to those skilled in the art and include but are not
limited to using reducing conditions such as a transition metal
(Fe, In, Zn) in the presence of HCl, hydrogenation in the presence
of a transition metal or transition metal catalyst.
[0229] From the amine compound G23, further transformations may be
carried out, such as but not limited to amide bond formation.
Methods to perform such transformation will be similar to those
described in General Synthesis 6.
[0230] In addition, an amine group may be introduced prior to
sulfonamide and benzisoxazole formation on the nitrile precursor G9
(when R.sup.1 or R.sup.2 or R.sup.3 or R.sup.4.dbd.NO.sub.2) in
General Synthesis 3, Scheme 3D.
[0231] General Synthesis 9
[0232] Scheme 9A illustrates the introduction of a nitrile group in
structure G25, as a substituent which is part or the Ar or on the
benzisoxazole moiety.
##STR00024##
[0233] The method for such transformation will be apparent to those
skilled in the art and include but are not limited to SnAr
displacement, or a transition metal catalysed coupling with a
suitable cyanide reagent. The group denoted by (X) in structure G24
may be, but not limited to, a halogen, triflate or mesylate and is
chosen to be suitable for the reaction employed.
[0234] Further Preferences
[0235] The following preferences may apply to all aspects of the
invention as described above, or may relate to a single aspect. The
preferences may be combined together in any combination.
[0236] R.sup.1, R.sup.2, R.sup.3 and R.sup.4
[0237] In some embodiments, at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 may be H. In some of these embodiments, one of
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are H. In other of these
embodiments, two of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are H. In
other of these embodiments, three of R.sup.1, R.sup.2, R.sup.3 and
R.sup.4 are H.
[0238] In some embodiments, at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 is not H.
[0239] In some embodiments, at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 may be C.sub.1-3 alkyl, optionally substituted
by:
hydroxy, C.sub.1-2 alkoxy, optionally substituted by one or more
fluoro groups
NH.sub.2,
[0240] phenyl, C.sub.5-6 heteroaryl, C.sub.1-4 alkyl carbamoyl,
acylamido, or one or more fluoro groups.
[0241] In these embodiments, at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 may be C.sub.1-3 alkyl. Further, in these
embodiments, the C.sub.1-3 alkyl group may be methyl, ethyl or
propyl. These groups may be unsubstituted. These groups may be
substituted by one or more fluoro groups, and may be
perfluorinated, e.g. CF.sub.3, C.sub.2F.sub.5. These groups may be
substituted by one, two, three, four or five fluoro groups. In some
embodiment, these groups may be substituted by one; one or two; or
one, two or three fluoro groups.
[0242] If the alkyl group is substituted, the substituent may be
selected from: [0243] (i) hydroxy; or [0244] (ii) unsubstituted
C.sub.1-2 alkoxy, i.e. methoxy, ethoxy; or C.sub.1-2 alkoxy
substituted by one or more fluoro groups, e.g. --OCH.sub.2F,
--OCH.sub.2CF.sub.3; or [0245] (iii) NH.sub.2; or [0246] (iv)
phenyl; or [0247] (v) C.sub.5-6 heteroaryl, e.g. N-pyrazolyl; or
[0248] (vi) C.sub.1-4 alkyl carbamoyl, e.g. NHC(O)Me; or [0249]
(vii) acylamido, e.g. NHCO.sub.2Me.
[0250] In some embodiments, at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 may be C.sub.1-3 alkoxy, optionally substituted
by C.sub.3-6 cycloalkyl or by one of more fluoro groups. In these
embodiments, the C.sub.1-3 alkoxy group may be methoxy, ethoxy or
propyloxy. These groups may be unsubstituted. These groups may be
substituted by one or more fluoro groups, and may be
perfluorinated, e.g. OCF.sub.3, OC.sub.2F.sub.5. These groups may
be substituted by one, two, three, four or five fluoro groups. In
some embodiment, these groups may be substituted by one; one or
two; or one, two or three fluoro groups. The alkoxy group may be
substituted by C.sub.3-6 cycloalkyl, e.g. cyclopropyl. Thus the
overall group may be OCH.sub.2 (cyclopropyl).
[0251] In some embodiments, at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 may be C.sub.3-6 cycloalkyl. In these
embodiments, the C.sub.3-6 cycloalkyl group may be cyclopropyl,
cyclobutyl, cyclopentyl or cyclohexyl. In particular, the C.sub.3-6
cycloalkyl group may be cyclopropyl.
[0252] In some embodiments, at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 may be halo. In these embodiments, the halo
group may be fluoro, chloro, bromo or iodo.
[0253] In some embodiments, at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 may be COR.sup.C, where R.sup.C is selected
from NR.sup.N1R.sup.N2, where R.sup.N1 and R.sup.N2 are
independently selected from H and methyl. In these embodiments, the
group may be selected from C(O)NH.sub.2, C(O)NHCH.sub.3 and
C(O)N(CH.sub.3).sub.2.
[0254] In some embodiments, at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 may be cyano, NH.sub.2, NO.sub.2. In some of
these embodiments at least one of R.sup.1, R.sup.2, R.sup.3 and
R.sup.4 may be cyano. In others of these embodiments at least one
of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 may be NH.sub.2. In others
of these embodiments at least one of R.sup.1, R.sup.2, R.sup.3 and
R.sup.4 may be NO.sub.2.
[0255] In some embodiments, at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 may be phenyl or C.sub.5-6 heteroaryl, which
groups are optionally substituted by methyl, cyano, hydroxy or
methoxy. In some of these embodiments at least one of R.sup.1,
R.sup.2, R.sup.3 and R.sup.4 may be phenyl. In others of these
embodiments at least one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4
may be C.sub.5-6 heteroaryl, for example oxazolyl, pyrazolyl,
triazolyl, pyridyl and pyrimidinyl. The phenyl or C.sub.5-6
heteroaryl group may be unsubstituted. In certain embodiments, the
phenyl group may be substituted by methyl, cyano or methoxy. In
certain embodiments, the C.sub.5-6 heteroaryl group may be
substituted by one or more methyl groups, such that the overall
group is, for example, dimethylpyrazolyl or N-methylpyrazolyl.
[0256] In some embodiments, R.sup.4 is methoxy.
[0257] In some embodiments, R.sup.4 is methoxy, R.sup.2 is
CH.sub.2OCH.sub.3 or CH.sub.2OCH.sub.2CH.sub.3 and R.sup.1 and
R.sup.3 are H.
[0258] In some embodiments, R.sup.4 is methoxy, R.sup.2 is phenyl,
optionally substituted by methyl or methoxy, and R.sup.1 and
R.sup.3 are H.
[0259] In some embodiments, R.sup.4 is methoxy, R.sup.2 is
C.sub.5-6 heteroaryl, optionally substituted by methyl.
[0260] In some embodiments, R.sup.4 is methoxy and R.sup.1, R.sup.2
and R.sup.3 are H.
[0261] In some embodiments, R.sup.4 is chloro, R.sup.2 is C.sub.1-3
alkyl or bromo, and R.sup.1 and R.sup.3 are H.
[0262] In some embodiments, R.sup.4 is chloro and R.sup.1, R.sup.2
and R.sup.4 are H.
[0263] In some embodiments, R.sup.3 is C.sub.1-3 alkyl and R.sup.1,
R.sup.2 and R.sup.4 are H.
[0264] Ar
[0265] Ar is selected from phenyl, napthyl and C.sub.5-10
heteroaryl groups, which may be unsubstituted or substituted.
[0266] In some embodiments, Ar is phenyl.
[0267] In some embodiments, Ar is napthyl.
[0268] In some embodiments, Ar is a C.sub.5-10 heteroaryl group.
The C.sub.5-10 heteroaryl group may be selected from: quinolinyl,
benzothiazolyl, quinoxalinyl, benzooxadiazolyl, benzothiadiazolyl,
benzofuran and benzotriazolyl. In certain of these embodiments, Ar
is quinolinyl or benzothiazolyl.
[0269] In some embodiments, Ar is the group:
##STR00025##
[0270] In some embodiments, a substituent for Ar is C.sub.1-4
alkyl, optionally substituted by hydroxy, C.sub.1-2 alkoxy,
NH.sub.2, C.sub.1-4 alkyl carbamoyl, or by one or more fluoro
groups. In these embodiments, the C.sub.1-4 alkyl group may be
methyl, ethyl, propyl or butyl. These groups may be unsubstituted.
These groups may be substituted by one or more fluoro groups, and
may be perfluorinated, e.g. CF.sub.3, C.sub.2F.sub.5. If the alkyl
group is substituted, the substituent may be selected from: [0271]
(i) hydroxy; or [0272] (ii) C.sub.1-2 alkoxy, i.e. methoxy, ethoxy;
or [0273] (iii) NH.sub.2; or [0274] (iv) C.sub.1-4 alkyl carbamoyl,
e.g. NHC(O)CH.sub.3.
[0275] In some embodiments, a substituent for Ar is C.sub.3-6
cycloalkyl. In these embodiments, the C.sub.3-6 cycloalkyl group
may be cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In
particular, the C.sub.3-6 cycloalkyl group may be cyclohexyl.
[0276] In some embodiments, a substituent for Ar is hydroxy; cyano;
NR.sup.N3R.sup.N4, where R.sup.N3 and R.sup.N4 are independently
selected from H and methyl; or acylamido. In some of these
embodiments, the substituent may be hydroxy. In other of these
embodiments, the substituent may be cyano. In other of these
embodiments, the substituent may be NR.sup.N3R.sup.N4, where
R.sup.N3 and R.sup.N4 are independently selected from H and
methyl--thus the substituent may be NH.sub.2, NHCH.sub.3 or
N(CH.sub.3).sub.2. In other of these embodiments, the substituent
may be acylamido, such as NHCO.sub.2CH.sub.3.
[0277] In some embodiments, a substituent for Ar is halo. In these
embodiments, the halo group may be fluoro, chloro, bromo or
iodo.
[0278] In some embodiments, a substituent for Ar is C.sub.1-3
alkoxy, optionally substituted by hydroxy, C(O)NH.sub.2, C.sub.3-6
cycloalkyl, phenyl, C.sub.5-6 heteroaryl, or by one of more fluoro
groups. In these embodiments, the C.sub.1-3 alkoxy group may be
methoxy, ethoxy or propyloxy. These groups may be unsubstituted.
These groups may be substituted by one or more fluoro groups, and
may be perfluorinated, e.g. OCF.sub.3, OC.sub.2F.sub.5. The alkoxy
group may be substituted by hydroxyl, such that the overall group
is, for example, OC.sub.2H.sub.4OH. The alkoxy group may be
substituted by C(O)NH.sub.2, such that the overall group is, for
example, OCH.sub.2C(O)NH.sub.2. The alkoxy group may be substituted
by C.sub.3-6 cycloalkyl, e.g. cyclopropyl, such that overall group
may be, for example, OCH.sub.2 (cyclopropyl). The alkoxy group may
be substituted by phenyl, such that the overall group is, for
example, benzyloxy. The alkoxy group may be substituted by
C.sub.5-6 heteroaryl, e.g. pyridyl, pyrazolyl, such that the
overall group is, for example, OCH.sub.2(N-methylpyrazolyl) or
OCH.sub.2 (methoxypyridyl).
[0279] In some embodiments, a substituent for Ar is phenoxy,
optionally substituted by fluoro. In some of these embodiments the
substituent may be phenoxy. In others of these embodiments, the
substituent may be OC.sub.6H.sub.4F.
[0280] In some embodiments, a substituent for Ar is phenyl or
C.sub.5-6 heteroaryl. In some of these embodiments, the substituent
is phenyl. In others of these embodiments, the substituent may be
C.sub.5-6 heteroaryl, such as oxazolyl or N-pyrazolyl.
[0281] In some embodiments, a substituent for Ar is SF.sub.5 or
SO.sub.2CH.sub.3. In some of these embodiments, the substituent is
SF.sub.5. In others of these embodiments, the substituent is
SO.sub.2Me.
[0282] In some embodiments, a substituent for Ar is
--(CH.sub.2).sub.n--Y--, where Y is O or CH.sub.2, and n is 2 or 3.
This substituent is particularly relevant when Ar is phenyl, and
forms a partially unsaturated fused ring with the phenyl. Thus, Ar
can be tetralinyl (i.e. fused cyclohexane), indanyl (i.e. fused
cyclopentane), chromanyl (i.e. fused tetrahydopyran) or
dihydrobenzofuranyl.
[0283] In some embodiments, a substituted for Ar is C.sub.1-4 alkyl
ester. In some of these embodiments, the substituent is
C(O)OCH.sub.3. In others of these embodiments, the substituent is
C(O)OC(CH.sub.3).sub.3.
[0284] Certain embodiments of Ar may be represented by the formula
(Ar-1):
##STR00026##
where Y is either N or C--R.sup.A4, and Z is either N or
C--R.sup.A5; and R.sup.A1, R.sup.A2, R.sup.A3, R.sup.A4 (if
present) and R.sup.A5 (if present) are independently selected from
H and the optional substituents for Ar.
[0285] In some embodiments, R.sup.A2 is ethyl.
[0286] In some embodiments, R.sup.A3 is selected from cycloalkyl;
phenoxy; phenyl; C.sub.5-6 heteroaryl; SF.sub.5; and
SO.sub.2CH.sub.3.
[0287] In some embodiments, Ar is 5-ethyl-2-methoxyphenyl.
[0288] In some embodiments, Ar is 5-CF.sub.3-2-methoxyphenyl.
[0289] In some embodiments, Ar is 2,6-dimethoxyphenyl.
[0290] In some embodiments, Ar is quinolinyl. These compounds may
show selective activity against HBO1.
[0291] In some embodiments, R.sup.4 is methoxy, R.sup.2 is selected
from CH.sub.2O CH.sub.3, CH.sub.2O CH.sub.2CH.sub.3 and optionally
substituted phenyl, and Ar is 2,6-dimethoxybenzene. These compounds
may show particular activity against MOZ and MORF. Compounds where
R.sup.2 is selected from CH.sub.2OCH.sub.3 and
CH.sub.2OCH.sub.2CH.sub.3 may show selective activity against MOZ
and MORF.
[0292] In some embodiments, for compounds where R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 are H, then Ar is not 4-aminophenyl.
[0293] In some embodiments, for compounds where R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 are H, then Ar is not
2,4,6-trimethylphenyl.
[0294] In some embodiments, for compounds where R.sup.1, R.sup.2
and R.sup.4 are H, and R.sup.3 is CF.sub.3, then Ar is not
2-(difluromethoxy)phenyl.
[0295] In some embodiments, for compounds where R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 are H, then Ar is not
4-fluoro-3-methyl-phenyl.
[0296] In some embodiments, for compounds where R.sup.1, R.sup.2
and R.sup.3 are H, and R.sup.4 is methoxy, then Ar is not
unsubstituted napthyl.
[0297] Compounds of particular interest include those of the
examples.
EXAMPLES
[0298] The following examples are provided solely to illustrate the
present invention and are not intended to limit the scope of the
invention, as described herein.
[0299] Acronyms
[0300] For convenience, many chemical moieties are represented
using well known abbreviations, including but not limited to,
methyl (Me), ethyl (Et), N-propyl (nPr), isopropyl (iPr), N-butyl
(nBu), tert-butyl (tBu), phenyl (Ph), benzyl (Bn), methoxy (MeO),
ethoxy (EtO), trimethylsilyl (TMS), and acetyl (Ac).
[0301] For convenience, many chemical compounds are represented
using well known abbreviations, including but not limited to,
methanol (MeOH), deuterated methanol (methanol-d.sub.4) ethanol
(EtOH), isopropanol (i-PrOH), ethyl acetate (EtOAc), acetic acid
(AcOH), acetonitrile (MeCN or ACN), dichloromethane (methylene
chloride, DCM), trifluoroacetic acid (TFA), N,N-dimethylformamide
(DMF), tetrahydrofuran (THF), dimethylsulfoxide (DMSO),
N-methyl-2-pyrrolidone (NMP), deuterated acetone (acetone-d.sub.6),
deuterated chloroform (CDCl.sub.3), deuterated dimethylsulfoxide
(DMSO-d.sub.6), 1,1'-bis(diphenylphosphino)ferrocene (dppf),
triethylamine (Et.sub.3N or TEA), N,N-diisopropylethylamine (DIPEA
or DIEA), 1,1'-bis(diphenylphosphino)ferrocene dichloropalladium
(II) (PdCl.sub.2(dppf)),
trans-dichlorobis(triphenylphosphine)palladium(II)
(PdCl.sub.2(PPh.sub.3).sub.2), tris(dibenzylideneacetone)
dipalladium(O) (Pd.sub.2(dba).sub.3),
tetrakis(triphenylphosphine)palladium(O) (Pd(PPh.sub.3).sub.4),
2,4-dimethoxybenzyl (DMB), petroleum ether (Pet. ether), lithium
bis(trimethylsilyl)amide (LHMDS or LiHMDS), potassium
bis(trimethylsilyl)amide (KHMDS), sodium bis(trimethylsilyl)amide
(NaHMDS), n-butyllithium (n-BuLi), N-bromosuccinimide (NBS),
N-chlorosuccinimide (NCS), pyridinium p-toluenesulfonate (PPTS),
azobisisobutyronitrile (AIBN), tetramethylethylenediamine (TMEDA),
tert-butyldimethylsilyl chloride (TBSCI), tetra-n-butylammonium
fluoride (TBAF), and diisopropyl azodicarboxylate (DIAD).
[0302] In addition, TLC refers to thin layer chromatography.
[0303] Other abbreviations: retention time (rt or R.sub.1),
minute(s) (min), hour(s) (h), room temperature (RT), concentrated
(conc.), atmosphere (atm), aqueous (aq.), saturated (sat.), eq.
(equivalent(s)).
GENERAL EXPERIMENTAL DETAILS
[0304] Unless otherwise stated the following generalizations apply.
.sup.1H NMR spectra were recorded on a Bruker Ultrashield Plus (400
MHz) or a Bruker AVANCE III (400 MHz). The multiplicity of a signal
is designated by the following abbreviations: s, singlet; d,
doublet; t, triplet; q, quartet; p, pentet; dd, doublet of
doublets; dt, doublet of triplets; tt, triplet of triplets; br,
broad; m, multiplet. All observed coupling constants, J, are
reported in Hertz (Hz). Exchangeable protons are not always
observed.
[0305] LCMS data was generated using either an Agilent 6100 Series
Single Quad (LCMS-A), an Agilent 1260 Infinity Series UPLC/MS
(LCMS-B), an Agilent 1200 (LCMS-C and LCMS-D), a Waters 2695
alliance (LCMS-E), an Agilent 6120 Single Quad (LCMS-F) or
mass-directed HPLC-MS. Chlorine isotopes are reported as .sup.35Cl,
Bromine isotopes are reported as either .sup.79Br or .sup.81Br or
both .sup.79Br/.sup.81Br.
[0306] LCMS Method A (LCMS-A):
[0307] Instrument: Agilent 6100 Series Single Quad LC/MS
[0308] Agilent 1200 Series HPLC
[0309] Pump: 1200 Series G1311A Quaternary pump
[0310] Autosampler: 1200 Series G1329A Thermostatted
Autosampler
[0311] Detector: 1200 Series G1314B Variable Wavelength
Detector
[0312] LC conditions:
[0313] Reverse Phase H PLC analysis
[0314] Column: Luna C8 (2) 5 .mu.m 50.times.4.6 mm 100 .ANG.
[0315] Column temperature: 30.degree. C.
[0316] Injection Volume: 5 .mu.L
[0317] Solvent A: Water 0.1% Formic Acid
[0318] Solvent B: MeCN 0.1% Formic Acid
[0319] Gradient: 5-100% solvent B over 10 min
[0320] Detection: 254 nm or 214 nm
[0321] MS conditions:
[0322] Ion Source: Quadrupole
[0323] Ion Mode: Multimode-ES
[0324] Drying gas temp: 300.degree. C.
[0325] Vaporizer temperature: 200.degree. C.
[0326] Capillary voltage (V): 2000 (positive)
[0327] Capillary voltage (V): 4000 (negative)
[0328] Scan Range: 100-1000
[0329] Step size: 0.1 sec
[0330] Acquisition time: 10 min
[0331] LCMS Method B (LCMS-B):
[0332] Instrument: Agilent 1260 Infinity Series UPLC/MS
[0333] Pump: 1260 Infinity G1312B Binary pump
[0334] Autosampler: 1260 Infinity G1367E 1260 HiP ALS
[0335] Detector: 1290 Infinity G4212A 1290 DAD
[0336] LC conditions:
[0337] Reverse Phase HPLC analysis
[0338] Column: Poroshell 120 EC-C18 2.7 .mu.m 50.times.3.0 mm
[0339] Column temperature: 35.degree. C.
[0340] Injection Volume: 1 .mu.L
[0341] Solvent A: Water 0.1% Formic Acid
[0342] Solvent B: MeCN 0.1% Formic Acid
[0343] Gradient: 5-100% solvent B over 3.8 min
[0344] Detection: monitored at 254 nm and 214 nm
[0345] MS conditions:
[0346] Ion Source: Quadrupole
[0347] Ion Mode: API-ES
[0348] Drying gas temp: 350.degree. C.
[0349] Capillary voltage (V): 3000 (positive)
[0350] Capillary voltage (V): 3000 (negative)
[0351] Scan Range: 100-1000
[0352] Step size: 0.1 sec
[0353] Acquisition time: 5 min
[0354] LCMS Method C (LCMS-C):
[0355] LC model: Agilent 1200
[0356] (Pump type: Binary Pump, Detector type: DAD)
[0357] MS model: Agilent G6110A Quadrupole
[0358] LC conditions:
[0359] Column: Xbridge-C18, 2.5 .mu.m, 2.1.times.30 mm
[0360] Column temperature: 30.degree. C.
[0361] Acquisition of wavelength: 214 nm, 254 nm
[0362] Mobile phase: A: 0.07% HCOOH aqueous solution, B: MeOH
[0363] MS conditions:
[0364] MS: Ion source: ES+ (or ES-) MS range: 50-900 m/z
[0365] Fragmentor: 60 Drying gas flow: 10 L/min
[0366] Nebulizer pressure: 35 psi Drying gas temperature:
350.degree. C.
[0367] Vcap: 3.5 kV
TABLE-US-00002 Gradient Table: Flow T A B (mL/min) (min) (%) (%)
0.5 0.0 70 30 0.5 0.2 70 30 0.5 1.8 5 95 0.5 2.4 5 95 0.5 2.6 70 30
0.5 3.5 70 30
[0368] Sample Preparation:
[0369] The sample was dissolved in methanol, the concentration
about 0.11-1 mg/mL, then filtered through syringe filter with 0.22
.mu.m. (Injection volume: 1-10 .mu.L)
[0370] LCMS Method D (LCMS-D):
[0371] LC model: Aqilent 1200
[0372] (Pump type: Binary Pump, Detector type: DAD)
[0373] MS model: Aqilent G6110A Quadrupole
[0374] LCMS conditions:
[0375] LC: Column: Xbridge-C18, 2.5 .mu.m, 2.1.times.30 mm
[0376] Column temperature: 30.degree. C.
[0377] Acquisition of wavelength: 214 nm, 254 nm
[0378] Mobile phase: A: 0.07% HCOOH aqueous solution, B: MeOH
[0379] MS conditions:
[0380] MS: Ion source: ES+ (or ES-) MS range: 50-900 m/z
[0381] Fragmentor: 60 Drying gas flow: 10 L/min
[0382] Nebulizer pressure: 35 psi Drying gas temperature:
350.degree. C.
[0383] Vcap: 3.5 kV
TABLE-US-00003 Gradient Table: Flow T A B (mL/min) (min) (%) (%)
0.5 0.0 70 30 0.5 0.3 70 30 0.5 0.6 50 50 0.5 0.9 40 60 0.5 1.2 30
70 0.5 3.2 5 95 0.5 3.5 5 95 0.5 4.0 70 30 0.5 5.0 70 30
[0384] Sample Preparation:
[0385] The sample was dissolved in methanol, the concentration
about 0.11-1 mg/mL, then filtered through the syringe filter with
0.22 .mu.m. (Injection volume: 1-10 .mu.L)
[0386] LCMS Method E (LCMS-E):
[0387] Equipment Information:
[0388] LC model: Waters 2695 alliance
[0389] (Pumptype: Quaternary Pump, Detector: 2996 Photodiode Array
Detector)
[0390] MS model: Micromass ZQ
[0391] LC conditions:
[0392] LC: Column: Xbridge-C18, 3.5 .mu.m, 2.1.times.50 mm
[0393] Column temperature: 30.degree. C.
[0394] Acquisition of wavelength: 214 nm, 254 nm
[0395] Mobile phase: A: 0.07% HCOOH aqueous solution, B: MeOH
[0396] MS conditions:
[0397] MS: Ion source: ES+ (or ES-) MS range: 50-900 m/z
[0398] Capillary: 3 kV Cone: 3 V Extractor: 3 V
[0399] Drying gas flow: 600 L/hr Cone: 50 L/hr
[0400] Desolvation temperature: 300.degree. C.
[0401] Source temperature: 100.degree. C.
TABLE-US-00004 Gradient Table: Flow T A B (mL/min) (min) (%) (%)
0.3 0.0 80 20 0.3 0.5 80 20 0.3 0.8 50 50 0.3 1.2 35 65 0.3 2.0 20
80 0.3 4.0 5 95 0.3 5.0 5 95 0.3 5.8 15 85 0.3 6.2 80 20 0.3 8.0 80
20
[0402] Sample Preparation:
[0403] The sample was dissolved in methanol, the concentration
about 0.11-1 mg/mL, then filtered through the syringe filter with
0.22 .mu.m. (Injection volume: 1-10 .mu.L)
[0404] LCMS Method F (LCMS-F)
[0405] Instrument: Agilent 6120 Series Single Quad LC/MS
[0406] Agilent 1200 Series HPLC
[0407] Pump: 1200 Series G1311A Quaternary pump
[0408] Autosampler: 1200 Series G1329A Thermostatted
Autosampler
[0409] Detector: 1200 Series G1314B Variable Wavelength
Detector
[0410] LC conditions:
[0411] Reverse Phase HPLC analysis
[0412] Column: Luna C.sub.8 (2) 5 .mu.m 50.times.4.6 mm 100
.ANG.
[0413] Column temperature: 30.degree. C.
[0414] Injection Volume: 1-10 .mu.L
[0415] Solvent A: Water 0.1% Formic Acid
[0416] Solvent B: MeCN 0.1% Formic Acid
[0417] Gradient: 0-95% solvent B over 10 min
[0418] Detection: 254 nm or 214 nm
[0419] MS conditions:
[0420] Ion Source: Quadrupole
[0421] Ion Mode: Multimode-ES & APCI
[0422] Drying gas temp: 250.degree. C.
[0423] Vaporizer temperature: 200.degree. C.
[0424] Capillary voltage (V): 4000 (positive)
[0425] Capillary voltage (V): 4000 (negative)
[0426] Scan Range: 100-1000
[0427] Step size: 0.1 sec
[0428] Acquisition time: 10 min
[0429] Preparative Mass-Directed HPLC
[0430] Instrument:
[0431] Waters ZQ 3100-Mass Detector
[0432] Waters 2545-Pump
[0433] Waters SFO System Fluidics Organizer
[0434] Waters 2996 Diode Array Detector
[0435] Waters 2767 Sample Manager
[0436] LC conditions:
[0437] Reverse Phase HPLC analysis
[0438] Column: XBridge TM C18 5 .mu.m 19.times.50 mm
[0439] Injection Volume 500 .mu.L
[0440] Solvent A: Water 0.1% Formic Acid
[0441] Solvent B: Acetonitrile 0.1% Formic Acid
[0442] Gradient: 25-100% B over 10 min
[0443] Flow rate: 19 mL/min
[0444] Detection: 100-600 nm
[0445] MS conditions:
[0446] Ion Source: Single-quadrupole
[0447] Ion Mode: ES positive
[0448] Source Temp: 150.degree. C.
[0449] Desolvation Temp: 350.degree. C.
[0450] Detection: Ion counting
[0451] Capillary (KV)-3.00
[0452] Cone (V): 30
[0453] Extractor (V): 3
[0454] RF Lens (V): 0.1
[0455] Scan Range: 100-1000 Amu
[0456] Scan Time: 0.5 sec
[0457] Acquisition time: 10 min
[0458] Gas Flow
[0459] Desolvation L/hour-650
[0460] Cone L/hour-100
[0461] Preparative HPLC (Prep. HPLC):
[0462] Instrument type: Varian 940-LC series;
[0463] Pump type: Quaternary Pump;
[0464] Detector type: Diode Array Detector
[0465] HPLC conditions:
[0466] Waters Sunfire prep C18 OBD, 5 .mu.m 19.times.100 mm column,
eluting with a gradient of MeOH in water with 0.07% TFA at a flow
rate of 15 mL/min. Acquisition wavelength 214 nm, 254 nm.
[0467] Analytical thin-layer chromatography was performed on Merck
silica gel 60 F254 aluminium-backed plates which were visualised
using fluorescence quenching under UV light or a basic KMnO.sub.4
dip or Ninhydrin dip.
[0468] Preparative thin-layer chromatography (preparative TLC or
prep. TLC) was performed using Tklst (China), grand grade: (HPTLC):
8.+-.2 .mu.m>80%; (TLC): 10-40 .mu.m. Type: GF254. Compounds
were visualised by UV (254 nm).
[0469] Column chromatography was performed using a Biotage Isolera
purification system using either Grace or RediSep.RTM. silica
cartridges or with Tklst (China), grand grade, 100-200 meshes
silica gel.
[0470] Microwave irradiation was achieved using a CEM Explorer SP
Microwave Reactor. Where necessary, anhydrous solvents were
purchased from Sigma-Aldrich or dried using conventional
methods.
[0471] Unless stated otherwise, acidification was done with
concentrated or aqueous solution of HCl.
[0472] Additional cartridges used are as follows:
[0473] Phase Separator:
[0474] Manufacturer: Biotage
[0475] Product: ISOLUTE.RTM. Phase Separator (3 mL unless otherwise
stated)
[0476] Si-Amine Cartridges:
[0477] Manufacturer: Biotage
[0478] Product: Isolute.RTM. NH2, 1 g/6 mL
[0479] Or
[0480] Manufacturer: Silicycle
[0481] Product: Si-amine 500 mg or 1 g
Synthesis of Intermediates
i) 6-(Methoxymethyl)-5-methylbenzo[d]isoxazol-3-amine I4
##STR00027##
[0482] a) Methyl 4-cyano-5-fluoro-2-methylbenzoate I1
[0483] A mixture of 4-bromo-2-fluoro-5-methylbenzonitrile (3.5 g,
16.4 mmol), Pd(dppf)Cl.sub.2.DCM (668 mg, 0.82 mmol) and Et.sub.3N
(5.0 g, 49.1 mmol) in MeOH (80 mL) was heated at 100.degree. C.
under a CO atmosphere (0.2 MPa) overnight. Additional
Pd(dppf)Cl.sub.2.DCM (340 mg, 0.4 mmol) was added and heating was
continued under a CO atmosphere (0.2 MPa) overnight. The catalyst
was removed by filtration, washed with MeOH and the filtrate was
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=20/1 to 10/1 to 5/1) to
give the title compound (2.4 g, 74%) as a yellow solid. LCMS-D:
R.sub.t 2.48 min; m/z 216.1 [M+Na].sup.+.
b) 2-Fluoro-4-(hydroxymethyl)-5-methylbenzonitrile I2
[0484] To a solution of methyl 4-cyano-5-fluoro-2-methylbenzoate I1
(2.4 g, 12.4 mmol) in anhydrous THF (20 mL) at RT under N.sub.2 was
added LiBH.sub.4 (2.0 M solution in THF, 12.4 mL, 24.8 mmol)
dropwise and the mixture was heated at reflux for 2 h. The reaction
was quenched with water (80 mL) and the mixture was extracted with
EtOAc (90 mL.times.3). The combined organic extracts were washed
with water (100 mL.times.3), brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=20/1 to 10/1 to 5/1) to give the title compound (1.6 g,
79%) as a yellow solid. LCMS-D: R.sub.t 1.43 min; m/z 166.1
[M+H].sup.+, 188.1 [M+Na].sup.+.
c) 2-Fluoro-4-(methoxymethyl)-5-methylbenzonitrile I3
[0485] To a solution of
2-fluoro-4-(hydroxymethyl)-5-methylbenzonitrile I2 (800 mg, 8.8
mmol) and iodomethane (3.4 g, 24.2 mmol) in DMF (12 mL) at
0.degree. C. was added NaH (60% w/w dispersion in oil, 379 mg, 9.7
mmol) and the mixture was stirred at 0.degree. C. for 30 min. Water
was added and the mixture was extracted with EtOAc (50 mL.times.3).
The combined organic extracts were washed with brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography (Pet.
ether/EtOAc=20/1 to 10/1 to 5/1) to give the title compound (660
mg, 76%) as a yellow solid. LCMS-D: R.sub.t 2.44 min; m/z 180.1
[M+H].sup.+, 202.1.1 [M+Na].sup.+.
d) 6-(Methoxymethyl)-5-methylbenzo[d]isoxazol-3-amine I4
[0486] To a solution of acetohydroxamic acid (792 mg, 10.6 mmol) in
anhydrous DMF (20 mL) at 0.degree. C. was added potassium
tert-butoxide (1.2 g, 10.6 mmol) and the mixture was stirred at RT
for 2 h. 2-Fluoro-4-(methoxymethyl)-5-methylbenzonitrile I3 (630
mg, 3.5 mmol) was then added and the mixture was heated at
60.degree. C. overnight. Water was added and the mixture was
extracted with EtOAc (80 mL.times.3). The combined organic extracts
were washed with brine, dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by column chromatography (DCM/MeOH=20/1 to 10/1) to give
the title compound (1.0 g, 77%) as a yellow solid. LCMS-D: R.sub.t
1.75 min; m/z 193.1 [M+H].sup.+.
ii) 4-Methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine I9
##STR00028##
[0487] a) 4-Bromo-2-fluoro-6-methoxybenzonitrile I5
[0488] To a solution of 4-bromo-2,6-difluorobenzonitrile (6.0 g,
27.5 mmol) in THF (100 mL) was added sodium methanolate (1.5 g,
55.0 mmol) and the mixture was stirred at RT for 48 h. Water was
added and the mixture was extracted with EtOAc (150 mL.times.3).
The combined organic extracts were washed with brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography (Pet.
ether/EtOAc=300/1 to 200/1) to give the title compound (4.3 g, 68%)
as a white solid. LCMS-D: R.sub.t 2.53 min; m/z 251.8/253.8
[M+Na].sup.+.
b) Methyl 4-cyano-3-fluoro-5-methoxybenzoate I6
[0489] A mixture of 4-bromo-2-fluoro-6-methoxybenzonitrile I5 (4.3
g, 18.7 mmol), Pd(dppf)Cl.sub.2.DCM (768 mg, 0.94 mmol) and
Et.sub.3N (5.7 g, 56.1 mmol) in MeOH (50 mL) was heated at
100.degree. C. under a CO atmosphere (0.2 MPa) overnight. The
catalyst was removed by filtration, washed with MeOH and the
filtrate was concentrated under reduced pressure. The residue was
purified by column chromatography (Pet. ether/EtOAc=200/1 to 50/1)
to give the title compound (2.9 g, 74%) as a white solid. LCMS-D:
R.sub.t 2.41 min; m/z 210.0 [M+H].sup.+, 232.0 [M+Na].sup.+.
c) 2-Fluoro-4-(hydroxymethyl)-6-methoxybenzonitrile I7
[0490] To a solution of LiBH.sub.4 (2.0 M solution in THF, 13.9 mL,
27.8 mmol) in anhydrous THF (60 mL) at RT under N.sub.2 was added a
solution of methyl 4-cyano-3-fluoro-5-methoxybenzoate 16 (2.9 g,
13.9 mmol) in anhydrous THF (10 mL) dropwise and the mixture was
heated at reflux for 1 h. The reaction was quenched with 1 M aq.
HCl and extracted with EtOAc (100 mL.times.3). The combined organic
extracts were washed with water (100 mL.times.3), brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give the
title compound (2.5 g, 100%) as a white solid. LCMS-D: R.sub.1 2.31
min; m/z 182.1 [M+H].sup.+, 204.1 [M+Na].sup.+.
d) 2-Fluoro-6-methoxy-4-(methoxymethyl)benzonitrile I8
[0491] To a solution of
2-fluoro-4-(hydroxymethyl)-6-methoxybenzonitrile I7 (2.7 g, 14.9
mmol) and iodomethane (10.6 g, 74.5 mmol) in DMF (100 mL) at
0.degree. C. was added NaH (60% w/w dispersion in oil, 1.2 g, 29.8
mmol) in small portions and the mixture was stirred at RT for 30
min. Water was added and the mixture was extracted with EtOAc (100
mL.times.3). The combined organic extracts were washed with brine,
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by column
chromatography (Pet. ether/EtOAc=100/1 to 5/1) to give the title
compound (2.2 g, 76%) as a yellow solid. LCMS-D: R.sub.t 2.22 min;
m/z 218.0 [M+Na].sup.+.
e) 4-Methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine I9
[0492] To a solution of acetohydroxamic acid (2.3 g, 30.8 mmol) in
anhydrous DMF (1500 mL) at RT was added potassium tert-butoxide
(3.5 g, 30.8 mmol) and the mixture was stirred at RT for 1 h.
2-Fluoro-6-methoxy-4-(methoxymethyl)benzonitrile I8 (2.0 g, 10.3
mmol) was then added and stirring was continued at RT overnight.
Water was added and the mixture was extracted with EtOAc. The
combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (DCM/MeOH=20/1 to
10/1 to 3/1) to give the title compound (580 mg, 27%) as a yellow
solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 6.92 (d, J=0.8
Hz, 1H), 6.65 (s, 1H), 5.91 (s, 2H), 4.48 (s, 2H), 3.90 (s, 3H),
3.32 (s, 3H, obscured by water peak). LCMS-D: R.sub.t 1.33 min; m/z
209.0 [M+H].sup.+.
iii) 4--Nitrobenzo[d]isoxazol-3-amine I10
##STR00029##
[0494] To a solution of 2-fluoro-6-nitrobenzonitrile (1.0 g, 6.17
mmol) in DMF/H.sub.2O (32 mL/32 mL) was added acetohydroxamic acid
(2.78 g, 37.0 mmol) and K.sub.2CO.sub.3 (10.23 g, 74.0 mmol) and
the mixture was heated at 70.degree. C. for 19 h. Water (200 mL)
was added and the mixture was extracted with EtOAc (100
mL.times.3). The combined organic extracts were washed with water,
brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=30/1 to 1/1) to give the
title compound (380 mg, 35%) as a yellow solid. LCMS-D: R.sub.t
2.82 min; m/z 180.1 [M+H].sup.+.
iv) 4-Methoxy-6-(1-methoxyethyl)benzo[d]isoxazol-3-amine I15
##STR00030##
[0495] a) 4-(1-Ethoxyvinyl)-2-fluoro-6-methoxybenzonitrile I11
[0496] To a solution of 4-bromo-2-fluoro-6-methoxybenzonitrile I5
(2.0 g, 8.7 mmol) in THF (40 mL) was added
tributyl(1-ethoxyvinyl)stannane (3.4 g, 9.6 mmol),
Pd(PPh.sub.3).sub.4 (201 mg, 0.174 mmol) and LiCl (1.15 g, 27.0
mmol) and the mixture was heated at reflux under N.sub.2 for 48 h.
The mixture was diluted with EtOAc and washed consecutively with
water, 5% aqueous ammonium hydroxide solution and brine. The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure. The residue was purified
by column chromatography (Pet. ether/EtOAc=200/1) to give the title
compound (1.6 g, 84%) as a light yellow solid. LCMS-C: R.sub.t 2.41
min; m/z 222.0 [M+H].sup.+.
b) 4-Acetyl-2-fluoro-6-methoxybenzonitrile I12
[0497] To a solution of
4-(1-ethoxyvinyl)-2-fluoro-6-methoxybenzonitrile I11 (1.0 g, 4.5
mmol) in THF (10 mL) was added 2 M aq. HCl (6.0 mL) and the mixture
was stirred at RT for 3 h. The mixture was diluted with diethyl
ether and washed with a saturated aqueous NaHCO.sub.3 solution and
water. The organic layer was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give the title
compound (710 mg, 81%) as a white solid. LCMS-C: R.sub.t 1.42 min;
m/z 194.0 [M+H].sup.+.
c) 2-Fluoro-4-(1-hydroxyethyl)-6-methoxybenzonitrile I13
[0498] To a solution of 4-acetyl-2-fluoro-6-methoxybenzonitrile I12
(700 mg, 3.6 mmol) in THF (30 mL) was added sodium borohydride (206
mg, 5.4 mmol) and the mixture was stirred at RT overnight. Water
was added and the mixture was extracted with EtOAc (30 mL.times.3).
The combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give the title compound (675 mg, 95%) as a colorless oil.
LCMS-C: R.sub.t 0.98 min; m/z 196.0 [M+H].sup.+.
d) 2-Fluoro-6-methoxy-4-(1-methoxyethyl)benzonitrile I14
[0499] To a solution of
2-fluoro-4-(1-hydroxyethyl)-6-methoxybenzonitrile I13 (670 mg, 3.4
mmol) and iodomethane (1.5 g, 10.3 mmol) in DMF (20 mL) at
0.degree. C. was added NaH (60% w/w dispersion in oil, 274 mg, 6.8
mmol) in small portions and the mixture was stirred at 0.degree. C.
for 2 h. Water was added and the mixture was extracted with EtOAc
(40 mL.times.3). The combined organic extracts were dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure to give the title compound (650 mg, 90%) as a light yellow
solid. LCMS-C: R.sub.t 1.95 min; m/z 210.0 [M+H].sup.+.
e) 4-Methoxy-6-(1-methoxyethyl)benzo[d]isoxazol-3-amine I15
[0500] To a solution of acetohydroxamic acid (698 mg, 9.3 mmol) in
anhydrous DMF (20 mL) at 0.degree. C. was added potassium
tert-butoxide (1.04 g, 9.3 mmol) and the mixture was stirred at RT
for 1 h. A solution of
2-fluoro-6-methoxy-4-(1-methoxyethyl)benzonitrile I14 (650 mg, 3.1
mmol) in anhydrous DMF (10 mL) was then added dropwise and the
mixture was stirred at RT overnight. Water was added and the
mixture was extracted with EtOAc (50 mL.times.3). The combined
organic extracts were dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by column chromatography (DCM/MeOH=300/1 to 200/1) to give
the title compound (130 mg, 19%) as a yellow solid. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 6.90 (s, 1H), 6.64 (s, 1H), 5.92
(s, 2H), 4.39 (q, J=6.4 Hz, 1H), 3.90 (s, 3H), 3.15 (s, 3H), 1.36
(d, J=6.4 Hz, 3H). LCMS-C: R.sub.t 0.73 min; m/z 223.0
[M+H].sup.+.
v) 4-Methoxy-6-phenylbenzo[d]isoxazol-3-amine I17
##STR00031##
[0501] a) 3-Fluoro-5-methoxy-[1,1'-biphenyl]-4-carbonitrile I16
[0502] To a solution of 4-bromo-2-fluoro-6-methoxybenzonitrile I5
(6.0 g, 26.1 mmol) and phenylboronic acid (6.36 g, 52.2 mmol) in
1,4-dioxane (200 mL) and water (50 mL) under N.sub.2 was added
Pd(PPh.sub.3).sub.4 (2.99 g, 2.66 mmol) and Na.sub.2CO.sub.3 (8.29
g, 78.2 mmol) and the mixture was heated at 100.degree. C.
overnight. Water was added and the mixture was extracted with
EtOAc. The combined organic extracts were washed with brine, dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by column chromatography
(Pet. ether/EtOAc=100/1 to 50/1) to give the title compound (5.45
g, 93%) as a yellow solid. LCMS-C: R.sub.t 2.48 min; m/z 228.0
[M+H].sup.+.
b) 4-Methoxy-6-phenylbenzo[d]isoxazol-3-amine I17
[0503] To a solution of acetohydroxamic acid (8.15 g, 23.98 mmol)
in anhydrous DMF (200 mL) at 0.degree. C. was added potassium
tert-butoxide (5.5 g, 24.0 mmol) and the mixture was stirred at RT
for 1 h. 3-Fluoro-5-methoxy-[1,1'-biphenyl]-4-carbonitrile I16
(5.45 g, 7.99 mmol) was then added and the mixture was heated at
60.degree. C. for 4 h. Water was added and the mixture was
extracted with EtOAc. The combined organic extracts were washed
with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=10/1 to 8/1 to 6/1) to give
the title compound (2.2 g, 38%) as a yellow solid. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 7.79-7.74 (m, 2H), 7.52-7.46 (m, 2H),
7.45-7.39 (m, 1H), 7.26 (d, J=1.1 Hz, 1H), 6.95 (s, 1H), 5.97 (s,
2H), 4.00 (s, 3H). LCMS-C: R.sub.t 2.15 min; m/z 241.0
[M+H].sup.+
vi) 3-(3-Amino-4-methoxybenzo[d]isoxazol-6-yl)phenol I19
##STR00032##
[0504] a)
3-Fluoro-3'-hydroxy-5-methoxy-[1,1'-biphenyl]-4-carbonitrile
I18
[0505] To a solution of 4-bromo-2-fluoro-6-methoxybenzonitrile I5
(650 mg, 2.8 mmol) and (3-hydroxyphenyl)boronic acid (1.2 g, 5.6
mmol) in 1,4-dioxane (40 mL) and water (10 mL) under N.sub.2 was
added Pd(PPh.sub.3).sub.4 (327 mg, 0.28 mmol) and Na.sub.2CO.sub.3
(899 mg, 8.5 mmol) and the mixture was heated at 100.degree. C.
overnight. Water was added and the mixture was extracted with
EtOAc. The combined organic extracts were washed with brine, dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by column chromatography
(Pet. ether/EtOAc=3/1) to give the title compound (687 mg, 94%) as
a yellow solid. LCMS-C: R.sub.t 2.07 min; m/z 244.0
[M+H].sup.+.
b) 3-(3-Amino-4-methoxybenzo[d]isoxazol-6-yl)phenol I19
[0506] To a solution of acetohydroxamic acid (636 mg, 8.5 mmol) in
anhydrous DMF (60 mL) at 0.degree. C. was added potassium
tert-butoxide (952 mg, 8.5 mmol) and the mixture was stirred at RT
for 1 h.
3-Fluoro-3'-hydroxy-5-methoxy-[1,1'-biphenyl]-4-carbonitrile I18
(687 mg, 2.8 mmol) was then added and the mixture was heated at
60.degree. C. for 4 h. Water was added and the mixture was
extracted with EtOAc. The combined organic extracts were washed
with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=1/1) to give the title
compound (282 mg, 39%) as a yellow solid. LCMS-C: R.sub.t 2.3 min;
m/z 257.0 [M+H].sup.+. [0507] vii)
6-(Ethoxymethyl)-4-methoxybenzo[d]isoxazol-3-amine I21
##STR00033##
[0507] a) 4-(Ethoxymethyl)-2-fluoro-6-methoxybenzonitrile I20
[0508] To a solution of
2-fluoro-4-(hydroxymethyl)-6-methoxybenzonitrile I7 (1.15 g, 6.4
mmol) and iodoethane (5.0 g, 31.7 mmol) in DMF (40 mL) at 0.degree.
C. was added NaH (60% w/w dispersion in oil, 508 mg, 12.7 mmol) in
small portions and the mixture was stirred at RT for 30 min. Water
was added and the mixture was extracted with EtOAc (100
mL.times.3). The combined organic extracts were washed with brine,
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by column
chromatography (Pet. ether/EtOAc=2/1) to give the title compound
(1.0 g, 79%) as a yellow solid. LCMS-C: R.sub.t 2.15 min; m/z 210.0
[M+H].sup.+.
b) 6-(Ethoxymethyl)-4-methoxybenzo[d]isoxazol-3-amine I21
[0509] To a solution of acetohydroxamic acid (1.1 g, 14.3 mmol) in
anhydrous DMF (50 mL) at RT was added potassium tert-butoxide (1.6
g, 14.3 mmol) and the mixture was stirred at RT for 1 h.
4-(Ethoxymethyl)-2-fluoro-6-methoxybenzonitrile I20 (1.0 g, 4.8
mmol) was then added and the mixture was stirred at RT overnight.
Water was added and the mixture was extracted with EtOAc. The
organic extracts were dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by column chromatography (DCM/MeOH=20/1 to 10/1 to 3/1) to
give the title compound (650 mg, 61%) as a yellow oil. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 6.92 (s, 1H), 6.65 (s, 1H), 5.91
(s, 2H), 4.53 (s, 2H), 3.89 (s, 3H), 3.51 (q, J=7.0 Hz, 2H), 1.17
(t, J=7.0 Hz, 3H). LCMS-C: R.sub.t 0.82 min; m/z 223.0
[M+H].sup.+.
viii) 6-Bromo-4-methoxybenzo[d]isoxazol-3-amine I22
##STR00034##
[0511] To a solution of acetohydroxamic acid (2.0 g, 26.1 mmol) in
anhydrous DMF (100 mL) at RT was added potassium tert-butoxide (2.9
g, 26.1 mmol) and the mixture was stirred at RT for 1 h.
4-Bromo-2-fluoro-6-methoxybenzonitrile I5 (2.0 g, 8.7 mmol) was
then added and the mixture was stirred at RT overnight. Water was
added and the mixture was extracted with EtOAc. The organic
extracts were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=2/1) to give the title
compound (296 mg, 14%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.32 (d, J=1.2 Hz, 1H), 6.90 (d, J=1.2 Hz,
1H), 6.04 (s, 2H), 3.92 (s, 3H). LCMS-C: R.sub.t 1.4 min; m/z 244.0
[M+H].sup.+.
ix) 7-Ethoxybenzo[d]isoxazol-3-amine I26
##STR00035##
[0512] a)
2-Fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonit-
rile I23
[0513] A mixture of 3-bromo-2-fluorobenzonitrile (3.0 g, 15.0
mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane)
(11.4 g, 45 mmol), potassium acetate (5.9 g, 60.0 mmol) and
Pd(dppf)Cl.sub.2 (2.2 g, 3.0 mmol) in DMSO (45 mL) and 1,4-dioxane
(15 mL) was heated at 105.degree. C. under N.sub.2 for 3 h. The
mixture was diluted with EtOAc (30 mL) and washed with water (30
mL.times.3). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered, concentrated under reduced pressure and
the residue was purified by column chromatography (Pet.
ether/EtOAc=20/1) to give the title compound (3.9 g, >100%) as a
white solid, which was used directly in the next step.
b) 2-Fluoro-3-hydroxybenzonitrile I24
[0514] To a solution of
2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile
I23 (1.9 g, 7.6 mol) in AcOH (19 mL) under N.sub.2 was added
H.sub.2O.sub.2 (30% aqueous solution, 1.9 mL) dropwise and the
mixture was stirred at RT for 2 h then poured into a mixture of
EtOAc and excess aqueous Na.sub.2SO.sub.3. The layers were then
separated and the organic layer was washed with water, brine, dried
over anhydrous Na.sub.2SO.sub.4, filtered, concentrated under
reduced pressure to give the title compound (650 mg, 62%) as an
off-white waxy solid. LCMS-D: R.sub.t 0.93 min; m/z 138.1
[M+H].sup.+.
c) 3-Ethoxy-2-fluorobenzonitrile I25
[0515] To a solution of 2-fluoro-3-hydroxybenzonitrile I24 (360 mg,
2.6 mmol) in DMF (30 mL) was added Cs.sub.2CO.sub.3 (4.3 g, 13.1
mmol) and iodoethane (1.0 g, 6.6 mmol) and the mixture was stirred
at RT overnight. The mixture was diluted with EtOAc (80 mL) and
washed with water (50 mL.times.3). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered, concentrated under reduced
pressure to give the title compound (220 mg, 51%) as a yellow
solid. LCMS-D: R.sub.t 2.31 min; m/z 166.1 [M+H].sup.+.
d) 7-Ethoxybenzo[d]isoxazol-3-amine I26
[0516] To a solution of acetohydroxamic acid (300 mg, 4.0 mmol) in
DMF (15 mL) at 0.degree. C. under N.sub.2 was added potassium
tert-butoxide (450 mg, 4.0 mmol) and the mixture was heated at
30.degree. C. for 1 h. A solution of 3-ethoxy-2-fluorobenzonitrile
I25 (220 mg, 1.3 mmol) in DMF (10 mL) was added and the mixture was
heated at 30.degree. C. overnight. EtOAc (80 mL) was added and the
mixture was washed with water (50 mL.times.3). The organic layer
was dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated
under reduced pressure to give the title compound (170 mg, 70%) as
a yellow solid. LCMS-D: R.sub.t 1.68 min; m/z 179.1
[M+H].sup.+.
x) 7-(Cyclopropylmethoxy)benzo[d]isoxazol-3-amine I28
##STR00036##
[0517] a) 3-(Cyclopropylmethoxy)-2-fluorobenzonitrile I27
[0518] To a solution of 2-fluoro-3-hydroxybenzonitrile I24 (360 mg,
2.6 mmol) in DMF (30 mL) was added Cs.sub.2CO.sub.3 (4.3 g, 13.1
mmol), KI (87 mg, 0.5 mmol) and (bromomethyl)cyclopropane (880 mg,
6.6 mmol) and the mixture was stirred at RT overnight. EtOAc (80
mL) was added and the mixture was washed with water (50
mL.times.3). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered, concentrated under reduced pressure to
give the title compound (150 mg, 30%) as a red solid. LCMS-D:
R.sub.t2.54 min; m/z 192.1 [M+H].sup.+.
b) 7-(Cyclopropylmethoxy)benzo[d]isoxazol-3-amine I28
[0519] Prepared from 3-(cyclopropylmethoxy)-2-fluorobenzonitrile
I27 according to the procedure described for
7-ethoxybenzo[d]isoxazol-3-amine I26, step d. LCMS-D: R.sub.t 2.23
min; m/z 205.1 [M+H].sup.+.
xi) 6-Ethoxybenzo[d]isoxazol-3-amine I32
##STR00037##
[0520] a) 2-Fluoro-4-((tetrahydro-2H-pyran-2-yl)oxy)benzonitrile
I29
[0521] To a solution of 2-fluoro-4-hydroxybenzonitrile (20 g, 145.9
mmol) and PPTS (733 mg, 2.9 mmol) in DCM (500 mL) under N.sub.2 was
added 3,4-dihydro-2H-pyran (24.5 g, 292 mmol) and the mixture was
stirred at RT overnight. The solvent was removed under reduced
pressure and the residue was purified by column chromatography
(Pet. ether/EtOAc=100/0 to 100/2) to give the title compound (27 g,
83%) as a white solid, which was used directly in the next
step.
b) 6-((Tetrahydro-2H-pyran-2-yl)oxy)benzo[d]isoxazol-3-amine
I30
[0522] To a solution of acetohydroxamic acid (13.7 g, 182.3 mmol)
in DMF (60 mL) at 0.degree. C. under N.sub.2 was added potassium
tert-butoxide (20.4 g, 182.3 mmol) and the mixture was stirred at
RT for 1 h. 2-Fluoro-4-((tetrahydro-2H-pyran-2-yl)oxy)benzonitrile
I29 (13.4 g, 60.8 mmol) was then added and the mixture was stirred
at RT overnight. EtOAc (500 mL) was added and the mixture was
washed with water (100 mL.times.5). The organic layer was dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by column chromatography
(Pet. ether/EtOAc=100/1 to 5/1) to give the title compound (12.1 g,
85%) as a white solid. LCMS-D: R.sub.t 2.31 min; m/z 235.1
[M+H].sup.+.
c) 3-Aminobenzo[d]isoxazol-6-ol I31
[0523] To a solution of
6-((tetrahydro-2H-pyran-2-yl)oxy)benzo[d]isoxazol-3-amine I30 (3.5
g, 15 mmol) in THF (50 mL) was added 2 M aq. HCl (20 mL) and the
mixture was stirred at RT for 3 h. The mixture was diluted with
EtOAc (300 mL) and washed with water (.times.2). The organic layer
was dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated
under reduced pressure to give the title compound (2.1 g, 94%) as a
white solid, which was used directly in the next step.
d) 6-Ethoxybenzo[d]isoxazol-3-amine I32
[0524] A mixture of 3-aminobenzo[d]isoxazol-6-ol I31 (300 mg, 2
mmol), Cs.sub.2CO.sub.3 (2.0 g, 6 mmol), KI (66 mg, 0.4 mmol) and
bromoethane (436 mg, 4 mmol) in DMF (30 mL) was heated at
50.degree. C. under N.sub.2 overnight. The mixture was diluted with
EtOAc (300 mL) and washed with water (100 mL.times.5). The organic
layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=100/1 to 5/1) to give the
title compound (270 mg, 76%) as a white solid. LCMS-D: R.sub.t 0.37
min; m/z 179.0 [M+H].sup.+.
xii) 6-(Cyclopropylmethoxy)benzo[d]isoxazol-3-amine I33
##STR00038##
[0526] Prepared from 3-aminobenzo[d]isoxazol-6-ol I31 according to
the procedure described for 6-ethoxybenzo[d]isoxazol-3-amine I32,
step d (395 mg, 97%). LCMS-D: R.sub.12.27 min; m/z 205.1
[M+H].sup.+.
xiii) 6-(1H-1,2,3-Triazol-1-yl)benzo[d]isoxazol-3-amine I36
##STR00039##
[0527] a) 4-Azido-2-fluorobenzonitrile I34
[0528] A mixture of 4-amino-2-fluorobenzonitrile (2.0 g, 14.7 mmol)
in water (4 mL), ACN (32 mL), and concentrated HCl (10 mL) was
stirred at RT under N.sub.2 overnight. NaNO.sub.2 (2.0 g, 29.4
mmol) was then added portion-wise and stirring was continued at RT
for 2 h. The mixture was cooled to 0.degree. C., NaN.sub.3 (1.9 g,
29.4 mmol) was added portion-wise and stirring was continued at RT
for 2 h. Water (50 mL) was added and most of the organic solvent
was removed under reduced pressure. The remaining aqueous mixture
was then extracted with DCM (50 mL.times.4) and the combined
organic extracts were washed with water, brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography (Pet.
ether/EtOAc=100/0 to 20/1) to give the title compound (1.5 g, 62%)
as a yellow solid, which was used directly in the next step.
b) 2-Fluoro-4-(1H-1,2,3-triazol-1-yl)benzonitrile I35
[0529] A mixture of 4-azido-2-fluorobenzonitrile I34 (500 mg, 3.1
mmol), ethynyltrimethylsilane (454 mg, 4.6 mmol) and CuI (704 mg,
3.7 mmol) in THF (50 mL) was heated at 50.degree. C. under N.sub.2
for 24 h. Additional ethynyltrimethylsilane (454 mg, 4.6 mmol) was
added and the mixture was heated at 50.degree. C. for a further 24
h, then concentrated under reduced pressure. The residue was
purified by column chromatography (Pet. ether/EtOAc=100/1 to 10/1)
to give
2-fluoro-4-(5-(trimethylsilyl)-1H-1,2,3-triazol-1-yl)benzonitrile
(410 mg), which was dissolved in a 1 M solution of TBAF in THF (50
mL) and heated at 45.degree. C. under N.sub.2 overnight. The
solvent was removed under reduced pressure and the residue was
purified by column chromatography (Pet. ether/EtOAc=50/1 to 2/1) to
give the title compound (200 mg, 34%) as a white solid, which was
used directly in the next step.
c) 6-(1H-1,2,3-Triazol-1-yl)benzo[d]isoxazol-3-amine I36
[0530] To a solution of acetohydroxamic acid (239 mg, 3.16 mmol) in
DMF (25 mL) at 0.degree. C. under N.sub.2 was added potassium
tert-butoxide (357 mg, 3.18 mmol) and the mixture was stirred at RT
for 2 h. A solution of
2-fluoro-4-(1H-1,2,3-triazol-1-yl)benzonitrile I35 (200 mg, 1.06
mmol) in DMF (15 mL) was then added and stirring was continued at
RT overnight. EtOAc (100 mL) was added and the mixture was washed
with water (.times.5). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=100/1 to 2/1) to give the title compound (150 mg, 70%)
as a white solid. LCMS-D: R.sub.t 0.47 min; m/z 202.1
[M+H].sup.+.
xiv) 6-(Pyrimidin-2-yl)benzo[d]isoxazol-3-amine I39
##STR00040##
[0531] a)
2-Fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonit-
rile I37
[0532] A mixture of 4-bromo-2-fluorobenzonitrile (1.0 g, 5.0 mmol),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (1.3 g,
5.0 mmol), potassium acetate (5.9 g, 20.0 mmol) and
Pd(dppf)Cl.sub.2 (2.0 g, 1.0 mmol) in DMSO (50 mL) and 1,4-dioxane
(10 mL) was heated at 105.degree. C. under N.sub.2 for 3 h. The
mixture was diluted with EtOAc (200 mL) and washed with water (100
mL.times.3). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered, concentrated under reduced pressure and
the residue was purified by column chromatography (Pet.
ether/EtOAc=100/0 to 50/1) to give the title compound (1.1 g, 89%)
as a white solid, which was used directly in the next step.
b) 2-Fluoro-4-(pyrimidin-2-yl)benzonitrile I38
[0533] To a solution of
2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile
I37 (464 mg, 2 mmol) and 2-bromopyrimidine (736 mg, 4 mmol) in
water (40 mL), toluene (40 mL) and i-PrOH (10 mL) under N.sub.2 was
added Pd(dppf)Cl.sub.2 (146 mg, 0.2 mmol) and
K.sub.3PO.sub.4.3H.sub.2O (1.33 g, 5.0 mmol) and the mixture was
heated at 85.degree. C. for 4 h. The mixture was diluted with EtOAc
(200 mL) and washed with water (50 mL). The organic layer was dried
over anhydrous Na.sub.2SO.sub.4, filtered, concentrated under
reduced pressure and the residue was purified by column
chromatography (Pet. ether/EtOAc=50/1 to 2/1) to give the title
compound (270 mg, 68%) as a white solid. LCMS-D: R.sub.t 2.38 min;
m/z 200.1 [M+H].sup.+.
c) 6-(Pyrimidin-2-yl)benzo[d]isoxazol-3-amine I39
[0534] To a solution of acetohydroxamic acid (306 mg, 4.07 mmol) in
DMF (20 mL) at 0.degree. C. under N.sub.2 was added potassium
tert-butoxide (457 mg, 4.07 mmol) and the mixture was heated at
30.degree. C. for 1 h. A solution of
2-fluoro-4-(pyrimidin-2-yl)benzonitrile I38 (270 mg, 1.36 mmol) in
DMF (10 mL) was then added and heating was continued at 30.degree.
C. overnight. The mixture was diluted with EtOAc (100 mL) and
washed with water (50 mL.times.3). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered, concentrated under reduced
pressure and the residue was purified by column chromatography
(Pet. ether/EtOAc=50/1 to 2/1) to give the title compound (200 mg,
69%) as a white solid. LCMS-D: R.sub.t 0.38 min; m/z 213.1
[M+H].sup.+, 235.1 [M+Na].sup.+.
xv) 5-Bromobenzo[d]isoxazol-3-amine I40
##STR00041##
[0536] To a solution of acetohydroxamic acid (23.7 g, 0.315 mol) in
DMF (800 mL) at 0.degree. C. under N.sub.2 was added t-BuOK (35.4
g, 0.315 mol) and the mixture was stirred at 15.degree. C. for 2 h.
5-Bromo-2-fluorobenzonitrile (21.0 g, 0.105 mol) was then added and
the mixture was stirred at RT overnight. The mixture was diluted
with EtOAc (1.5 L) and washed with water (400 mL.times.4). The
organic layer was washed with brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered, concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=50/1 to 3/1) to give the title compound (19 g, 86%) as
a white solid. LCMS-D: R.sub.t 2.13 min; m/z 212.9/214.9
[M+H].sup.+.
xvi) 4-Bromobenzo[d]isoxazol-3-amine I41
##STR00042##
[0538] To a solution of acetohydroxamic acid (11.25 g, 0.15 mol) in
DMF (220 mL) at 0.degree. C. under N.sub.2 was added t-BuOK (16.8
g, 0.15 mol) and the mixture was stirred at 25.degree. C. for 1 h.
A solution of 2-bromo-6-fluorobenzonitrile (10.0 g, 0.05 mol) in
DMF (80 mL) was then added dropwise and stirring was continued at
25.degree. C. overnight. The mixture was diluted with water (200
mL) and extracted with EtOAc (400 mL). The organic extract was
washed with water (400 mL.times.3), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give the title compound (7.0 g, 66%) as a light red solid.
LCMS-D: R.sub.t 2.05 min; m/z 212.9/214.9 [M+H].sup.+.
xvii) 4-(Trifluoromethyl)benzo[d]isoxazol-3-amine I42
##STR00043##
[0540] To a solution of acetohydroxamic acid (2.25 g, 30 mmol) in
DMF (80 mL) at 0.degree. C. under N.sub.2 was added t-BuOK (3.37 g,
30 mmol) and the mixture was heated at 30.degree. C. for 1 h. A
solution of 2-fluoro-6-(trifluoromethyl)benzonitrile (1.89 g, 10
mmol) in DMF (20 mL) was then added and heating was continued at
30.degree. C. overnight. The mixture was partitioned between EtOAc
(300 mL) and water (100 mL), the layers were separated and the
organic layer was washed with water (100 mL.times.3), brine, dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by column chromatography
(Pet. ether/EtOAc=50/1 to 1/1) to give the title compound (1.3 g,
64%) as a white solid LCMS-D: R.sub.t 2.19 min; m/z 203.0
[M+H].sup.+.
xviii) 5-Bromo-4-chlorobenzo[d]isoxazol-3-amine I44
##STR00044##
[0541] a) 3-Bromo-2-chloro-6-fluorobenzonitrile I43
[0542] To a solution of 2-chloro-6-fluorobenzonitrile (1.0 g, 6.4
mmol) in trifluoromethanesulfonic acid (10 mL) at 0.degree. C.
under N.sub.2 was added NBS (1.1 g, 6.4 mmol) and the mixture was
stirred at RT overnight. The mixture was poured onto ice and
extracted with EtOAc (30 mL.times.2). The combined organic extracts
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=200/1 to 100/1) to give the
title compound (705 mg, 47%) as a white solid, which was used
directly in the next step.
b) 5-Bromo-4-chlorobenzo[d]isoxazol-3-amine I44
[0543] To a solution of acetohydroxamic acid (5.1 g, 67.8 mmol) in
DMF (150 mL) at 0.degree. C. under N.sub.2 was added t-BuOK (7.6 g,
6.4 mmol) and the mixture was stirred at RT for 2 h.
3-Bromo-2-chloro-6-fluorobenzonitrile I43 (5.3 g, 22.6 mmol) was
then added and the mixture was stirred at RT overnight. The mixture
was diluted with EtOAc (500 mL) and washed with water (.times.3),
brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=50/1 to 2/1) to give the
title compound (3.1 g, 52%) as a white solid, which was used
directly in the next step.
xix) 5-Bromo-4-methoxybenzo[d]isoxazol-3-amine I48
##STR00045##
[0544] a) 3-Bromo-6-fluoro-2-methoxybenzoic acid I45
[0545] To a solution of diisopropylamine (5.4 g, 53.7 mmol) in THF
(150 mL) at -78.degree. C. under N.sub.2 was added n-BuLi (2.5 M
solution in hexanes, 23.4 mL, 58.5 mmol) dropwise and the mixture
was stirred at -78.degree. C. for 1 h. The resulting mixture was
added dropwise to a solution of 1-bromo-4-fluoro-2-methoxybenzene
(10.0 g, 48.8 mmol) in THF (50 mL) at -78.degree. C. and stirring
was continued for 90 min. CO.sub.2 was bubbled through the mixture
for 20 min with stirring at -78.degree. C., then allowed to warm to
RT and stirred for 15 min. The reaction mixture was adjusted to
pH=1 with HCl and the mixture was diluted with water and extracted
with DCM (500 mL). The combined organic extracts were washed with
brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (DCM/MeOH=100/1 to 30/1) to give the title
compound (8.0 g, 66%) as a colorless oil. LCMS-D: R.sub.t 2.12 min;
m/z 248.9/250.9 [M+H].sup.+.
b) 3-Bromo-6-fluoro-2-methoxybenzamide I46
[0546] A mixture of 3-bromo-6-fluoro-2-methoxybenzoic acid I45 (8.0
g, 32.1 mmol) and SOCl.sub.2 (30 mL) was heated at 85.degree. C.
for 3 h. The mixture was concentrated under reduced pressure and
the residue was dissolved in DCM (5 mL) and added to conc.
NH.sub.4OH (20 mL) at 0.degree. C. dropwise. The mixture was
allowed to warm to RT, stirred for 20 min then extracted with DCM
(50 mL.times.3). The combined organic extracts were washed with
water, brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=50/1 to 1/1) to give the
title compound (6.8 g, 80%) as a white solid. LCMS-E: R.sub.t 2.24
min; m/z 247.8/249.8 [M+H].sup.+
c) 3-Bromo-6-fluoro-2-methoxybenzonitrile I47
[0547] A mixture of 3-bromo-6-fluoro-2-methoxybenzamide I46 (6.8 g,
25.6 mmol) and SOCl.sub.2 (30 mL) was heated at 80.degree. C.
overnight, then concentrated under reduced pressure. The residue
was partitioned between water and EtOAc, the phases were separated
and the organic layer was washed with water, brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography (Pet.
ether/EtOAc=100/1 to 20/1) to give the title compound (3.5 g, 55%)
as a colorless oil, which was used directly in the next step.
d) 5-Bromo-4-methoxybenzo[d]isoxazol-3-amine I48
[0548] To a solution of acetohydroxamic acid (3.4 g, 45.7 mmol) in
DMF (150 mL) at 0.degree. C. under N.sub.2 was added t-BuOK (5.1 g,
45.7 mmol) and the mixture was stirred at RT for 90 min. A solution
of 3-bromo-6-fluoro-2-methoxybenzonitrile I47 (3.5 g, 15.2 mmol) in
DMF (30 mL) was then added and the mixture was heated at 70.degree.
C. overnight. The mixture was diluted with EtOAc (1000 mL) and
washed with water (.times.3), brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=50/1 to 3/1) to give the title compound (3.2 g, 86%) as
a white solid. LCMS-D: R.sub.t 2.24 min; m/z 243.0/244.9
[M+H].sup.+.
xx) 4-(Methoxymethyl)benzo[d]isoxazol-3-amine I51
##STR00046##
[0549] a) 2-(Bromomethyl)-6-chlorobenzonitrile I49
[0550] A mixture of 2-chloro-6-methylbenzonitrile (2.0 g, 13.2
mmol), NBS (2.5 g, 13.8 mmol) and AIBN (660 mg, 4.0 mmol) in
CCl.sub.4 (60 mL) was heated at 85.degree. C. under N.sub.2
overnight. The mixture was concentrated under reduced pressure and
the residue was purified by column chromatography (Pet.
ether/EtOAc=100/1 to 50/1) to give the title compound (1.7 g, 37%)
as a white solid, which was used directly in the next step.
b) 2-Chloro-6-(methoxymethyl)benzonitrile I50
[0551] Sodium metal (115 mg, 4.8 mmol) was dissolved in MeOH (5 mL)
and THF (5 mL) and the mixture was stirred at RT for 20 min.
2-(Bromomethyl)-6-chlorobenzonitrile I49 (560 mg, 2.4 mmol) was
then added and the mixture was stirred at RT for 5 h. The solvent
was removed under reduced pressure and the residue was purified by
column chromatography (Pet. ether/EtOAc=50/1) to give the title
compound (340 mg, 77%) as a colorless oil. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.75-7.70 (m, 3H), 4.82 (s, 2H), 3.35 (s,
3H).
c) 4-(Methoxymethyl)benzo[d]isoxazol-3-amine I51
[0552] To a solution of acetohydroxamic acid (422 mg, 5.6 mmol) in
DMF (25 mL) at -78.degree. C. under N.sub.2 was added t-BuOK (630
mg, 5.6 mmol) and the mixture was stirred at 0.degree. C. for 1 h.
2-Chloro-6-(methoxymethyl)benzonitrile I50 (340 mg, 1.9 mmol) was
then added and the mixture was stirred at RT overnight, then heated
at 85.degree. C. overnight. The mixture was diluted with water (70
mL) and extracted with EtOAc (100 mL.times.2). The combined organic
extracts were washed with water (200 mL.times.3), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOA=5/1 to 3/1) to give the title compound (105 mg, 31%) as
a light yellow oil. LCMS-D: R.sub.t 1.57 min; m/z 179.1
[M+H].sup.+.
xxi) 4-Ethoxybenzo[d]isoxazol-3-amine I53
##STR00047##
[0553] a) 2-Ethoxy-6-fluorobenzonitrile I52
[0554] A mixture of 2-fluoro-6-hydroxybenzonitrile (2.0 g, 14.6
mmol), K.sub.2CO.sub.3 (6.04 g, 43.8 mmol) and bromoethane (2.38 g,
21.9 mmol) in DMF (4 mL) was stirred at RT under N.sub.2 overnight.
The mixture was diluted with EtOAc (300 mL), washed with water (100
mL.times.5), brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=100/1 to 5/1) to give the
title compound (1.6 g, 67%) as a white solid. LCMS-E: R.sub.t 5.24
min; m/z 166.1 [M+H].sup.+.
b) 4-Ethoxybenzo[d]isoxazol-3-amine I53
[0555] To a solution of acetohydroxamic acid (2.18 g, 29 mmol) in
DMF (40 mL) at 0.degree. C. under N.sub.2 was added t-BuOK (3.26 g,
29 mmol) and the mixture was stirred at RT for 1 h.
2-Ethoxy-6-fluorobenzonitrile I52 (1.6 g, 9.7 mmol) was then added
and the mixture was stirred at RT overnight. The mixture was
diluted with DCM (80 mL), washed with water (60 mL.times.4), dried
over Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography (Pet.
ether/EtOAc=5/1) to give the title compound (240 mg, 15%) as a
white solid. LCMS-E: R.sub.t 5.05 min; m/z 179.0 [M+H].sup.+.
xxii) 5-Methoxybenzo[d]isoxazol-3-amine I54
##STR00048##
[0557] To a solution of acetohydroxamic acid (1.49 mg, 19.8 mmol)
in DMF (35 mL) at 0.degree. C. under N.sub.2 was added t-BuOK (2.23
mg, 19.8 mmol) and the mixture was heated at 30.degree. C. for 1 h.
A solution of 2-fluoro-5-methoxybenzonitrile (1.0 g, 6.6 mmol) in
DMF (5 mL) was then added and the mixture was heated at 30.degree.
C. overnight. The mixture was diluted with water (70 mL) and
extracted with EtOAc (100 mL.times.3). The combined organic
extracts were washed with water (200 mL.times.3) then dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure to give the title compound (110 mg, 11%) as a yellow
solid, which was used directly in the next step.
xxiii) 5-Ethoxybenzo[d]isoxazol-3-amine I57
##STR00049##
[0558] a) 2-Fluoro-5-hydroxybenzonitrile I55
[0559] A mixture of 2-fluoro-5-methoxybenzonitrile (1.7 g, 1.2
mmol) and pyridine HCl (17 g) was heated at 80.degree. C. under
N.sub.2 for 5 h, then diluted with DCM (40 mL) and washed with 2 M
aq. HCl (8 mL) and water (2.times.40 mL). The organic layer was
extracted with an aqueous K.sub.2CO.sub.3 solution (50 mL.times.2)
and the combined aqueous extracts were washed with DCM (70
mL.times.2), then adjusted to pH 3-4 with 2 M aq. HCl and extracted
with DCM (80 mL.times.3). The combined organic extracts were dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give the title compound (430 mg, 28%) as an
off-white solid, which was used directly in the next step.
b) 5-Ethoxy-2-fluorobenzonitrile I56
[0560] To a solution of 2-fluoro-5-hydroxybenzonitrile I55 (430 mg,
3.1 mmol) in DMF (15 mL) was added K.sub.2CO.sub.3 (1.3 g, 9.4
mmol) and the mixture was stirred at RT under N.sub.2 for 30 min.
Bromoethane (512 mg, 4.7 mmol) was then added and stirring was
continued at RT overnight. The mixture was diluted with water (70
mL) and extracted with EtOAc (100 mL.times.2). The combined organic
extracts were washed with water (200 mL.times.3), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by silica gel chromatography
(Pet. ether/EtOAc=30/1 to 20/1) to give the title compound (480 mg,
92%) as a white solid. LCMS-D: R.sub.t 2.44 min; m/z 166.0
[M+H].sup.+ 188.0 [M+Na].sup.+.
c) 5-Ethoxybenzo[d]isoxazol-3-amine I57
[0561] To a solution of acetohydroxamic acid (645 mg, 8.7 mmol) in
DMF (35 mL) at 0.degree. C. under N.sub.2 was added t-BuOK (978 mg,
8.7 mmol) and the mixture was heated at 30.degree. C. for 1 h. A
solution of 5-ethoxy-2-fluorobenzonitrile I56 (480 mg, 2.9 mmol) in
DMF (5 mL) was then added and the mixture was heated at 30.degree.
C. overnight. The mixture was diluted with water (60 mL) and
extracted with EtOAc (80 mL.times.2). The combined organic extracts
were washed with water (150 mL.times.2), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give the title compound (400 mg, 77%) as a light yellow solid.
LCMS-D: R.sub.t 2.02 min; m/z 179.1 [M+H].sup.+.
xxiv) 6-(3,5-Dimethyl-1H-pyrazol-1-yl)benzo[d]isoxazol-3-amine
I59
##STR00050##
[0562] a) 2-Chloro-4-(3,5-dimethyl-1H-pyrazol-1-yl) benzonitrile
I58
[0563] A mixture of 3,5-dimethyl-1H-pyrazole (5 g, 0.052 mol), NaH
(60% dispersion in oil, 2.6 g, 0.065 mol) in DMF (50 mL) was
stirred at RT for 1 h. A solution of 2-chloro-4-fluorobenzonitrile
(6.74 g, 0.043 mol) in DMF (50 mL) was then added and stirring was
continued at RT for 1 h. The reaction was quenched with water and
the mixture was extracted with EtOAc. The organic extract was
concentrated under reduced pressure to give the title compound
(11.0 g, 92%) as a yellow solid. LCMS-D: R.sub.t 2.58 min; m/z
232.1 [M+H].sup.+.
b) 6-(3,5-Dimethyl-1H-pyrazol-1-yl)benzo[d]isoxazol-3-amine I59
[0564] To a solution of acetohydroxamic acid (972 mg, 12.9 mmol) in
DMF (20 mL) at 0.degree. C. under N.sub.2 was added t-BuOK (1.45 g,
12.9 mmol) and the mixture was heated at 30.degree. C. for 1 h.
2-Chloro-4-(3,5-dimethyl-1H-pyrazol-1-yl) benzonitrile I58 (1 g,
4.3 mmol) was then added and the mixture was heated at 60.degree.
C. for 5 h. The mixture was diluted with water and extracted with
EtOAc. The organic extract was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by preparative TLC (MeOH/DCM=1/20) to give
the title compound (150 mg, 15%) as a white solid. LCMS-D: R.sub.t
2.22 min; m/z 229.1 [M+H].sup.+.
xxv) 5-Methylbenzo[d]isoxazol-3-amine I60
##STR00051##
[0566] To a solution of acetohydroxamic acid (8.33 g, 0.11 mol) in
DMF (200 mL) was added t-BuOK (12.5 g, 0.11 mol) and the mixture
was stirred at RT for 1 hour. 2-Fluoro-5-methylbenzonitrile (5 g,
0.37 mol) was then added and the mixture was heated at 60.degree.
C. overnight. The mixture was diluted with water and extracted with
EtOAc. The organic extract was concentrated under reduced pressure
and the residue was purified by a silica gel chromatography
(DCM/MeOH=200/1 to 50/1) to give the title compound (3.0 g, 55%) as
a white solid. LCMS-D: R.sub.t 1.75 min, m/z 149.0 [M+H].sup.+.
xxvi) 6-(Methoxymethyl)benzo[d]isoxazol-3-amine I62
##STR00052##
[0567] a) 2-Fluoro-4-(methoxymethyl)benzonitrile I61
[0568] A mixture of Mel (2.0 g, 13.2 mmol) and NaH (60% suspension
in oil, 790 mg, 19.8 mmol) in THF (50 mL) was stirred at 0.degree.
C. for 10 min, then 2-fluoro-4-(hydroxymethyl)benzonitrile (2.0 g,
13.2 mmol) was added and the mixture was stirred at RT for 2 h. The
reaction was quenched with water and the mixture was extracted with
EtOAc. The organic extract was concentrated under reduced pressure
and the residue was purified by silica gel chromatography (Pet.
ether/EtOAc=100/1 to 20/1) to give the title compound (1.7 g, 78%)
as a white solid. LCMS-D: R.sub.t 2.01 min; m/z 166.0 [M+H].sup.+
187.9 [M+Na].sup.+.
b) 6-(Methoxymethyl)benzo[d]isoxazol-3-amine I62
[0569] To a solution of acetohydroxamic acid (1.5 g, 9.1 mmol) in
DMF (50 mL) was added t-BuOK (3.06 g, 27.2 mmol) and the mixture
was stirred at RT for 1 h. 2-Fluoro-4-(methoxymethyl)benzonitrile
I61 (1.5 g, 9.1 mmol) was then added and the mixture was heated at
40.degree. C. overnight. The mixture was diluted with water and
extracted with EtOAc. The organic extract was concentrated under
reduced pressure and the residue was purified by silica gel
chromatography (Pet. ether/EtOAc=100/1 to 10/1) to give the title
compound (1 g, 62%) as a yellow solid. LCMS-D: R.sub.t 0.95 min,
m/z 179.0 [M+H].sup.+.
xxvii) 5-(Trifluoromethoxy)benzo[d]isoxazol-3-amine I63
##STR00053##
[0571] To a solution of acetohydroxamic acid (2.2 g, 0.029 mol) in
DMF (50 mL) was added t-BuOK (3.28 g, 0.029 mol) and the mixture
was stirred RT for 1 hour.
2-Fluoro-5-(trifluoromethoxy)benzonitrile (2 g, 9.75 mmol) was then
added and the mixture was heated at 60.degree. C. overnight. The
mixture was diluted with water and extracted with EtOAc. The
organic extract was concentrated under reduced pressure to give the
title compound (1.6 g, 75%) as a yellow solid. LCMS-D: R.sub.t 2.43
min; m/z 219.0 [M+H].sup.+.
xxviii) 5-Methyl-6-(oxazol-2-yl)benzo[d]isoxazol-3-amine I65
##STR00054##
[0572] a) 2-Fluoro-5-methyl-4-(oxazol-2-yl)benzonitrile I64
[0573] To a solution of oxazole (90 mg, 1.31 mmol) in THF (10 mL)
at -70.degree. C. under N.sub.2 was added n-BuLi (2.5 M solution in
hexane, 1.1 mL, 2.66 mmol) and the mixture was stirred for 10 min.
Solid ZnCl.sub.2 (380 mg, 2.79 mmol) was added and the mixture was
allowed to warm to RT. 4-Bromo-2-fluoro-5-methylbenzonitrile (200
mg, 0.93 mmol) was added and the mixture was heated at 60.degree.
C. overnight. The solvent was removed under reduced pressure and
the residue was purified by silica gel chromatography (Pet.
ether/EtOAc 10/1) to give the title compound (50 mg, 27%) as a
white solid. LCMS-D: R.sub.t 2.51 min; m/z 203.0 [M+H].sup.+.
b) 5-Methyl-6-(oxazol-2-yl)benzo[d]isoxazol-3-amine I65
[0574] To a solution of acetohydroxamic acid (189 mg, 2.52 mmol) in
DMF (10 mL) at 0.degree. C. under N.sub.2 was added t-BuOK (377 mg,
3.26 mmol) and the mixture was stirred at 0.degree. C. for 1 h.
2-Fluoro-5-methyl-4-(oxazol-2-yl)benzonitrile I64 (170 mg, 0.84
mmol) was then added and the mixture was heated at 50.degree. C.
overnight. The mixture was diluted with water and extracted with
EtOAc. The organic extract was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by preparative TLC (DCM/MeOH=10/1) to give
the title compound (90 mg, 50%) as a white solid. LCMS-D: R.sub.t
2.10 min; m/z 216.0 [M+H].sup.+.
xxix) 7-Bromobenzo[d]isoxazol-3-amine I66
##STR00055##
[0576] To a solution of acetohydroxamic acid (3.75 g, 0.05 mol) in
DMF (60 mL) at 0.degree. C. was added t-BuOK (5.6 g, 0.05 mol) and
the mixture was stirred at RT for 1 h. A solution of
3-bromo-2-fluorobenzonitrile (5.0 g, 0.025 mol) in DMF (90 mL) was
then added dropwise and stirred was continued at RT overnight. The
mixture was diluted with DCM (300 mL), washed with water (250
mL.times.4), brine, dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give the title compound
(4.0 g, 63%) as a white solid. LCMS-D: R.sub.t 2.09 min, m/z
213.0/215.0 [M+H].sup.+.
xxx) 7-(Pyrimidin-2-yl)benzo[d]isoxazol-3-amine I68
##STR00056##
[0577] a) 2-Fluoro-3-(pyrimidin-2-yl)benzonitrile I67
[0578] A mixture of
2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile
I23 (1.5 g, 6.1 mmol), 2-bromopyrimidine (1.9 g, 12.0 mmol),
Pd(dppf)Cl.sub.2 (1.3 g, 1.8 mmol) and K.sub.3PO.sub.4 (6.5 g, 24.2
mmol) in water (60 mL), toluene (60 mL) and i-PrOH (15 mL) was
heated at 85.degree. C. under N.sub.2 for 4 h. The mixture was
diluted with EtOAc (50 mL) and washed with water (80 mL.times.3).
The organic layer was dried over anhydrous Na.sub.2SO.sub.4,
filtered, concentrated under reduced pressure and the residue was
purified by column chromatography (Pet. ether/EtOAc=4/1) to give
the title compound (450 mg, 38%) as a light yellow solid. LCMS-E:
R.sub.t 4.82 min; m/z 199.9 [M+H].sup.+.
b) 7-(Pyrimidin-2-yl)benzo[d]isoxazol-3-amine I68
[0579] To a solution of acetohydroxamic acid (243 mg, 3.2 mmol) in
DMF (15 mL) at 0.degree. C. under N.sub.2 was added t-BuOK (363 mg,
3.2 mmol) and the mixture was stirred for 1 h. A solution of
2-fluoro-3-(pyrimidin-2-yl)benzonitrile I67 (400 mg, 1.6 mmol) in
DMF (5 mL) was then added dropwise and the mixture was stirred at
RT overnight. The mixture was diluted with EtOAc (80 mL) and washed
with water (60 mL.times.3). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure to give the title compound (230 mg, 67%) as a yellow
solid. LCMS-D: R.sub.t 0.80 min, m/z 213.1 [M+H].sup.+.
xxxi) 6-O H-Pyrazol-1-yl)benzo[d]isoxazol-3-amine I70
##STR00057##
[0580] a) 2-Fluoro-4-(1H-pyrazol-1-yl)benzonitrile I69
[0581] A mixture of 4-bromo-2-fluorobenzonitrile (400 mg, 2.0
mmol), 1H-pyrazole (177 mg, 2.6 mmol), CuI (381 mg, 2.0 mmol),
K.sub.3PO.sub.4 (849 mg, 4.0 mmol) and
(1S,2S)--N.sup.1,N.sup.2-dimethylcyclohexane-1,2-diamine (28 mg,
0.2 mmol) in DMF (20 mL) was heated at 100.degree. C. in a
microwave for 1 h. The mixture was partitioned between EtOAc (200
mL) and water (100 mL), the layers were separated and the organic
layer was washed with water (.times.3), brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=100/1 to 20/1) to give the title compound (120 mg, 32%)
as a white solid. LCMS-D: R.sub.t2.20 min, m/z 188.1
[M+H].sup.+.
b) 6-(1H-Pyrazol-1-yl)benzo[d]isoxazol-3-amine I70
[0582] To a solution of acetohydroxamic acid (215 mg, 2.9 mmol) in
DMF (25 mL) at 0.degree. C. was added t-BuOK (322 mg, 2.9 mmol) and
the mixture was heated 30.degree. C. for 2 h.
2-Fluoro-4-(1H-pyrazol-1-yl)benzonitrile I69 (120 mg, 0.64 mmol)
was then added and the mixture was heated at 30.degree. C.
overnight. The mixture was partitioned between EtOAc (100 mL) and
water (50 mL), the layers were separated and the organic layer was
washed with water (.times.3), brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=50/1 to 2/1) to give the title compound (72 mg, 57%) as
a white solid, which was used directly in the next step.
xxxii) 6-(2H-1,2,3-Triazol-2-yl)benzo[d]isoxazol-3-amine I72
##STR00058##
[0583] a) 2-Chloro-4-(2H-1,2,3-triazol-2-yl)benzonitrile I71
[0584] A mixture of 2H-1,2,3-triazole (553 mg, 8.0 mmol) and NaH
(60% dispersion in oil, 192 mg, 4.8 mmol) in DMF (20 mL) was
stirred at 0.degree. C. for 30 min, then a solution of
2-chloro-4-fluorobenzonitrile (622 mg, 4.0 mmol) in DMF (10 mL) was
added. The mixture was stirred at 0.degree. C. for 2 h then allowed
to warm to RT and stirred for 2 h. The mixture was partitioned
between EtOAc (300 mL) and water (100 mL), the layers were
separated and the organic layer was washed with water (.times.3),
brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=100/1 to 20/1) to give the
title compound (200 mg, 24%) as a white solid. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 8.31-8.28 (m, 3H), 8.19-8.14 (m 2H).
b) 6-(2H-1,2,3-Triazol-2-yl)benzo[d]isoxazol-3-amine I72
[0585] To a solution of acetohydroxamic acid (221 mg, 2.9 mmol) in
DMF (25 mL) at 0.degree. C. was added t-BuOK (330 mg, 2.9 mmol) and
the mixture was heated at 30.degree. C. for 2 h.
2-Chloro-4-(2H-1,2,3-triazol-2-yl)benzonitrile I71 (200 mg, 0.98
mmol) was then added and the mixture was heated at 30.degree. C.
overnight. The mixture was partitioned between EtOAc (100 mL) and
water (50 mL), the layers were separated and the organic layer was
washed with water (.times.3), brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=50/1 to 2/1) to give the title compound (90 mg, 46%) as
a white solid. LCMS-D: R.sub.t 1.93 min, m/z 202.1 [M+H].sup.+.
xxxiii) 6-(Pyridin-2-yl)benzo[d]isoxazol-3-amine I74
##STR00059##
[0586] a) 2-Fluoro-4-(pyridin-2-yl)benzonitrile I73
[0587] A mixture of
2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile
I37 (494 mg, 2.0 mmol), 2-bromopyridine (948 mg, 6.0 mmol),
Pd(dppf)Cl.sub.2 (293 mg, 0.4 mmol) and K.sub.3PO.sub.4.3H.sub.2O
(2.66 g, 10.0 mmol) in H.sub.2O (40 mL), toluene (40 mL) and i-PrOH
(10 mL) was heated at 85.degree. C. under N.sub.2 for 4 h. The
mixture was partitioned between EtOAc (200 mL) and water (30 mL),
the layers were separated and the organic layer was washed with
water, brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=30/1 to 10/1) to give the
title compound (190 mg, 48%) as a white solid. LCMS-D: R.sub.t 2.32
min, m/z 199.1 [M+H].sup.+.
b) 6-(Pyridin-2-yl)benzo[d]isoxazol-3-amine I74
[0588] To a solution of acetohydroxamic acid (216 mg, 2.88 mmol) in
DMF (50 mL) at 0.degree. C. was added t-BuOK (323 mg, 2.88 mmol)
and the mixture was stirred at RT for 1 h. A solution of
2-fluoro-4-(pyridin-2-yl)benzonitrile I73 (190 mg, 0.96 mmol) in
DMF (10 mL) was then added and the mixture was stirred at RT
overnight. The mixture was partitioned between EtOAc (200 mL) and
water (50 mL), the layers were separated and the organic layer was
washed with water (50 mL.times.3), brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (DCM/MeOH=100/0
to 100/1) to give the title compound (105 mg, 52%) as a white
solid. LCMS-D: R.sub.t 0.83 min, m/z 212.1 [M+H].sup.+.
xxxiv) 6-Bromobenzo[d]isoxazol-3-amine I75
##STR00060##
[0590] To a solution of acetohydroxamic acid (13.7 g, 182 mmol) in
DMF (60 mL) at 0.degree. C. was added t-BuOK (20.5 g, 182 mmol) and
the mixture was stirred at 0.degree. C. for 1 h. A solution of
4-bromo-2-fluorobenzonitrile (12.2 g, 60.8 mmol) in DMF (30 mL) was
then added and the mixture was stirred at RT overnight. The mixture
was partitioned between EtOAc (500 mL) and water (200 mL), the
layers were separated and the organic layer was washed with water
(.times.2), brine, dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure. The residue was purified
by column chromatography (Pet. ether/EtOAc=100/1 to 30/1) to give
the title compound (8.1 g, 63%) as a white solid. LCMS-D: R.sub.t
2.34 min; m/z 213.0/215.0 [M+H].sup.+.
xxxv) 4-Methoxy-6-(pyridin-2-yl)benzo[d]isoxazol-3-amine I77
##STR00061##
[0591] a) 2-Fluoro-6-methoxy-4-(pyridin-2-yl)benzonitrile I76
[0592] A mixture of 4-bromo-2-fluoro-6-methoxybenzonitrile I5 (244
mg, 1.06 mmol), pyridin-2-ylboronic acid (195 mg, 1.59 mmol), CuCl
(105 mg, 1.06 mmol), Pd(OAc).sub.2 (24 mg, 0.106 mmol), XPhos (100
mg, 0.212 mmol) and Cs.sub.2CO.sub.3 (1.38 g, 4.24 mmol) in DMF
(10.6 mL) was heated under a nitrogen atmosphere in a 20 mL sealed
tube at 100.degree. C. for 16 h. The reaction was repeated a
further three times on the same scale and the four reactions were
quenched with a saturated aqueous NH.sub.4Cl, combined and
extracted with EtOAc (80 mL.times.3). The combined organic extracts
were washed with water, dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by column chromatography (Pet. ether/EtOAc=10/1 to 5/1) to
give the title compound (190 mg, 20%) as a yellow solid. LCMS-C:
R.sub.t 2.13 min; m/z 229.0 [M+H].sup.+.
b) 4-Methoxy-6-(pyridin-2-yl)benzo[d]isoxazol-3-amine I77
[0593] To a solution of acetohydroxamic acid (178 mg, 2.37 mmol) in
anhydrous DMF (20 mL) at 0.degree. C. was added potassium
tert-butoxide (266 mg, 2.37 mmol) and the mixture was stirred at
0.degree. C. for 1 h.
2-Fluoro-6-methoxy-4-(pyridin-2-yl)benzonitrile I76 (180 mg, 0.79
mmol) was then added and the mixture was heated at 40.degree. C.
overnight. Water was added and the mixture was extracted with
EtOAc. The combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (DCM/MeOH=200/1
to 100/1 to 60/1) to give the title compound (70 mg, 37%) as a
yellow solid. LCMS-C: R.sub.t 0.52 min; m/z 242.0 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.73-8.68 (m, 1H),
8.14-8.09 (m, 1H), 7.95-7.88 (m, 1H), 7.70 (d, J=1.0 Hz, 1H), 7.46
(s, 1H), 7.44-7.38 (m, 1H), 6.01 (s, 2H), 4.01 (s, 3H).
xxxvi) 4-Methoxy-7-phenylbenzo[d]isoxazol-3-amine I80
##STR00062##
[0594] a) 3-Bromo-2-fluoro-6-methoxybenzonitrile I78
[0595] A solution of Br.sub.2 (507 mg, 3.2 mmol) in CCl.sub.4 (4.0
mL) was added to a solution of 2-fluoro-6-methoxybenzonitrile (480
mg, 3.2 mmol) and Fe (8.0 mg, 0.1 mmol) in CCl.sub.4 (4.0 mL) at
-10.degree. C. over a period of 30 min and the mixture was then
allowed to warm to RT and stirred overnight. The mixture was
partitioned between water and EtOAc, the layers were separated and
the organic layer was washed with a saturated aqueous
Na.sub.2SO.sub.3 solution (.times.2), brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated to give the title
compound (580 mg, 80%) as a white solid. LCMS-C: R.sub.t 2.12 min;
m/z 229.9 [M+H].sup.+.
b) 2-Fluoro-4-methoxy-[1,1'-biphenyl]-3-carbonitrile I79
[0596] To a solution of 3-bromo-2-fluoro-6-methoxybenzonitrile I78
(600 mg, 2.6 mmol), phenylboronic acid (636 mg, 5.2 mmol) and
Na.sub.2CO.sub.3 (829 mg, 7.8 mmol) in 1,4-dioxane (40 mL) and
water (10 mL) under N.sub.2 was added Pd(PPh.sub.3).sub.4 (300 mg,
0.26 mmol) and the mixture was heated at 100.degree. C. overnight.
The mixture was partitioned between water and EtOAc, the layers
were separated and the organic layer was washed with water, brine,
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by column
chromatography (Pet. ether/EtOAc=10/1 to 3/1) to give the title
compound (538 mg, 90%) as a white solid. LCMS-C: R.sub.t 2.43 min;
m/z 228.0 [M+H].sup.+.
c) 4-Methoxy-7-phenylbenzo[d]isoxazol-3-amine I80
[0597] To a solution of acetohydroxamic acid (533 mg, 7.11 mmol) in
anhydrous DMF (30 mL) at RT was added potassium tert-butoxide (797
mg, 7.11 mmol) and the mixture was stirred at RT for 1 h.
2-Fluoro-4-methoxy-[1,1'-biphenyl]-3-carbonitrile I79 (538 mg, 2.37
mmol) was then added and the mixture was heated at 60.degree. C.
overnight. Water was added and the mixture was extracted with EtOAc
(30 mL.times.3). The combined organic extracts were dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography (Pet.
ether/EtOAc=10/1 to 5/1) to give the title compound (413 mg, 72%)
as an orange solid. LCMS-C: R.sub.t 1.33 min; m/z 209.0
[M+H].sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.84-7.79
(m, 2H), 7.61 (d, J=8.1 Hz, 1H), 7.46 (t, J=7.7 Hz, 2H), 7.38-7.32
(m, 1H), 6.67 (d, J=8.2 Hz, 1H), 4.00 (s, 3H).
xxxvii)
4-Methoxy-7-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-amine
I82
##STR00063##
[0598] a)
2-Fluoro-6-methoxy-3-(1-methyl-1H-pyrazol-4-yl)benzonitrile I81
[0599] To a solution of 3-bromo-2-fluoro-6-methoxybenzonitrile I78
(720 mg, 3.31 mmol),
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(1.30 g, 6.26 mmol) and Na.sub.2CO.sub.3 (995 mg, 9.39 mmol) in
1,4-dioxane (50 mL) and water (10 mL) under N.sub.2 was added
Pd(PPh.sub.3).sub.4 (358 mg, 0.30 mmol) and the mixture was heated
at 100.degree. C. overnight. Water was added and the mixture was
extracted with EtOAc. The combined organic extracts were washed
with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (DCM/MeOH=100/1) to give the title compound
(485 mg, 63%) as a white solid. LCMS-C: R.sub.t 1.26 min; m/z 232.0
[M+H].sup.+.
b) 4-Methoxy-7-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-amine
I82
[0600] To a solution of acetohydroxamic acid (474 mg, 6.24 mmol) in
anhydrous DMF (20 mL) at RT was added potassium tert-butoxide (700
mg, 6.24 mmol) and the mixture was stirred at RT for 1 h.
2-Fluoro-6-methoxy-3-(1-methyl-1H-pyrazol-4-yl)benzonitrile I81
(485 mg, 2.04 mmol) was then added and the mixture was heated at
60.degree. C. overnight. Water was added and the mixture was
extracted with EtOAc (30 mL.times.3). The combined organic extracts
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (DCM/MeOH=50/1) to give the title compound
(225 mg, 45%) as a yellow solid. LCMS-C: R.sub.t 0.46 min; m/z
245.0 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.19
(s, 1H), 7.95 (s, 1H), 7.69 (d, J=8.2 Hz, 1H), 6.74 (d, J=8.2 Hz,
1H), 6.00 (s, 2H), 3.91 (s, 3H), 3.90 (s, 3H).
xxxviii)
5-Chloro-4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine
I83
##STR00064##
[0602] To a solution of
4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine I9 (300 mg, 1.4
mmol) in DMF (10 mL) was added NCS (192 mg, 1.4 mmol) and the
mixture was heated at 50.degree. C. for 2 h. The mixture was
diluted with EtOAc (100 mL) and washed with H.sub.2O (40
mL.times.3). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=7/1 to 5/1) to give the title compound (190 mg, 54%) as
a white solid. LCMS-C: R.sub.t 1.21 min; m/z 242.9 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.32 (s, 1H), 6.19 (s,
2H), 4.55 (s, 2H), 3.93 (s, 3H), 3.41 (s, 3H).
xxxix) 4-Methoxy-6-(oxazol-2-yl)benzo[d]isoxazol-3-amine I86
##STR00065##
[0603] a) 2-(Tributylstannyl)oxazole I84
[0604] To a solution of oxazole (500 mg, 7.25 mmol) in THF (15 mL)
at -78.degree. C. under N.sub.2 was added n-BuLi (2.5 M solution in
hexanes, 2.9 mL, 7.32 mmol) dropwise and the mixture was stirred at
-78.degree. C. for 30 min. Tributylchlorostannane (1.96 mL, 7.25
mmol) was then added and the mixture was allowed to warm to RT and
stirred for 1 h. The solvent was removed under reduced pressure and
residue was taken up in hexanes (50 mL). The resulting precipitate
was removed by filtration and the filtrate was concentrated under
reduced pressure to give the title compound (2.0 g, 77%) as
colorless oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.20 (s,
1H), 7.20 (s, 1H), 1.59-1.49 (m, 6H), 1.31-1.26 (m, 6H), 1.16-1.10
(m, 6H), 0.83 (t, J=7.3 Hz, 9H).
b) 2-Fluoro-6-methoxy-4-(oxazol-2-yl)benzonitrile I85
[0605] To a solution of 4-bromo-2-fluoro-6-methoxybenzonitrile I5
(305 mg, 1.33 mmol) in 1,4-dioxane (25 mL) was added
2-(tributylstannyl)oxazole 184 (1.43 g, 3.98 mmol) and
Pd(PPh.sub.3).sub.4 (154 mg, 0.133 mmol) and the mixture was heated
at 90.degree. C. overnight. The mixture was diluted with water and
extracted with EtOAc. The combined organic extracts were washed
with water, brine, dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated. The residue was purified by column chromatography
(Pet. ether/EtOAc=8/1) to give the title compound (370 mg, 96%) as
a white solid. LCMS-C: R.sub.t 1.86 min; m/z 218.9 [M+H].sup.+.
c) 4-Methoxy-6-(oxazol-2-yl)benzo[d]isoxazol-3-amine I86
[0606] To a solution of acetohydroxamic acid (382 mg, 5.09 mmol) in
DMF (25 mL) at 0.degree. C. was added potassium tert-butoxide (570
mg, 5.09 mmol) and the mixture was stirred at RT for 1 h.
2-Fluoro-6-methoxy-4-(oxazol-2-yl)benzonitrile I85 (370 mg, 1.7
mmol) was then added and the mixture was heated at 60.degree. C.
for 2 h. The mixture was diluted with EtOAc and washed with water,
brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated to give the title compound (100 mg, 26%) as a yellow
solid. LCMS-C: R.sub.t 0.57 min; m/z 232.0 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 8.29 (s, 1H), 7.56 (s, 1H),
7.47-7.42 (m, 1H), 7.27 (s, 1H), 6.09 (s, 2H), 4.00 (s, 3H).
xl) 6-(Oxazol-2-yl)benzo[d]isoxazol-3-amine I90
##STR00066##
[0607] a) 4-Cyano-3-fluorobenzoyl chloride I87
[0608] To a solution of 4-cyano-3-fluorobenzoic acid (1.0 g, 6.1
mmol) in DCM (20 mL) at 0.degree. C. was added DMF (0.1 mL) and
oxalyl chloride (1.86 g, 12.1 mmol) dropwise and the mixture was
stirred at RT overnight. The solvent was removed under reduced
pressure to give the title compound (1.2 g) as a white solid, which
was used directly in the next step without further
purification.
b) 4-Cyano-N-(2,2-dimethoxyethyl)-3-fluorobenzamide I88
[0609] To a solution of 4-cyano-3-fluorobenzoyl chloride I87 (1.1
g, 6.06 mmol) and Et.sub.3N (1.84 g, 18 mmol) in DCM (20 mL) at
0.degree. C. was added 2,2-dimethoxyethanamine (955 mg, 9.1 mmol)
and the mixture was stirred for 2 h. The mixture was poured into
water and extracted with EtOAc. The organic extract was washed with
water and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure. The residue was purified
by column chromatoyraphy (Pet. ether/EtOAc=1/1, v/v) to give the
title compound (1.2 g, 78%) as a white solid. LCMS-D: R.sub.t 1.55
min, m/z 274.9 [M+Na].sup.+.
c) 3-Amino-N-(2,2-dimethoxyethyl)benzo[d]isoxazole-6-carboxamide
I89
[0610] To a solution of acetohydroxamic acid (448 mg, 5.95 mmol) in
DMF (30 mL) at 0.degree. C. was added t-BuOK (889 mg, 7.92 mmol)
portion-wise and the mixture was stirred for 1 h.
4-Cyano-N-(2,2-dimethoxyethyl)-3-fluorobenzamide I88 (500 mg, 1.98
mmol) was then added and the mixture was heated at 40.degree. C.
overnight. The mixture was poured into water and extracted with
EtOAc. The organic layer was washed with water and brine, dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by column chromatography
(Pet. ether/EtOAc=1/1, v/v) to give the title compound (380 mg,
72%) as a yellow solid. LCMS-D: R.sub.t 0.59 min, m/z 287.9
[M+Na].sup.+.
d) 6-(Oxazol-2-yl)benzo[d]isoxazol-3-amine I90
[0611] A mixture of
3-amino-N-(2,2-dimethoxyethyl)benzo[d]isoxazole-6-carboxamide 189
(240 mg, 0.9 mmol) and P.sub.2O.sub.5 (193 mg, 1.36 mmol) in
methanesulfonic acid (10 mL) was heated at 150.degree. C. under
microwave irradiation for 30 min. The mixture was poured into
water, made basic with aquesous KOH and extracted with EtOAc. The
organic extract was washed with brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by prep. TLC (DCM/MeOH=10/1, v/v) to give
the title compound (50 mg, 28%) as a solid. LCMS-D: R.sub.t 1.57
min, m/z 201.9 [M+H].sup.+.
xli) 5-Ethyl-1-methyl-2-oxo-1,2-dihydropyridine-3-sulfonyl chloride
I94
##STR00067##
[0612] a) 5-Ethyl-2-methoxypyridine I91
[0613] To a solution of 5-bromo-2-methoxypyridine (10.2 g, 54.25
mmol) in THF (200 mL) at -78.degree. C. under N.sub.2 was added
n-BuLi (2.5 M solution in hexane, 24.0 mL, 60.0 mmol) dropwise and
the mixture was stirred at -78.degree. C. for 1.5 h. Iodoethane
(12.7 g, 81.4 mmol) was then added dropwise and the mixture was
stirred at -78.degree. C. for 20 min, then warmed to RT and stirred
for 30 min. The reaction was quenched with water (5 mL) and the
solvent was removed under reduced pressure. The residue was
dissolved in DCM, washed with water, brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=100/1 to 5/1) to give the title compound (2.8 g, 38%)
as a colorless oil. LCMS-D: R.sub.t 1.80 min; m/z 138.1
[M+H].sup.+.
b) 5-Ethylpyridin-2(1H)-one I92
[0614] A solution of 5-ethyl-2-methoxypyridine 191 (1.6 g, 11.66
mmol) in conc. HCl (30 mL) was heated at 100.degree. C. overnight.
The mixture was concentrated under reduced pressure and the residue
was purified by column chromatography (DCM/MeOH=20/1) to give the
title compound (800 mg, 56%) as a white solid, which was used
directly in the next step.
5-Ethyl-1-methylpyridin-2(1H)-one I93
[0615] A mixture of 5-ethylpyridin-2(1H)-one 192 (800 mg, 6.5
mmol), K.sub.2CO.sub.3 (1.8 g, 13 mmol) and iodomethane (1.85 g, 13
mmol) in MeOH (20 mL) was heated at 50.degree. C. under N.sub.2
overnight. The solvent was removed under reduced pressure and the
residue was dissolved in DCM (100 mL), washed with water, brine,
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by column
chromatography (DCM/MeOH=40/1) to give the title compound (500 mg,
56%) as a colorless oil. LCMS-D: R.sub.t 0.68 min; m/z 138.1
[M+H].sup.+.
d) 5-Ethyl-1-methyl-2-oxo-1,2-dihydropyridine-3-sulfonyl chloride
I94
[0616] A mixture of chlorosulfonic acid (6 mL) and
5-ethyl-1-methylpyridin-2(1H)-one 193 (0.6 g, 4.37 mmol) was heated
at 150.degree. C. under N.sub.2 for 3 h, then allowed to cool to RT
and poured onto ice (100 g). The mixture was extracted with DCM (50
mL.times.3) and the combined organic extracts were washed twice
with ice-cold water, dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by column chromatography (DCM/MeOH=100/1 to 20/1) to give
the title compound (200 mg, 12%) as a yellow solid. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.87-7.84 (m, 1H), 7.71 (s, 1H),
3.47 (s, 3H), 2.39 (q, J=7.6 Hz, 2H), 1.09 (t, J=7.5 Hz, 3H).
LCMS-D: R.sub.t 1.58 min; m/z 236.0 [M+H].sup.+.
xlii) 5-Bromo-2,3-dihydrobenzofuran-7-sulfonyl chloride I95
##STR00068##
[0618] 5-Bromo-2,3-dihydrobenzofuran (2.0 g, 10 mmol) was added
slowly to chlorosulfonic acid (6 mL) at -5.degree. C. and the
mixture was stirred at -5.degree. C. for 30 min. The mixture was
poured into ice-cold water (100 mL) and extracted with EtOAc (180
mL.times.2). The combined organic extracts were washed with water
(250 mL.times.3), dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure. The residue was purified
by column chromatography (Pet. ether/EtOAc=15/1) to give the title
compound (1.45 g, 48%) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.80-7.76 (m, 1H), 7.63-7.58 (m, 1H), 4.91 (m,
2H), 3.35 (m, 2H). LCMS-D: R.sub.t2.74 min; m/z 318.8/320.8
[M+Na].sup.+.
xliii) 4,6-Dimethoxy-2,3-dihydro-1H-indene-5-sulfonyl chloride
I98
##STR00069##
[0619] a) 5,7-Dimethoxy-2,3-dihydro-1H-inden-1-one I96
[0620] A mixture of 3-(3,5-dimethoxyphenyl)propanoic acid (5 g,
23.8 mmol) and methanesulfonic acid (24 mL) was heated at
90.degree. C. for 10 min then allowed to cool to RT and poured into
water. The mixture was adjusted to pH 9 with 10 M aq. KOH and
extracted with EtOAc (.times.5). The combined organic extracts were
washed with water, brine, dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give the title
compound (3.5 g, 76%) as a white solid. LCMS-D: R.sub.t 1.76 min;
m/z 193.1 [M+H].sup.+.
b) 4,6-Dimethoxy-2,3-dihydro-1H-indene I97
[0621] A mixture of 5,7-dimethoxy-2,3-dihydro-1H-inden-1-one 196
(3.0 g, 15.6 mmol) and triethylsilane (7.3 g, 62.4 mmol) in TFA (20
mL) was stirred at RT under N.sub.2 for 11 h. The mixture was
concentrated under reduced pressure and the residue was purified by
column chromatography (Pet. ether/EtOAc=20/1) to give the title
compound (2.0 g, 72%) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 6.42 (s, 1H), 6.29 (d, J=2.0 Hz, 1H), 3.81 (s,
3H), 3.80 (s, 3H), 2.89 (t, J=7.5 Hz, 2H), 2.80 (t, J=7.3 Hz, 2H),
2.13-2.02 (m, 2H).
c) 4,6-Dimethoxy-2,3-dihydro-1H-indene-5-sulfonyl chloride I98
[0622] To a solution of 4,6-dimethoxy-2,3-dihydro-1H-indene 197 (1
g, 5.6 mmol) and TMEDA (0.72 g, 6.17 mmol) in n-hexane (20 mL) at
-70.degree. C. was added n-BuLi (2.5 M in hexane, 2.5 mL, 6.17
mmol) dropwise and the mixture was allowed to warm to 0.degree. C.
and stirred for 2 h. The mixture was then re-cooled to -65.degree.
C., bubbled with SO.sub.2 gas for 20 min, then allowed to warm
slowly to 10.degree. C. The resulting precipitate was collected by
filtration and washed with dry diethyl ether. The solid was
suspended in n-hexane (20 mL), cooled to 0.degree. C. and
SO.sub.2Cl.sub.2 (0.83 g, 6.2 mmol) was added dropwise. The mixture
was stirred at 0.degree. C. under N.sub.2 for 1 h, then filtered.
The filter cake was dissolved in diethyl ether and washed with
water, brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give the title compound (550
mg, 35%) as a white solid. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 6.72 (s, 1H), 3.95 (s, 3H), 3.94 (s, 3H), 2.96 (q, J=7.6
Hz, 4H), 2.18-2.08 (m, 2H).
xliv) 2-Methoxy-5-phenoxybenzenesulfonyl chloride I100
##STR00070##
[0623] a) 2-Methoxy-5-phenoxybenzenesulfonic acid I99
[0624] To a solution of 1-methoxy-4-phenoxybenzene (2 g, 10 mmol)
in DCM (15 mL) at 0.degree. C. was added a solution of
chlorosulfonic acid (0.35 mL) in DCM (10 mL) dropwise and the
mixture was stirred at 0.degree. C. for 15 min. The mixture was
poured slowly into ice-cold water (100 mL) and then concentrated
under reduced pressure. The residue was rinsed with DCM (100
mL.times.2) and dried to give the title compound (560 mg, 40%) as
an off-white solid. LCMS-D: R.sub.t 0.62 min; m/z 281.0
[M+H].sup.+.
b) 2-Methoxy-5-phenoxybenzenesulfonyl chloride I100
[0625] A mixture of 2-methoxy-5-phenoxybenzenesulfonic acid I99
(250 mg, 0.9 mmol) and PCl.sub.5 (284 mg, 1.3 mmol) in POCl.sub.3
(3 mL) was heated at 90.degree. C. under N.sub.2 for 1 h. The
mixture was then added slowly to ice-cold water (20 mL) and
extracted with DCM (15 mL.times.2). The combined organic extracts
were washed with brine, dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give the title
compound (51 mg, 18%) as a yellow oil. LCMS-D: R.sub.t 2.72 min;
m/z 295.0 [M-Cl+OCH.sub.3].sup.+, 317.0
[M-Cl+OCH.sub.3+Na].sup.+.
xlv) 2,6-Dimethoxy-3-(trifluoromethyl)benzenesulfonyl chloride
I102
##STR00071##
[0626] a) 2,4-Dimethoxy-1-(trifluoromethyl)benzene I101
[0627] To a mixture of (2,4-dimethoxyphenyl)boronic acid (3.0 g,
16.5 mmol), CF.sub.3SO.sub.2Na (18.0 g, 115.4 mmol), Cu(OAc).sub.2
(748 mg, 4.1 mmol), imidazole (281 mg, 4.1 mmol), 2,4,6-collidine
(3.0 g, 33.0 mmol) and NH.sub.4Cl (11.3 g, 206.0 mmol) in water
(16.5 mL) and DCM (100 mL) at 0.degree. C. was added t-BuOOH (3.6
mL, 4.1 mmol) dropwise and the mixture was stirred at RT for 16 h.
The layers were then separated and the aqueous layer was extracted
with DCM. The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet. ether) to
give the title compound (900 mg, 27%) as an oil. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.42-7.36 (m, 1H), 6.45-6.37 (m, 2H), 3.79
(s, 3H), 3.76 (s, 3H).
b) 2,6-Dimethoxy-3-(trifluoromethyl)benzenesulfonyl chloride
I102
[0628] To a solution of 2,4-dimethoxy-1-(trifluoromethyl)benzene
I101 (1.5 g, 7.3 mmol) and TMEDA (0.93 g, 8.0 mmol) in n-hexane (30
mL) at -78.degree. C. under N.sub.2 was added n-BuLi (2.5 M in
hexane, 3.2 mL, 8.0 mmol) dropwise and the mixture was stirred at
0.degree. C. for 1 h. SO.sub.2 gas was then bubbled through the
mixture at -78.degree. C. for 20 minutes and then allowed to warm
to 0.degree. C. and stirred for 1 h. The resulting precipitate was
collected by filtration and washed with hexane. The filter cake was
suspended in n-hexane (30 mL), cooled to 0.degree. C. and
SO.sub.2Cl.sub.2 (1.1 g, 8.0 mmol) was added dropwise. The mixture
was stirred at 0.degree. C. for 1 h and the solids were collected
by filtration and washed with cold n-hexane. The filter cake was
dissolved in ether and washed with water. The aqueous phase was
extracted with ether and the combined organic layers were dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by column chromatography
(Pet. ether/EtOAc=5/1) to give the title compound (1.5 g, 68%) as a
yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.90-7.84
(m, 1H), 6.98-6.92 (m, 1H), 4.09 (s, 3H), 4.04 (s, 3H).
xlvi) 3-Ethyl-2,6-dimethoxybenzenesulfonyl chloride I106
##STR00072##
[0629] a) 2,4-Dimethoxy-1-vinylbenzene I103
[0630] A suspension of 1-bromo-2,4-dimethoxybenzene (4.0 g, 18.4
mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (3.4 g, 22.1
mmol), Pd(dppf)Cl.sub.2.DCM (753 mg, 0.92 mmol) and K.sub.2CO.sub.3
(7.6 g, 55.2 mmol) in 1,4-dioxane (40 mL) and water (10 mL) was
heated at 90.degree. C. under N.sub.2 overnight. The mixture was
filtered through a pad of Celite and rinsed with EtOAc. The
filtrate was diluted with water and extracted with EtOAc (60
mL.times.3). The combined organic extracts were washed with brine,
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by column
chromatography (Pet. ether) to give the title compound (2.6 g, 86%)
as a yellow oil. LCMS-D: R.sub.t 2.28 min; m/z 165.0
[M+H].sup.+.
b) Ethyl-2,4-dimethoxybenzene I104
[0631] To a solution of 2,4-dimethoxy-1-vinylbenzene I103 (2.6 g,
15.8 mmol) in EtOAc (50 mL) was added 10% Pd/C (300 mg) and the
mixture was stirred at RT under a H2 atmosphere overnight. The
catalyst was removed by filtration through Celite and rinsed with
EtOAc. The filtrate was concentrated under reduced pressure to give
the title compound (2.0 g, 83%) as a yellow oil. LCMS-D: R.sub.t
2.39 min; m/z 167.1 [M+H].sup.+.
3-Ethyl-2,6-dimethoxybenzenesulfonic acid I105
[0632] Prepared from ethyl-2,4-dimethoxybenzene I104 according to
the procedure described for 2,6-dimethoxybenzenesulfonyl chloride
I111. The product obtained was found to be mostly
3-ethyl-2,6-dimethoxybenzenesulfonic acid. LCMS-D: R.sub.t 2.36
min; m/z 247.0 [M+H].sup.+.
d) 3-Ethyl-2,6-dimethoxybenzenesulfonyl chloride I106
[0633] A mixture of 3-ethyl-2,6-dimethoxybenzenesulfonic acid I105
(300 mg, 1.22 mmol) and thionyl chloride (6 mL) was heated at
95.degree. C. for 3 h then concentrated under reduced pressure to
give the title compound (322 mg, 100%) as a brown oil, which was
used directly in the next step.
xlvii) 7-Methoxyquinoline-8-sulfonyl chloride I107
##STR00073##
[0635] Chlorosulfonic acid (1.8 g, 15.7 mmol) was added dropwise to
7-methoxyquinoline (500 mg, 3.14 mmol) at 0.degree. C. and the
mixture was heated at 100.degree. C. for 1 h. The mixture allowed
to cool to RT, poured onto ice and then neutralised with a
saturated aqueous NaHCO.sub.3 solution. The mixture was extracted
with EtOAc (30 mL.times.3) and the combined organic extracts were
washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give the title compound
(280 mg, 34%) as a yellow solid. LCMS-D: R.sub.t 0.27 min; m/z
239.9 [M-Cl+H.sub.2O].sup.+
xlviii) 6-Methoxy-2,3-dihydro-1H-indene-5-sulfonyl chloride
I108
##STR00074##
[0637] To a solution of 5-methoxy-2,3-dihydro-1H-indene (3.1 g,
20.9 mmol) in DCM (40 mL) at -5.degree. C. under N.sub.2 was added
chlorosulfonic acid (6.5 g, 62.8 mmol) dropwise and the mixture was
stirred at -5.degree. C. for 40 min. The reaction was quenched with
ice water (20 mL) and the mixture was extracted with EtOAc (30
mL.times.2). The combined organic extracts were washed with water
(50 mL.times.3), dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure. The residue was purified
by column chromatography (Pet. ether/EtOAc=20/1) to give the title
compound (3.1 g, 60%) as a white solid, which was used directly in
the next step. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.77 (d,
J=1.1 Hz, 1H), 6.98 (s, 1H), 4.02 (s, 3H), 2.99 (t, J=7.5 Hz, 2H),
2.91 (t, J=7.4 Hz, 2H), 2.19-2.10 (m, 2H).
xlix) 3-Methoxy-5,6,7,8-tetrahydronaphthalene-2-sulfonyl chloride
I109
##STR00075##
[0639] Prepared from 6-methoxy-1,2,3,4-tetrahydronaphthalene and
chlorosulfonic according to the procedure described for
6-methoxy-2,3-dihydro-1H-indene-5-sulfonyl chloride I108 and used
directly in the next step without purification.
l) 4-Bromo-2-methoxybenzenesulfonyl chloride I110
##STR00076##
[0641] 1-Bromo-3-methoxybenzene (15.0 g, 80 mmol) was added slowly
to chlorosulfonic acid (16 mL) at -5.degree. C. and the mixture was
stirred at -5.degree. C. for 5 min. The mixture was poured into
ice-cold water (50 mL) and extracted with EtOAc (80 mL.times.2).
The combined organic extracts were washed with water (150
mL.times.3), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=50/1) to give the title
compound (2.6 g, 17%) as yellow solid, which was used directly in
the next step.
li) 2,6-Dimethoxybenzenesulfonyl chloride I111
##STR00077##
[0643] To a solution of 1,3-dimethoxybenzene (5.0 g, 36 mmol) and
TMEDA (4.6 g, 39.8 mmol) in n-hexane (100 mL) at 0.degree. C. under
N.sub.2 was added n-BuLi (2.5 M solution in hexanes, 16.0 mL, 39.8
mmol) dropwise while keeping the internal reaction temperature
below 5.degree. C. The mixture was stirred at 0.degree. C. for 20
min then cooled to -78.degree. C. and bubbled with SO.sub.2 gas for
20 min. The mixture was then allowed to warm slowly to 10.degree.
C. and the resulting precipitate was collected by filtration and
washed with dry diethyl ether. The solid was suspended in n-hexane
(100 mL), cooled to 0.degree. C. and a solution of SO.sub.2Cl.sub.2
(4.9 g, 36 mmol) in n-hexane (20 mL) was added dropwise while
keeping the internal temperature below 3.degree. C. The mixture was
then stirred at 0.degree. C. for 1 h and the solids were collected
by filtration and washed with cold n-hexane. The solids were then
partitioned between diethyl ether and water, the layers were
separated and the aqueous layer was further extracted with diethyl
ether. The combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give the title compound (4.0 g, 47%) as a white solid. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.54 (t, J=8.4 Hz, 1H), 6.66 (d,
J=8.4 Hz, 2H), 3.97 (s, 6H).
lii) 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
##STR00078##
[0645] 1-Ethyl-4-methoxybenzene (5.0 g, 37 mmol) was added dropwise
to chlorosulfonic acid (20 mL) at 0.degree. C. and the mixture was
stirred at RT for 2 h then poured onto ice and extracted with EtOAc
(50 mL.times.3). The combined organic extracts were dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography (Pet.
ether/EtOAc=100/1 to 30/1) to give the title compound (4.6 g, 53%)
as a white solid. LCMS-D: R.sub.t 2.70 min; m/z 256.9
[M+Na].sup.+.
liii) 2,4-Dimethoxy-[1,1'-biphenyl]-3-sulfonyl chloride I114
##STR00079##
[0646] a) 2,4-Dimethoxy-1,1'-biphenyl I113
[0647] A suspension of 1-bromo-2,4-dimethoxybenzene (5.0 g, 23.0
mmol), phenylboronic acid (3.4 g, 27.6 mmol), Pd(PPh.sub.3).sub.4
(1.3 g, 1.15 mmol) and potassium carbonate (7.3 g, 69.0 mmol) in
1,4-dioxane (30 mL) and water (6 mL) was heated at 90.degree. C.
under N.sub.2 for 16 h. The mixture was filtered through a pad of
Celite and washed with EtOAc. The filtrate was diluted with water
and extracted with EtOAc (30 mL.times.3). The combined organic
extracts were dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=100/1 to 10/1) to give the
title compound (2.8 g, 57%) as a yellow oil. LCMS-D: R.sub.t2.46
min; m/z 215.0 [M+H].sup.+.
b) 2,4-Dimethoxy-[1,1'-biphenyl]-3-sulfonyl chloride I114
[0648] To a solution of 2,4-dimethoxy-1,1'-biphenyl I113 (1.0 g,
4.70 mmol) and TMEDA (601 mg, 5.20 mmol) in n-hexane (40 mL) at
0.degree. C. under N.sub.2 was added n-BuLi (2.5 M solution in
hexanes, 2.1 mL, 5.20 mmol) dropwise while keeping the internal
reaction temperature below 5.degree. C. The mixture was stirred at
0.degree. C. for 20 min then cooled to -70.degree. C. and bubbled
with SO.sub.2 gas for 20 min. The mixture was then allowed to warm
slowly to 10.degree. C. and the resulting precipitate was collected
by filtration and washed with dry diethyl ether. The solid was
suspended in n-hexane (40 mL), cooled to 0.degree. C. and a
solution of SO.sub.2Cl.sub.2 (634 mg, 4.7 mmol) in n-hexane (5 mL)
was added dropwise while keeping the internal temperature below
3.degree. C. The mixture was then stirred at 0.degree. C. for 1 h
and the solids were collected by filtration and washed with cold
n-hexane. The solids were then partitioned between diethyl ether
and water, the layers were separated and the aqueous layer was
further extracted with diethyl ether. The combined organic extracts
were dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give the title compound (590 mg, 40%) as a
white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.48-7.35
(m, 4H), 7.34-7.21 (m, 2H), 6.87 (m, 1H), 3.76 (s, 3H), 3.29 (s,
3H).
liv) 3,5-Dimethoxy-[1,1'-biphenyl]-4-sulfonyl chloride I116
##STR00080##
[0649] a) 3,5-Dimethoxy-1,1'-biphenyl I115
[0650] A suspension of 1-bromo-3,5-dimethoxybenzene (5.0 g, 23.0
mmol), phenylboronic acid (2.8 g, 23.0 mmol), Pd(dppf)Cl.sub.2
(0.57 g, 0.69 mmol) and potassium carbonate (4.8 g, 34.6 mmol) in
1,4-dioxane (80 mL) and water (20 mL) was heated at 90.degree. C.
under N.sub.2 for 4 h. The mixture was diluted with water,
extracted with EtOAc and the combined organic extracts were dried
over Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography (Pet.
ether/EtOAc=500/1 to 200/1 to 100/1) to give the title compound
(5.2 g, 100%) as a white solid. LCMS-C: R.sub.t 2.47 min; m/z 215.0
[M+H].sup.+.
b) 3,5-Dimethoxy-[1,1'-biphenyl]-4-sulfonyl chloride I116
[0651] Prepared from 3,5-dimethoxy-1,1'-biphenyl I115 according to
the procedure described for
2,4-dimethoxy-[1,1'-biphenyl]-3-sulfonyl chloride I114. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.62-7.55 (m, 2H), 7.54-7.44 (m, 3H),
6.81 (s, 2H), 4.04 (s, 6H).
lv)
4-Methoxy-6-((2,2,2-trifluoroethoxy)methyl)benzo[d]isoxazol-3-amine
I118
##STR00081##
[0652] a)
2-Fluoro-6-methoxy-4((2,2,2-trifluoroethoxy)methyl)benzonitrile
I117
[0653] To a solution of
2-fluoro-4-(hydroxymethyl)-6-methoxybenzonitrile I7 (500 mg, 2.76
mmol) in dry THF (50 mL) at 0.degree. C. under N.sub.2 was added
NaH (60% w/w dispersion in oil, 331 mg, 8.28 mmol) followed by
2,2,2-trifluoroethyl trifluoromethanesulfonate (1.9 g, 8.28 mmol)
and the mixture was stirred at 0.degree. C. for 1 h, then allowed
to warm to RT and stirred overnight. The mixture was diluted with
water and extracted with EtOAc. The organic extract was washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The reaction was repeated two times using
2-fluoro-4-(hydroxymethyl)-6-methoxybenzonitrile I7 (100 mg, 0.55
mmol) and the three batches were combined and purified by column
chromatography (Pet. ether/EtOAc=5/1 to 2/1) to give the title
compound (577 mg, 57%) as a white solid. LCMS-C: R.sub.t 2.43 min;
m/z 263.9 [M+H].sup.+.
b)
4-Methoxy-6-((2,2,2-trifluoroethoxy)methyl)benzo[d]isoxazol-3-amine
I118
[0654] A suspension of acetohydroxamic acid (86 mg, 1.14 mmol) and
t-BuOK (128 mg, 1.14 mmol) in anhydrous DMF (10 mL) was stirred at
RT for 1 h.
2-Fluoro-6-methoxy-4-((2,2,2-trifluoroethoxy)methyl)benzonitrile
I117 (100 mg, 0.38 mmol) was then added and the mixture was stirred
at RT overnight. The mixture was diluted with water and extracted
with EtOAc. The organic extract was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The reaction was scaled up accordingly using
2-fluoro-6-methoxy-4-((2,2,2-trifluoroethoxy)methyl)benzonitrile
I117 (400 mg, 1.52 mmol) and the two batches were combined and
purified by column chromatography (Pet. ether/EtOAc=20/1 to 5/1) to
give the title product (350 mg, 67%) as a yellow solid. LCMS-C:
R.sub.t 2.08 min; m/z 277.0 [M+H].sup.+. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 6.96 (s, 1H), 6.68 (s, 1H), 5.94 (s, 2H),
4.74 (s, 2H), 4.13 (q, J=9.4 Hz, 2H), 3.90 (s, 3H).
lvi) 6-(Difluoromethoxy)-4-methoxybenzo[d]isoxazol-3-amine I121
##STR00082##
[0655] a) 4-(Difluoromethoxy)-2,6-difluorobenzonitrile I119
[0656] To a suspension of KOH (22.0 g, 392 mmol) in acetonitrile
(30 mL) and water (30 mL) at -20.degree. C. was added
2,6-difluoro-4-hydroxybenzonitrile (3.1 g, 20.0 mmol) portion-wise
followed by diethyl (bromodifluoromethyl)phosphonate (10.0 g, 37.4
mmol) and the mixture was stirred at RT overnight. Water was added
and the mixture was extracted with EtOAc (50 mL.times.3). The
combined organic extracts were washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give the title compound (4.0 g, 97%) as a colorless oil. LCMS-C:
R.sub.t 2.11 min; m/z 205.9 [M+H].sup.+.
b) 4-(Difluoromethoxy)-2-fluoro-6-methoxybenzonitrile I120
[0657] To a solution of
4-(difluoromethoxy)-2,6-difluorobenzonitrile I119 (2.52 g, 12.3
mmol) in dry THF (30 mL) was added NaOMe (1.32 g, 24.57 mmol)
portion-wise and the mixture was warmed at 40.degree. C. overnight.
Water was added and the mixture was extracted with EtOAc (30
mL.times.3). The combined organic extracts were washed with brine,
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by column chromatography
(Pet. ether/EtOAc=10/1 to 5/1) to give the title compound (663 mg,
25%) as a white solid. LCMS-C: R.sub.t 2.11 min; m/z 217.9
[M+H].sup.+.
c) 6-(Difluoromethoxy)-4-methoxybenzo[d]isoxazol-3-amine I121
[0658] A suspension of acetohydroxamic acid (680 mg, 9.15 mmol) and
t-BuOK (1.03 g, 9.15 mmol) in anhydrous DMF (50 mL) was stirred at
RT for 1 h.
5-(Difluoromethoxy)-1-fluoro-2-isocyano-3-methoxybenzene 1120 (663
mg, 3.05 mmol) was then added and the mixture was stirred at RT
overnight. The mixture was diluted with water and extracted with
EtOAc (30 mL.times.3). The combined organic extracts were washed
with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by column
chromatography (Pet. ether/EtOAc=10/1 to 8/1) to give the title
compound (186 mg, 26%) as light orange solid. LCMS-C: R.sub.t 1.14
min; m/z 231.0 [M+H].sup.+.
lviii) 5-Methyl-6-(pyridin-2-yl)benzo[d]isoxazol-3-amine I124
##STR00083##
[0659] a)
2-Fluoro-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl-
)benzonitrile I122
[0660] A mixture of 4-bromo-2-fluoro-5-methylbenzonitrile (500 mg,
2.34 mmol),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (1.78
g, 2.34 mmol), potassium acetate (918 mg, 9.36 mmol) and
Pd(dppf)Cl.sub.2 (188 mg, 0.23 mmol) in 1,4-dioxane (20 mL) was
heated at reflux under N.sub.2 for 3 h. The mixture was diluted
with water, extracted with EtOAc (300 mL) and the organic layer was
washed with water (50 mL.times.3), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give the title
compound (1.56 g, 88%), which was used in the next step without
further purification. LCMS-C: R.sub.t 2.75 min; m/z 262.0
[M+H].sup.+.
b) 2-Fluoro-5-methyl-4-(pyridin-2-yl)benzonitrile I123
[0661] To a solution of
2-fluoro-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonit-
rile I122 (1.32 g, 5.1 mmol) and 2-bromopyridine (1.69 g, 7.65
mmol) in 1,4-dioxane (50 mL) and water (10 mL) under N.sub.2 was
added Pd(PPh.sub.3).sub.4 (589 mg, 0.5 mmol) and Na.sub.2CO.sub.3
(2.16 g, 20.4 mmol) and the mixture was heated at 100.degree. C.
for 3 h. Water was added and the mixture was extracted with EtOAc.
The combined organic extracts were washed with brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography (Pet.
ether/EtOAc=8/1 to 3/1) to give the title compound (440 mg, 88%) as
a red solid. LCMS-C: R.sub.t 1.09 min; m/z 213.0 [M+H].sup.+.
c) 5-Methyl-6-(pyridin-2-yl)benzo[d]isoxazol-3-amine I124
[0662] A suspension of acetohydroxamic acid (255 mg, 3.39 mmol) and
t-BuOK (381 mg, 3.39 mmol) in anhydrous DMF (30 mL) was stirred at
0.degree. C. for 1 h.
2-Fluoro-5-methyl-4-(pyridin-2-yl)benzonitrile I123 (240 mg, 1.13
mmol) was then added and the mixture was allowed to warm to RT and
stirred overnight. Water was added and the mixture was extracted
with EtOAc (50 mL.times.3). The combined organic extracts were
washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=8/1 to 3/1) to give the
title compound (180 mg, 73%) as a white solid. LCMS-C: R.sub.t 0.50
min; m/z 226.0 [M+H].sup.+.
lix) 7-Bromo-5-methylbenzo[d]isoxazol-3-amine I129
##STR00084##
[0663] a) 3-Bromo-2-fluoro-5-methylbenzoic acid I125
[0664] To a solution of 2-bromo-1-fluoro-4-methylbenzene (10.0 g,
53 mmol) and diisopropylamine (5.9 g, 58 mmol) in anhydrous THF
(200 mL) at -78.degree. C. under N.sub.2 was added n-BuLi (2.5 M
solution in hexanes, 25.6 mL, 64.0 mmol) dropwise and the mixture
stirred at -78.degree. C. for 1 h. Excess solid CO.sub.2 (dry ice)
was added and stirring was continued at -78.degree. C. for 3 h. The
mixture was diluted with water (500 mL) and extracted with EtOAc
(500 mL). The organic layer was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give the title compound (12.3 g, 100%) as a brown solid, which
was used in the next step without further purification. LCMS-C:
R.sub.t2.03 min; m/z 232.8 [M+H].sup.+.
b) 3-Bromo-2-fluoro-5-methylbenzoyl chloride I126
[0665] To a solution of 3-bromo-2-fluoro-5-methylbenzoic acid I125
(12.3 g, 53 mmol) and DMF (4 drops) in DCM (100 mL) at RT under
N.sub.2 was added oxalyl chloride (13.0 g, 106 mmol) dropwise and
the mixture was stirred for 2 h. The mixture was concentrated under
reduced pressure to give the title compound (14.0 g, 100%) as a
brown solid, which was used in the next step without further
purification.
c) 3-Bromo-2-fluoro-5-methylbenzamide I127
[0666] A solution of 3-bromo-2-fluoro-5-methylbenzoyl chloride I126
(14.0 g, 53 mmol) in DCM (100 mL) was added dropwise to a 30%
aqueous ammonium hydroxide solution (100 mL) and the mixture was
stirred for 2 h. The mixture was diluted with EtOAc (200 mL),
washed with water (200 mL.times.3), brine and the organic layer was
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give the title compound (12.0 g, 97%) as a
brown solid, which was used in the next step without further
purification. LCMS-C: R.sub.t 1.01 min; m/z 231.9 [M+H].sup.+.
d) 3-bromo-2-fluoro-5-methylbenzonitrile I128
[0667] A solution of 3-bromo-2-fluoro-5-methylbenzamide I127 (10.0
g, 43.0 mmol) and thionyl chloride (15.4 g, 129 mmol) in DMF (100
mL) was heated at 100.degree. C. for 3 h. The mixture was diluted
with EtOAc (200 mL) and washed with water (400 mL.times.5), brine
and the organic layer was dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give the title compound (5.0
g, 54%) as a brown solid, which was used in the next step without
further purification. LCMS-C: R.sub.t 2.50 min; m/z 213.9
[M+H].sup.+.
e) 7-Bromo-5-methylbenzo[d]isoxazol-3-amine I129
[0668] A suspension of acetohydroxamic acid (5.27 g, 70.2 mmol) and
t-BuOK (7.88 g, 70.2 mmol) in anhydrous DMF (200 mL) was stirred at
0.degree. C. for 1 h. 3-Bromo-2-fluoro-5-methylbenzonitrile I128
(5.0 g, 23.4 mmol) was then added and the mixture was allowed to
warm to RT and stirred overnight. The mixture was diluted with
EtOAc (300 mL), washed with water (600 mL.times.4), brine and the
organic layer was dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=10/1) to give the title
compound (2.8 g, 52%) as a yellow solid. LCMS-C: R.sub.t 0.50 min;
m/z 226.9 [M+H].sup.+.
SYNTHESIS OF EXAMPLES
Examples 1-45 (Table A)
##STR00085##
[0670] Method AA
[0671] LiHMDS (1 M in THF, 445 .mu.L, 0.445 mmol) was added to a
solution of 4-chlorobenzo[d]isoxazol-3-amine (50 mg, 0.297 mmol) in
THF (3 mL) and stirred at room temperature for 10 minutes. The
sulfonyl chloride (0.445 mmol) was added and the reaction was
stirred for 16 hours at room temperature. The volatiles were
reduced to approximately 1 mL before DCM (3 mL) and water (3 mL)
were added and the mixture was stirred for 10 minutes. The mixture
was passed through a phase separator, the organic fraction was then
loaded onto a 1 g Si-amine cartridge (Biotage) and the cartridge
was washed with MeOH (6 mL), the product was then eluted with a HCl
solution (2 M, 1:1 methanol:1,4-dioxane 6 mL). The HCl washings
were then evaporated in vacuo to yield the desired product.
[0672] Method AB
[0673] LiHMDS (1 M in THF, 445 .mu.L, 0.445 mmol) was added to a
solution of 4-chlorobenzo[d]isoxazol-3-amine (50 mg, 0.297 mmol) in
THF (3 mL) and stirred at room temperature for 10 minutes. The
sulfonyl chloride (0.445 mmol) was added and the reaction was
stirred for 16 hours at room temperature. The volatiles were
reduced to approximately 1 mL before DCM (3 mL) and water (3 mL)
were added and the mixture was stirred for 10 minutes. The mixture
was passed through a phase separator, the organic fraction was then
loaded onto a 1 g Si-amine cartridge (Biotage) and the cartridge
was washed with MeOH (6 mL), the product was then eluted with a HCl
solution (2 M, 1:1 methanol:1,4-dioxane 6 mL). The HCl washings
were then evaporated in vacuo to yield the crude product which was
loaded onto silica gel and purified by silica gel column
chromatography (Biotage Isolera, SiO.sub.2 cartridge, 0-100% EtOAc
in petroleum benzine 40-60.degree. C.) to yield the desired
product.
[0674] Method AC
[0675] LiHMDS (1 M in THF, 445 .mu.L, 0.445 mmol) was added to a
solution of 4-chlorobenzo[d]isoxazol-3-amine (50 mg, 0.297 mmol) in
THF (3 mL) and stirred at room temperature for 10 minutes. The
sulfonyl chloride (0.445 mmol) was added and the reaction stirred
for 16 hours at room temperature. The volatiles were reduced to
approximately 1 mL before DCM (3 mL) and water (3 mL) were added
and the mixture was stirred for 10 minutes. The mixture was passed
through a phase separator, the organic fraction was then loaded
onto a 1 g Si-amine cartridge (Biotage) and the cartridge was
washed with MeOH (10 mL), the product was then eluted with a
methanolic HCl solution (.about.1.25 M, 10 mL). The HCl washings
were then evaporated in vacuo to yield the desired product.
[0676] Method AD
[0677] A solution of 4-chlorobenzo[d]isoxazol-3-amine (50 mg, 0.298
mmol) and the sulfonyl chloride (2 eq., 0.595 mmol) in pyridine
(1.5 mL) was irradiated in the microwave for 2 hours at 100.degree.
C. Upon cooling, the reaction mixture was loaded onto silica gel
and purified using silica gel column chromatography (Biotage
Isolera, 24 g SiO.sub.2 cartridge, 0-100% EtOAc in petroleum
benzine 40-60.degree. C.) to yield the desired product.
[0678] Method AE
[0679] A suspension of the sulfonyl chloride (2 eq., 1.19 mmol) and
4-chlorobenzo[d]isoxazol-3-amine (100 mg, 0.593 mmol) in pyridine
(1.5 mL) was irradiated in the microwave for 2 hours at 100.degree.
C. Upon cooling, the mixture was loaded onto silica gel and
purified using silica gel column chromatography (Biotage Isolera,
24 g SiO.sub.2 cartridge, 0-100% EtOAc in petroleum benzine
40-60.degree. C. then 0-40% MeOH in EtOAc) to yield the desired
product.
[0680] Method AF
[0681] A suspension of the sulfonyl chloride (2 eq., 1.19 mmol) and
4-chlorobenzo[d]isoxazol-3-amine (100 mg, 0.593 mmol) in pyridine
(1 mL) was irradiated in the microwave for 1 hour at 80.degree. C.
Upon cooling, the mixture was loaded onto silica gel and purified
using silica gel column chromatography (Biotage Isolera, 24 g
SiO.sub.2 cartridge, 0-100% EtOAc in petroleum benzine
40-60.degree. C.) to yield the desired product.
TABLE-US-00005 TABLE A Analytical Structure Name data Method 1
##STR00086## N-(4-chlorobenzo [d]isoxazol-3-yl)-2- (methoxymethyl)
benzenesulfonamide LCMS-A: rt 6.538 min, m/z 352.1 [M - H].sup.-
Method AB 2 ##STR00087## N-(4-chlorobenzo [d]isoxazol-3-yl)-3-
(methoxymethyl) benzenesulfonamide LCMS-A: rt 6.374 min, m/z 353.1
[M + H].sup.+ Method AA 3 ##STR00088## 3,4-dichloro-N-(4-
chlorobenzo[d]isoxazol-3-yl)- 2-methoxy benzenesulfonamide LCMS-A:
rt 6.751 min, m/z 409.0 [M + H].sup.+ Method AA 4 ##STR00089##
N-(4-chlorobenzo [d]isoxazol-3-yl)-2- phenoxybenzene sulfonamide
LCMS-A: rt 6.745 min, m/z 401.1 [M + H].sup.+ Method AA 5
##STR00090## N-(4-chlorobenzo [d]isoxazol-3-yl)-5-fluoro-2-
methoxybenzene sulfonamide LCMS-A: rt 6.360 min, m/z 357.1 [M +
H].sup.+ Method AA 6 ##STR00091## N-(4-chlorobenzo
[d]isoxazol-3-yl)-2-methyl- 2H-benzo[d][1,2,3]triazole-4-
sulfonamide LCMS-A: rt 6.200 min, m/z 364.1 [M + H].sup.+ Method AA
7 ##STR00092## N-(4-chlorobenzo [d]isoxazol-3-
yl)benzo[d][1,2,5]thiadiazole- 4-sulfonamide LCMS-A: rt 6.346 min,
m/z 367.0 [M + H].sup.+ Method AA 8 ##STR00093## N-(4-chlorobenzo
[d]isoxazol-3-yl)-2- hydroxybenzo[d]oxazole-6- sulfonamide LCMS-A:
rt 5.856 min, m/z 364.0 [M - H].sup.- Method AA 9 ##STR00094##
4-(benzyloxy)-N-(4- chlorobenzo[d]isoxazol-3- yl)benzenesulfonamide
LCMS-A: rt 6.764 min, m/z 415.1 [M + H].sup.+ Method AA 10
##STR00095## N-(4-chlorobenzo [d]isoxazol-3-yl)-2,5-
bis(2,2,2-trifluoroethoxy) benzenesulfonamide LCMS-A: rt 6.785 min,
m/z 505.0 [M + H].sup.+ Method AA 11 ##STR00096## N-(4-chlorobenzo
[d]isoxazol-3-yl)-2-(oxazol-5- yl)benzenesulfonamide LCMS-A: rt
6.322 min, m/z 376.1 [M + H].sup.+ Method AA 12 ##STR00097## methyl
2-(N-(4- chlorobenzo[d]isoxazol-3- yl)sulfamoyl)benzoate LCMS-A: rt
6.484 min, m/z 367.1 [M + H].sup.+ Method AA 13 ##STR00098##
N-(4-chlorobenzo [d]isoxazol-3-yl)-3-methyl-2- oxo-2,3-
dihydrobenzo[d]oxazole-6- sulfonamide LCMS-A: rt 6.103 min, m/z
378.0 [M - H].sup.- Method AA 14 ##STR00099## N-(4-chlorobenzo
[d]isoxazol-3-yl)-3,4-dihydro- 2H-benzo[b][1,4]dioxepine-7-
sulfonamide LCMS-A: rt 6.362 min, m/z 381.1 [M + H].sup.+ Method AA
15 ##STR00100## N-(4-chlorobenzo [d]isoxazol-3-yl)benzo
[d]thiazole-4-sulfonamide LCMS-A: rt 6.212 min, m/z 366.0 [M +
H].sup.+ Method AA 16 ##STR00101## N-(4-chlorobenzo [d]isoxazol-3-
yl)benzo[c][1,2,5]oxadiazole- 4-sulfonamide LCMS-A: rt 6.973 min,
m/z 351.0 [M + H].sup.+ Method AA 17 ##STR00102##
N-(4-chlorobenzo[d]isoxazol- 3-yl)-4-(4-fluorophenoxy)
benzenesulfonamide LCMS-A: rt 6.781 min, m/z 419.1 [M + H].sup.+
Method AA 18 ##STR00103## N-(4-chlorobenzo
[d]isoxazol-3-yl)-2-methyl-5- (2-methyloxazol-5-
yl)benzenesulfonamide LCMS-A: rt 6.410 min, m/z 404.1 [M + H].sup.+
Method AA 19 ##STR00104## N-(4-chlorobenzo
[d]isoxazol-3-yl)-5-(isoxazol- 5-yl)-2- methylbenzenesulfonamide
LCMS-A: rt 6.542 min, m/z 390.1 [M + H].sup.+ Method AA 20
##STR00105## N-(4-chlorobenzo [d]isoxazol-3-yl)-2-methyl-5-
(1H-pyrazol-1- yl)benzenesulfonamide LCMS-A: rt 6.493 min, m/z
389.1 [M + H].sup.+ Method AA 21 ##STR00106## N-(4-chlorobenzo
[d]isoxazol-3-yl)-4- (methylsulfonyl) benzenesulfonamide LCMS-A: rt
6.317 min, m/z 385.0 [M - H].sup.- Method AA 22 ##STR00107##
N-(4-chlorobenzo [d]isoxazol-3-yl)-4- isopropylbenzene sulfonamide
LCMS-A: rt 6.745 min, m/z 351.1 [M + H].sup.+ Method AA 23
##STR00108## N-(4-chlorobenzo [d]isoxazol-3-yl)-4-
(trifluoromethoxy)benzenesul- fonamide LCMS-A: rt 6.763 min, m/z
391.0 [M - H].sup.- Method AA 24 ##STR00109## N-(4-chlorobenzo
[d]isoxazol-3-yl)-4- isopropoxybenzene sulfonamide LCMS-A: rt 6.620
min, m/z 367.1 [M + H].sup.+ Method AA 25 ##STR00110##
4-(tert-butyl)-N-(4- chlorobenzo[d]isoxazol-3-
yl)benzenesulfonamide LCMS-A: rt 6.826 min, m/z 365.1 [M + H].sup.+
Method AA 26 ##STR00111## 4-chloro-N-(4- chlorobenzo[d]isoxazol-3-
yl)benzenesulfonamide LCMS-A: rt 6.614 min, m/z 343.0, 345.0 [M +
H].sup.+ Method AA 27 ##STR00112## N-(4-chlorobenzo
[d]isoxazol-3-yl)-4- methylbenzenesulfonamide LCMS-A: rt 6.447 min,
m/z 321.0 [M - H].sup.- Method AC 28 ##STR00113## N-(4-chlorobenzo
[d]isoxazol-3-yl)-4-(1H- pyrazol-1- yl)benzenesulfonamide LCMS-A:
rt 6.321 min, m/z 375.1 [M + H].sup.+ Method AC 29 ##STR00114##
N-(4-chlorobenzo [d]isoxazol-3-yl)-2,6- difluorobenzene sulfonamide
LCMS-A: rt 6.657 min, m/z 343.0 [M - H].sup.- Method AC 30
##STR00115## N-(4-chlorobenzo [d]isoxazol-3-yl)-2-
fluorobenzenesulfonamide LCMS-A: rt 6.389 min, m/z 327.0 [M +
H].sup.+ Method AC 31 ##STR00116## 3-chloro-N-(4-
chlorobenzo[d]isoxazol-3-yl)- 4- methylbenzenesulfonamide LCMS-A:
rt 6.808 min, m/z 355.0, 357.0 [M - H].sup.- Method AC 32
##STR00117## 4-bromo-N-(4- chlorobenzo[d]isoxazol-3-yl)-
2-(trifluoromethoxy) benzenesulfonamide LCMS-A: rt 7.440 min, m/z
468.9, 470.9 [M - H].sup.- Method AC 33 ##STR00118##
N-(4-chlorobenzo [d]isoxazol-3-yl)-2-methoxy- 4,5-
dimethylbenzenesulfonamide LCMS-A: rt 6.538 min, m/z 367.1 [M +
H].sup.+ Method AC 34 ##STR00119## N-(4-chlorobenzo
[d]isoxazol-3-yl)-4- (difluoromethoxy) benzenesulfonamide LCMS-A:
rt 6.487 min, m/z 373.0 [M - H].sup.- Method AC 35 ##STR00120##
N-(4-chlorobenzo [d]isoxazol-3-yl)-4- (pentafluoro-.lamda..sup.6-
sulfanyl)benzene- sulfonamide LCMS-A: rt 7.021 min; m/z 435.0 [M +
H].sup.+ Method AD 36 ##STR00121## N-(4-chlorobenzo
[d]isoxazol-3-yl)-3- (pentafluoro-.lamda..sup.6- sulfanyl)benzene-
sulfonamide LCMS-A: rt 7.219 min; m/z 435.0 [M + H].sup.+ Method AD
37 ##STR00122## N-(4-chlorobenzo [d]isoxazol-3-yl)-5-isopropyl-
2-methoxy-4- methylbenzenesulfonamide LCMS-A: rt 7.537 min; m/z
395.1 [M + H].sup.+ Method AD 38 ##STR00123## N-(4-chlorobenzo
[d]isoxazol-3-yl)-2- (difluoromethoxy) benzenesulfonamide LCMS-A:
rt 6.695 min; m/z 375.0 [M + H].sup.+ Method AD 39 ##STR00124##
N-(4-chlorobenzo [d]isoxazol-3-yl)-2,3-
dihydrobenzo[b][1,4]dioxine- 5-sulfonamide LCMS-A: rt 6.620 min;
m/z 367.0 [M + H].sup.+ Method AD 40 ##STR00125##
5-(tert-butyl)-N-(4- chlorobenzo[d]isoxazol-3-yl)- 2-methoxy-
benzenesulfonamide LCMS-B: rt 3.876 min, m/z 395.2 [M + H].sup.+
Method AD 41 ##STR00126## N-(4-chlorobenzo
[d]isoxazol-3-yl)-5-isopropyl- 2-methoxybenzene- sulfonamide
LCMS-A: rt 6.684 min, m/z 381.1 [M + H].sup.+ Method AD 42
##STR00127## N-(4-chlorobenzo [d]isoxazol-3-yl)-5,6,7,8-
tetrahydronaphthalene-2- sulfonamide LCMS-B: rt 3.859 min, m/z
363.1 [M + H].sup.+ Method AD 43 ##STR00128## N-(4-chlorobenzo
[d]isoxazol-3-yl)-[1,1'- biphenyl]-3-sulfonamide LCMS-B: rt 3.823
min, m/z 385.1 [M + H].sup.+ Method AD 44 ##STR00129##
N-(4-chlorobenzo [d]isoxazol-3-yl)-2,3-dihydro-
1H-indene-5-sulfonamide LCMS-B: rt 3.785 min, m/z 349.1 [M +
H].sup.+ Method AE 45 ##STR00130## 5-bromo-N-(4-
chlorobenzo[d]isoxazol-3-yl)- 2- methoxybenzenesulfonamide LCMS-B:
rt 3.752 min, m/z 417.0, 419.0 [M + H].sup.+ Method AF
Examples 46-71 (Table D)
##STR00131##
[0683] Method BA
[0684] A mixture of benzo[d]isoxazol-3-amine and a sulfonyl
chloride in pyridine (1 mL) was stirred at room temperature for 16
hours. The reaction was concentrated and diluted with 5% aqueous
HCl (1 mL) and sonicated for a minimum of 30 minutes. The resulting
precipitate was collected by filtration or DCM extraction
(2.times.1 mL) and purified using silica gel column chromatography
(EtOAc/petroleum benzine 40-60.degree. C. gradient) or preparative
mass-directed HPLC to give the desired product. See Table B for
reaction components and amounts used as well as purification
conditions.
[0685] Method BB
[0686] A mixture of benzo[d]isoxazol-3-amine and a sulfonyl
chloride in pyridine (0.5 mL) was stirred at room temperature for
64 hours. The reaction was concentrated and diluted with 5% aqueous
HCl (1 mL) and sonicated for a minimum of 30 minutes. The resulting
precipitate was collected by filtration and a portion of the crude
material (50 mg or less) was purified by preparative mass-directed
HPLC to give the desired product. See Table B for reaction
components and amounts used.
[0687] Method BC
[0688] A mixture of benzo[d]isoxazol-3-amine and a sulfonyl
chloride in pyridine (0.5 mL) was stirred at room temperature for
16 hours. The reaction was concentrated and diluted with 5% aqueous
HCl (1 mL) and sonicated for a minimum of 30 minutes. The resulting
precipitate was collected by filtration and a portion of the crude
material (50 mg or less) was purified by mass directed preparative
HPLC to give the desired product. See Table B for reaction
components and amounts used.
TABLE-US-00006 TABLE B Benzo[d] isoxazol-3- Benzenesulfonyl
chloride amine Method/Work- Mass Moles Mass Moles up/Purification
Product Structure and name (g) (mmol) (g) (mmol) method 46
##STR00132## 0.143 0.602 0.030 0.23 Method BA/Filtration/ Column
chromatography 0-45% gradient 47 ##STR00133## 0.150 0.682 0.031
0.23 Method BA/Filtration/ Column chromatography 0-40% gradient 48
##STR00134## 0.148 0.563 0.033 0.25 Method BA/Filtration/ Column
chromatography 0-40% gradient 49 ##STR00135## 0.140 0.561 0.031
0.23 Method BA/Filtration/ Column chromatography 0-40% gradient 50
##STR00136## 0.142 0.535 0.030 0.22 Method BA/Filtration/ Column
chromatography 0-100% gradient 51 ##STR00137## 0.156 0.611 0.033
0.24 Method BA/Filtration/ Column chromatography 0-100% gradient 52
##STR00138## 0.036 0.180 0.028 0.21 Method BA/DCM extraction/Column
chromatography 0-50% gradient 53 ##STR00139## 0.074 0.360 0.032
0.24 Method BA/DCM extraction/Column chromatography 0-50% gradient
54 ##STR00140## 0.148 0.54 0.034 0.25 Method BA/Filtration/ Column
chromatography 0-100% gradient 55 ##STR00141## 0.158 0.58 0.030
0.22 Method BA/Filtration/ Column chromatography 0-100% gradient 56
##STR00142## 0.063 0.27 0.030 0.22 Method BA/DCM extraction/Column
chromatography 0-100% gradient 57 ##STR00143## 0.170 0.68 0.027
0.20 Method BA/Filtration/ Column chromatography 0-40% gradient.
Triturated minimal acetone, product contained approx. 10% sulfonic
acid by-product 58 ##STR00144## 0.101 0.488 0.029 0.21 Method
BA/Filtration/ mass-directed HPLC 59 ##STR00145## 0.105 0.466 0.033
0.25 Method BB 60 ##STR00146## 0.171 0.698 0.032 0.24 Method BB 61
##STR00147## 0.100 0.640 0.032 0.24 Method BB 62 ##STR00148## 0.170
0.694 0.031 0.23 Method BB 63 ##STR00149## 0.107 0.454 0.033 0.25
Method BC 64 ##STR00150## 0.120 0.553 0.029 0.22 Method BC 65
##STR00151## 0.103 0.447 0.032 0.24 Method BC 66 ##STR00152## 0.111
0.490 0.030 0.23 Method BC 67 ##STR00153## 0.1025 0.396 0.033 0.25
Method BC 68 ##STR00154## 0.111 0.432 0.033 0.25 Method BB
[0689] Method CA
[0690] A mixture of benzo[d]isoxazol-3-amine and a sulfonyl
chloride in pyridine (1 mL) was stirred at room temperature for 16
hours when a second portion of benzenesulfonyl chloride was added
and stirred for an additional 64 hours. The reaction was
concentrated and diluted with 5% aqueous HCl (1 mL) and sonicated
for a minimum of 30 minutes. The resulting precipitate was
collected by filtration and purified either by preparative
mass-directed HPLC (up to 50 mg of crude material) or by silica gel
column chromatography (0-40% EtOAc/petroleum benzine 40-60.degree.
C.) to give the desired product. See Table C for reaction
components and amounts used as well as purification conditions.
TABLE-US-00007 TABLE C Benzo[d] Benzenesulfonyl chloride (added in
2 portions) isoxazol-3-amine Method/ Mass Moles Mass Moles
Purification Product Structure and name Portion (g) (mmol) (g)
(mmol) method 69 ##STR00155## 1st 2nd 0.117 0.103 0.463 0.408 0.060
0.45 Method CA/Mass- directed HPLC 70 ##STR00156## 1st 2nd 0.092
0.088 0.36 0.35 0.052 0.39 Method CA/column chromatog- raphy 71
##STR00157## 1st 2nd 0.103 0.101 0.398 0.390 0.065 0.48 Method
CA/column chromatog- raphy
TABLE-US-00008 TABLE D Structure Name Analytical 46 ##STR00158## N-
(benzo[d]isoxazol- 3-yl)-2,5- dimethoxybenzene- sulfonamide LCMS-B:
rt 3.560 min; m/z 335.1 [M + H].sup.+; .sup.1H NMR (400 MHz,
acetone-d.sub.6) .delta. 10.01 (br s, 1H), 8.14-8.06 (m, 1H), 7.64
(ddd, J = 8.3, 7.0, 1.2 Hz, 1H), 7.55 (dt, J = 8.5, 0.8 Hz, 1H),
7.43-7.36 (m, 2H), 7.17 (dd, J = 9.1, 2.9 Hz, 1H), 7.13 (d, J = 8.8
Hz, 1H), 3.82 (s, 3H), 3.77 (s, 3H) 47 ##STR00159## N-
(benzo[d]isoxazol- 3-yl)-2-methoxy- 5-methyl benzene- sulfonamide
LCMS-B: rt 3.585 min; m/z 319.1 [M + H].sup.+; .sup.1H NMR (400
MHz, acetone-d.sub.6) .delta. 9.94 (br s, 1H), 8.14-8.06 (m, 1H),
7.70-7.65 (m, 1H), 7.63 (ddd, J = 8.3, 7.0, 1.2 Hz, 1H), 7.54 (dt,
J = 8.5, 0.8 Hz, 1H), 7.43- 7.36 (m, 2H), 7.08 (d, J = 8.5 Hz, 1H),
3.84 (s, 3H), 2.28 (s, 3H). 48 ##STR00160## N- (benzo[d]isoxazol-
3-yl)-5-(tert- butyl)-2-methoxy- benzene- sulfonamide LCMS-B: rt
3.806 min; m/z 361.2 [M + H].sup.+; .sup.1H NMR (400 MHz,
acetone-d.sub.6) .delta. 10.00 (br s, 1H), 8.13-8.08 (m, 1H), 7.91
(d, J = 2.5 Hz, 1H), 7.66-7.60 (m, 2H), 7.55- 7.51 (m, 1H), 7.38
(ddd, J = 8.0, 7.0, 0.9 Hz, 1H), 7.10 (d, J = 8.7 Hz, 1H), 3.83 (s,
3H), 1.26 (s, 9H) 49 ##STR00161## N- (benzo[d]isoxazol-
3-yl)-5-isopropyl- 2-methoxy- benzene- sulfonamide LCMS-B: rt 3.744
min; m/z 347.2 [M + H].sup.+; .sup.1H NMR (400 MHz,
acetone-d.sub.6) .delta. 9.97 (br s, 1H), 8.12-8.08 (m, 1H), 7.75
(d, J = 2.4 Hz, 1H), 7.63 (ddd, J = 8.4, 7.0, 1.2 Hz, 1H), 7.53
(dt, J = 8.5, 0.9 Hz, 1H), 7.48 (ddd, J = 8.5, 2.4, 0.6 Hz, 1H),
7.38 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H), 7.10 (d, J = 8.6 Hz, 1H),
3.84 (s, 3H), 2.95-2.87 (m, 1H), 1.18 (d, J = 6.9 Hz, 6H) 50
##STR00162## methyl 3-(N- (benzo[d] isoxazol-3- yl)sulfamoyl)-4-
methoxy- benzoate LCMS-A: rt 6.024 min; m/z 363.1 [M + H].sup.+;
.sup.1H NMR (400 MHz, acetone-d.sub.6) .delta. 10.24 (br s, 1H),
8.55 (d, J = 2.2 Hz, 1H), 8.24 (dd, J = 8.8, 2.2 Hz, 1H), 8.09 (dt,
J = 8.1, 1.0 Hz, 1H), 7.66 (ddd, J = 8.4, 7.0, 1.2 Hz, 1H), 7.57
(dt, J = 8.5, 0.8 Hz, 1H), 7.42 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H),
7.35 (d, J = 8.8 Hz, 1H), 4.00 (s, 3H), 3.89 (s, 3H) 51
##STR00163## N- (benzo[d]isoxazol- 3-yl)-5-chloro-2- methoxy-4-
methylbenzene- sulfonamide LCMS-B: rt 3.738 min; m/z 353.1/355.1 [M
+ H].sup.+; .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta.
8.10-8.06 (m, 1H), 7.81 (s, 1H), 7.65 (ddd, J = 8.3, 7.0, 1.2 Hz,
1H), 7.58-7.54 (m, 1H), 7.43-7.37 (m, 1H), 7.21 (s, 1H), 3.87 (s,
3H), 2.38 (s, 3H) 52 ##STR00164## N- (benzo[d]isoxazol- 3-yl)-3-
ethylbenzene- sulfonamide LCMS-B: rt 3.707 min; m/z 303.2 [M +
H].sup.+; .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta. 8.02- 7.97
(m, 1H), 7.85-7.82 (m, 1H), 7.82- 7.78 (m, 1H), 7.68-7.62 (m, 1H),
7.59- 7.55 (m, 1H), 7.54-7.45 (m, 2H), 7.41- 7.35 (m, 1H), 2.70 (q,
J = 7.6 Hz, 2H)*, 1.19 (t, J = 7.6 Hz, 3H). * Partially overlapping
with water peak 53 ##STR00165## N- (benzo[d]isoxazol- 3-yl)-4-
ethylbenzene- sulfonamide LCMS-B: rt 3.690 min; m/z 303.1 [M +
H].sup.+; .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta. 8.03- 7.98
(m, 1H), 7.93-7.89 (m, 2H), 7.65 (ddd, J = 8.2, 7.0, 1.1 Hz, 1H),
7.60-7.55 (m, 1H), 7.46-7.41 (m, 2H), 7.41-7.36 (m, 1H), 2.71 (q, J
= 7.6 Hz, 2H)*, 1.21 (t, J = 7.6 Hz, 3H). * Partially overlapping
with water peak 54 ##STR00166## N- (benzo[d]isoxazol-
3-yl)-2-methoxy- 5-(1H-pyrazol-1- yl)benzene- sulfonamide LCMS-B:
rt 3.554 min; m/z 371.1 [M + H].sup.+; .sup.1H NMR (400 MHz,
acetone-d.sub.6) .delta. 8.36 (d, J = 2.8 Hz, 1H), 8.33-8.31 (m,
1H), 8.13- 8.07 (m, 1H), 8.05 (dd, J = 9.0, 2.8 Hz, 1H), 7.68-7.61
(m, 2H), 7.56-7.52 (m, 1H), 7.40 (ddd, J = 8.0, 7.1, 0.8 Hz, 1H),
7.33 (d, J = 9.0 Hz, 1H), 6.49 (dd, J = 2.5, 1.7 Hz, 1H), 3.93 (s,
3H). 55 ##STR00167## N- (benzo[d]isoxazol- 3-yl)-5-(isoxazol-
5-yl)-2- methoxybenzene- sulfonamide LCMS-B: rt 3.516 min; m/z
372.1 [M + H].sup.+; .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta.
8.46 (d, J = 1.9 Hz, 1H), 8.36 (d, J = 2.3 Hz, 1H), 8.13-8.07 (m,
2H), 7.63 (ddd, J = 8.3, 7.1, 1.2 Hz, 1H), 7.56-7.52 (m, 1H), 7.43-
7.36 (m, 2H), 6.90 (d, J = 1.9 Hz, 1H), 3.97 (s, 3H) 56
##STR00168## N- (benzo[d]isoxazol- 3-yl)-2,5- diethylbenzene-
sulfonamide LCMS-B: rt 3.801 min; m/z 331.2 [M + H].sup.+; .sup.1H
NMR (400 MHz, acetone-d.sub.6) .delta. 8.01- 7.97 (m, 1H),
7.95-7.92 (m, 1H), 7.66- 7.61 (m, 1H), 7.57-7.52 (m, 1H), 7.45-
7.41 (m, 1H), 7.40-7.34 (m, 2H), 3.10 (q, J = 7.5 Hz, 2H)*, 2.66
(q, J = 7.5 Hz, 2H)*, 1.22 (t, J = 7.5 Hz, 3H), 1.17 (t, J = 7.6
Hz, 3H). * Partially overlapping with water peak 57 ##STR00169## N-
(benzo[d]isoxazol- 3-yl)-2-ethoxy-5- ethylbenzene- sulfonamide
LCMS-B: rt 3.763 min; m/z 347.2 [M + H].sup.+; .sup.1H NMR (400
MHz, acetone-d.sub.6) .delta. 9.78 (br s, 1H), 8.13-8.08 (m, 1H),
7.74(d, J = 2.3 Hz, 1H), 7.63 (ddd, J = 8.3, 7.0, 1.2 Hz, 1H),
7.56-7.52 (m, 1H), 7.44-7.41 (m, 1H), 7.38 (ddd, J = 8.0, 7.1, 0.8
Hz, 1H), 7.09(d, J = 8.5 Hz, 1H), 4.16 (q, J = 7.0 Hz, 2H), 1.29
(t, J = 7.0 Hz, 3H), 1.16 (t, J = 7.6 Hz, 3H). NB approx. 10%
sulfonic acid impurity 58 ##STR00170## N- (benzo[d]isoxazol-
3-yl)-2- methoxybenzene- sulfonamide HPLC-MS: rt 5.11 min; m/z
305.2 [M + H].sup.+; .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta.
10.35- 9.70 (br s, 1H), 8.12-8.08 (m, 1H), 7.90- 7.86 (dd, J = 7.9,
1.7 Hz, 1H), 7.66-7.58 (m, 2H), 7.56-7.52 (m, 1H), 7.41-7.36 (ddd,
J = 8.0, 7.0, 0.9 Hz, 1H), 7.21-7.17 (dd, J = 8.4, 0.8 Hz, 1H),
7.09-7.03 (m, 1H), 3.90-3.87 (s, 3H). 59 ##STR00171## N-
(benzo[d]isoxazol- 3-yl)-3-chloro-2- methylbenzene- sulfonamide
HPLC-MS: rt 6.38 min; m/z 323.22/325.17 [M + H].sup.+; .sup.1H NMR
(400 MHz, acetone-d.sub.6) .delta. 11.10-10.01 (br s, 1H),
8.15-8.10 (dd, J = 8.0, 1.0 Hz, 1H), 8.02-7.97 (m, 1H), 7.74-7.69
(m, 1H), 7.68-7.62 (ddd, J = 8.3, 7.0, 1.2 Hz, 1H), 7.59-7.54 (m,
1H), 7.46-7.41 (m, 1H), 7.41-7.37 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H),
2.77-2.74 (s, 3H) 60 ##STR00172## N- (benzo[d]isoxazol- 3-yl)-2,3-
dichlorobenzene- sulfonamide HPLC-MS: rt 6.27 min; m/z
343.13/345.15/347.10 [M + H].sup.+; .sup.1H NMR (400 MHz,
acetone-d.sub.6) .delta. 8.25-8.22 (dd, J = 8.0, 1.5 Hz, 1H),
8.06-8.02 (m, 1H), 7.93-7.90 (dd, J = 8.1, 1.5 Hz, 1H), 7.69- 7.64
(ddd, J = 8.3, 7.0, 1.2 Hz, 1H), 7.64- 7.59 (m, 1H), 7.59-7.55 (m,
1H), 7.44- 7.38 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H) 61 ##STR00173## N-
(benzo[d]isoxazol- 3-yl)-3,4- dichlorobenzene- sulfonamide HPLC-MS:
rt 6.62 min; m/z 343.13/345.15/347.10 [M + H].sup.+; .sup.1H NMR
(400 MHz, acetone-d.sub.6) .delta. 8.19-8.17 (d, J = 2.1 Hz, 1H),
7.99-7.94 (m, 2H), 7.85- 7.82 (d, J = 8.5 Hz, 1H), 7.70-7.65 (ddd,
J = 8.2, 7.0, 1.2 Hz, 1H), 7.62-7.58 (m, 1H), 7.43-7.37 (ddd, J =
8.0, 7.0, 0.9 Hz, 1H) 62 ##STR00174## N- (benzo[d]isoxazol-
3-yl)-2,4- dichlorobenzene- sulfonamide HPLC-MS: rt 6.42 min; m/z
343.13/345.15/347.16 [M + H].sup.+; .sup.1H NMR (400 MHz,
acetone-d.sub.6) .delta. 8.26-8.22 (d, J = 8.6 Hz, 1H), 8.06-8.02
(m, 1H), 7.75- 7.72 (d, J = 2.0 Hz, 1H), 7.70-7.63 (m, 2H),
7.60-7.56 (m, 1H), 7.44-7.38 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H) 63
##STR00175## N- (benzo[d]isoxazol- 3-yl)-5-ethyl-2- methoxy-
benzenesulfon- amide HPLC-MS: rt 5.39 min; m/z 333.2 [M + H].sup.+;
.sup.1H NMR (400 MHz, acetone-d.sub.6) .delta. 10.19-9.73 (br s,
1H), 8.13-8.08 (ddd, J = 8.1, 1.0, 1.0 Hz, 1H), 7.73-7.69 (d, J =
2.3 Hz, 1H), 7.66-7.60 (ddd, J = 8.3, 7.0, 1.2 Hz, 1H), 7.56-7.52
(ddd, J = 8.5, 0.8, 0.8 Hz, 1H), 7.47-7.42 (dd, J = 8.5, 2.3 Hz,
1H), 7.42-7.36 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H), 7.13-7.07 (d, J =
8.5 Hz, 1H), 3.86-3.82 (s, 3H), 2.65-2.56 (q, J = 7.6 Hz, 2H),
1.18-1.12 (t, J = 7.6 Hz, 3H) 64 ##STR00176## N- (benzo[d]isoxazol-
3-yl)-2,3-dihydro- 1H-indene-5- sulfonamide HPLC-MS: rt 5.45 min;
m/z 315.2 [M + H].sup.+; .sup.1H NMR (400 MHz, acetone-d.sub.6)
.delta. 10.80-9.43 (br s, 1H), 8.04-7.99 (ddd, J = 8.1, 1.0, 1.0
Hz, 1H), 7.84-7.80 (m, 1H), 7.78-7.73 (m, 1H), 7.68-7.62 (ddd, J =
8.3, 7.0, 1.2 Hz, 1H), 7.60-7.54 (ddd, J = 8.5, 0.8, 0.8 Hz, 1H),
7.42-7.35 (m, 2H), 2.97-2.88 (t, J = 7.5 Hz, 4H), 2.12- 2.05 (dd, J
= 15.0, 7.6 Hz, 2H). 65 ##STR00177## N- (benzo[d]isoxazol-
3-yl)-5,6,7,8- tetrahydro- naphthalene-2- sulfonamide HPLC-MS: rt
5.57 min; m/z 329.5 [M + H].sup.+; .sup.1H NMR (400 MHz,
acetone-d.sub.6) .delta. 10.44-9.90 (br s, 1H), 8.03-7.98 (ddd, J =
8.1, 1.1, 1.1 Hz, 1H), 7.69-7.63 (m, 3H), 7.60-7.55 (ddd, J = 8.5,
0.8, 0.8 Hz, 1H), 7.42-7.36 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H),
7.25-7.20 (d, J = 8.3 Hz, 1H), 2.81- 2.76 (m, 4H), 1.83-1.73 (m,
4H). 66 ##STR00178## N- (benzo[d]isoxazol- 3- yl)naphthalene-2-
sulfonamide HPLC-MS: rt 5.46 min; m/z 325.1 [M + H].sup.+; .sup.1H
NMR (400 MHz, acetone-d.sub.6) .delta. 11.03-9.58 (br s, 1H),
8.66-8.62 (m, 1H), 8.15-8.11 (m, 1H), 8.11-8.08 (d, J = 8.9 Hz,
1H), 8.04-7.98 (m, 3H), 7.73- 7.60 (m, 3H), 7.56-7.52 (ddd, J =
8.5, 0.8, 0.8 Hz, 1H), 7.41-7.35 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H).
67 ##STR00179## N- (benzo[d]isoxazol- 3-yl)-3-(5-methyl-
1,3,4-oxadiazol-2- yl)benzene- sulfonamide HPLC-MS: rt 5.00 min;
m/z 357.7 [M + H].sup.+; .sup.1H NMR (400 MHz, acetone-d.sub.6)
.delta. 8.62-8.61 (dd, J = 1.6, 1.6 Hz, 1H), 8.29- 8.26 (ddd, J =
7.8, 1.4, 1.4 Hz, 1H), 8.23- 8.19 (ddd, J = 8.0, 1.8, 1.1 Hz, 1H),
7.99- 7.96 (ddd, J = 8.1, 0.9, 0.9 Hz, 1H), 7.85- 7.80 (dd, J =
7.9, 7.9 Hz, 1H), 7.67- 7.63 (ddd, J = 8.3, 7.0, 1.2 Hz, 1H), 7.58-
7.55 (m, 1H), 7.41-7.36 (ddd, J = 7.9, 7.0, 0.8 Hz, 1H), 2.63-2.60
(s, 3H). 68 ##STR00180## N- (benzo[d]isoxazol- 3-yl)-5-
bromopyridine-2- sulfonamide LCMS-B: rt 3.583 min; m/z 354/356 [M +
H].sup.+; .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta.
11.19-10.23 (br s, 1H), 8.81-8.78 (dd, J = 2.3, 0.7 Hz, 1H),
8.38-8.34 (dd, J = 8.4, 2.3 Hz, 1H), 8.15-8.10 (dd, J = 8.4, 0.7
Hz, 1H), 8.06-8.01 (ddd, J = 8.1, 1.0, 1.0 Hz, 1H), 7.69-7.64 (ddd,
J = 8.3, 7.0, 1.2 Hz, 1H), 7.59-7.56 (ddd, J = 8.5, 0.8, 0.8 Hz,
1H), 7.43-7.38 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H). 69 ##STR00181## N-
(benzo[d]isoxazol- 3-yl)-[1,1'- biphenyl]-4- sulfonamide HPLC-MS:
rt 6.62 min; m/z 351.19 [M + H].sup.+; .sup.1H NMR (400 MHz,
acetone-d.sub.6) .delta. 8.11-8.06 (m, 2H), 8.04-8.00 (m, 1H),
7.90-7.85 (m, 2H), 7.76-7.68 (m, 2H), 7.66 (ddd, J = 8.3, 7.0, 1.2
Hz, 1H), 7.58 (dt, J = 8.5, 0.8 Hz, 1H), 7.53-7.47 (m, 2H),
7.46-7.37 (m, 2H). 70 ##STR00182## N- (benzo[d]isoxazol-
3-yl)-[1,1'- biphenyl]-3- sulfonamide LCMS-B: rt 3.754 min; m/z
351.1 [M + H].sup.+; .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta.
8.29-8.26 (m, 1H), 8.03-8.00 (m, 1H), 7.97 (dddd, J = 9.0, 7.8,
1.8, 0.9 Hz, 2H), 7.72-7.63 (m, 4H), 7.58 (dt, J = 8.5, 0.8 Hz,
1H), 7.54-7.48 (m, 2H), 7.46-7.37 (m, 2H) 71 ##STR00183## N-
(benzo[d]isoxazol- 3-yl)-4- cyclohexyl- benzenesulfonamide LCMS-B:
rt 3.999 min; m/z 357.1 [M + H].sup.+; .sup.1H NMR (400 MHz,
acetone-d.sub.6) .delta. 8.02-7.98 (m, 1H), 7.93-7.89 (m, 2H), 7.65
(ddd, J = 8.3, 7.0, 1.2 Hz, 1H), 7.59- 7.55 (m, 1H), 7.47-7.43 (m,
2H), 7.38 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H), 2.66-2.58 (m, 2H),
1.85-1.78 (m, 4H), 1.76-1.69 (m, 1H), 1.50-1.34 (m, 4H), 1.32-1.24
(m, 1H).
Examples 72-88 (Table E)
##STR00184##
[0692] Method EA
[0693] NaH (60% in mineral oil, 49 mg, 1.22 mmol) was added to a
solution of 4-methoxybenzo[d]isoxazol-3-amine (50 mg, 0.305 mmol)
in THF (3.0 mL) and stirred at room temperature for 10 minutes. The
sulfonyl chloride (1 eq., 0.305 mmol) was added and the reaction
was stirred for 16 hours. The volatiles were reduced to
approximately 1 mL before DCM (3 mL) and water (3 mL) were added
and the mixture was stirred for 10 minutes. The mixture was passed
through a phase separator, the organic fraction was then loaded
onto a 1 g Si-amine cartridge (Biotage) and the cartridge was
washed with MeOH (6 mL), the product was then eluted with a HCl
solution (1.25 M in methanol, 6 mL). The HCl washings were
evaporated in vacuo to yield the desired product.
[0694] Method EB
[0695] NaH (60% in mineral oil, 61 mg, 1.52 mmol) was added to a
solution of 4-methoxybenzo[d]isoxazol-3-amine (50 mg, 0.305 mmol)
in DMF (3.0 mL) and stirred at room temperature for 10 minutes. The
sulfonyl chloride (1 eq., 0.305 mmol) was added and the reaction
was stirred for 16 hours. The resultant mixture was loaded onto
silica gel and purified by column chromatography (0-100% petroleum
benzine 40-60.degree. C. then 0-60% MeOH in EtOAc) to yield the
desired product.
[0696] Method EC
[0697] NaH (60% in mineral oil, 22 mg, 0.914 mmol) was added to a
solution of 4-methoxybenzo[d]isoxazol-3-amine (50 mg, 0.305 mmol)
in DMF (5 mL) and stirred at room temperature for 10 minutes. The
sulfonyl chloride (1 eq., 0.305 mmol) was added and the reaction
was stirred for 16 hours. The resultant mixture was quenched with
water (3 mL), stirred for 10 minutes at room temperature then
loaded onto silica gel and purified by column chromatography
(0-100% petroleum benzine 40-60.degree. C. then 0-60% MeOH in
EtOAc) to yield the desired product.
[0698] Method ED
[0699] NaH (60% in mineral oil, 5 or 10 eq.) was added to a
solution of 4-methoxybenzo[d]isoxazol-3-amine (100 mg, 0.609 mmol)
in THF (5.0 mL) and stirred at room temperature for 10 minutes. The
sulfonyl chloride (1 eq., 0.609 mmol) was added and the reaction
was stirred for 16 hours. The resultant mixture was loaded onto
silica gel and purified by column chromatography (0-100% petroleum
benzine 40-60.degree. C. then 0-60% MeOH in EtOAc) to yield the
desired product.
[0700] Method EF
[0701] NaH (60% in mineral oil, 122 mg, 3.05 mmol) was added to a
solution of 4-methoxybenzo[d]isoxazol-3-amine (100 mg, 0.609 mmol)
in THF (5.0 mL) and stirred at room temperature for 10 minutes. The
sulfonyl chloride (1 eq., 0.609 mmol) was added and the reaction
was stirred for 16 hours. The resultant mixture was loaded onto
silica gel and purified by column chromatography (0-100% petroleum
benzine 40-60.degree. C. then 0-60% MeOH in EtOAc) and the isolated
solid was sonicated in MeOH (1 mL) and collected by filtration to
yield the desired product.
[0702] Method EG
[0703] NaH (60% in mineral oil, 122 mg, 3.05 mmol) was added to a
solution of 4-methoxybenzo[d]isoxazol-3-amine (100 mg, 0.609 mmol)
in THF (5.0 mL) and stirred at room temperature for 10 minutes. The
sulfonyl chloride (1 eq., 0.609 mmol) was added and the reaction
was stirred for 16 hours at room temperature. The volatiles were
reduced to approximately 1 mL before DCM (3 mL) and water (3 mL)
were cautiously added and stirred for 10 minutes. The mixture was
passed through a phase separator, the organic fraction was then
loaded onto a 1 g Si-amine cartridge (Biotage) and the cartridge
was washed with MeOH (6 mL), the product was then eluted with a HCl
solution (2 M, 1:1 methanol:1,4-dioxane, 6 mL). The HCl washings
were then evaporated in vacuo to yield the desired product.
[0704] Method EH
[0705] A suspension of 4-methoxybenzo[d]isoxazol-3-amine (48 mg,
0.29 mmol) and NaH (60% in mineral oil, 0.117 mg, 2.93 mmol) in DMF
(10 mL) was stirred at room temperature for 10 minutes before being
cooled to -78.degree. C. To this cooled mixture the sulfonyl
chloride (1.5 eq., 0.439 mmol) was added and the mixture was
stirred at -78.degree. C. for 1 hour then warmed to room
temperature and stirred for 16 hours. The reaction mixture was
loaded onto silica gel and purified by column chromatography
(Biotage Isolera, 24 g SiO.sub.2 cartridge, 0-100% EtOAc in
petroleum benzine 40-60.degree. C. then 0-40% MeOH in EtOAc) to
give a solid which was suspended in diethyl ether (25 mL) and
sonicated for 5 minutes. The solid was collected by filtration and
air dried to give the desired product.
[0706] Method EI
[0707] A mixture of 4-methoxybenzo[d]isoxazol-3-amine (0.035 g,
0.21 mmol) and a sulfonyl chloride (1.05 eq., 0.22 mmol) in
pyridine (1 mL) was stirred at room temperature for 16 hours. The
reaction was concentrated and diluted with 5% aqueous HCl (1 mL)
and sonicated for a minimum of 30 minutes. Extraction with DCM
(2.times.1 mL) and purification using silica gel column
chromatography (0-100% EtOAc in petroleum benzine 40-60.degree. C.)
gave the desired product.
[0708] Method EJ
[0709] To a solution of 4-methoxybenzo[d]isoxazol-3-amine (1 eq.)
in THF (3 mL) was added LiHMDS (1M in THF, 1.5 eq.). After 10
minutes of stirring, the sulfonyl chloride (1.5 eq.) was added and
the reaction was left to stir for 17 hours, open to air. The THF
was removed in vacuo, then DCM (3 mL) and water (3 mL) were added
and stirred for 10 minutes. After separation of the layers, the
organic fraction was loaded onto a 1 g Si-amine cartridge
(Biotage). The cartridge was washed with MeOH (6 mL), then stripped
with 1.25 M HCl in MeOH (6 mL). The HCl washings were then
evaporated in vacuo to yield the desired product.
TABLE-US-00009 TABLE E Structure and yield (where applicable) Name
Analytical data Method 72 ##STR00185## 2-methoxy-N-(4-
methoxybenzo[d] isoxazol-3-yl)-4,6- dimethylbenzene- sulfonamide
LCMS-A: rt 6.474 min, m/z 363.1 [M + H].sup.+; .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.51- 9.42 (s, 1H), 7.61-7.51 (t, J =
8.2 Hz, 1H), 7.19-7.10 (d, J = 8.4 Hz, 1H), 6.90-6.87 (s, 1H),
6.87-6.82 (d, J = 8.0 Hz, 1H), 6.80-6.74 (s, 1H), 3.99-3.93 (s,
3H), 3.84- 3.78 (s, 3H), 2.58-2.54 (s, 3H), 2.31-2.25 (s, 3H).
Method EA 73 ##STR00186## N-(4-methoxy benzo[d]isoxazol-
3-yl)-3-methyl- quinoline-8- sulfonamide LCMS-A: rt 6.469 min, m/z
= 370.1 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
= 8.94 (d, J = 2.1 Hz, 1H), 8.37 (dd, J = 7.3, 1.3 Hz, 1H), 8.33
(s, 1H), 8.25 (d, J = 8.0 Hz, 1H), 7.76 (t, J = 7.8 Hz, 1H), 7.50
(t, J = 8.2 Hz, 1H), 7.10 (d, J = 8.4 Hz, 1H), 6.79 (d, J = 8.1 Hz,
1H), 3.89 (s, 3H), 2.52 (s, 3H). Method EB 74 ##STR00187##
5-methoxy-N-(4- methoxybenzo[d] isoxazol-3-yl) quinoline-8-
sulfonamide LCMS-A: rt 6.461 min, m/z 386.1 [M + H].sup.+; .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. = 9.08 (dd, J = 4.3, 1.7 Hz,
1H), 8.62 (dd, J = 8.5, 1.7 Hz, 1H), 8.42 (d, J = 8.4 Hz, 1H), 7.67
(dd, J = 8.5 Hz, 4.3, 1H), 7.49 (t, J = 8.2 Hz, 1H), 7.22 (d, J =
8.5 Hz, 1H), 7.08 (d, J = 8.4 Hz, 1H), 6.78 (d, J = 8.0 Hz, 1H),
4.08 (s, 3H), 3.94 (s, 3H) Method EC 75 ##STR00188##
4-methoxy-N-(4- methoxybenzo[d] isoxazol-3- yl)quinoline-8-
sulfonamide LCMS-A: rt 6.253 min, m/z 386.1 [M + H].sup.+; .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. = 8.93 (d, J = 5.4 Hz, 1H),
8.52-8.37 (m, 2H), 7.75 (t, J = 7.8 Hz, 1H), 7.50 (t, J = 8.3 Hz,
1H), 7.22 (d, J = 5.3 Hz, 1H), 7.10 (d, J = 8.6 Hz, 1H), 6.80 (d, J
= 8.1 Hz, 1H), 4.08 (s, 3H), 3.93 (s, 3H) Method EC 76 ##STR00189##
2,4-dimethoxy-N- (4-methoxybenzo [d]isoxazol-3- yl)benzene-
sulfonamide LCMS-A: rt 6.188 min, m/z = 365.1 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. = 9.76 (s, 1H),
7.80-7.73 (m, 1H), 7.55 (t, J = 8.2 Hz, 1H), 7.15 (d, J = 8.4 Hz,
1H), 6.84 (d, J = 8.0 Hz, 1H), 6.73-6.62 (m, 2H), 3.92 (s, 3H),
3.84 (s, 3H), 3.79 (s, 3H) Method ED 10 eq. NaH used 77
##STR00190## N-(4-methoxy benzo[d]isoxazol- 3-yl)-7-methyl-
quinoline-8- sulfonamide LCMS-A: rt 6.419 min, m/z = 370.2 [M +
H].sup.+ Method EF 78 ##STR00191## N-(4-methoxy- benzo[d]isoxazol-
3-yl)-6-methyl- quinoline-8- sulfonamide LCMS-A: rt 6.332 min, m/z
= 370.1 [M + H].sup.+ Method ED 5 eq. NaH used 79 ##STR00192##
N-(4-methoxy- benzo[d]isoxazol- 3-yl)-4-methyl- quinoline-8-
sulfonamide LCMS-A: rt 6.325 min, m/z = 370.1 [M + H].sup.+ Method
ED 5 eq. NaH used 80 ##STR00193## N-(4-methoxy- benzo[d]isoxazol-
3-yl)-2-methyl- quinoline-8- sulfonamide LCMS-A: rt 6.349 min, m/z
= 370.1 [M + H].sup.+ Method ED 5 eq. NaH used 81 ##STR00194##
N-(4-methoxy- benzo[d]isoxazol- 3-yl)benzofuran- 7-sulfonamide
LCMS-A: rt 6.225 min, m/z = 345.1 [M + H].sup.+ Method EG 82
##STR00195## N-(4-methoxy- benzo[d]isoxazol- 3-yl)chromane-8-
sulfonamide LCMS-A: rt 6.282 min, m/z = 361.1 [M + H].sup.+ Method
EG 83 ##STR00196## N-(4-methoxy- benzo[d]isoxazol-
3-yl)quinoline-8- sulfonamide LCMS-A: rt 6.198 min; m/z = 356.2 [M
+ H].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. = 9.08 (dd,
J = 4.3, 1.7 Hz, 1H), 8.57 (dd, J = 8.4, 1.6 Hz, 1H), 8.46 (dd, J =
7.4, 1.4 Hz, 1H), 8.36 (d, J = 8.0 Hz, 1H), 7.81 (t, J = 7.8 Hz,
1H), 7.73 (dd, J = 8.3, 4.3 Hz, 1H), 7.50 (t, J = 8.2 Hz, 1H), 7.10
(d, J = 8.4 Hz, 1H), 6.78 (d, J = 8.0 Hz, Method EH 1H), 3.88 (s,
3H) 84 ##STR00197## N-(4-methoxy- benzo[d]isoxazol- 3-yl)-5,6,7,8-
tetrahydronaphtha- lene-2- sulfonamide LCMS-A: rt 6.955 min; m/z
359.1 [M + H].sup.+; .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta.
7.81- 7.76 (m, 2H), 7.55-7.49 (m, 1H), 7.23 (d, J = 7.7 Hz, 1H),
7.06 (d, J = 8.4 Hz, 1H), 6.79 (d, J = 8.0 Hz, 1H), 3.96 (s, 3H),
1.81-1.77 (m, 4H). NB: solvent obscuring aliphatic 2 .times.
CH.sub.2 Method EI 85 ##STR00198## 2,4,6-trimethoxy- N-(4-methoxy-
benzo[d]isoxazol- 3-yl)benzene- sulfonamide LCMS-A: rt 6.095 min;
m/z 395.1 [M + H].sup.+ Method EJ 86 ##STR00199## 2,6-dimethoxy-N-
(4-methoxy- benzo[d]isoxazol- 3-yl)benzene- sulfonamide LCMS-A: rt
5.991 min; m/z 365.1 [M + H].sup.+ Method EJ 87 ##STR00200##
2-methoxy-N-(4- methoxybenzo[d] isoxazol-3-yl)-5- (trifluoromethyl)
benzenesulfonamide LCMS-B: rt 3.379 min; m/z 403.0 [M + H].sup.+
Method EJ 88 ##STR00201## N-(4-methoxy- benzo[d]isoxazol-
3-yl)-2-(methoxy methyl)benzene- sulfonamide LCMS-B: rt 3.344 min;
m/z 349.1 [M + H].sup.+ Method EJ
Examples 89-147 (Table F)
[0710] Method FA
##STR00202##
[0711] To a solution of the amine (0.5 mmol, 1.0 eq.) in anhydrous
THF (10 mL) at -78.degree. C. under N.sub.2 was added LiHMDS (1 M
solution in THF, 3 eq.) dropwise and the mixture was stirred at
-78.degree. C. for 30 min. A solution of the sulfonyl chloride (1.5
eq.) in anhydrous THF (2.0 mL) was then added dropwise and the
mixture was allowed to warm to RT and stirred overnight. Water was
added and the mixture was extracted with EtOAc. The combined
organic extracts were washed with brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography or prep. TLC to
give the title compound. Variations to above conditions have been
noted in Table F.
[0712] Method FB
##STR00203##
[0713] To a solution of the amine (0.3 mmol, 1.0 eq.) in pyridine
(5 mL) under N.sub.2 was added the sulfonyl chloride (2.0 eq.) and
the mixture was heated at 100.degree. C. overnight. The reaction
was quenched with 1 M aq. HCl, water was then added and the mixture
was extracted with EtOAc. The combined organic extracts were washed
with brine, anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by column
chromatography or prep. TLC to give the title compound. Variations
to above conditions have been noted in Table F.
[0714] Method FC
##STR00204##
[0715] To a solution of the amine (0.5 mmol, 1.0 eq.) in anhydrous
THF (10 mL) at -20.degree. C. under N.sub.2 was added n-BuLi (2.5 M
in hexanes, 1.5 eq.) dropwise and the mixture was stirred at
-20.degree. C. for 1 h. A solution of the sulfonyl chloride (1.5
eq.) in anhydrous THF (2.0 mL) was then added dropwise and the
mixture was allowed to warm to RT and stirred overnight. Water was
added and the mixture was extracted with EtOAc. The combined
organic extracts were washed with brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified prep. TLC (DCM/MeOH=20/1) to give the
title compound.
[0716] Method FD
##STR00205##
[0717] To a solution of the amine (0.5 mmol, 1.0 eq.) in anhydrous
THF (10 mL) at -78.degree. C. under N.sub.2 was added KHMDS (1 M
solution in THF, equivalents specified in Table F) dropwise and the
mixture was stirred at -78.degree. C. or 0.degree. C. for 30 min to
1 h (specified in Table F). A solution of the sulfonyl chloride
(equivalents specified in Table F) in anhydrous THF (2.0 mL) was
then added dropwise and the mixture was allowed to warm to RT and
stirred overnight. Water was added and the mixture was extracted
with EtOAc. The combined organic extracts were washed with brine,
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by column
chromatography or prep. TLC to give the title compound.
TABLE-US-00010 TABLE F Intermediates Name and structure Analytical
(if applicable) Method Notes 89 ##STR00206## LCMS-C: R.sub.t 2.18
min, m/z 442.9 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6)
.delta. 9.80 (s, 1H), 7.54-7.48 (m, 2H), 7.05 (s, 1H), 6.78 (d, J =
8.4 Hz, 2H), 3.92 (s, 3H), 3.76 (s, 6H). 2,6-Dimethoxy-
benzenesulfonyl chloride I111 6-Bromo-4- methoxybenzo
[d]isoxazol-3- amine I22 FA 4 eq. LiHMDS used. Prep. TLC (DCM/MeOH
= 100/1) N-(6-Bromo-4- methoxybenzo[d]isoxazol- 3-yl)-2,6-
dimethoxybenzene- sulfonamide 90 ##STR00207## LCMS-C: R.sub.t 1.97
min, m/z 304.9 [M + H].sup.+; .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.21 (d, J = 7.2 Hz, 2H), 7.82 (s, 1H), 7.61-7.54 (m, 3H),
7.43 (t, J = 8.0 Hz, 1H), 7.02 FA Stirred 2 h before adding
sulfonyl chloride. Prep. TLC (DCM/MeOH = 100/1) N-(4- (d, J = 8.0
Hz, 1H), Methoxybenzo[d]isoxazol- 6.59 (d, J = 7.6 Hz, 3- 1H), 3.91
(s, 3H). yl)benzenesulfonamide 91 ##STR00208## LCMS-C: R.sub.t 2.33
min, m/z 485.0 [M + H].sup.+; .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.26 (s, 1H), 7.52 (d, J = 6.8 Hz, 2H), 7.46-7.42 (m, 3H),
6.98 (s, 1H), 6.73 (s, 2H), 6.65 (s, 1H), 4.51 (s, 2H), 4.04 (s,
3H), 3,5- Dimethoxy- [1,1'-biphenyl]- 4-sulfonyl chloride I116
4-Methoxy-6- (methoxymeth- yl)benzo[d]isox- azol-3-amine I9 FA 5
eq. LiHMDS used, stirred 2 h before adding sulfonyl chloride.
Column chromatog- raphy (Pet. ether/EtOAc = 3,5-Dimethoxy-N-(4-
3.94 (s, 6H), 3.41 2/1) methoxy-6- (s, 3H). (methoxymethyl)benzo
[d]isoxazol-3-yl)-[1,1'- biphenyl]-4-sulfonamide 92 ##STR00209##
LCMS-C: R.sub.t 2.19 min, m/z 457.0 [M + H].sup.+; .sup.1H NMR (400
MHz, DMSO- d.sub.6) .delta. 9.64 (s, 1H), 9.62 (s, 1H), 7.53 (t, J
= 8.8 Hz, 1H), 7.35 (s, 1H), 7.30 (t, J = 8.0 Hz, 1H),
2,6-Dimethoxy- benzenesulfo- nyl chloride I111 3-(3-Amino-4-
methoxybenzo [d]isoxazol-6- yl)phenol I19 FA 8 eq. LiHMDS used,
stirred 1 h before adding sulfonyl chloride. Prep. TLC (DCM/MeOH =
N-(6-(3- 7.19 (d, J = 7.6 Hz, 30/1) Hydroxyphenyl)-4- 1H), 7.12 (s,
1H), methoxybenzo[d]isoxa- 7.01 (s, 1H), 6.85- zol-3-yl)-2,6- 6.78
(m, 3H), 4.01 dimethoxybenzene- (s, 3H), 3.79 (s, sulfonamide 6H).
93 ##STR00210## LCMS-C: R.sub.t 1.94 min, m/z 423.0 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 9.56 (s, 1H), 7.51 (t,
J = 8.4 Hz, 1H), 7.08 (s, 1H), 6.80 (d, J = 8.4 Hz, 2H), 6.78 (s,
1H), 4.45 (q, J = 6.4 Hz, 1H), 3.92 (s, 3H), 3.78 (s, 6H), 3.17
2,6-Dimethoxy- benzenesulfo- nyl chloride I111 4-Methoxy-6- (1-
methoxyethyl) benzo[d]isoxazol- 3-amine I15 FA Stirred 2 h before
adding sulfonyl chloride. Prep. TLC (DCM/MeOH = 75/1)
2,6-Dimethoxy-N-(4- (s, 3H), 1.37 (d, J = methoxy-6-(1- 6.4 Hz,
3H). methoxyethyl)benzo[d] isoxazol-3- yl)benzenesulfonamide 94
##STR00211## LCMS-C: R.sub.t 2.31 min, m/z 393.0 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 9.57 (s, 1H), 7.57 (t,
J = 8.0 Hz, 1H), 7.46 (d, J = 8.8 Hz, 1H), 7.16 (d, J = 8.8 Hz,
1H), 6.93 (d, J = 8.8 Hz, 3-Ethyl-2,6- dimethoxy- benzenesulfonyl
chloride I106 FA Stirred 3 h before adding sulfonyl chloride. Prep.
TLC (DCM/MeOH = 75/1) 3-Ethyl-2,6-dimethoxy- 1H), 6.85 (d, J =
N-(4- 8.0 Hz, 1H), 3.92 methoxybenzo[d]isoxazol- (s, 3H), 3.78 (s,
3- 3H), 3.75 (s, 3H), yl)benzenesulfonamide 2.60 (q, J = 7.2 Hz,
2H), 1.15 (t, J = 7.2 Hz, 3H). 95 ##STR00212## LCMS-C: R.sub.t 2.34
min, m/z 441.0 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6)
.delta. 9.60 (s, 1H), 7.79 (d, J = 6.8 Hz, 2H), 7.58-7.44 (m, 4H),
7.17 (d, J = 8.0 Hz, 1H), 6.99 (s, 2H), 6.87 (d, J = 7.6 Hz, 1H),
3.95 3,5- Dimethoxy- [1,1'-biphenyl]- 4-sulfonyl chloride I116 FA
3.3 eq. LiHMDS used, stirred at -70.degree. C. for 3 h before
adding 1.8 eq. sulfonyl chloride. Purification: Precipitated from
DCM 3,5-Dimethoxy-N-(4- (s, 3H), 3.88 (s, with Pet.
methoxybenzo[d]isoxazol- 6H). ether 3-yl)-[1,1'-biphenyl]-
4-sulfonamide 96 ##STR00213## LCMS-C: R.sub.t 2.42 min, m/z 441.0
[M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 9.84
(s, 1H), 7.59-7.54 (m, 2H), 7.48-7.42 (m, 4H), 7.39-7.35 (m, 1H),
7.18 (d, J = 8.0 Hz, 1H), 7.07 (d, J = 8.8 Hz, 1H), 6.87 (d, J =
8.0 Hz, 1H), 3.91 (s, 3H), 3.81 2,4- Dimethoxy- [1,1'-biphenyl]-
3-sulfonyl chloride I114 FA 2 eq. LiHMDS used and stirred 3 h
before adding sulfonyl chloride. Prep. TLC (DCM/MeOH = 50/1)
2,4-Dimethoxy-N-(4- (s, 3H), 3.34 (s, methoxybenzo[d]isoxazol- 3H).
3-yl)-[1,1'-biphenyl]- 3-sulfonamide 97 ##STR00214## LCMS-D:
R.sub.t 2.38 min, m/z 393.1 [M + H].sup.+; .sup.1H NMR (400 MHz,
DMSO- d.sub.6) .delta. 11.3 (s, 1H), 7.81 (s, 1H), 7.48- 7.45 (m,
2H), 6.75- 6.73 (m, 2H), 4.51 (s, 2H), 3.73 (s, 6H), 3.35 (s, 3H),
2,6-Dimethoxy- benzenesulfonyl chloride I111 6-(Methoxy methyl)-5-
methylbenzo[d] isoxazol-3- amine I4 FA 2 eq. LiHMDS used, stirred
2.5 h before adding 3 eq. sulfonyl chloride. Purified by prep-HPLC
2,6-Dimethoxy-N-(6- 2.31 (s, 3H). (methoxymethyl)-5-
methylbenzo[d]isoxazol 3- yl)benzenesulfonamide 98 ##STR00215##
LCMS-D: R.sub.t 1.79 min, m/z 385.8 [M + H].sup.+; .sup.1H NMR (400
MHz, DMSO- d.sub.6) .delta. 10.6 (s, 1H), 8.98 (dd, J = 4.4, 1.6
Hz, 1H), 8.46 (dd, J = 8.0, 1.6 Hz, 1H), 8.31 (d, J = 5.6 Hz, 1H),
7.55 7-Methoxy quinoline-8- sulfonyl chloride I107 FA LiHMDS added
at -60.degree. C. and stirred at 0.degree. C. for 1 h before adding
sulfonyl chloride. Prep. TLC 7-Methoxy-N-(4- (dd, J = 8.0, 4.0
(DCM/MeOH = methoxybenzo[d]isoxazol- Hz, 1H), 7.52 (d, J = 20/1)
3-yl)quinoline-8- 6.0 Hz, 1H), 7.50 sulfonamide (t, J = 8.0 Hz,
1H), 7.08 (d, J = 8.4 Hz, 1H), 6.81 (d, J = 8.0 Hz, 1H), 4.06 (s,
3H), 3.99 (s, 3H). 99 ##STR00216## LCMS-D: R.sub.t 2.91 min, m/z
427.1 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta.
7.79 (d, J = 8.8 Hz, 2H), 7.56 (t, J = 8 . 4 Hz, 1H), 7.15-6.99 (m,
7H), 6.82 (d, J = 8.0 Hz, 1H), 3.90 (s, 3H), 3.76 (s, 3H).
2-Methoxy-5- phenoxybenzene- sulfonyl chloride I100 FA 1.1 eq.
LiHMDS and 1.05 eq. sulfonyl chloride used. Product not purified
2-Methoxy-N-(4- methoxybenzo[d]isoxazol- 3-yl)-5- phenoxybenzene-
sulfonamide 100 ##STR00217## LCMS-D: R.sub.t 2.61 min, m/z 419.0 [M
+ H].sup.+; .sup.1H NMR (400 MHz, methanol-d.sub.4) .delta. 8.01
(d, J = 0.8 Hz, 1H), 7.59-7.42 (m, 3H), 6.71 (d, J = 8.8 Hz, 2H),
3.82 2,6-Dimethoxy- benzenesulfonyl chloride I111 5-
(Trifluoromethoxy) benzo[d]isox- azol-3-amine I63 FA Added LiHMDS
at -60.degree. C. and stirred at 0.degree. C. for 1 h before adding
sulfonyl chloride. 2,6-Dimethoxy-N-(5- (s, 6H). Prep. TLC
(trifluoromethoxy)benzo (Pet. [d]isoxazol-3- ether/EtOAc =
yl)benzenesulfonamide 5/1) 101 ##STR00218## LCMS-D: R.sub.t 2.64
min, m/z 433.1 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6)
.delta. 10.5 (s, 1H), 7.92 (d, J = 8.8 Hz, 1H), 7.59 (t, J = 8.0
Hz, 1H), 7.20-7. 14 (m, 2H), 6.85 (d, J = 8.4 Hz, 1H), 3.87
2,6-Dimethoxy- 3-(trifluoro- methyl)benzene- sulfonyl chloride I102
FA Added 2 eq. LiHMDS at 0.degree. C. Prep. TLC (Pet. ether/EtOAc =
1/1) 2,6-Dimethoxy-N-(4- (s, 6H), 3.85 (s, methoxybenzo[d]isoxa-
3H). zol-3-yl)-3- (trifluoromethyl)benzene- sulfonamide 102
##STR00219## LCMS-D: R.sub.t 2.42 min, m/z 415.9 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 11.5 (s, 1H), 8.32 (s,
1H), 7.07 (t, J = 8.4 Hz, 2H), 7.50-7.30 (m, 2H), 2,6-Dimethoxy-
benzenesulfonyl chloride I111 5-Methyl-6- (oxazol-2-
yl)benzo[d]isox- azol-3-amine I65 FA 1.5 eq. LiHMDS used. Prep. TLC
(Pet. ether/EtOAc = 1/2) 2,6-Dimethoxy-N-(5- 6.76 (d, J = 8.4 Hz,
methyl-6-(oxazol-2- 2H), 3.75 (s, 6H), yl)benzo[d]isoxazol-3- 2.70
(s, 3H). yl)benzenesulfonamide 103 ##STR00220## LCMS-D: R.sub.t
2.22 min, m/z 378.9 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO-
d.sub.6) .delta. 11.4 (s, 1H), 8.04 (d, J = 8.4 Hz, 1H), 7.50-7.45
(m, 2H), 7.31 (d, J = 2,6-Dimethoxy- benzenesulfonyl chloride I111
6-(Methoxy methyl)benzo [d]isoxazol-3- amine I62 FA Added LiHMDS at
-78.degree. C. and stirred at 0.degree. C. for 30 min before adding
sulfonyl 2,6-Dimethoxy-N-(6- 8.4 Hz, 1H), 6.75 chloride.
(methoxymethyl)benzo (d, J = 8.8 Hz, 2H), Prep. TLC [d]isoxazol-3-
4.55 (s, 2H), 3.73 (Pet. yl)benzenesulfonamide (s, 6H), 3.32 (s,
ether/EtOAc = 3H). 2/1) 104 ##STR00221## LCMS-D: R.sub.t 2.40 min,
m/z 348.9 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6)
.delta. 11.3 (s, 1H), 7.83 (s, 1H), 7.49- 7.44 (m, 3H), 6.75 (d, J
= 8.4 Hz, 2H), 2,6-Dimethoxy- benzenesulfonyl chloride I111
5-Methyl benzo[d]isoxa- zol-3-amine I60 FA Added LiHMDS at
-60.degree. C. and stirred at 0.degree. C. for 1 h before adding
sulfonyl 2,6-Dimethoxy-N-(5- 3.74 (s, 6H), 2.40 chloride.
methylbenzo[d]isoxazol- (s, 3H). Prep. TLC 3- (DCM/MeOH =
yl)benzenesulfonamide 20/1) 105 ##STR00222## LCMS-D: R.sub.t 2.44
min, m/z 429.1 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6)
.delta. 11.5 (s, 1H), 8.16 (d, J = 8.8 Hz, 1H), 7.74 (d, J = 1.6
Hz, 1H), 7.57 (dd, J = 8.8, 2.0 2,6-Dimethoxy- benzenesulfonyl
chloride I111 6-(3,5- Dimethyl-1H- pyrazol-1- yl)benzo[d]isox-
azol-3-amine I59 FC N-(6-(3,5-Dimethyl-1H- Hz, 1H), 7.48 (t, J =
pyrazol-1- 8.8 Hz, 1H), 6.75 yl)benzo[d]isoxazol-3- (d, J = 8.4 Hz,
2H), yl)-2,6- 6.13 (s, 1H), 3.75 dimethoxybenzene- (s, 6H), 2.36
(s, sulfonamide 3H), 2.19 (s, 3H). 106 ##STR00223## LCMS-D: R.sub.t
2.58 min, m/z 405.1 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO-
d.sub.6) .delta. 9.40 (s, 1H), 7.58 (t, J = 8.0 Hz, 1H), 7.16 (d, J
= 8.0 Hz, 1H), 6.86- 6.84 (m, 2H), 3.96 (s, 3H), 3.80 (s, 4,6-
Dimethoxy- 2,3-dihydro- 1H-indene-5- sulfonyl chloride I98 FA Added
1.33 eq. LiHMDS at -78.degree. C. and stirred at 0.degree. C. for 1
h before adding 0.67 eq. sulfonyl chloride. Column
4,6-Dimethoxy-N-(4- 3H), 3.75 (s, 3H), chromatog-
methoxybenzo[d]isoxa- 2.88-2.84 (m, 4H), raphy (Pet.
zol-3-yl)-2,3-dihydro-1H- 2.02-1.99 (m, 2H). ether/EtOAc =
indene-5-sulfonamide 1/1) 107 ##STR00224## LCMS-D: R.sub.t 2.34
min, m/z 368.0 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6)
.delta. 10.5 (s, 1H), 7.98 (s, 2H), 7.70- 7.62 (m, 2H), 7.45 (d, J
= 7.2 Hz, 1H), 3.50 (s, 3H), 2.46 (q, J = 7.2 Hz, 2H), 5-Ethyl-1-
methyl-2-oxo- 1,2- dihydropyridine- 3-sulfonyl chloride I94 FA
Added 2.1 eq. LiHMDS At -78.degree. C. and stirred at 0.degree. C.
for 1 h before adding 2 eq. sulfonyl chloride. Column N-(4- 1.13
(t, J = 7.2 Hz, chromatog- Chlorobenzo[d]isoxazol- 3H). raphy
3-yl)-5-ethyl-1-methyl- (DCM/MeOH = 2-oxo-1,2- 50/1)
dihydropyridine-3- sulfonamide 108 ##STR00225## LCMS-D: R.sub.t
2.82 min, m/z 441.0 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO-
d.sub.6) .delta. 10.8 (s, 1H), 7.86 (d, J = 8.8 Hz, 1H), 7.62 (d, J
= 1.2 Hz, 1H), 7.50- 7.43 (m, 2H), 7.17 5-Ethyl-2- methoxybenzene-
sulfonyl chloride I112 5-Bromo-4- methoxybenzo [d]isoxazol-3- amine
I48 FD Added 2 eq. KHMDS, stirred at 0.degree. C. for 1 h before
adding 2 eq. sulfonyl chloride. Column N-(5-Bromo-4- (d, J = 8.8
Hz, 1H), Chromatog- methoxybenzo[d]isoxa- 3.91 (s, 3H), 3.79 raphy
(Pet. zol-3-yl)-5-ethyl-2- (s, 3H), 2.64 (q, J = ether/EtOAc =
methoxybenzenesulfon- 7.6 Hz, 2H), 1.17 50/1 to amide (t, J = 7.6
Hz, 3H). 3/1) 109 ##STR00226## LCMS-D: R.sub.t 2.78 min, m/z 377.1
[M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 11.5 (s, 1H), 7.69 (d,
J = 2.0 Hz, 1H), 7.50-7.44 (m, 3H), 7.20 (dd, J = 9.8, 2.4 Hz, 1H),
7.10 (d, J = 8.8 Hz, 1H), 4.06 (q, J = 7.4 Hz, 2H), 3.74 5-Ethyl-2-
methoxybenzene- sulfonyl chloride I112 5-Ethoxy benzo[d]isoxa-
zol-3-amine I57 FB Column chromatog- raphy (Pet. ether/EtOAc = 4/1
to 3/1) N-(5- (s, 3H), 2.63 (q, J = Ethoxybenzo[d]isoxazol- 7.6 Hz,
2H), 1.38 3-yl)-5-ethyl-2- (t, J = 6.8 Hz, 3H),
methoxybenzenesulfon- 1.16 (t, J = 7.6 Hz, amide 3H). 110
##STR00227## LCMS-D: R.sub.t 2.66 min, m/z 377.1 [M + H].sup.+;
5-Ethyl-2- methoxybenzene- sulfonyl chloride I112 4-(Methoxy
methyl)benzo [d]isoxazol-3- amine I51 FB Prep. TLC (Pet.
ether/EtOAc = 2/1) 5-Ethyl-2-methoxy-N- (4- (methoxymethyl)benzo
[d]isoxazol-3- yl)benzenesulfonamide 111 ##STR00228## LCMS-D:
R.sub.t 2.65 min, m/z 363.1 [M + H].sup.+; .sup.1H NMR (400 MHz,
DMSO- d.sub.6) .delta. 11.5 (s, 1H), 7.69 (d, J = 2.0 Hz, 1H),
7.52-7.44 (m, 3H), 7.23 (dd, J = 9.8, 2.4 Hz, 1H), 7.10 (d, J = 8.8
Hz, 1H), 3.79 (s, 3H), 5-Ethyl-2- methoxybenzene- sulfonyl chloride
I112 5-Methoxy benzo[d]isoxazol- 3-amine I54 FB Prep. TLC
5-Ethyl-2-methoxy-N- 3.75 (s, 3H), 2.63 (5- (q, J = 7.6 Hz, 2H),
methoxybenzo[d]isoxazol- 1.16 (t, J = 7.6 Hz, 3- 3H).
yl)benzenesulfonamide 112 ##STR00229## LCMS-E: R.sub.t 5.57 min m/z
376.7 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta.
9.68 (s, 1H), 7.67 (d, J = 2.0 Hz, 1H), 7.56-7.45 (m, 2H),
7.16-7.08 (m, 2H), 6.83 (d, J = 8.0 Hz, 1H), 4.19 (q, J = 7.2 Hz,
2H), 5-Ethyl-2- methoxybenzene- sulfonyl chloride I112 4-Ethoxy
benzo[d]isoxazol- 3-amine I53 FA Added 1.5 eq. LiHMDS at
-78.degree. C. and stirred at 0.degree. C. for 1 h before adding
1.5 eq. sulfonyl chloride. Column chromatog- N-(4- 3.63 (s, 3H),
2.64 raphy (Pet. Ethoxybenzo[d]isoxazol- (q, J = 7.6 Hz, 2H),
ether/EtOAc = 3-yl)-5-ethyl-2- 1.31 (t, J = 7.4 Hz, 3/1)
methoxybenzenesulfon- 3H), 1.17 (t, J = 7.6 amide Hz, 3H). 113
##STR00230## LCMS-D: R.sub.t 3.07 min, m/z 437.0 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 11.6 (s, 1H), 8.25 (s,
1H), 7.78 (dd, J = 8.8, 2.0 Hz, 1H), 7.62-7.56 (m, 2H), 6.85 (s,
3-Methoxy- 5,6,7,8- tetrahydronaph- thalene-2- sulfonyl chloride
I109 5-Bromo benzo[d]isoxazol- 3-amine I40 FA Added 2 eq. LiHMDS at
-78.degree. C. and stirred at 0.degree. C. for 1 h before adding 2
eq. sulfonyl chloride. N-(5- 1H), 3.69 (s, 3H), Column
Bromobenzo[d]isoxazol- 2.73-2.69 (m, 4H), chromatog-
3-yl)-3-methoxy- 1.71 (m, 4H). raphy 5,6,7,8- (DCM/MeOH =
tetrahydronaphthalene- 200/1 to 2-sulfonamide 100/1) 114
##STR00231## LCMS-D: R.sub.t 2.82 min, m/z 401.0 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 8.02 (d, J = 2.0 Hz,
1H), 7.73- 7.76 (m, 3H), 7.43- 7.35 (m, 2H), 2.49 FA Added 1 eq.
LiHMDS at -78.degree. C. and stirred at 0.degree. C. for 1 h before
adding 0.67 eq. sulfonyl 5-Bromo-N-(4- (s, 3H). chloride.
chlorobenzo[d]isoxazol- Column 3-yl)-2- chromatog-
methylbenzenesulfon- raphy (Pet. amide ether/EtOAc = 6/1) 115
##STR00232## LCMS-D: R.sub.t 2.78 min, m/z 401.0 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 8.09 (d, J = 2.0 Hz,
1H), 7.88 (d, J = 8.4 Hz, 1H), 7.74-7.65 (m, 3H), 7.48 (d, J = 7.6
Hz, 1H), 3.64 (s, 3H). FD Added 0.67 eq. KHMDS, stirred at
0.degree. C. for 1 h before adding 0.67 eq. sulfonyl chloride.
Column chromatog- Methyl 4-chloro-2-(N- raphy (Pet. (4- ether/EtOAc
= chlorobenzo[d]isoxazol- 1/1) 3-yl)sulfamoyl)benzoate 116
##STR00233## LCMS-D: R.sub.t 2.56 min, m/z 363.1 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 9.96 (s, 1H), 7.66 (d,
J = 1.6 Hz, 1H), 7.57 (t, J = 8.0 Hz, 1H), 7.48-7.46 (m, 1H),
7.12-7.11 5-Ethyl-2- methoxybenzene- sulfonyl chloride I112 FA
Added 2 eq. LiHMDS at -78.degree. C. and stirred at 0.degree. C.
for 1 h before adding 2 eq. sulfonyl chloride. 5-Ethyl-2-methoxy-N-
(m, 2H), 6.84 (d, J = Column (4- 8.0 Hz, 1H), 3.87 chromatog-
methoxybenzo[d]isoxazol- (s, 3H), 3.75 (s, raphy 3- 3H), 2.64 (q, J
= (DCM/MeOH = yl)benzenesulfonamide 7.6 Hz, 2H), 1.17 30/1) (t, J =
7.6 Hz, 3H). 117 ##STR00234## LCMS-D: R.sub.t 2.59 min, m/z 363.0
[M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta.
7.74-7.69 (m, 1H), 7.54-7.40 (m, 3H), 7.31 (d, J = 7.0 Hz, 1H). FA
Added 1 eq. LiHMDS at -78.degree. C. and stirred at 0.degree. C.
for 1 h before adding 1 eq. sulfonyl N-(4- chloride.
Chlorobenzo[d]isoxazol- Column 3-yl)-2,3,4- chromatog-
trifluorobenzenesulfon- raphy amide (DCM/MeOH = 100/1 to 30/1) 118
##STR00235## LCMS-D: R.sub.t 2.62 min, m/z 374.1 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 8.81 (d, J = 2.4 Hz,
1H), 8.34- 8.25 (m, 3H), 7.76- 7.61 (m, 3H), 7.43 (d, J = 8.8 Hz,
1H), 2.50 (s, 3H). FA Added 1.5 eq. LiHMDS at -78.degree. C. and
stirred at 0.degree. C. for 1 h before adding 1.5 eq. sulfonyl
chloride. Column chromatog- raphy N-(4- (DCM/MeOH =
Chlorobenzo[d]isoxazol- 20/1) 3-yl)-3- methylquinoline-8-
sulfonamide 119 ##STR00236## LCMS-D: R.sub.t 2.69 min, m/z 379.0 [M
+ H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 10.4 (s,
1H), 7.75-7.63 (m, 2H), 7.58 (s, 1H), 7.47 (d, J = 7.2 Hz, 1H),
7.11 (s, 1H), 3.69 6-Methoxy-2,3- dihydro-1H- indene-5- sulfonyl
chloride I108 FD Added 0.67 eq. KHMDS, stirred at 0.degree. C. for
1 h before adding 0.67 eq. sulfonyl chloride. Column N-(4- (s, 3H),
2.93 (t, J = chromatog- Chlorobenzo[d]isoxazol- 7.2 Hz, 2H), 2.84
raphy (Pet. 3-yl)-6-methoxy-2,3- (t, J = 7.2 Hz, 2H), ether/EtOAc =
dihydro-1H-indene-5- 2.07-2.00 (m, 2H). 1/1) sulfonamide 120
##STR00237## LCMS-D: R.sub.t 2.86 min, m/z 393.1 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 10.4 (s, 1H),
7.73-7.64 (m, 2H), 7.48 (d, J = 7.6 Hz, 1H), 7.45 (s, 1H), 6.91 (s,
1H), 3.67 3-Methoxy- 5,6,7,8- tetrahydronaph- thalene-2- sulfonyl
chloride I109 FA Added 1 eq. LiHMDS at -78.degree. C. and stirred
at 0.degree. C. for 1 h before adding 1 eq. sulfonyl chloride.
N-(4- (s, 3H), 2.78 (m, Column Chlorobenzo[d]isoxazol- 2H), 2.67
(m, 2H), chromatog- 3-yl)-3-methoxy- 1.72 (m, 4H). raphy (Pet.
5,6,7,8- ether/EtOAc = tetrahydronaphthalene- 50/1 to 2-sulfonamide
2/1) 121 ##STR00238## LCMS-D: R.sub.t 2.74 min, m/z 401.1 [M +
H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 10.3 (s,
1H), 8.14 (t, J = 6.4 Hz, 1H), 7.91-7.88 (m, 2H), 7.55 (d, J = 2.4
Hz, 1H), 7.51 5-Ethyl-2- methoxybenzene- sulfonyl chloride I112 4-
(Trifluoromethyl) benzo[d]isoxa- zol-3-amine I42 FA Added 2 eq.
LiHMDS at -78.degree. C. and stirred at 0.degree. C. for 1 h before
adding 2 eq. sulfonyl chloride. 5-Ethyl-2-methoxy-N- (dd, J = 8.8,
2.4 Column (4- Hz, 1H), 7.20 (d, J = chromatog-
(trifluoromethyl)benzo[d] 8.4 Hz, 1H), 3.82 raphy isoxazol-3- (s,
3H), 2.63 (q, J = (DCM/MeOH = yl)benzenesulfonamide 7.6 Hz, 2H),
1.17 20/1) (t, J = 7.6 Hz, 3H). 122 ##STR00239## LCMS-D: R.sub.t
2.30 min, m/z 361.0 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO-
d.sub.6) .delta. 11.0 (s, 1H), 9.09 (d, J = 1.4 Hz, 1H), 9.08 (d, J
= 1.4 Hz, 1H), 8.47- 8.43 (m, 2H), 8.05 (t, J = 8.0 Hz, 1H),
7.72-7.62 (m, 2H), FA 2 eq. LiHMDS used. Purification: dissolved in
2 M aq. NaOH, washed with EtOAc, then acidified to pH 2 and N-(4-
7.42 (d, J = 7.8 Hz, extracted Chlorobenzo[d]isoxazol- 1H). with
DCM 3-yl)quinoxaline-5- sulfonamide 123 ##STR00240## LCMS-D:
R.sub.t 2.41 min, m/z 360.0 [M + H].sup.+; .sup.1H NMR (400 MHz,
DMSO- d.sub.6) .delta. 8.93 (dd, J = 4.3, 1.8 Hz, 1H), 8.58 (dd, J
= 8.4, 1.8 Hz, 1H), 8.40- 8.35 (m, 2H), 7.79 (t, J = 7.8 Hz, 1H),
7.73-7.60 (m, FA 2 eq. LiHMDS used. Purification: dissolved in 2 M
aq NaOH, washed with EtOAc, then acidified to pH 2 and N-(4- 3H),
7.41 (d, J = extracted chlorobenzo[d]isoxazol- 7.4 Hz, 1H). with
DCM 3-yl)quinoline-8- sulfonamide 124 ##STR00241## LCMS-D: R.sub.t
2.60 min, m/z 359.0 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO-
d.sub.6) .delta. 8.73 (t, J = 8.4 Hz, 1H), 8.31- 8.24 (m, 2H), 8.14
(d, J = 7.8 Hz, 1H), 7.74-7.63 (m, 5H), 7.46 (d, J = 7.2 Hz, FA 2
eq. LiHMDS used. Column chromatog- raphy (Pet. ether/EtOAc = 50/1
to 2/1 N-(4- 1H). Chlorobenzo[d]isoxazol- 3-yl)naphthalene-1-
sulfonamide 125 ##STR00242## LCMS-D: R.sub.t 2.48 min, m/z 400.1 [M
+ H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 11.9 (s,
1H), 8.96 (s, 1H), 8.26- 8.23 (m, 2H), 8.04- 8.01 (m, 2H), 7.73
5-Ethyl-2- methoxybenzene- sulfonyl chloride I112 6-(1H-1,2,3-
Triazol-1- yl)benzo[d]isox- azol-3-amine FB Column chromatog- raphy
(DCM/MeOH = 100/0 to 50/1) N-(6-(1H-1,2,3-Triazol- (d, J = 2.0 Hz,
1H), I36 1-yl)benzo[d]isoxazol-3- 7.49 (dd, J = 8.4, yl)-5-ethyl-2-
2.0 Hz, 1H), 7.11 methoxybenzenesulfon- (d, J = 8.4 Hz, 1H), amide
3.73 (s, 3H), 2.65 (q, J = 8.0 Hz, 2H), 1.23 (t, J = 8.0 Hz, 3H).
126 ##STR00243## LCMS-D:R.sub.t 2.65 min, m/z 411.1 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 11.8 (s, 1H), 8.98 (d,
J = 4.8 Hz, 2H), 8.43 (d, J = 8.4 Hz, 1H), 8.48 5-Ethyl-2-
methoxybenzene- sulfonyl chloride I112 6-(Pyrimidin-2-
yl)benzo[d]isox- azol-3-amine I39 FB Prep. TLC (DCM/MeOH = 50/1)
5-Ethyl-2-methoxy-N- (s, 1H), 8.20 (d, J = (6-(pyrimidin-2- 8.4 Hz,
1H), 7.73 yl)benzo[d]isoxazol-3- (d, J = 2.0 Hz, 1H),
yl)benzenesulfonamide 7.55 (t, J = 4.8 Hz, 1H), 7.48 (dd, J = 8.4,
2.0 Hz, 1H), 7.11 (d, J = 8.8 Hz, 1H), 3.72 (s, 3H), 2.65 (q, J =
7.2 H, 2H), 1.17 (t, J = 7.2 Hz, 3H). 127 ##STR00244## LCMS-D:
R.sub.t 2.66 min, m/z 428.9 [M + H].sup.+; .sup.1H NMR (400 MHz,
DMSO- d.sub.6) .delta. 11.2 (s, 1H), 7.75-7.65 (m, 3H), 7.58 (d, J
= 2.0 Hz, 1H), 7.48 (d, J = 7.2 Hz, 1H), 4.57 5-Bromo-2,3-
dihydrobenzo- furan-7-sulfonyl chloride I95 FA Added LiHMDS at
-78.degree. C. and stirred at 0.degree. C. for 1 h before adding
sulfonyl chloride. Column 5-Bromo-N-(4- (t, J = 8.8 Hz, 2H),
chromatog- chlorobenzo[d]isoxazol- 3.26 (t, J = 8.8 Hz, raphy (Pet.
3-yl)-2,3- 2H). ether/EtOAc = dihydrobenzofuran-7- 5/1) sulfonamide
128 ##STR00245## LCMS-E: R.sub.t 5.81 min, m/z 444.7 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 10.7 (s, 1H), 8.02 (d,
J = 8.8 Hz, 1H), 7.74 (d, J = 8.8 Hz, 1H), 7.60 (d, J = 2.4 Hz,
1H), 5-Ethyl-2- methoxybenzene- sulfonyl chloride I112 5-Bromo-4-
chlorobenzo[d] isoxazol-3- amine I44 FD Added 1 eq. KHMDS, stirred
at -78.degree. C. for 30 min before adding 2 eq. sulfonyl chloride.
Column N-(5-Bromo-4- 7.50-7.48 (m, 1H), chromatog-
chlorobenzo[d]isoxazol- 7.16 (d, J = 8.4 Hz, raphy (Pet.
3-yl)-5-ethyl-2- 1H), 3.66 (s, 3H), ether/EtOAc =
methoxybenzenesulfon- 2.63 (q, J = 7.6 Hz, 50/1 to amide 2H), 1.17
(t, J = 7.6 2/1) Hz, 3H). 129 ##STR00246## LCMS-D: R.sub.t 2.69
min, m/z 410.9 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6)
.delta. 10.4 (s, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.65-7.56 (m, 3H),
7.49 (dd, J = 5-Ethyl-2- methoxybenzene- sulfonyl chloride I112 4-
Bromobenzo[d] isoxazol-3- amine I41 FA 1.5 eq. LiHMDS used. Column
chromatog- raphy (Pet. ether/EtOAc = 2/1 to 1/1) N-(4- 8.4, 2.4 Hz,
1H), Bromobenzo[d]isoxazol- 7.15 (d, J = 8.4 Hz, 3-yl)-5-ethyl-2-
1H), 3.68 (s, 3H),
methoxybenzenesulfon- 2.60 (q, J = 7.6 Hz, amide 2H), 1.16 (t, J =
7.6 Hz, 3H). 130 ##STR00247## LCMS-D: R.sub.t 2.77 min, m/z 400.1
[M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 11.9
(s, 1H), 8.24-8.17 (m, 3H), 8.12-8.10 (m, 2H), 7.70 (s, 1H), 7.48
(d, J = 8.4 Hz, 1H), 7.11 (d, J = 8.4 Hz, 5-Ethyl-2-
methoxybenzene- sulfonyl chloride I112 6-(2H-1,2,3- Triazol-2-
yl)benzo[d]isox- azol-3-amine I72 FB 3 eq. sulfonyl chloride used.
Prep. TLC (DCM/MeOH = 50/1) N-(6-(2H-1,2,3-Triazol- 1H), 3.76 (s,
3H), 2-yl)benzo[d]isoxazol-3- 2.65 (q, J = 7.6 Hz, yl)-5-ethyl-2-
2H), 1.17 (t, J = 7.6 methoxybenzenesulfon- Hz, 3H). amide 131
##STR00248## LCMS-D: R.sub.t 2.61 min, m/z 399.1 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 11.8 (s, 1H), 8.66 (d,
J = 2.4 Hz, 1H), 8.15 (d, J = 8.4 Hz, 1H), 8.07 5-Ethyl-2-
methoxybenzene- sulfonyl chloride I112 6-(1H-Pyrazol- 1-yl)benzo
[d]isoxazol-3- amine I70 FB 3 eq. sulfonyl chloride used. Column
chromatog- raphy (DCM/MeOH = N-(6-(1H-Pyrazol-1- (d, J = 2.0 Hz,
1H), 50/1) yl)benzo[d]isoxazol-3- 7.97 (dd, J = 8.8, yl)-5-ethyl-2-
2.0 Hz, 1H), 7.83 methoxybenzenesulfon- (d, J = 1.2 Hz, 1H), amide
7.72 (d, J = 2.0 Hz, 1H), 7.48 (dd, J = 8.4, 2.0 Hz, 1H), 7.11 (d,
J = 8.4 Hz, 1H), 6.62 (d, J = 1.2 Hz, 1H), 3.73 (s, 3H), 2.65 (q, J
= 7.6 Hz, 2H), 1.18 (t, J = 7.6 Hz, 3H). 132 ##STR00249## LCMS-D:
R.sub.t 2.67 min, m/z 377.1 [M + H].sup.+; .sup.1H NMR (400 MHz,
DMSO- d.sub.6) .delta. 11.6 (s, 1H), 7.69 (d, J = 2.0 Hz, 1H), 7.57
(d, J = 7.6 Hz, 1H), 7.46 (dd, J = 8.8, 2.0 Hz, 1H), 7.26 (d, J =
8.0 Hz, 1H), 7.15 5-Ethyl-2- methoxybenzene- sulfonyl chloride I112
7-Ethoxybenzo [d]isoxazol-3- amine I26 FB Column chromatog- raphy
(Pet. ether/EtOAc = 5/1) N-(7- (d, J = 8.0 Hz, 1H),
Ethoxybenzo[d]isoxazol- 7.08 (d, J = 8.4 Hz, 3-yl)-5-ethyl-2- 1H),
4.21 (q, J = methoxybenzenesulfon- 6.8 Hz, 2H), 3.72 amide (s, 3H),
3.62 (q, J = 7.6 Hz, 2H), 1.36 (t, J = 6.8 Hz, 3H), 1.16 (t, J =
7.6 Hz, 3H). 133 ##STR00250## LCMS-D: R.sub.t 2.87 min, m/z 403.1
[M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 11.6
(s, 1H), 7.69 (d, J = 1.6 Hz, 1H), 7.56 (d, J = 8.0 Hz, 1H), 7.46
5-Ethyl-2- methoxybenzene- sulfonyl chloride I112 7- (Cyclopropyl-
methoxy)benzo [d]isoxazol-3- FB Column chromatog- raphy (DCM/MeOH =
200/1) N-(7- (d, J = 8.0 Hz, 1H), amine I28 (Cyclopropylmethoxy)
7.24 (t, J = 8.0 Hz, benzo[d]isoxazol-3-yl)-5- 1H), 7.14 (d, J =
ethyl-2- 7.6 Hz, 1H), 7.09 methoxybenzenesulfon- (d, J = 8.4 Hz,
1H), amide 3.99 (d, J = 7.2 Hz, 2H), 3.72 (s, 3H), 2.63 (q, J = 7.6
Hz, 2H), 1.30 (m, 1H), 1.16 (t, J = 7.6 H, 3H), 0.59-0.54 (m, 2H),
0.35-0.30 (m, 2H). 134 ##STR00251## LCMS-D: R.sub.t 2.89 min, m/z
403.1 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta.
11.5 (s, 1H), 7.87 (d, J = 8.8 Hz, 1H), 7.68 (d, J = 2.4 Hz, 1H),
7.46 (dd, J = 8.4, 2.0 5-Ethyl-2- methoxybenzene- sulfonyl chloride
I112 6-(Cyclopropyl methoxy)benzo [d]isoxazol-3- amine I33 FB
Purified by prep. HPLC N-(6- Hz, 1H), 7.09-7.07
(Cyclopropylmethoxy) (m, 2H), 6.96 (dd, J = benzo[d]isoxazol-3-yl)-
8.8, 2.0 Hz, 1H), 5-ethyl-2- 3.88 (d, J = 6.8 Hz,
methoxybenzenesulfon- 2H), 3.73 (s, 3H), amide 2.63 (q, J = 7.6 Hz,
2H), 1.30 (m, 1H), 1.16 (t, J = 7.6 Hz, 3H), 0.60-0.55 (m, 2H),
0.25-0.22 (m, 2H). 135 ##STR00252## LCMS-D: R.sub.t 2.83 min, m/z
410.9 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta.
11.8 (s, 1H), 8.25 (s, 1H), 8.78 (d, J = 8.8 Hz, 1H), 7.71 (s, 1H),
7.62 5-Ethyl-2- methoxybenzene sulfonyl chloride I112 5-Bromo
benzo[d]isoxa- zol-3-amine I40 FA 2 eq. LiHMDS and 2 eq. sulfonyl
chloride used. Prep. TLC (Pet. N-(5- (d, J = 8.8 Hz, 1H),
ether/EtOAc = Bromobenzo[d]isoxazol- 7.48 (d, J = 8.0 Hz, 2/1)
3-yl)-5-ethyl-2- 1H), 7.11 (d, J = methoxybenzenesulfon- 8.4 Hz,
1H), 3.72 amide (s, 3H), 3.64 (q, J = 7.2 Hz, 2H), 1.17 (t, J = 7.2
Hz, 3H). 136 ##STR00253## LCMS-D: R.sub.t 2.52 min, m/z 411.1 [M +
H].sup.+; .sup.1H NMR (400 MHz, methanol- d.sub.4/CDCl.sub.3)
.delta. 8.87 (d, J = 4.8 Hz, 2H), 8.51 (d, J = 7.6 Hz, 1H),
8.15-8.13 (m, 1H), 7.76 (d, J = 5-Ethyl-2- methoxybenzene- sulfonyl
chloride I112 7-(Pyrimidin-2- yl)benzo[d]isox- azol-3-amine I68 FB
Prep. TLC (Pet. ether/EtOAc = 1/1) 5-Ethyl-2-methoxy-N- 2.4 Hz,
1H), 7.49 (7-(pyrimidin-2- (t, J = 8.0 Hz, 1H),
yl)benzo[d]isoxazol-3- 7.37-7.33 (m, 2H), yl)benzenesulfonamide
6.95 (d, J = 8.8 Hz, 1H), 3.81 (s, 3H), 2.61 (q, J = 7.6 Hz, 2H),
1.18 (t, J = 7.6 Hz, 3H). 137 ##STR00254## LCMS-D: R.sub.t 2.85
min, m/z 411.0 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6)
.delta. 11.9 (s, 1H), 8.08 (d, J = 8.0 Hz, 1H), 7.89 (d, J = 7.2
Hz, 1H), 7.71 5-Ethyl-2- methoxybenzene- sulfonyl chloride I112
7-Bromobenzo [d]isoxazol-3- amine I66 FB 3 eq. sulfonyl chloride
used. Prep. TLC (Pet. ether/EtOAc = 5/1) N-(7- (d, J = 2.0 Hz, 1H),
Bromobenzo[d]isoxazol- 7.48 (d, J = 8.4 Hz, 3-yl)-5-ethyl-2- 1H),
7.34 (d, J = methoxybenzenesulfon- 7.6 Hz, 1H), 7.10 amide (d, J =
8.4 Hz, 1H), 3.71 (s, 3H), 2.64 (q, J = 7.6 Hz, 2H), 1.17 (t, J =
7.6 Hz, 3H). 138 ##STR00255## LCMS-D: R.sub.t 2.93 min, m/z 411.0
[M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 11.8
(s, 1H), 8.08-7.99 (m, 2H), 7.70 (d, J = 2.0 Hz, 1H), 7.58 (dd, J =
5-Ethyl-2- methoxybenzene- sulfonyl chloride I112 6-Bromobenzo
[d]isoxazol-3- amine I75 FB 1.8 eq. sulfonyl chloride used. Column
chromatog- raphy (Pet. ether/EtOAc = N-(6- 8.8, 1.6 Hz, 1H), 100/0
to Bromobenzo[d]isoxazol- 7.48 (dd, J = 8.8, 5/1) 3-yl)-5-ethyl-2-
2.0 Hz, 1H), 7.10 methoxybenzenesulfon- (d, J = 8.4 Hz, 1H), amide
3.71 (s, 3H), 2.64 (q, J = 7.6 Hz, 2H), 1.17 (t, J = 7.6 Hz, 3H).
139 ##STR00256## LCMS-D: R.sub.t 2.76 min, m/z 377.1 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 11.5 (s, 1H), 7.88(d,
J = 8.8 Hz, 1H), 7.66 (d, J = 2.0 Hz, 1H), 7.47- 5-Ethyl-2-
methoxybenzene- sulfonyl chloride I112 6-Ethoxy benzo[d]isoxa-
zol-3-amine I32 FB Column chromatog- raphy (Pet. ether/EtOAc =
100/0 to 5/1) N-(6- 7.44 (m, 1H), 7.11- Ethoxybenzo[d]isoxazol-
7.08 (m, 1H), 6.94 3-yl)-5-ethyl-2- (dd, J = 8.8, 2.0
methoxybenzenesulfon- Hz, 1H), 4.11 (q, J = amide 7.2 Hz, 2H), 3.73
(s, 3H), 2.63 (q, J = 7.6 Hz, 2H), 1.36 (t, J = 7.2 Hz, 3H), 1.17
(t, J = 7.6 Hz, 3H). 140 ##STR00257## LCMS-D: R.sub.t 2.63 min, m/z
410.1 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta.
11.8 (s, 1H), 8.74-8.69 (m, 1H), 8.26 (s, 1H), 8.18-8.09 (m,
5-Ethyl-2- methoxybenzene- sulfonyl chloride I112 6-(Pyridin-2-
yl)benzo[d]isox- azol-3-amine I74 FB Column chromatog- raphy
(DCM/MeOH = 100/0 to 100/1) 5-Ethyl-2-methoxy-N- 3H), 7.98-7.90
(6-(pyridin-2- (m, 1H), 7.72 (d, J = yl)benzo[d]isoxazol-3- 2.3 Hz,
1H), 7.50 yl)benzenesulfonamide 7.39 (m, 2H), 7.13-7.06 (m, 1H),
3.73 (s, 3H), 2.61 (q, J = 7.6 Hz, 2H), 1.15 (t, J = 7.6 Hz, 3H).
141 ##STR00258## LCMS-D: R.sub.t 2.58 min, m/z 382.9 [M + H].sup.+;
4-Bromo-2- methoxybenzene- sulfonyl chloride I110 FB 1.2 eq.
sulfonyl chloride used. Column chromatog- raphy (Pet. ether/EtOAc =
5/1 to 2/1) N-(benzo[d]isoxazol-3- yl)-4-bromo-2-
methoxybenzenesulfon- amide 142 ##STR00259## LCMS-C: R.sub.t 2.39
min, m/z 441.0 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6)
.delta. 9.62 (s, 1H), 7.90-7.69 (m, 3H), 7.59-7.32 (m, 4H), 6.97
(d, J = 8.4 Hz, 1H), 6.86- 6.73 (m, 2H), 2,6-Dimethoxy-
benzenesulfonyl chloride I111 4-Methoxy-7- phenylbenzo[d]
isoxazol-3- amine I80 FA 4 eq. LiHMDS used, stirred 2 h before
adding sulfonyl chloride. Prep. TLC (DCM/MeOH = 100/1)
2,6-Dimethoxy-N-(4- 3.98 (s, 3H), 3.79 methoxy-7- (s, 6H).
phenylbenzo[d]isoxazol- 3- yl)benzenesulfonamide 143 ##STR00260##
LCMS-C: R.sub.t 1.89 min, m/z 445.0 [M + H].sup.+; .sup.1H NMR (400
MHz, DMSO- d.sub.6) .delta. 9.58 (s, 1H), 8.22 (s, 1H), 7.96 (s,
1H), 7.80 (d, J = 8.2 Hz, 1H), 7.50 (t, J = 8.5 Hz, 1H), 6.88 (d, J
= 8.3 Hz, 2,6-Dimethoxy- benzenesulfonyl chloride I111 4-Methoxy-7-
(1-methyl-1H- pyrazol-4- yl)benzo[d]isox- azol-3-amine I82 FA 4 eq.
LiHMDS used, stirred 2 h before adding sulfonyl chloride. Prep. TLC
(DCM/MeOH = 30/1) 2,6-Dimethoxy-N-(4- 1H), 6.78 (d, J =
methoxy-7-(1-methyl- 8.5 Hz, 2H), 3.95 1H-pyrazol-4- (s, 3H), 3.88
(s, yl)benzo[d]isoxazol-3- 3H), 3.79 (s, 6H). yl)benzenesulfonamide
144 ##STR00261## LCMS-C: R.sub.t 1.81 min, m/z 431.9 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 9.81 (s, 1H), 8.33 (d,
J = 0.9 Hz, 1H), 7.73 (d, J = 0.9 Hz, 1H), 7.51 (t, J = 8.5 Hz,
1H), 7.48 (d, J = 0.8 Hz, 1H), 7.38 (d, J = 2,6-Dimethoxy-
benzenesulfonyl chloride I111 4-Methoxy-6- (oxazol-2-
yl)benzo[d]isox- azol-3-amine I86 FA Stirred 2 h before adding
sulfonyl chloride. Prep. TLC (DCM/MeOH = 75/1) 2,6-Dimethoxy-N-(4-
1.0 Hz, 1H), 6.79 methoxy-6-(oxazol-2- (d, J = 8.5 Hz, 2H),
yl)benzo[d]isoxazol-3- 4.01 (s, 3H), 3.78 yl)benzenesulfonamide (s,
6H). 145 ##STR00262## LCMS-C: R.sub.t 2.27 min, m/z 442.9 [M +
H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 10.4 (s,
1H), 7.56-7.49 (m, 2H), 6.80 (d, J = 8.5 Hz, 2H), 4.58 (s, 2H),
3.93 (s, 3H), 3.78 (s, 6H), 3.41 (s, 3H). 2,6-Dimethoxy-
benzenesulfonyl chloride I111 5-Chloro-4- methoxy-6-
(methoxymethyl) benzo[d]isoxa- zol-3-amine I83 FA 1.5 eq. LiHMDS
used, stirred 2 h before adding 2 eq. sulfonyl chloride. Prep. TLC
(DCM/MeOH = 100/1) N-(5-Chloro-4-methoxy- 6- (methoxymethyl)benzo
[d]isoxazol-3-yl)-2,6- dimethoxybenzenesulfon- amide 146
##STR00263## LCMS-C: R.sub.t 1.89 min, m/z 442.0 [M + H].sup.+;
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 9.67 (s, 1H),
8.75-8.68 (m, 1H), 8.15 (d, J = 8.0 Hz, 1H), 7.98- 7.90 (m, 1H),
7.86 (s, 1H), 7.59 (s 1H), 7.51 (t, J = 8.5 2,6-Dimethoxy-
benzenesulfonyl chloride I111 4-Methoxy-6- (pyridin-2-
yl)benzo[d]isox- azol-3-amine I77 FA 4 eq. LiHMDS used and stirred
1 h before adding sulfonyl chloride. Prep. TLC (DCM/MeOH = 50/1)
2,6-Dimethoxy-N-(4- Hz, 1H), 7.47- methoxy-6-(pyridin-2- 7.41 (m,
1H), 6.79 yl)benzo[d]isoxazol-3- (d, J = 8.6 Hz, 2H),
yl)benzenesulfonamide 4.02 (s, 3H), 3.79 (s, 6H). 147 ##STR00264##
LCMS-C: R.sub.t 2.37 min, m/z 401.8 [M + H].sup.+; .sup.1H NMR (400
MHz, DMSO- d.sub.6) .delta. 11.6 (s, 1H), 8.32 (s, 1H), 8.19- 8.18
(m, 1H), 8.07 (s, 1H), 7.96 (d, J = 2,6-Dimethoxy- benzenesulfonyl
chloride I111 6-(Oxazol-2- yl)benzo[d]isox- azol-3-amine I90 FB
Prep. TLC (Pet. ether/EtOAc = 1/1)
2,6-Dimethoxy-N-(6- 8.4 Hz, 1H), 7.45- (oxazol-2- 7.40 (m, 2H),
6.73 yl)benzo[d]isoxazol-3- (d, J = 8.4 Hz, 2H),
yl)benzenesulfonamide 3.71 (s, 6H).
Example 148:
N-(benzo[d]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonamide 148
##STR00265##
[0719] A solution of 2,4-dimethoxybenzenesulfonyl chloride (0.18 g,
0.75 mmol) and benzo[d]isoxazol-3-amine (0.10 g, 0.75 mmol) in
pyridine (1 mL) was irradiated in the microwave at 110.degree. C.
for 2 hours. The resultant mixture was loaded onto silica gel and
the product purified twice by column chromatography (4 g SiO.sub.2
cartridge, 0-45% EtOAc in petroleum benzine 40-60.degree. C. then 4
g SiO.sub.2 cartridge, 0-35% EtOAc in petroleum benzine
40-60.degree. C.) to yield two batches (78 mg and 5 mg) of the
title compound (total mass 83 mg, 33% yield) as white solids.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.11 (d, J=8.05 Hz, 1H),
7.79 (s, 1H), 7.70 (d, J=8.81 Hz, 1H), 7.57-7.50 (m, 1H), 7.47-7.40
(m, 1H), 7.37-7.29 (m, 1H), 6.50 (d, J=2.27 Hz, 1H), 6.42 (dd,
J=2.25, 8.81 Hz, 1H), 3.98 (s, 3H), 3.81 (s, 3H). LCMS-B: rt 3.20
min, m/z=356.8 [M+Na].sup.+, 334.8 [M+H].sup.+.
Example 149:
N-(benzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 149
##STR00266##
[0721] A solution of 2,6-dimethoxybenzene-1-sulfonyl chloride I111
(0.088 g, 0.37 mmol) and benzo[d]isoxazol-3-amine (0.050 g, 0.37
mmol) in pyridine (1 mL) was irradiated in the microwave at
110.degree. C. for 2 hours, then at 120.degree. C. for 2 hours. The
reaction mixture was loaded onto silica and purified by column
chromatography (12 g SiO.sub.2 cartridge, 0-35% EtOAc in petroleum
benzine 40-60.degree. C.) to give the title compound (3.9 mg, 3.1%
yield) as a white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
8.30 (s, 1H), 8.17 (dt, J=1.04, 8.15 Hz, 1H), 7.55-7.47 (m, 1H),
7.47-7.34 (m, 2H), 7.34-7.28 (m, 1H), 6.60 (d, J=8.52 Hz, 2H), 3.91
(s, 6H). LCMS-B: rt 3.13 min, m/z=334.8 [M+H].sup.+.
Example 150:
N-(5-chlorobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
150
##STR00267##
[0722] a) 5-Chlorobenzo[d]isoxazol-3-amine A1
[0723] Potassium tert-butoxide (793 mg, 7.07 mmol) was added to a
suspension of acetohydroxamic acid (531 mg, 7.07 mmol) in DMF (10
mL) and stirred at room temperature for 30 minutes.
5-Chloro-2-fluorobenzonitrile (1.00 g, 6.43 mmol) was added and the
reaction heated to 50.degree. C. for 1 hour. Upon cooling, the
reaction mixture was diluted with an aqueous saturated solution of
NaCl (15 mL), the aqueous layer was extracted with EtOAc
(3.times.100 mL), the organics were combined, dried
(Na.sub.2SO.sub.4), filtered and the volatiles were removed in
vacuo. The residue was loaded onto silica gel and the product
purified by column chromatography (Biotage Isolera, 40 g SiO.sub.2
cartridge, 0-40% EtOAc in petroleum benzine 40-60.degree. C.) to
yield the title compound as a white solid (507 mg, 47%). .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta.=7.94 (dd, J=2.1, 0.6, 1H),
7.59-7.48 (m, 1H), 6.51 (s, 1H).
b)
N-(5-chlorobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
150
[0724] A suspension of 5-ethyl-2-methoxybenzene-1-sulfonyl chloride
(150 mg, 0.639 mmol) and 5-chlorobenzo[d]isoxazol-3-amine A1 (108
mg, 0.639 mmol) in pyridine (1.5 mL) was irradiated in the
microwave at 110.degree. C. for 2 hours. A 10 M aqueous solution of
KOH (1 mL) was added and the resultant mixture was stirred for 4
hours at room temperature. The reaction mixture was loaded onto
silica gel and the product purified by column chromatography
(0-100% EtOAc in petroleum benzine 40-60.degree. C.) to yield the
title compound as a white solid (53 mg, 23%). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta.=8.11 (t, J=1.4, 1H), 7.71 (d, J=2.3, 1H),
7.68 (d, J=1.4, 2H), 7.48 (dd, J=8.5, 2.3, 1H), 7.10 (d, J=8.6,
1H), 3.72 (s, 3H), 2.62 (q, J=7.6, 2H), 1.16 (t, J=7.6, 3H).
LCMS-A: rt 6.637 min; m/z 367.0 [M+H].sup.+.
Example 151: N-(4-chlorobenzo[d]isoxazol-3-yl)benzenesulfonamide
151
##STR00268##
[0725] a)
N-(4-chlorobenzo[d]isoxazol-3-yl)-N-(phenylsulfonyl)benzenesulfo-
namide A2
[0726] A solution of 4-chlorobenzo[d]isoxazol-3-amine (50 mg, 0.298
mmol) and benzenesulfonyl chloride (2 eq., 0.595 mmol) in pyridine
(1.5 mL) was irradiated in the microwave for 2 hours at 100.degree.
C. Upon cooling, the reaction mixture was loaded onto silica gel
and purified using silica gel column chromatography (Biotage
Isolera, 24 g SiO.sub.2 cartridge, 0-100% EtOAc in petroleum
benzine 40-60.degree. C.) to yield the title compound. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta.=7.97 (d, J=8.6, 1H), 7.90-7.77 (m,
6H), 7.71-7.64 (m, 7H), 7.57 (d, J=7.7, 1H).
b) N-(4-chlorobenzo[d]isoxazol-3-yl)benzenesulfonamide 151
[0727] A suspension of
N-(4-chlorobenzo[d]isoxazol-3-yl)-N-(phenylsulfonyl)benzenesulfonamide
A2 (50 mg, 0.11 mmol) in THF (10 mL) and 10M KOH aqueous solution
(1 mL) was stirred at room temperature for 16 hours. The reaction
mixture was diluted with water (25 mL) and the aqueous layer
extracted with EtOAc (3.times.50 mL), the combined organics were
washed with brine (25 mL) dried (Na.sub.2SO.sub.4), filtered and
concentrated under reduced pressure. The resulting gum was
dissolved in a minimum amount of acetone before petroleum benzine
40-60.degree. C. (50 mL) was added and the precipitate was filtered
and air dried to give the title compound as a tan solid (10 mg,
29%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.=7.89-7.80 (m,
2H), 7.41-7.30 (m, 4H), 7.26 (dd, J=8.3, 0.8, 1H), 7.10 (dd, J=7.5,
0.8, 1H). LCMS-A: rt 6.334 min, m/z 307.0 [M-H].sup.-.
Example 152:
5-Ethyl-N-(7-fluorobenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide
152
##STR00269##
[0728] a) 7-Fluorobenzo[d]isoxazol-3-amine A3
[0729] Potassium tert-butoxide (887 mg, 7.91 mmol) was added to a
suspension of acetohydroxamic acid (594 mg, 7.91 mmol) in DMF (10
mL) and the reaction was stirred at room temperature for 30
minutes. 2,3-Difluorobenzonitrile (1.00 g, 7.19 mmol) was added and
the reaction was heated to 50.degree. C. for 1 hour. Upon cooling,
the reaction mixture was diluted with an aqueous saturated solution
of NaCl (15 mL), the aqueous layer was extracted with EtOAc
(3.times.100 mL) the organics were combined, dried
(Na.sub.2SO.sub.4) and filtered and the volatiles were removed in
vacuo. The resulting gum was loaded onto silica gel and purified by
column chromatography (Biotage Isolera, 24 g SiO.sub.2 cartridge,
0-100% EtOAc in petroleum benzine 40-60.degree. C.) to yield the
title compound as a white solid (303 mg, 27%). .sup.1H NMR (400
MHz, CDCl.sub.3) .delta.=7.33 (dd, J=7.6, 1.3, 1H), 7.30-7.26 (m,
1H), 7.26-7.19 (m, 1H), 4.45 (s, 2H). LCMS-B: rt 3.371 min, m/z
153.2 [M+H].sup.+.
b)
5-Ethyl-N-(7-fluorobenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide
152
[0730] A solution of 7-fluorobenzo[d]isoxazol-3-amine A3 (100 mg,
0.657 mmol) and 2-methoxy-5-ethylsulfonyl chloride I112 (154 mg,
0.657 mmol) in pyridine (2 mL) was irradiated in the microwave for
2 hours at 100.degree. C. Upon cooling, the reaction mixture was
loaded onto silica gel and purified by column chromatography
(Biotage Isolera, 24 g SiO.sub.2 cartridge, 0-100% EtOAc in
petroleum benzine 40-60.degree. C.) to yield the title compound as
a white solid (127 mg, 55%). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta.=7.91 (dd, J=8.1, 0.8, 1H), 7.73 (d, J=2.3, 1H), 7.57 (dd,
J=11.9, 8.0, 1H), 7.48 (dd, J=8.5, 2.3, 1H), 7.38 (td, J=8.0, 4.1,
1H), 7.10 (d, J=8.6, 1H), 3.73 (s, 3H), 2.63 (q, J=7.6, 2H), 1.16
(t, J=7.6, 3H). LCMS-A: rt 6.429 min, m/z 351.1 [M+H].sup.+.
Example 153:
N-(4-Chloro-5-methylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfon-
amide 153
##STR00270##
[0731] a) 4-Chloro-5-methylbenzo[d]isoxazol-3-amine A4
[0732] Potassium tert-butoxide (728 mg, 6.49 mmol) was added to a
suspension of acetohydroxamic acid (487 mg, 6.49 mmol) in DMF (10
mL) and stirred at room temperature for 30 minutes.
2-Chloro-6-fluoro-3-methylbenzonitrile (1.00 g, 5.90 mmol) was
added and the reaction heated to 50.degree. C. for 1 hour. Upon
cooling, the reaction mixture was diluted with an aqueous saturated
solution of NaCl (15 mL), the aqueous layer was extracted with
EtOAc (3.times.100 mL), the organics were combined, dried
(Na.sub.2SO.sub.4), filtered and the volatiles were removed in
vacuo. The resultant solid was sonicated in acetone (10 mL) before
petroleum benzine 40-60.degree. C. (50 mL) was added, the
precipitate was collected by filtration and air dried to yield the
product as a white solid (524 mg, 49%). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta.=7.51 (d, J=8.5, 1H), 7.38 (d, J=8.5, 1H),
6.15 (s, 2H), 2.38 (s, 3H). LCMS-B: rt 3.562 min, m/z 183.1
[M+H].sup.+.
b)
N-(4-Chloro-5-methylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulf-
onamide 153
[0733] A solution of 4-chloro-5-methylbenzo[d]isoxazol-3-amine A4
(100 mg, 0.548 mmol) and 2-methoxy-5-ethylsulfonyl chloride I112
(129 mg, 0.548 mmol) in pyridine (2 mL) was irradiated in the
microwave for 2 hours at 100.degree. C. Upon cooling, the reaction
mixture was added to water, the precipitate was removed by
filtration and the filtrate was loaded onto silica gel and purified
by column chromatography (Biotage Isolera, 24 g SiO.sub.2
cartridge, 0-100% EtOAc in petroleum benzine 40-60.degree. C.) to
give the title compound as a white solid (33 mg, 16%). .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta.=10.48 (s, 1H), 7.69-7.57 (m, 3H),
7.48 (dd, J=8.5, 2.3, 1H), 7.15 (d, J=8.5, 1H), 3.68 (s, 3H), 2.60
(q, J=7.5, 2H), 2.42 (s, 3H), 1.15 (t, J=7.6, 3H). LCMS-A: rt 6.665
min, m/z 381.1 [M+H].sup.+.
Example 154:
N-(4-chlorobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
154
##STR00271##
[0735] A mixture of 5-ethyl-2-methoxybenzenesulfonyl chloride I112
(0.414 g, 1.77 mmol) and 4-chlorobenzo[d]isoxazol-3-amine (0.225 g,
1.34 mmol) in pyridine (2.0 mL) was stirred at 30.degree. C. for 40
hours under a nitrogen atmosphere. The reaction was concentrated,
then sonicated for 2 hours with aqueous HCl (5%) and the resulting
precipitate collected. The precipitate was purified using silica
gel column chromatography (0-100% ethyl acetate/petroleum benzine
40-60.degree. C.) to give the title compound as two fractions (A
and B) with a combined yield of 0.060 g, 12% yield. Fraction A:
Yield 0.038 g. .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta. 8.88
(br s, 1H), 7.75 (d, J=2.3 Hz, 1H), 7.66 (dd, J=8.5, 7.6 Hz, 1H),
7.57 (dd, J=8.5, 0.6 Hz, 1H), 7.49 (dd, J=8.5, 2.3 Hz, 1H), 7.42
(dd, J=7.6, 0.6 Hz, 1H), 7.14 (d, J=8.5 Hz, 1H), 3.91 (s, 3H), 2.66
(q, J=7.6 Hz, 2H), 1.21 (t, J=7.6 Hz, 3H). LCMS-B: rt 3.766 min;
m/z 367.1/369.1 [M+H].sup.+.
[0736] Fraction B: Yield 0.021 g. .sup.1H NMR (400 MHz,
acetone-d.sub.6) .delta. 8.88 (br s, 1H), 7.75 (d, J=2.3 Hz, 1H),
7.65 (dd, J=8.5, 7.6 Hz, 1H), 7.59-7.55 (m, 1H), 7.49 (dd, J=8.5,
2.3 Hz, 1H), 7.44-7.40 (m, 1H), 7.14 (d, J=8.5 Hz, 1H), 3.90 (s,
3H), 2.66 (q, J=7.6 Hz, 2H), 1.21 (t, J=7.6 Hz, 3H). LCMS-B: rt
3.755 min; m/z 367.1/369.1 [M+H].sup.+
Example 155:
N-(4-chlorobenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide
155
##STR00272##
[0738] A mixture of 4-chlorobenzo[d]isoxazol-3-amine (0.034 g,
0.200 mmol) and 2-methoxybenzenesulfonyl chloride (0.092 g, 0.450
mmol) in pyridine (1.0 mL) and triethylamine (0.1 mL) was stirred
at room temperature for 16 hours. The reaction was concentrated and
diluted with 5% aqueous HCl (1 mL) and sonicated for a minimum of
30 minutes. The resulting precipitate was collected by filtration
and purified using preparative mass-directed HPLC to give the title
compound. .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta. 7.94-7.91
(dd, J=7.8, 1.7 Hz, 1H), 7.69-7.63 (m, 2H), 7.60-7.57 (dd, J=8.5,
0.7 Hz, 1H), 7.44-7.42 (dd, J=7.6, 0.7 Hz, 1H), 7.25-7.22 (m, 1H),
7.16-7.11 (m, 1H), 3.94-3.94 (s, 3H). HPLC-MS: rt 6.02 min; m/z
339.16/341.18 [M+H].sup.+.
Example 156:
N-(4-fluorobenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide
156
##STR00273##
[0740] A mixture of 4-fluorobenzo[d]isoxazol-3-amine (0.032 g, 0.21
mmol) and 2-methoxybenzenesulfonyl chloride (0.109 g, 0.529 mmol)
in pyridine (1.0 mL) and triethylamine (0.1 mL) was stirred at room
temperature for 16 hours. The reaction was concentrated and diluted
with 5% aqueous HCl (1 mL) and sonicated for a minimum of 30
minutes. The resulting precipitate was collected by filtration and
purified using preparative mass-directed HPLC to give the title
compound. .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta. 9.73-9.45
(br s, 1H), 7.92-7.88 (dd, J=7.9, 1.7 Hz, 1H), 7.70-7.61 (m, 2H),
7.44-7.40 (d, J=8.5 Hz, 1H), 7.25-7.21 (d, J=8.3 Hz, 1H), 7.13-7.07
(m, 2H), 3.95-3.91 (s, 3H). HPLC-MS: rt 5.72 min; m/z 323.16
[M+H].sup.+.
Example 157:
N-(6-bromobenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide
157
##STR00274##
[0742] A mixture of 6-bromobenzo[d]isoxazol-3-amine I75 (0.039 g,
0.180 mmol) and 2-methoxybenzenesulfonyl chloride (0.101 g, 0.490
mmol) in pyridine (1.0 mL) and triethylamine (0.1 mL) was stirred
at room temperature for 16 hours. The reaction was concentrated and
diluted with 5% aqueous HCl (1 mL) and sonicated for a minimum of
30 minutes. The resulting precipitate was collected by filtration
and purified using preparative mass-directed HPLC to give the title
compound. .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta. 8.06-8.02
(dd, J=8.6, 0.5 Hz, 1H), 7.90-7.87 (dd, J=7.9, 1.7 Hz, 1H),
7.85-7.83 (dd, J=1.6, 0.5 Hz, 1H), 7.64-7.59 (ddd, J=8.4, 7.4, 1.8
Hz, 1H), 7.58-7.54 (dd, J=8.6, 1.6 Hz, 1H), 7.21-7.18 (dd, J=8.4,
0.8 Hz, 1H), 7.10-7.05 (m, 1H), 3.88-3.85 (s, 3H). HPLC-MS: rt 6.32
min; m/z 383.1/385.2 [M+H].sup.+.
Example 158:
N-(6-chlorobenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide
158
##STR00275##
[0744] A mixture of 6-chlorobenzo[d]isoxazol-3-amine (0.033 g,
0.200 mmol) and 2-methoxybenzenesulfonyl chloride (0.095 g, 0.460
mmol) were stirred in pyridine (1.0 mL) and triethylamine (0.1 mL)
at room temperature for 16 hours. The reaction was concentrated and
diluted with 5% aqueous HCl (1 mL) and sonicated for a minimum of
30 min. The resulting precipitate was collected by filtration and
purified using mass directed preparative HPLC to give the title
compound. .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta. 8.12-8.07
(dd, J=8.6, 0.5 Hz, 1H), 7.91-7.87 (dd, J=7.9, 1.7 Hz, 1H),
7.68-7.65 (dd, J=1.7, 0.5 Hz, 1H), 7.64-7.58 (ddd, J=8.4, 7.4, 1.8
Hz, 1H), 7.44-7.39 (dd, J=8.6, 1.7 Hz, 1H), 7.21-7.17 (m, 1H),
7.10-7.05 (m, 1H), 3.88-3.86 (s, 3H). HPLC-MS: rt 6.26 min; m/z
339.16/341.18 [M+H]+.
Example 159:
N-(4-chlorobenzo[d]isoxazol-3-yl)isoquinoline-8-sulfonamide 159
##STR00276##
[0746] A solution of 4-chlorobenzo[d]isoxazol-3-amine (0.050 g,
0.30 mmol) in anhydrous THF (2 mL) was cooled to -78.degree. C.
under a nitrogen atmosphere. A solution of lithium
bis(trimethylsilyl)amide (1.0 M in THF, 0.59 mL, 0.59 mmol) was
cautiously added before the mixture was stirred at 0.degree. C. for
1 hour. The mixture was cooled to -78.degree. C., a solution of
8-isoquinolinesulfonyl chloride (0.068 g, 0.30 mmol) in anhydrous
THF (1 mL) was added and the mixture was allowed to warm to room
temperature. After stirring for 3 hours, TLC indicated only the
presence of starting material. The mixture was cooled to
-78.degree. C., sodium hydride (60% dispersion in mineral oil,
0.059 g, 1.5 mmol) was added and the mixture was returned to room
temperature and stirred overnight. Water (10 mL) was added and the
pH was adjusted to -3 with aq. HCl (2 M). The aqueous phase was
extracted with DCM (3.times.20 mL), the organics were combined,
dried (MgSO.sub.4) and the solvent removed in vacuo. The solid
residue was purified by column chromatography (Biotage Isolera, 12
g SiO.sub.2 cartridge, 0-100% EtOAc in petroleum benzine
40-60.degree. C., then 0-40% MeOH in EtOAc) to give the title
compound as a white solid (0.026 g, 24%). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 10.31 (s, 1H), 8.53 (d, J=5.8 Hz, 1H),
8.28-8.21 (m, 1H), 8.09 (d, J=8.2 Hz, 1H), 7.92 (d, J=5.7 Hz, 1H),
7.89-7.82 (m, 1H), 7.39 (t, J=7.9 Hz, 1H), 7.31 (d, J=8.3 Hz, 1H),
7.17 (d, J=7.5 Hz, 1H). LCMS-A: rt 5.43 min; m/z 360.1
[M+H].sup.+.
Example 160:
N-(7-iodo-4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-yl)-2,6-dimethoxy-
benzenesulfonamide 160
##STR00277##
[0747] a)
7-iodo-4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine A5
[0748] A portion of
4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine I9 (0.121 g,
0.581 mmol) was dissolved in N,N-dimethylformamide (2 mL) and then
N-iodosuccinimide (0.131 g, 0.581 mmol) was added. Upon completion
of addition, the reaction mixture was heated at 50.degree. C. for 2
h. At the conclusion of this period, the reaction mixture was
poured over ice and then diluted with EtOAc (15 mL). The resulting
mixture was washed with H.sub.2O (3.times.8 mL) and brine (8 mL),
dried over Na.sub.2SO.sub.4 and filtered. The volatiles were
removed under reduced pressure and the residue purified twice by
column chromatography (12 g SiO.sub.2 cartridge, 0-35% EtOAc in
petroleum benzine 40-60.degree. C. then 12 g SiO.sub.2 cartridge,
0-25% EtOAc in petroleum benzine 40-60.degree. C.) to give the
title compound (0.038 g, 20% yield) as an off-while solid. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 6.78 (s, 1H), 4.73 (s, 2H), 4.54
(s, 2H), 3.96 (s, 3H), 3.50 (s, 3H). LCMS-A: rt 3.26 min, m/z 334.7
[M+H].sup.+.
b)
N-(7-iodo-4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-yl)-2,6-dimetho-
xybenzenesulfonamide 160
[0749] A solution of 7-iodo-4-methoxy-6-(methoxymethyl)
benzo[d]isoxazol-3-amine A5 (0.024 g, 0.099 mmol) and
2,6-dimethoxybenzenesulfonyl chloride I111 (0.023 g, 0.099 mmol) in
pyridine (0.5 mL) was irradiated in the microwave at 110.degree. C.
for 2 h. The reaction mixture was cooled to room temperature and
wet-loaded onto a silica cartridge. The residue was purified by
column chromatography (12 g SiO.sub.2 cartridge, 0-70% EtOAc in
petroleum benzine 40-60.degree. C.) to give the title compound
(0.032 g, 53% yield) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.38 (t, J=8.5 Hz, 1H), 6.86 (s, 1H), 6.58 (d,
J=8.5 Hz, 2H), 4.52 (s, 2H), 4.04 (s, 3H), 3.88 (s, 6H), 3.51 (s,
3H). LCMS-A: rt 5.86 min, m/z 534.6 [M+H].sup.+.
Example 161:
N-(7-chloro-4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-yl)-2,6-dimetho-
xybenzenesulfonamide 161
##STR00278##
[0750] a)
7-Chloro-4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine A6
[0751] 4-Methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine I9
(0.150 g, 0.720 mmol) was dissolved in N,N-dimethylformamide (2 mL)
and then N-chlorosuccinimide (96 mg, 0.72 mmol) was added. Upon
completion of addition, the reaction mixture was heated at
50.degree. C. for 2 hours. At the conclusion of this period, the
reaction mixture was poured over ice and then diluted with EtOAc
(15 mL). The resulting mixture was washed with H.sub.2O (3.times.8
mL) and brine (8 mL), dried over Na.sub.2SO.sub.4 and filtered. The
volatiles were removed under reduced pressure and the residue
purified by column chromatography (12 g SiO.sub.2 cartridge, 0-40%
EtOAc in petroleum benzine 40-60.degree. C.) to give the title
compound (0.0240 g, 14% yield) as a white solid. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 6.77 (s, 1H), 4.63 (d, J=0.6 Hz, 2H), 3.97
(s, 3H), 3.49 (s, 3H).
b)
N-(7-chloro-4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-yl)-2,6-dimet-
hoxybenzene-sulfonamide 161
[0752] A solution of 7-chloro-4-methoxy-6-(methoxymethyl)
benzo[d]isoxazol-3-amine A6 (0.024 g, 0.099 mmol) and
2,6-dimethoxybenzenesulfonyl chloride I111 (0.023 g, 0.099 mmol) in
pyridine (0.5 mL) was irradiated in the microwave at 110.degree. C.
for 2 hours. The reaction mixture was cooled to room temperature
and wet-loaded onto a silica cartridge. The residue was purified by
column chromatography (12 g SiO.sub.2 cartridge, 0-100% EtOAc in
petroleum benzine 40-60.degree. C.) to give the title compound
(0.0094 g, 21% yield) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.20 (s, 1H), 7.39 (t, J=8.5 Hz, 1H), 6.85 (s,
1H), 6.59 (d, J=8.5 Hz, 2H), 4.61 (d, J=0.6 Hz, 2H), 4.04 (s, 3H),
3.88 (s, 6H), 3.49 (s, 3H). LCMS-F: rt 6.39 min, m/z 442.8
[M+H].sup.+.
Example 162:
5-methoxy-N-(4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-yl)quinoline-8-
-sulfonamide 162
##STR00279##
[0754] A solution of
4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine I9 (0.0500 g,
0.240 mmol) and 5-methoxyquinoline-8-sulfonyl chloride (0.0619 g,
0.240 mmol) in pyridine (0.500 mL) was irradiated in the microwave
at 110.degree. C. for 2 hours. The reaction was cooled to room
temperature and added to DCM (10 mL). The organic layer was washed
with 0.5 M HCl (10 mL) and the layers separated by phase separation
cartridge. The collected organic layers was dried in vacuo and the
residue purified by column chromatography (12 g SiO.sub.2
cartridge, 0-80% EtOAc in petroleum benzine 40-60.degree. C.) to
give the title compound (0.0110 g, 11% yield) as an off-white
solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.98 (dd, J=4.3,
1.8 Hz, 1H), 8.64-8.53 (m, 2H), 7.45 (dd, J=8.5, 4.3 Hz, 1H), 6.91
(d, J=8.4 Hz, 1H), 6.87 (d, J=0.9 Hz, 1H), 6.54 (s, 1H), 4.44 (s,
2H), 4.05 (s, 3H), 4.03 (s, 3H), 3.36 (s, 3H). LCMS-B: rt 3.49 min,
m/z=429.8 [M+H].sup.+.
Example 163:
2-hydroxy-6-methoxy-N-(4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-yl)b-
enzenesulfonamide 163
##STR00280##
[0756] A solution of
4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine I9 (0.0440 g,
0.211 mmol) and 2,6-dimethoxybenzenesulfonyl chloride I111 (0.0500
g, 0.211 mmol) in pyridine (0.500 mL) was irradiated in the
microwave at 120.degree. C. for 2 hours then at 120.degree. C. for
1 hour. The reaction was cooled to room temperature and added to
DCM (10 mL). The organics were washed with 1M HCl (2.times.10 mL)
then dried over MgSO.sub.4. The crude material was purified twice
by silica gel chromatography (24 g SiO.sub.2 cartridge, 0-85% EtOAc
in petroleum benzine 40-60.degree. C. then 12 g SiO.sub.2
cartridge, 0-75% EtOAc in petroleum benzine 40-60.degree. C.) to
give the title compound (1.5 mg). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 9.62 (s, 1H), 8.14 (s, 1H), 7.33 (t, J=8.4 Hz, 1H), 6.99
(q, J=0.9 Hz, 1H), 6.70-6.61 (m, 2H), 6.37 (dd, J=8.3, 1.0 Hz, 1H),
4.51 (s, 1H), 4.03 (s, 3H), 3.87 (s, 3H), 3.42 (s, 3H). LCMS-A: rt
3.54 min, m/z 394.8 [M+H].sup.+
Example 164:
6-methoxy-N-(6-(methoxymethyl)-5-methylbenzo[d]isoxazol-3-yl)pyridine-3-s-
ulfonamide 164
##STR00281##
[0758] A solution of 6-methoxypyridine-3-sulfonyl chloride (0.0540
g, 0.260 mmol) and
6-(methoxymethyl)-5-methylbenzo[d]isoxazol-3-amine I4 (0.050 g,
0.26 mmol) in pyridine (0.500 mL) was irradiated in the microwave
at 120.degree. C. for 2 hours. The reaction was cooled to room
temperature then taken up in DCM and washed (.times.2) with 1M HCl.
The organic layer was dried in vacuo then wet-loaded onto silica
gel and the product purified by column chromatography (24 g
SiO.sub.2 cartridge, 0-80% EtOAc in petroleum benzine 40-60.degree.
C.) to give the title compound (15.6 mg, 17% yield) as a colourless
oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.63 (dd, J=0.71,
2.62 Hz, 1H), 7.99-7.89 (m, 2H), 7.72 (s, 1H), 7.55 (s, 1H), 6.75
(dd, J=0.71, 8.94 Hz, 1H), 4.54 (s, 2H), 3.95 (s, 3H), 3.50 (s,
3H), 2.39 (s, 3H). LCMS-F: rt 6.39 min, m/z 348.1 [M+H].sup.+.
Example 165:
N-(6-(methoxymethyl)-5-methylbenzo[d]isoxazol-3-yl)pyridine-3-sulfonamide
165
##STR00282##
[0760] A solution of pyridine-3-sulfonyl chloride (0.0462 g, 0.260
mmol) and 6-(methoxymethyl)-5-methylbenzo[d]isoxazol-3-amine I4
(0.050 g, 0.26 mmol) in pyridine (0.500 mL) was irradiated in the
microwave at 120.degree. C. for 2 hours. The reaction was cooled to
room temperature then wet-loaded onto silica gel and the product
purified by column chromatography (24 g SiO.sub.2 cartridge, 0-80%
EtOAc in petroleum benzine 40-60.degree. C.) to give the title
compound (0.0220 g, 25% yield) as a yellow solid. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 9.09 (s, 1H), 8.83 (s, 1H), 8.18 (d,
J=8.09 Hz, 1H), 7.70 (s, 1H), 7.57 (s, 1H), 7.46 (s, 1H), 4.54 (s,
2H), 3.50 (s, 3H), 2.40 (s, 3H). LCMS-F: rt 6.12 min m/z 334.1
[M+H].sup.+, 332.0 [M-H].sup.-.
Example 166:
2,4-dimethoxy-N-(6-(methoxymethyl)-5-methylbenzo[d]isoxazol-3-yl)benzenes-
ulfonamide 166
##STR00283##
[0762] A solution of 2,4-dimethoxybenzenesulfonyl chloride (0.052
g, 0.22 mmol) and
6-(methoxymethyl)-5-methylbenzo[d]isoxazol-3-amine I4 (0.042 g,
0.22 mmol) in pyridine (0.500 mL) was irradiated in the microwave
at 110.degree. C. for 2 hours. The resulting mixture was loaded
onto silica gel and the product purified by column chromatography
(4 g SiO.sub.2 cartridge, 0-45% EtOAc in petroleum benzine
40-60.degree. C.) to yield the title compound (0.0413 g, 48% yield)
as a white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.96
(s, 1H), 7.83 (s, 1H), 7.71 (d, J=8.33 Hz, 1H), 7.47 (s, 1H), 6.48
(d, J=1.82 Hz, 1H), 6.42 (d, J=8.36 Hz, 1H), 4.50 (s, 2H), 3.95 (s,
3H), 3.80 (s, 3H), 3.47 (s, 3H), 2.37 (s, 3H). LCMS-A: rt 5.78 min,
m/z=392.8 [M+H].sup.+, 414.7 [M+Na].sup.+.
Example 167:
N-(6-(hydroxymethyl)benzo[d]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonamid-
e 167
##STR00284##
[0763] a) Methyl 3-aminobenzo[d]isoxazole-6-carboxylate A9
[0764] To a solution of ethanehydroxamic acid (0.629 g, 8.37 mmol)
in DMF (5 mL) was added potassium 2-methylpropan-2-olate (0.94 g,
8.4 mmol) and the reaction was stirred for 30 minutes. Methyl
4-cyano-3-fluorobenzoate (1.0 g, 5.6 mmol) was added followed by
DMF (2 mL) and the reaction was stirred for a further 2 hours at
room temperature. The reaction was diluted with ethyl acetate (50
mL) and water (50 mL), the aqueous layer was extracted with ethyl
acetate and the combined organic layers were washed with water,
dried, filtered and concentrated. The crude material was purified
by silica gel chromatography (12 g SiO.sub.2 cartridge, 0-50% EtOAc
in petroleum benzine 40-60.degree. C.) to give the title compound
(0.55 g, 51% yield) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.12 (d, J=1.02 Hz, 1H), 7.96 (dd, J=1.24, 8.29
Hz, 1H), 7.59 (dd, J=0.77, 8.25 Hz, 1H), 3.98 (s, 3H). LCMS: rt
2.97 min, m/z 193.0 [M+H].sup.+.
b) Methyl
3-((2,4-dimethoxyphenyl)sulfonamido)benzo[d]isoxazole-6-carboxyl-
ate A10
[0765] A solution of 2,4-dimethoxybenzene-1-sulfonyl chloride (0.67
g, 2.8 mmol) and methyl 3-aminobenzo[d]isoxazole-6-carboxylate A9
(0.55 g, 2.8 mmol) in pyridine (4 mL) was irradiated in the
microwave at 130.degree. C. for 3 hours. The reaction was cooled to
room temperature then diluted with DCM (40 mL). The organics were
washed with 1M HCl (40 mL) and the aqueous layer back-extracted
with DCM (2.times.40 mL). The combined organic layers were dried in
vacuo and the residue purified twice by column chromatography (24 g
SiO.sub.2 cartridge, 0-35% EtOAc in petroleum benzine 40-60.degree.
C.) to give two batches of the title compound (0.369 g, impure and
0.0310 g, 2.8% yield, >95% purity) as white solids. .sup.1H NMR
(400 MHz, methanol-d.sub.4) .delta. 8.13-8.06 (m, 2H), 7.97 (dd,
J=1.25, 8.47 Hz, 1H), 7.86-7.80 (m, 1H), 6.58 (dq, J=2.29, 4.60 Hz,
2H), 3.95 (s, 3H), 3.83 (s, 3H), 3.79 (s, 3H). LCMS: rt 3.26 min,
m/z 392.8 [M+H].sup.+, 415.8 [M+Na].sup.+.
c)
N-(6-(hydroxymethyl)benzo[d]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonam-
ide 167
[0766] To a suspension of lithium aluminium hydride powder (0.0758
g, 2.00 mmol) in anhydrous THF (4 mL) under nitrogen was added a
solution of methyl 3-((2,4-dimethoxyphenyl)sulfonamido)benzo
[d]isoxazole-6-carboxylate A10 (impure, 0.392 g, 0.500 mmol) in THF
(8 mL). The mixture was stirred overnight at room temperature. The
reaction was quenched under nitrogen by the dropwise addition of
wet THF followed by 1 mL of water. After the evolution of gas
ceased, a solution of 0.5 M aqueous HCl was added and the aqueous
layer extracted with ethyl acetate (3.times.20 mL). The combined
organic layers were washed with water, brine, dried over MgSO.sub.4
and the solvent removed in vacuo. The crude residue was purified by
column chromatography (24 g SiO.sub.2 cartridge, 0-100% EtOAc in
petroleum benzine 40-60.degree. C.) to give the title compound
(0.191 g, 100% yield) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.08 (dd, J=0.78, 8.30 Hz, 1H), 7.77 (s, 1H),
7.69 (d, J=8.82 Hz, 1H), 7.47 (t, J=1.03 Hz, 1H), 7.30 (dd, J=1.34,
8.29 Hz, 1H), 6.49 (d, J=2.26 Hz, 1H), 6.42 (dd, J=2.30, 8.79 Hz,
1H), 4.84 (s, 2H), 3.98 (s, 3H), 3.80 (s, 3H). LCMS-B: rt 3.02 min,
m/z 364.8 [M+H].sup.+, 386.8 [M+Na].sup.+.
Example 168:
3-((5-methoxyquinoline)-8-sulfonamido)-5-methylbenzo[d]isoxazole-6-carbox-
amide 168
##STR00285##
[0767] a) Methyl 3-amino-5-methylbenzo[d]isoxazole-6-carboxylate
A11
[0768] To a solution of ethanehydroxamic acid (0.126 g, 1.69 mmol)
in DMF (2 mL) was added potassium 2-methylpropan-2-olate (0.19 g,
1.7 mmol) and the reaction was stirred for 30 minutes. Methyl
4-cyano-5-fluoro-2-methylbenzoate (0.22 g, 1.1 mmol) was added
followed by DMF (3 mL) and the reaction was stirred for a further 2
hours at 40.degree. C. The reaction was diluted with ethyl acetate
(50 mL) and water (50 mL). The aqueous layer was extracted with
ethyl acetate and the combined organic layers were washed with
water, dried, filtered and concentrated. The crude material was
purified by silica gel chromatography (12 g SiO.sub.2 cartridge,
0-50% EtOAc in petroleum benzine 40-60.degree. C.) to give the
title compound (0.11 g, 48% yield) as a white solid. .sup.1H NMR
(400 MHz, methanol-d.sub.4) .delta. 7.85 (s, 1H), 7.64 (t, J=0.79
Hz, 1H), 3.92 (s, 3H), 2.62 (d, J=0.85 Hz, 3H). LCMS: rt 3.02 min,
m/z 207.0 [M+H].sup.+.
b) Methyl
3-((5-methoxyquinoline)-8-sulfonamido)-5-methylbenzo[d]isoxazole-
-6-carboxylate hydrochloride salt A 12
[0769] A solution of 5-methoxyquinoline-8-sulfonyl chloride (0.12
g, 0.48 mmol) and methyl
3-amino-5-methylbenzo[d]isoxazole-6-carboxylate A11 (0.10 g, 0.48
mmol) in pyridine (3 mL) was irradiated in the microwave at
110.degree. C. for 2 hours. The reaction was irradiated in the
microwave at 110.degree. C. for a further 1.5 hours. The resultant
mixture was loaded onto silica gel and the product purified by
column chromatography (4 g SiO.sub.2 cartridge, 0-45% EtOAc in
petroleum benzine 40-60.degree. C.). The product was purified
further by solid phase extraction (1 g, Si-amine, 3 void volumes of
MeOH followed by 4 void volumes of methanolic HCl). The acidic
elutes were collected and dried in vacuo to give the title compound
(17.7 mg, 7.9% yield) as a pale yellow solid. .sup.1H NMR (400 MHz,
methanol-d.sub.4) .delta. 9.40-9.33 (m, 2H), 8.77 (d, J=8.66 Hz,
1H), 8.14-8.05 (m, 1H), 7.91 (s, 1H), 7.80-7.73 (m, 1H), 7.47 (d,
J=8.73 Hz, 1H), 4.23 (s, 3H), 3.91 (s, 3H), 2.62 (s, 3H). LCMS: rt
3.35 min, m/z 427.8 [M+H].sup.+.
c)
3-((5-Methoxyquinoline)-8-sulfonamido)-5-methylbenzo[d]isoxazole-6-carb-
oxamide 168
[0770] To a 15 mL heavy walled pressure tube equipped with magnetic
stir bar under nitrogen was added methyl
3-((5-methoxyquinoline)-8-sulfonamido)-5-methylbenzo[d]isoxazole-6-carbox-
ylate hydrochloride salt A12 (15.0 mg, 0.0323 mmol), ammonia
solution (2.0 M in methanol, 0.50 mL, 1.0 mmol) and calcium
dichloride (3.59 mg, 0.0323 mmol). The reaction vessel was sealed
and heated at 80.degree. C. for 3 days. The solvent was removed
under a stream of air and ammonia solution (7.0 M in methanol, 0.50
mL, 3.5 mmol) and calcium dichloride (3.6 mg, 0.032 mmol) were
added. The reaction vessel was sealed and heated at 80.degree. C.
for 24 hours. The reaction was cooled to room temperature and the
solvent removed in vacuo. The compound was purified by column
chromatography (4 g SiO.sub.2 cartridge, 0-100% EtOAc in petroleum
benzine 40-60.degree. C.) to give the title compound (0.00180 g,
13% yield) as a white solid. .sup.1H NMR (400 MHz,
methanol-d.sub.4) .delta. 9.01 (dd, J=1.78, 4.24 Hz, 1H), 8.65 (dd,
J=1.78, 8.52 Hz, 1H), 8.43 (d, J=8.47 Hz, 1H), 7.73 (d, J=12.46 Hz,
2H), 7.57 (dd, J=4.28, 8.54 Hz, 1H), 7.08 (d, J=8.50 Hz, 1H), 4.09
(s, 3H), 2.57 (s, 3H). LCMS-B: rt 3.15 min, m/z 413.8
[M+H].sup.+.
Example 169:
N-(6-cyano-5-methylbenzo[d]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonamide
169
##STR00286##
[0771] a) 3-Amino-5-methylbenzo[d]isoxazole-6-carbonitrile A 13
[0772] To a solution of ethanehydroxamic acid (0.176 g, 2.34 mmol)
in DMF (5 mL) was added potassium 2-methylpropan-2-olate (0.26 g,
2.3 mmol) and the reaction was stirred for 30 minutes.
2-Fluoro-5-methyl-terephthalonitrile (0.25 g, 1.6 mmol) was added
followed by DMF (3 mL) and the reaction was stirred overnight at
room temperature. The reaction was diluted with ethyl acetate (50
mL) and water (50 mL), the aqueous layer was extracted with ethyl
acetate and the combined organic layers were washed with water,
dried, filtered and concentrated. The crude material was purified
by silica gel chromatography (12 g SiO.sub.2 cartridge, 0-50% EtOAc
in petroleum benzine 40-60.degree. C.) to give the title compound
(0.16 g, 59% yield) as an off-white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.72 (s, 1H), 7.47 (t, J=0.86 Hz, 1H), 2.65 (d,
J=0.86 Hz, 3H). LCMS-B: rt 2.90 min, m/z 174.0 [M+H].sup.+.
b)
N-(6-cyano-5-methylbenzo[d]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonami-
de 169
[0773] A solution of 2,4-dimethoxybenzene-1-sulfonyl chloride (0.22
g, 0.92 mmol) and 3-amino-5-methyl-1,2-benzoxazole-6-carbonitrile
A13 (0.16 g, 0.92 mmol) in pyridine (2.5 mL) was irradiated in the
microwave at 130.degree. C. for 2 hours. The reaction sat at room
temperature for 50 minutes, then irradiated in the microwave at
130.degree. C. for a further 2 hours. The reaction was cooled to
room temperature then diluted with DCM (40 mL). The organics were
washed with 1M HCl (40 mL) and the aqueous layer back extracted
twice with DCM (2.times.40 mL). The combined organic layers were
dried in vacuo and the residue loaded onto silica gel and the
product purified by column chromatography (24 g SiO.sub.2
cartridge, 0-45% EtOAc in petroleum benzine 40-60.degree. C.) to
give a yellow solid. The solid was dissolved in warm MeOH and DCM
and purified by solid phase extraction (1 g Si-amine, 3 void
volumes of MeOH followed by 3 void volumes of .about.1.25 M
methanolic ammonia). The acidic elute was dried in vacuo to give a
white solid. The solid was taken up in MeOH and the MeOH removed in
vacuo (repeated.times.3). The residue was repurified by column
chromatography (24 g SiO.sub.2 cartridge, 45% EtOAc in petroleum
benzine 40-60.degree. C.) to give two batches of the title compound
(22 and 78 mg, total mass 100 mg, 29% yield) as off white solids.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.09 (s, 1H), 7.82 (s,
1H), 7.72 (s, 1H), 7.66 (d, J=8.80 Hz, 1H), 6.52 (d, J=2.25 Hz,
1H), 6.44 (dd, J=2.24, 8.85 Hz, 1H), 3.98 (s, 3H), 3.82 (s, 3H),
2.67 (s, 3H). LCMS-B: rt 3.30 min, m/z=373.8 [M+H].sup.+, 371.9
[M-H].sup.-.
Example 170:
2,4-dimethoxy-N-(5-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide
170
##STR00287##
[0775] A solution of 2,4-dimethoxybenzene-1-sulfonyl chloride
(0.0799 g, 0.337 mmol) and 5-methylbenzo[d]isoxazol-3-amine I60
(0.050 g, 0.34 mmol) in pyridine (1 mL) was irradiated in the
microwave at 110.degree. C. for 2 hours. The resultant mixture was
loaded onto silica gel and the product purified by column
chromatography (12 g SiO.sub.2 cartridge, 0-45% EtOAc in petroleum
benzine 40-60.degree. C.) to yield the title compound (55.0 mg, 42%
yield) as a white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.86-7.82 (m, 1H), 7.72 (d, J=8.81 Hz, 1H), 7.37-7.28 (m, 2H), 6.49
(d, J=2.26 Hz, 1H), 6.43 (dd, J=2.29, 8.82 Hz, 1H), 3.96 (s, 3H),
3.80 (s, 3H), 2.47 (s, 3H). LCMS-A: rt 5.66 min, m/z 348.8
[M+H].sup.+, 347.1 [M-H].sup.-.
Example 171:
N-(6-bromo-5-methylbenzo[d]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonamide
171
##STR00288##
[0776] a) 6-Bromo-5-methylbenzo[d]isoxazol-3-amine A 14
[0777] To a solution of ethanehydroxamic acid (0.263 g, 3.50 mmol)
in N,N-dimethylformamide (5 mL) was added t-BuOK (393 mg, 3.50
mmol) and the reaction was stirred for 30 minutes.
4-Bromo-2-fluoro-5-methylbenzonitrile (0.50 g, 2.3 mmol) was added
to the reaction which was stirred for a further 2 hours at room
temperature. The reaction was diluted with ethyl acetate (50 mL)
and water (50 mL), the aqueous layer was extracted with ethyl
acetate and the combined organic layers were washed with water,
dried, filtered and concentrated. The crude material was purified
by silica gel chromatography (12 g SiO.sub.2 cartridge, 0-50% EtOAc
in petroleum benzine 40-60.degree. C.) to give the title compound
(0.30 g, 56% yield) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.68 (s, 1H), 7.37 (s, 1H), 4.35 (br s, 2H),
2.49 (s, 3H). LCMS-B: rt 3.18 min, m/z 229.8 [M+H].sup.+.
b)
N-(6-bromo-5-methylbenzo[d]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonami-
de 171
[0778] A solution of 2,4-dimethoxybenzenesulfonyl chloride (0.052
g, 0.22 mmol) and 6-bromo-5-methylbenzo[d]isoxazol-3-amine A14
(0.050 g, 0.22 mmol) in pyridine (1 mL) was irradiated twice in the
microwave at 110.degree. C. for 2 hours, then at 130.degree. C. for
2 hours. The resultant mixture was loaded onto silica gel and the
product purified by column chromatography (4 g SiO.sub.2 cartridge,
0-45% EtOAc in petroleum benzine 40-60.degree. C.) to give the
title compound (102 mg, quantitative yield) as a white solid.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.98-7.94 (m, 1H),
7.70-7.65 (m, 2H), 6.50 (d, J=2.24 Hz, 1H), 6.43 (dd, J=2.26, 8.82
Hz, 1H), 3.97 (s, 3H), 3.81 (s, 3H), 2.51 (d, J=0.87 Hz, 4H).
LCMS-A: rt 6.08 min, m/z 426.9 [M+H].sup.+.
Example 172:
N-(6-(Ethoxymethyl)-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenes-
ulfonamide 172
##STR00289##
[0780] To a solution of
6-(ethoxymethyl)-4-methoxybenzo[d]isoxazol-3-amine I21 (250 mg,
1.13 mmol) in anhydrous THF (25 mL) at -78.degree. C. under N.sub.2
was added LiHMDS (1 M solution in THF, 4.5 mL, 4.5 mmol) dropwise
and the mixture was stirred at -78.degree. C. for 2 h. A solution
of 2,6-dimethoxybenzenesulfonyl chloride I111 (400 mg, 1.69 mmol)
in anhydrous THF (2 mL) was then added dropwise and the mixture was
allowed to warm to RT and stirred overnight. The mixture was
acidified to pH 4-5 with 2 M aqueous HCl and extracted with EtOAc.
The combined organic extracts were washed with water, brine, dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by prep. TLC
(DCM/MeOH=100/1) to give the title compound (170 mg, 96% purity) as
a white solid. Further purification by prep. HPLC gave the title
compound (60 mg, 100% purity, 13% yield). LCMS-C: Rt 2.08 min; m/z
423.0 [M+H].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.53
(s, 1H), 7.51 (t, J=8.4 Hz, 1H), 7.09 (s, 1H), 6.78-6.76 (m, 3H),
4.55 (s, 2H), 3.91 (s, 3H), 3.77 (s, 6H), 3.54 (q, J=6.8 Hz, 2H),
1.19 (t, J=6.8 Hz, 3H).
Example 173:
2,6-Dimethoxy-N-(4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-yl)benzene-
sulfonamide 173
##STR00290##
[0782] To a solution of
4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine I9 (3.0 g, 14.4
mmol) in anhydrous THF (200 mL) at -78.degree. C. under N.sub.2 was
added LiHMDS (1 M solution in THF, 43.2 mL, 43.2 mmol) dropwise and
the mixture was stirred at -78.degree. C. for 2 h. A solution of
2,6-dimethoxybenzenesulfonyl chloride I111 (5.1 g, 21.6 mmol) in
anhydrous THF (10 mL) was then added dropwise and the mixture was
allowed to warm to RT and stirred overnight. The mixture was
acidified to pH 4-5 with 2 M aqueous HCl and extracted with EtOAc.
The combined organic extracts were washed with water, brine, dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The reaction was repeated using
4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine I9 (2.0 g, 9.6
mmol) in 150 mL of THF and the two batches were combined and
purified by column chromatography (Pet. ether/EtOAc=8/1 to 2/1) to
give the title compound (4.1 g, 42%) as a white solid. LCMS-C:
R.sub.t 1.96 min; m/z 409.0 [M+H].sup.+; .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.58 (s, 1H), 7.52 (t, J=8.4 Hz, 1H), 7.09
(s, 1H), 6.78 (d, J=8.4 Hz, 2H), 6.76 (s, 1H), 4.51 (s, 2H), 3.91
(s, 3H), 3.77 (s, 6H), 3.33 (s, 3H).
Example 174: 2,6-Di methoxy-N-(4-methoxy-6-phenyl
benzo[d]isoxazol-3-yl)benzenesulfonamide 174
##STR00291##
[0784] To a solution of 4-methoxy-6-phenylbenzo[d]isoxazol-3-amine
I17 (2.5 g, 10.4 mmol) in anhydrous THF (60 mL) at -78.degree. C.
under N.sub.2 was added LiHMDS (1 M solution in THF, 31.0 mL, 31.0
mmol) dropwise and the mixture was stirred at -78.degree. C. for 2
h. A solution of 2,6-dimethoxybenzenesulfonyl chloride I111 (3.7 g,
15.6 mmol) in anhydrous THF (20 mL) was then added dropwise and the
mixture was allowed to warm to 0.degree. C. and stirred overnight.
Water was added and the mixture was washed with EtOAc (50
mL.times.2). The aqueous layer was acidified to pH 3 with 1 M
aqueous HCl and extracted with EtOAc (50 mL.times.3). The combined
organic extracts were washed with brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (DCM/MeOH=300/1)
and further purified by column chromatography (DCM/MeOH=200/1) to
give the title compound (1.5 g, 33%) as a white solid. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.61 (s, 1H), 7.80 (d, J=7.2 Hz,
2H), 7.50-7.44 (m, 5H), 7.09 (s, 1H), 6.80 (d, J=8.8 Hz, 2H), 4.02
(s, 3H), 3.79 (s, 6H); LCMS-C: R.sub.t 2.46 min; m/z 441.0
[M+H].sup.+.
Example 175:
3-Chloro-2,6-dimethoxy-N-(4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-y-
l)benzenesulfonamide 175
##STR00292##
[0786] To a solution of
2,6-dimethoxy-N-(4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-yl)benzene-
sulfonamide 173 (50 mg, 0.123 mmol) in DMF (10 mL) was added NCS
(14 mg, 0.123 mmol) and the mixture was heated at 50.degree. C. for
2 h. The mixture was then diluted with EtOAc (150 mL) and washed
with water, brine, dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure. The residue was purified
by prep-TLC (DCM/MeOH=120/1) to give the title compound (15 mg,
27%) as a white solid. LCMS-C: R.sub.t 2.21 min; m/z 441.0
[M+H].sup.+. .sup.1H NMR (400 MHz, methanol-d.sub.4) .delta. 7.56
(d, J=9.1 Hz, 1H), 7.01 (s, 1H), 6.91 (d, J=9.1 Hz, 1H), 6.76 (s,
1H), 4.55 (s, 2H), 3.99 (s, 6H), 3.76 (s, 3H), 3.41 (s, 3H).
Example 176:
2,6-Dimethoxy-N-(4-methoxy-6-(2-methoxyphenyl)benzo[d]isoxazol-3-yl)benze-
nesulfonamide 176
##STR00293##
[0788] A mixture of
N-(6-bromo-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamid-
e 89 (30 mg, 0.068 mmol), (2-methoxyphenyl)boronic acid (21 mg,
0.135 mmol), Pd(PPh.sub.3).sub.4 (9 mg, 0.007 mmol) and
Na.sub.2CO.sub.3 (22 mg, 0.203 mmol) in 1,4-dioxane (4 mL) and
water (1 mL) was heated at 100.degree. C. under N.sub.2 overnight.
The mixture was diluted with water and extracted with EtOAc. The
combined organic extracts were washed with brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by prep. TLC (Pet.
ether/EtOAc=1/2) to give the title compound (10 mg, 31%) as a white
solid. LCMS-C: R.sub.t 2.36 min, m/z 471.0 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.60 (s, 1H), 7.51 (t, J=8.5 Hz,
1H), 7.44-7.37 (m, 2H), 7.22 (s, 1H), 7.16 (d, J=8.8 Hz, 1H), 7.05
(t, J=7.4 Hz, 1H), 6.90 (s, 1H), 6.79 (d, J=8.6 Hz, 2H), 3.94 (s,
3H), 3.81 (s, 6H), 3.79 (s, 3H).
Example 177:
2,6-Dimethoxy-N-(4-methoxy-6-(3-methoxyphenyl)benzo[d]isoxazol-3-yl)benze-
nesulfonamide 177
##STR00294##
[0790] A mixture of
N-(6-bromo-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamid-
e 89 (50 mg, 0.113 mmol), (3-methoxyphenyl)boronic acid (35 mg,
0.226 mmol), Pd(PPh.sub.3).sub.4 (14 mg, 0.011 mmol) and
Na.sub.2CO.sub.3 (36 mg, 0.339 mmol) in 1,4-dioxane (8 mL) and
water (2 mL) was heated at 100.degree. C. under N.sub.2 overnight.
The mixture was allowed to cool to RT, adjusted to pH 4-5 then
diluted with water and extracted with EtOAc. The combined organic
extracts were washed with brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by prep. TLC (DCM/MeOH=50/1) to give the
title compound (8 mg, 15%) as a white solid. LCMS-C: R.sub.t 2.35
min; m/z 471.0 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.63 (s, 1H), 7.51 (t, J=8.5 Hz, 1H), 7.46 (d, J=1.0 Hz,
1H), 7.41 (t, J=7.9 Hz, 1H), 7.37-7.29 (m, 2H), 7.07 (s, 1H),
7.03-6.98 (m, 1H), 6.79 (d, J=8.5 Hz, 2H), 4.02 (s, 3H), 3.84 (s,
3H), 3.79 (s, 6H).
Example 178:
5-Ethyl-2-methoxy-N-(7-phenylbenzo[d]isoxazol-3-yl)benzenesulfonamide
178
##STR00295##
[0792] A suspension of
N-(7-bromobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
137 (100 mg, 0.24 mmol), phenylboronic acid (60 mg, 0.48 mmol),
Pd(dppf)Cl.sub.2 (18 mg, 0.024 mmol) and K.sub.3PO.sub.4.3H.sub.2O
(260 mg, 0.97 mmol) in toluene (5 mL), isopropanol (2 mL) and water
(5 mL) was heated at 100.degree. C. under N.sub.2 for 2 h. The
mixture was allowed to cool to RT, diluted with EtOAc and washed
with water (25 mL.times.3). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by preparative TLC (Pet.
ether/EtOAc=2/1) to give the title compound (55 mg, 55%) as a white
solid. LCMS-D: R.sub.t 3.06 min; m/z 409.1 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 11.8 (s, 1H), 8.08-8.02 (m, 1H),
7.89-7.78 (m, 3H), 7.72 (d, J=2.3 Hz, 1H), 7.55-7.40 (m, 5H),
7.13-7.08 (m, 1H), 3.75 (s, 3H), 2.61 (q, J=7.5 Hz, 2H), 1.15 (t,
J=7.5 Hz, 3H).
Example 179:
5-Ethyl-2-methoxy-N-(6-phenylbenzo[d]isoxazol-3-yl)benzenesulfonamide
179
##STR00296##
[0794] A mixture of
N-(6-bromobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
138 (120 mg, 0.3 mmol), Pd(dppf)Cl.sub.2 (45 mg, 0.06 mmol),
phenylboronic acid (150 mg, 1.2 mmol) and K.sub.3PO.sub.4.3H.sub.2O
(399 mg, 1.5 mmol) in water (10 mL), toluene (10 mL) and
isopropanol (5 mL) was heated at 85.degree. C. under N.sub.2 for 4
h. The mixture was allowed to cool to RT, diluted with water (200
mL) and extracted with diethyl ether (200 mL.times.3). The combined
organic extracts were washed with water, brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography (Pet.
ether/EtOAc=100/1 to 5/1) to give the title compound (60 mg, 50%)
as a white solid. LCMS-D: R.sub.t 3.10 min; m/z 409.1 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.7 (s, 1H), 8.14-8.08
(m, 1H), 7.87 (s, 1H), 7.79-7.74 (m, 2H), 7.73-7.67 (m, 2H),
7.54-7.39 (m, 4H), 7.12-7.07 (m, 1H), 3.74 (s, 3H), 2.61 (q, J=7.5
Hz, 2H), 1.15 (t, J=7.6 Hz, 3H).
Example 180:
N-(4-Chlorobenzo[d]isoxazol-3-yl)-2,3-dihydrobenzofuran-7-sulfonamide
180
##STR00297##
[0796] To a solution of
5-bromo-N-(4-chlorobenzo[d]isoxazol-3-yl)-2,3-dihydrobenzofuran-7-sulfona-
mide 127 (100 mg, 0.23 mmol) in THF (10 mL) was added 10% Pd/C (20
mg) and KOAc (20 mg, 0.28 mmol) and the mixture was stirred at
40.degree. C. under a H.sub.2 atmosphere for 2 h then at RT
overnight. The mixture was filtered and the filtrate was
concentrated under reduced pressure. The residue was purified by
prep. TLC (DCM/MeOH=30/1) to give the title compound (22 mg 27%) as
a light yellow solid. LCMS-D: R.sub.t 2.32 min; m/z 351.0
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.9 (s,
1H), 7.76-7.62 (m, 2H), 7.53-7.43 (m, 3H), 6.96 (t, J=7.7 Hz, 1H),
4.50 (t, J=8.8 Hz, 2H), 3.22 (t, J=8.8 Hz, 2H).
Example 181:
N-(4-Chlorobenzo[d]isoxazol-3-yl)-5-ethyl-2,3-dihydrobenzofuran-7-sulfona-
mide 181
##STR00298##
[0797]
a)N-(4-Chlorobenzo[d]isoxazol-3-yl)-5-vinyl-2,3-dihydrobenzofuran-7-
-sulfonamide A15
[0798] To a solution of
5-bromo-N-(4-chlorobenzo[d]isoxazol-3-yl)-2,3-dihydrobenzofuran-7-sulfona-
mide 127 (200 mg, 0.47 mmol) in 1,4-dioxane (20 mL), EtOH (10 mL)
and H.sub.2O (10 mL) was added K.sub.2CO.sub.3 (206 mg, 1.86 mmol),
4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (140 mg, 0.93 mmol)
and Pd(PPh.sub.3).sub.4 (54 mg, 0.047 mmol) and the mixture was
heated at 90.degree. C. under a N.sub.2 atmosphere overnight. The
solvents were removed under reduced pressure and the residue was
partitioned between DCM (50 mL), water (45 mL) and 2 M aq. HCl (5
mL). The layers were separated and the organic layer was washed
with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography (Pet. ether/EtOAc=3/1) to give the
title compound (120 mg 70%) as a yellow solid. LCMS-D: R.sub.t 0.40
min; m/z 377.0 [M+H].sup.+.
b)
N-(4-chlorobenzo[d]isoxazol-3-yl)-5-ethyl-2,3-dihydrobenzofuran-7-sulfo-
namide 181
[0799] To a solution of
N-(4-chlorobenzo[d]isoxazol-3-yl)-5-vinyl-2,3-dihydrobenzofuran-7-sulfona-
mide A15 (120 mg, 0.32 mmol) in MeOH (15 mL) was added 10% Pd/C (24
mg) and the mixture was stirred at RT under a H.sub.2 atmosphere
overnight. The mixture was filtered and the filtrate was
concentrated under reduced pressure. The residue was purified by
prep. TLC (DCM/MeOH=30/1) to give the title compound (33 mg, 27%)
as a white solid. LCMS-D: R.sub.t 2.59 min; m/z 379.1 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.8 (s, 1H), 7.75-7.70
(m, 1H), 7.69-7.63 (m, 1H), 7.47-7.43 (m, 1H), 7.37 (s, 1H), 7.29
(s, 1H), 4.46 (t, J=8.7 Hz, 2H), 3.18 (t, J=8.7 Hz, 2H), 2.56 (q,
J=7.6 Hz, 2H), 1.13 (t, J=7.6 Hz, 3H).
Example 182:
N-(Benzo[d]isoxazol-3-yl)-4-ethyl-2-methoxybenzenesulfonamide
182
##STR00299##
[0800] a)
N-(Benzo[d]isoxazol-3-yl)-2-methoxy-4-vinylbenzenesulfonamide A
16
[0801] To a solution of
N-(benzo[d]isoxazol-3-yl)-4-bromo-2-methoxybenzenesulfonamide 141
(200 mg, 0.52 mmol) in toluene (16 mL), water (8 mL) and
isopropanol (8 mL) was added K.sub.2CO.sub.3 (288 mg, 2.09 mmol),
4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (160 mg, 1.04 mmol)
and Pd(PPh.sub.3).sub.4 (60 mg, 0.052 mmol) and the mixture was
heated at 90.degree. C. under a N.sub.2 atmosphere for 2 h. The
mixture was diluted with water (50 mL) and 2 M aq. HCl (20 mL) and
extracted with EtOAc (80 mL.times.2). The combined organic extracts
were washed with brine, dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by silica gel chromatography (Pet. ether/EtOAc=5/1) to
give the title compound (150 mg, 76%) as a light yellow solid.
LCMS-D: R.sub.t 2.47 min; m/z 331.0 [M+H].sup.+, 353.0
[M+Na].sup.+.
b) N-(Benzo[d]isoxazol-3-yl)-4-ethyl-2-methoxybenzenesulfonamide
182
[0802] To a solution of
N-(benzo[d]isoxazol-3-yl)-2-methoxy-4-vinylbenzenesulfonamide A16
(80 mg, 0.24 mmol) in MeOH (10 mL) was added 10% Pd/C (16 mg) and
the mixture was stirred at 25.degree. C. overnight under a H2
atmosphere. The mixture was filtered and the filtrate was
concentrated under reduced pressure. The residue was purified by
prep. TLC (Pet. ether/EtOAc=5/1) to give the title compound (20 mg,
25%) as a light yellow solid. LCMS-D: R.sub.t 2.55 min; m/z 333.1
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.7 (s,
1H), 8.07 (d, J=8.0 Hz, 1H), 7.79 (d, J=8.0 Hz, 1H), 7.66-7.55 (m,
2H), 7.44-7.31 (m, 1H), 7.01 (s, 1H), 6.96 (d, J=8.4 Hz, 1H), 3.76
(s, 3H), 2.63 (q, J=7.6 Hz, 2H), 1.17 (t, J=7.6 Hz, 3H).
Example 183:
N-(Benzo[d]isoxazol-3-yl)-3-methoxy-[1,1'-biphenyl]-4-sulfonamide
183
##STR00300##
[0804] To a solution of
N-(benzo[d]isoxazol-3-yl)-4-bromo-2-methoxybenzenesulfonamide 141
(100 mg, 0.26 mmol) in toluene (7 mL), water (7 mL) and isopropanol
(2.5 mL) was added K.sub.2CO.sub.3 (144 mg, 10 mmol), phenylboronic
acid (64 mg, 0.52 mmol) and Pd(PPh.sub.3).sub.4 (30 mg, 0.026 mmol)
and the mixture was heated at 90.degree. C. under a N.sub.2
atmosphere for 2 h. The mixture was diluted with water (50 mL) and
2 M aq. HCl (10 mL) and extracted with EtOAc (70 mL.times.2). The
combined organic extracts were washed with brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by silica gel chromatography
(Pet. ether/EtOAc=100/1 to 5/1) to give the title compound (55 mg,
46%) as a light yellow solid. LCMS-D: R.sub.t 2.79 min; m/z 381.0
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.9 (s,
1H), 8.10 (d, J=8.0 Hz, 1H), 7.98 (d, J=8.0 Hz, 1H), 7.79-7.72 (m,
2H), 7.66-7.58 (m, 2H), 7.54-7.35 (m, 6H), 3.89 (s, 3H).
Example 184:
3-(5-Ethyl-2-methoxyphenylsulfonamido)-N-methylbenzo[d]isoxazole-7-carbox-
amide 184
##STR00301##
[0805] a)
N-(7-Bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5--
ethyl-2-methoxybenzenesulfonamide A 17
[0806] To a solution of
N-(7-bromobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
137 (2.3 g, 5.5 mmol), (2,4-dimethoxyphenyl)methanol (1.4 g, 8.4
mmol) and PPh.sub.3 (3.6 g, 14.2 mmol) in THF (400 mL) at 0.degree.
C. under N.sub.2 was added DIAD (3.3 g, 16.4 mmol) and the mixture
was stirred at RT over the weekend. The mixture was concentrated
under reduced pressure and the residue was purified by silica gel
chromatography (Pet. ether/EtOAc=5/1) to give the title compound
(1.0 g, 32%) as a light blue solid. LCMS-D: R.sub.t 3.35 min; m/z
583.1/585.1 [M+Na].sup.+.
b) Methyl 3-(N-((2,4-dimethoxybenzyl)
oxy)-5-ethyl-2-methoxyphenylsulfonamido)
benzo[d]isoxazole-7-carboxylate A18
[0807] To a solution of
N-(7-bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-m-
ethoxybenzenesulfonamide A17 (250 mg, 1.34 mmol) in MeOH (5 mL) and
DMF (45 mL) was added Et.sub.3N (675 mg, 6.68 mmol) and
Pd(dppf)Cl.sub.2 (98 mg, 0.13 mmol) and the mixture was heated at
80.degree. C. under a CO atmosphere overnight. The solvent was
removed under reduced pressure and the residue was diluted with
EtOAc (50 mL), washed with water (50 mL.times.3), brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by silica gel chromatography
(Pet. ether/EtOAc=4/1) to give the title compound (210 mg, 31%) as
a white solid. LCMS-D: R.sub.t 3.10 min; m/z 541.2 [M+H].sup.+.
c)
3-(N-((2,4-Dimethoxybenzyl)oxy)-5-ethyl-2-methoxyphenylsulfonamido)-N-m-
ethylbenzo[d]isoxazole-7-carboxamide A 19
[0808] A mixture of methyl
3-(N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxyphenylsulfonamido)
benzo[d]isoxazole-7-carboxylate A18 (20 mg, 0.037 mmol) and
CH.sub.3NH.sub.2 (33% solution in EtOH, 4 mL) was heated at
100.degree. C. in a sealed tube for 30 min. The solvent was removed
under reduced pressure to give the title compound (20 mg, 100%),
which was used in the next step without further purification.
LCMS-D: R.sub.t 2.86 min; m/z 540.2 [M+H].sup.+.
d)
3-(5-Ethyl-2-methoxyphenylsulfonamido)-N-methylbenzo[d]isoxazole-7-carb-
oxamide 184
[0809] A mixture of 3-(N-((2,4-di
methoxybenzyl)oxy)-5-ethyl-2-methoxyphenyl
sulfonamido)-N-methylbenzo[d]isoxazole-7-carboxamide A19 (40 mg,
0.07 mmol) and TFA (2 mL) was stirred at RT for 3 h, then
concentrated under reduced pressure. The residue was purified by
preparative TLC to give the title compound (18 mg, 64%) as a white
solid. LCMS-D: R.sub.t 2.35 min; m/z 390.1 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 11.9 (s, 1H), 8.31-8.11 (m, 2H),
8.00-7.90 (m, 1H), 7.72 (s, 1H), 7.52-7.41 (m, 2H), 7.14-7.04 (m,
1H), 3.73 (s, 3H), 2.81 (s, 3H), 2.62 (q, J=8.0, 7.6 Hz, 2H), 1.15
(t, J=7.9 Hz, 3H).
Example 185:
3-(5-Ethyl-2-methoxyphenylsulfonamido)-N,N-dimethylbenzo[d]isoxazole-7-ca-
rboxamide 185
##STR00302##
[0810] a) Methyl
3-(5-ethyl-2-methoxyphenylsulfonamido)benzo[d]isoxazole-7-carboxylate
A20
[0811] A mixture of methyl
3-(N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxyphenylsulfonamido)
benzo[d]isoxazole-7-carboxylate A18 (150 mg, 0.28 mmol) and TFA (5
mL) was stirred at RT for 3 h then concentrated under reduced
pressure. The residue was purified by silica gel chromatography
(Pet. ether/EtOAc=3/1) to give the title compound (80 mg, 74%) as a
white solid. LCMS-D: R.sub.t 2.63 min; m/z 391.1 [M+H].sup.+, 413.1
[M+Na].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.9 (s,
1H), 8.39-8.32 (m, 1H), 8.19-8.12 (m, 1H), 7.72 (d, J=2.2 Hz, 1H),
7.56-7.44 (m, 2H), 7.12-7.05 (m, 1H), 3.89 (s, 3H), 3.71 (s, 3H),
2.62 (q, J=7.5 Hz, 2H), 1.16 (t, J=7.6 Hz, 3H).
b)
3-((5-Ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-7-carboxylic
acid A21
[0812] To a suspension of methyl
3-(5-ethyl-2-methoxyphenylsulfonamido)
benzo[d]isoxazole-7-carboxylate A20 (200 mg, 0.5 mmol) in MeOH (10
mL) and THF (10 mL) was added 2 M aq. NaOH (1.28 mL) and the
mixture was stirred at RT overnight. The solvent was removed under
reduced pressure and the residue was diluted with water and
adjusted to pH 2-3. The resulting precipitate was collected by
filtration to give the title compound (144 mg, 75%) as an off-white
solid. LCMS-D: R.sub.t 2.43 min; m/z 377.1 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 13.5 (s, 1H), 11.8 (s, 1H),
8.35-8.29 (m, 1H), 8.15-8.09 (m, 1H), 7.72 (d, J=2.3 Hz, 1H),
7.53-7.44 (m, 2H), 7.12-7.05 (m, 1H), 3.71 (s, 3H), 2.62 (q, J=7.6
Hz, 2H), 1.16 (t, J=7.6 Hz, 3H).
c)
3-(5-Ethyl-2-methoxyphenylsulfonamido)benzo[d]isoxazole-7-carbonyl
chloride A22
[0813] A mixture of
3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-7-carboxylic
acid A21 (30 mg, 0.08 mmol) and SOCl.sub.2 (5 mL) was heated at
85.degree. C. under a N.sub.2 atmosphere for 3 h, then concentrated
under reduced pressure to give the title compound (31 mg, 100%),
which was used directly in the next step without further
purification.
d)
3-(5-Ethyl-2-methoxyphenylsulfonamido)-N,N-dimethylbenzo[d]isoxazole-7--
carboxamide 185
[0814] To a solution of
3-(5-ethyl-2-methoxyphenylsulfonamido)benzo[d]isoxazole-7-carbonyl
chloride A22 (31 mg, 0.08 mmol) in THF (1 mL) was added
dimethylamine (40% solution in water, 2 mL) dropwise and the
mixture was stirred at RT for 1 h. The mixture concentrated under
reduced pressure and the residue was diluted with water, adjusted
to pH 2-3 and extracted with DCM (30 mL.times.3). The combined
organic extracts were washed with brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by prep. TLC to give the title compound
(18 mg, 56%) as a white solid. LCMS-D: R.sub.t 2.36 min; m/z 404.1
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.9 (s,
1H), 8.21-8.06 (m, 1H), 7.78-7.56 (m, 2H), 7.45 (s, 2H), 7.17-6.97
(m, 1H), 3.71 (s, 3H), 3.01 (s, 3H), 2.80 (s, 3H), 2.61 (m, 2H),
1.15 (m, 3H).
Example 186:
5-Ethyl-N-(7-(hydroxymethyl)benzo[d]isoxazol-3-yl)-2-methoxybenzenesulfon-
amide 186
##STR00303##
[0816] A mixture of
3-(5-ethyl-2-methoxyphenylsulfonamido)benzo[d]isoxazole-7-carboxylic
acid A21 (30 mg, 0.08 mmol) and BH.sub.3.THF (1 M solution in THF,
3 mL, 3 mmol) was stirred at RT under a N.sub.2 atmosphere for 5 h.
The reaction was quenched with water and most of the THF was
removed under reduced pressure. The residue was adjusted to pH 2-3
and extracted with DCM (15 mL.times.3). The combined organic
extracts were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by a
silica gel chromatography (DCM/MeOH=30/1) to give the title
compound (20 mg, 71%) as a light red solid. LCMS-D: R.sub.t 2.35
min; m/z 363.1 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 11.7 (s, 1H), 7.96-7.89 (m, 1H), 7.70 (d, J=2.3 Hz, 1H),
7.61-7.55 (m, 1H), 7.46 (dd, J=8.5, 2.3 Hz, 1H), 7.34 (t, J=7.6 Hz,
1H), 7.12-7.05 (m, 1H), 5.39 (t, J=5.7 Hz, 1H), 4.70 (d, J=5.4 Hz,
2H), 3.73 (s, 3H), 2.61 (q, J=7.2 Hz, 2H), 1.16 (t, J=7.6 Hz,
3H).
Example 187:
5-Ethyl-2-methoxy-N-(7-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide
187
##STR00304##
[0818] To a suspension of
N-(7-bromobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
137 (150 mg, 0.36 mmol) in 1,4-dioxane (18 mL) and water (4.5 mL)
was added K2003 (150 mg, 1.09 mmol), methylboronic acid (45 mg,
0.73 mmol) and Pd(dppf)Cl.sub.2 (27 mg, 0.036 mmol) and the mixture
was heated at 90.degree. C. under a N.sub.2 atmosphere for 4 h. The
mixture was concentrated under reduced pressure and the residue was
partitioned between EtOAc (50 mL), water (40 mL) and 1 M aq. HCl
(15 mL). The layers were separated and the organic layer was washed
with 0.5 M aq. HCl (40 mL.times.2), brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by preparative HPLC to give the title
compound (20 mg, 16%) as a white solid. LCMS-D: R.sub.t 2.25 min;
m/z 347.1 [M+H].sup.+, 369.1 [M+Na].sup.+. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 11.7 (s, 1H), 7.89-7.82 (m, 1H), 7.70 (d,
J=2.3 Hz, 1H), 7.50-7.38 (m, 2H), 7.25 (t, J=7.6 Hz, 1H), 7.12-7.04
(m, 1H), 3.73 (s, 3H), 2.60 (q, J=7.6 Hz, 2H), 2.39 (s, 3H), 1.15
(t, J=7.6 Hz, 3H).
Example 188:
5-Ethyl-N-(7-ethylbenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide
188
##STR00305##
[0819] a)
N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxy-N-(7-vinylbenzo[d-
]isoxazol-3-yl)benzenesulfonamide A23
[0820] To a suspension of
N-(7-bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-m-
ethoxybenzenesulfonamide A17 (200 mg, 0.36 mmol) in 1,4-dioxane (15
mL) and water (3 mL) was added
4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (110 mg, 0.71
mmol), K.sub.2CO.sub.3 (148 mg, 1.07 mmol) and Pd(dppf)Cl.sub.2 (26
mg, 0.036 mmol) and the mixture was heated at 90.degree. C. under a
N.sub.2 atmosphere for 4 h. The mixture was concentrated under
reduced pressure and the residue was diluted with EtOAc (30 mL) and
washed with water (25 mL.times.3). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by a silica gel chromatography
(Pet. ether/EtOAc=2/1) to give the title compound (95 mg, 52%) as a
white solid. LCMS-D: R.sub.t 3.28 min; m/z 509.0 [M+H].sup.+.
b)
5-Ethyl-2-methoxy-N-(7-vinylbenzo[d]isoxazol-3-yl)benzenesulfonamide
A24
[0821] A mixture of N-((2,4-di
methoxybenzyl)oxy)-5-ethyl-2-methoxy-N-(7-vinylbenzo[d]isoxazol-3-yl)benz-
enesulfonamide A23 (95 mg, 0.18 mmol) and TFA (4 mL) was stirred at
RT for 3 h then concentrated under reduced pressure. The residue
was purified by a silica gel chromatography (Pet. ether/EtOAc=2/1)
to give the title compound (40 mg, 61%) as a white solid. LCMS-D:
R.sub.t 2.78 min; m/z 359.1 [M+H].sup.+
c)
5-Ethyl-N-(7-ethylbenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide
188
[0822] To a solution of
5-ethyl-2-methoxy-N-(7-vinylbenzo[d]isoxazol-3-yl)benzenesulfonamide
A24 (35 mg, 0.098 mmol) in EtOAc (5 mL) was added 10% Pd/C (7 mg)
and the mixture was stirred at RT under a H2 atmosphere overnight.
The mixture was filtered, the filtrate was concentrated and the
residue was purified by silica gel chromatography (Pet.
ether/EtOAc=4/1) to give the title compound (30 mg, 85%) as a white
solid. LCMS-D: R.sub.t 2.83 min; m/z 361.1 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 11.7 (s, 1H), 7.90-7.83 (m, 1H),
7.70 (d, J=2.3 Hz, 1H), 7.50-7.40 (m, 2H), 7.28 (t, J=7.6 Hz, 1H),
7.12-7.05 (m, 1H), 3.73 (s, 3H), 2.80 (q, J=7.5 Hz, 2H), 2.61 (q,
J=7.6 Hz, 2H), 1.22 (t, J=7.5 Hz, 3H), 1.15 (t, J=7.6 Hz, 3H).
Example 189:
N-(7-Cyclopropylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamid-
e 189
##STR00306##
[0823] a)
N-(7-Cyclopropylbenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)o-
xy)-5-ethyl-2-methoxybenzenesulfonamide A25
[0824] To a suspension of
N-(7-bromobenzo[d]isoxazol-3-yl)-N-((2,4-diethylbenzyl)oxy)-5-ethyl-2-met-
hoxybenzenesulfonamide A17 (200 mg, 0.36 mmol) in 1,4-dioxane (15
mL) and H.sub.2O (3 mL) was added cyclopropylboronic acid (61 mg,
0.71 mmol), K.sub.2CO.sub.3 (148 mg, 1.07 mmol) and
Pd(dppf)Cl.sub.2 (26 mg, 0.036 mmol) and the mixture was heated at
90.degree. C. under a N.sub.2 atmosphere for 4 h. The mixture was
concentrated under reduced pressure and the residue was diluted
with EtOAc (30 mL) and washed with water (25 mL.times.3). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure. The residue was purified
by silica gel chromatography (Pet. ether/EtOAc=3/1) to give the
title compound (140 mg, 75%) as a white solid. LCMS-D: R.sub.t 3.34
min; m/z 523.2 [M+H].sup.+.
b)
N-(7-Cyclopropylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonam-
ide 189
[0825] A mixture of
N-(7-cyclopropylbenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-eth-
yl-2-methoxybenzenesulfonamide A25 (140 mg, 0.27 mmol) and TFA (6
mL) was stirred at RT for 3 h then concentrated under reduced
pressure. The residue was purified by silica gel chromatography
(Pet. ether/EtOAc=2/1) to give the title compound (80 mg, 81%) as a
white solid. LCMS-D: R.sub.t 2.86 min; m/z 373.1 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.7 (s, 1H), 7.81 (dd,
J=7.0, 2.1 Hz, 1H), 7.70 (d, J=2.3 Hz, 1H), 7.50-7.43 (m, 1H),
7.27-7.19 (m, 2H), 7.12-7.06 (m, 1H), 3.73 (s, 3H), 2.61 (q, J=7.6
Hz, 2H), 2.17-2.10 (m, 1H), 1.15 (t, J=7.5 Hz, 3H), 1.04-0.97 (m,
2H), 0.90-0.84 (m, 2H).
Example 190:
N-(7-Cyclohexylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
190
##STR00307##
[0826]
a)N-(7-(cyclohex-1-en-1-yl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxyb-
enzyl)-5-ethyl-2-methoxybenzenesulfonamide A26
[0827] To a suspension of
N-(7-bromobenzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-2-methox-
ybenzenesulfonamide A17 (200 mg, 0.36 mmol) in 1,4-dioxane (15 mL)
and water (3 mL) was added cyclohex-1-en-1-ylboronic acid (90 mg,
0.71 mmol), Pd(dppf)Cl.sub.2 (26 mg, 0.036 mmol) and
K.sub.2CO.sub.3 (148 mg, 1.07 mmol) and the mixture was heated at
90.degree. C. under N.sub.2 for 4 h. The mixture was concentrated
under reduced pressure and the residue was diluted with EtOAc (30
mL) and washed with water (25 mL.times.3). The organic layer was
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by column
chromatography (Pet. ether/EtOAc=3/1) to give the title compound
(150 mg, 75%) as a white solid, which was used directly in the next
step.
b)
N-(7-(Cyclohex-1-en-1-yl)benzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzen-
esulfonamide A27
[0828] A mixture of
N-(7-(cyclohex-1-en-1-yl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-
-ethyl-2-methoxybenzenesulfonamide A26 (150 mg, 0.26 mmol) and TFA
(7 mL) was stirred at RT for 3 h then concentrated under reduced
pressure. The residue was purified by silica gel chromatography
(Pet. ether/EtOAc=2/1) to give the title compound (65 mg, 60%) as a
white solid. LCMS-D: R.sub.t 3.64 min; m/z 413 [M+H].sup.+.
c)
N-(7-Cyclohexylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonami-
de 190
[0829] A mixture of
N-(7-(cyclohex-1-en-1-yl)benzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenes-
ulfonamide A27 (65 mg, 0.15 mmol) and 10% Pd/C (13 mg) in EtOAc (10
mL) was stirred at RT under a H2 atmosphere for 3 h. The mixture
was filtered and the filtrate was concentrated under reduced
pressure to give the title compound (40 mg, 61%) as a white solid.
LCMS-D: R.sub.t 3.34 min; m/z 415.2 [M+H].sup.+. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 11.7 (s, 1H), 7.87-7.82 (m, 1H), 7.70
(d, J=2.3 Hz, 1H), 7.49-7.41 (m, 2H), 7.28 (t, J=7.6 Hz, 1H),
7.12-7.06 (m, 1H), 3.72 (s, 3H), 2.62 (q, J=7.6 Hz, 2H), 1.85-1.75
(m, 4H), 1.75-1.67 (m, 1H), 1.61-1.48 (m, 2H), 1.44-1.20 (m, 4H),
1.15 (t, J=7.6 Hz, 3H).
Example 191:
5-Ethyl-2-methoxy-N-(7-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-yl)be-
nzenesulfonamide 191
##STR00308##
[0830] a)
N-((2,4-Dimethoxybenzyl)oxy)-5-ethyl-2-methoxy-N-(7-(1-methyl-1H-
-pyrazol-4-yl)benzo[d]isoxazol-3-yl)benzenesulfonamide A29
[0831] To a mixture of
N-(7-bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-m-
ethoxybenzenesulfonamide A17 (150 mg, 0.27 mmol) in 1,4-dioxane (15
mL) and H.sub.2O (3 mL) was added K.sub.2CO.sub.3 (110 mg, 0.80
mmol), (1-methyl-1H-pyrazol-4-yl)boronic acid (67 mg, 0.33 mmol)
and Pd(dppf)Cl.sub.2 (20 mg, 0.027 mmol) and the mixture was heated
at 90.degree. C. under a N.sub.2 atmosphere overnight. The mixture
was diluted with 0.5 M aq. HCl (30 mL) and the organic solvent was
mostly removed under reduced pressure. The remaining aqueous
mixture was extracted with DCM (40 mL.times.3) and the combined
organic extracts were dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by silica gel chromatography (DCM/MeOH=100/1) to give the
title compound (80 mg, 53%) as a light yellow solid, which was used
directly in the next step.
b)
5-Ethyl-2-methoxy-N-(7-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-yl)-
benzenesulfonamide 191
[0832] A mixture of
N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxy-N-(7-(1-methyl-1H-pyrazol--
4-yl)benzo[d]isoxazol-3-yl)benzenesulfonamide A29 (80 mg, 0.14
mmol) and TFA (3 mL) was stirred at RT for 3 h then concentrated
under reduced pressure. The residue was purified by silica gel
chromatography (DCM/MeOH=80/1) to give the title compound (50 mg,
86%) as a light yellow solid. LCMS-D: R.sub.t 2.59 min; m/z 413.1
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.8 (s,
1H), 8.34 (s, 1H), 8.06 (s, 1H), 7.91-7.84 (m, 2H), 7.72 (d, J=2.3
Hz, 1H), 7.50-7.43 (m, 1H), 7.41-7.32 (m, 1H), 7.13-7.06 (m, 1H),
3.90 (s, 3H), 3.74 (s, 3H), 2.62 (q, J=7.6 Hz, 2H), 1.16 (t, J=7.5
Hz, 3H).
Example 192:
5-Ethyl-2-methoxy-N-(7-(pyrimidin-5-yl)benzo[d]isoxazol-3-yl)benzenesulfo-
namide 192
##STR00309##
[0834] To a suspension of
N-(7-bromobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
137 (200 mg, 0.49 mmol) in toluene (16 mL), water (8 mL) and
isopropanol (8 mL) was added pyrimidin-5-ylboronic acid (181 mg,
1.46 mmol), K.sub.3PO.sub.4 (518 mg, 1.95 mmol) and
Pd(dppf)Cl.sub.2 (36 mg, 0.049 mmol) and the mixture was heated at
90.degree. C. under a N.sub.2 atmosphere overnight. The mixture was
adjusted to pH 5-6 and extracted with EtOAc (20 mL.times.3). The
combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by silica gel chromatography
(DCM/MeOH=40/1) to give the title compound (18 mg, 9%) as a brown
solid. LCMS-D: R.sub.t 2.49 min; m/z 411.1 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 11.9 (s, 1H), 9.34-9.18 (m, 2H),
8.17 (d, J=8.1 Hz, 1H), 8.06 (d, J=7.4 Hz, 1H), 7.73 (s, 1H),
7.62-7.42 (m, 3H), 7.17-7.06 (m, 1H), 3.75 (s, 3H), 2.62 (q, J=7.6
Hz, 2H), 1.16 (t, J=7.6 Hz, 3H).
Example 193:
5-Ethyl-2-methoxy-N-(6-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide
193
##STR00310##
[0835] a)
N-(6-Bromobenzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-
-2-methoxybenzenesulfonamide A30
[0836] To a solution of
N-(6-bromobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
138 (4.1 g, 10.0 mmol), (2,4-dimethoxyphenyl)methanol (2.5 g, 15.0
mmol) and PPh.sub.3 (6.6 g, 25.0 mmol) in THF (100 mL) at 0.degree.
C. under N.sub.2 was added DIAD (4.0 g, 20.0 mmol) and the mixture
was stirred at RT overnight. The mixture was concentrated under
reduced pressure and the residue was purified by silica gel
chromatography (Pet. ether/EtOAc=50/1 to 5/1) to give the title
compound (4.0 g, 71%) as a white solid, which was used directly in
the next step.
b)
N-(2,4-Dimethoxybenzyl)-5-ethyl-2-methoxy-N-(6-methylbenzo[d]isoxazol-3-
-yl)benzenesulfonamide A31
[0837] A mixture of N-(6-bromobenzo[d]isoxazol-3-yl)-N-(2,4-di
methoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A30 (120 mg,
0.214 mmol), CH.sub.3B(OH).sub.2 (64 mg, 1.07 mmol),
Pd(dppf)Cl.sub.2 (31 mg, 0.428 mmol) and K2003 (148 mg, 1.07 mmol)
in 1,4-dioxane (10 mL) and water (2 mL) was heated at 90.degree. C.
under N.sub.2 overnight. The solvent was removed under reduced
pressure and the residue was purified by column chromatography
(Pet. ether/EtOAc=50/1 to 5/1) to give the title compound (72 mg,
68%) as a white solid, which was used directly in the next
step.
c)
5-Ethyl-2-methoxy-N-(6-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide
193
[0838] A mixture of
N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxy-N-(6-methylbenzo[d]isoxazol-3-y-
l)benzenesulfonamide A31 (72 mg, 0.145 mmol) and TFA (3 mL) was
stirred at RT for 3 h then concentrated under reduced pressure. The
residue was purified by column chromatography (Pet.
ether/EtOAc=50/1 to 3/1) to give the title compound (45 mg, 90%) as
a white solid. LCMS-D: R.sub.t 2.76 min; m/z 347.1 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.6 (s, 1H), 7.94-7.87
(m, 1H), 7.68 (d, J=2.3 Hz, 1H), 7.49-7.42 (m, 1H), 7.39 (s, 1H),
7.23-7.16 (m, 1H), 7.13-7.05 (m, 1H), 3.72 (s, 3H), 2.60 (q, J=7.5
Hz, 2H), 2.43 (s, 3H), 1.14 (t, J=7.5 Hz, 3H).
Example 194:
5-Ethyl-N-(6-ethylbenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide
194
##STR00311##
[0839] a)
N-(2,4-Dimethoxybenzyl)-5-ethyl-N-(6-ethylbenzo[d]isoxazol-3-yl)-
-2-methoxybenzenesulfonamide A32
[0840] A mixture of
N-(6-bromobenzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-2-methox-
ybenzenesulfonamide A30 (200 mg, 0.356 mmol), ethylboronic acid
(132 mg, 1.78 mmol), Pd(dppf)Cl.sub.2 (52 mg, 0.071 mmol) and
K.sub.2CO.sub.3 (246 mg, 1.78 mmol) in 1,4-dioxane (20 mL) and
water (4 mL) was heated at 90.degree. C. under N.sub.2 overnight.
The solvent was removed under reduced pressure and the residue was
partitioned between DCM (200 mL) and water (50 mL). The layers were
separated and the organic layer was washed with water, brine, dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by column chromatography
(Pet. ether/EtOAc=100/1 to 5/1) to give the title compound (120 mg,
67%) as a white solid, which was used directly in the next
step.
b)
5-Ethyl-N-(6-ethylbenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide
194
[0841] A mixture of
N-(2,4-dimethoxybenzyl)-5-ethyl-N-(6-ethylbenzo[d]isoxazol-3-yl)-2-methox-
ybenzenesulfonamide A32 (120 mg, 0.24 mmol) and TFA (5 mL) was
stirred at RT overnight then concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=50/1 to 3/1) to give the title compound (80 mg, 94%) as
a white solid. LCMS-D: R.sub.t 2.84 min; m/z 361.1 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.6 (s, 1H), 7.97-7.90
(m, 1H), 7.69 (d, J=2.3 Hz, 1H), 7.49-7.37 (m, 2H), 7.26-7.19 (m,
1H), 7.11-7.04 (m, 1H), 3.72 (s, 3H), 2.73 (q, J=7.6 Hz, 2H), 2.60
(q, J=7.6 Hz, 2H), 1.20 (t, J=7.6 Hz, 3H), 1.14 (t, J=7.6 Hz,
3H).
Example 195:
N-(6-Cyclopropylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamid-
e 195
##STR00312##
[0843] A mixture of
N-(6-bromobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
138 (206 mg, 0.5 mmol), cyclopropylboronic acid (215 mg, 2.5 mmol),
Pd(dppf)Cl.sub.2 (73 mg, 0.1 mmol) and K.sub.2CO.sub.3 (345 mg, 2.5
mmol) in 1,4-dioxane (20 mL) and water (4 mL) was heated at
90.degree. C. under N.sub.2 overnight. The solvent was removed
under reduced pressure and the residue was purified by column
chromatography (Pet. ether/EtOAc=50/1 to 3/1) to give the title
compound (80 mg, 43%) as a white solid. LCMS-D: R.sub.t 2.86 min;
m/z 373.1 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
11.6 (s, 1H), 7.91-7.85 (m, 1H), 7.68 (d, J=2.3 Hz, 1H), 7.48-7.42
(m, 1H), 7.27 (s, 1H), 7.12-7.04 (m, 2H), 3.72 (s, 3H), 2.60 (q,
J=7.6 Hz, 2H), 2.10-2.00 (m, 1H), 1.14 (t, J=7.5 Hz, 3H), 1.07-0.99
(m, 2H), 0.83-0.75 (m, 2H).
Example 196:
5-Ethyl-2-methoxy-N-(6-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-yl)be-
nzenesulfonamide 196
##STR00313##
[0844] a)
N-(2,4-Dimethoxybenzyl)-5-ethyl-2-methoxy-N-(6-(1-methyl-1H-pyra-
zol-4-yl)benzo[d]isoxazol-3-yl)benzenesulfonamide A33
[0845] A mixture of N-(6-bromobenzo[d]isoxazol-3-yl)-N-(2,4-di
methoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A30 (120 mg,
0.214 mmol), (1-methyl-1H-pyrazol-4-yl)boronic acid (54 mg, 0.428
mmol), Pd(dppf)Cl.sub.2 (31 mg, 0.043 mmol) and K.sub.2CO.sub.3
(148 mg, 1.07 mmol) in 1,4-dioxane (10 mL) and water (2 mL) was
heated at 90.degree. C. under N.sub.2 overnight. The solvent was
removed under reduced pressure and the residue was partitioned
between DCM (150 mL) and water (50 mL). The layers were separated
and the organic layer was washed with water, brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography (Pet.
ether/EtOAc=50/1 to 5/1) to give the title compound (90 mg, 75%) as
a brown solid. LCMS-D: R.sub.t 3.05 min; m/z 563.0 [M+H].sup.+.
b)
5-Ethyl-2-methoxy-N-(6-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-yl)-
benzenesulfonamide 196
[0846] A mixture of
N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxy-N-(6-(1-methyl-1H-pyrazol-4-yl)
benzo[d]isoxazol-3-yl)benzenesulfonamide A33 (90 mg, 0.16 mmol) and
TFA (5 mL) was stirred at RT overnight then concentrated under
reduced pressure. The residue was purified by column chromatography
(Pet. ether/EtOAc=50/1 to 5/1) to give the title compound (30 mg,
46%) as a brown solid. LCMS-D: R.sub.t 2.57 min; m/z 413.2
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.6 (s,
1H), 8.29 (s, 1H), 8.06-7.95 (m, 2H), 7.78 (s, 1H), 7.70 (d, J=2.3
Hz, 1H), 7.64-7.56 (m, 1H), 7.50-7.42 (m, 1H), 7.13-7.05 (m, 1H),
3.87 (s, 3H), 3.71 (s, 3H), 2.61 (q, J=7.6 Hz, 2H), 1.15 (t, J=7.5
Hz, 3H).
Example 197:
5-Ethyl-2-methoxy-N-(6-(pyrimidin-5-yl)benzo[d]isoxazol-3-yl)benzenesulfo-
namide 197
##STR00314##
[0848] A mixture of
N-(6-bromobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
138 (100 mg, 0.24 mmol), pyrimidin-5-ylboronic acid (45 mg, 0.36
mmol), Pd(PPh.sub.3).sub.4 (28 mg, 0.024 mmol) and K.sub.2CO.sub.3
(166 mg, 1.2 mmol) in toluene (8 mL), water (8 mL) and isopropanol
(2 mL) was heated at 90.degree. C. under N.sub.2 for 4 h. 2 M aq.
NaOH (15 mL) was added and the mixture was stirred at RT for 20
min. The mixture was washed with EtOAc (20 mL.times.2) then
adjusted to pH 2 with conc. HCl and extracted with DCM (50
mL.times.2). The combined organic extracts were washed with water,
brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
silica gel chromatography (DCM/MeOH=30/1) to give the title
compound (15 mg, 15%) as a white solid. LCMS-D: R.sub.t 2.97 min;
m/z 411.0 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
11.8 (s, 1H), 9.24 (s, 3H), 8.21-8.15 (m, 1H), 8.11 (s, 1H),
7.85-7.78 (m, 1H), 7.72 (d, J=2.3 Hz, 1H), 7.49-7.42 (m, 1H),
7.13-7.05 (m, 1H), 3.73 (s, 3H), 2.61 (q, J=7.6 Hz, 2H), 1.15 (t,
J=7.6 Hz, 3H).
Example 198:
5-Ethyl-2-methoxy-N-(4-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide
198
##STR00315##
[0849] a)
N-(4-Bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5--
ethyl-2-methoxybenzenesulfonamide A34
[0850] To a solution of
N-(4-bromobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
129 (2.1 g, 5.1 mmol), (2,4-dimethoxyphenyl)methanol (1.3 g, 7.7
mmol) and PPh.sub.3 (3.35 g, 12.8 mmol) in THF (200 mL) at
0.degree. C. under N.sub.2 was added DIAD (3.1 g, 15.3 mmol) and
the mixture was stirred at 0.degree. C. for 1 h then allowed to
warm to RT and stirred overnight. The mixture was concentrated
under reduced pressure and the residue was purified by column
chromatography (Pet. ether/EtOAc=5/1) to give the title compound
(1.2 g, 42%) as a white solid. LCMS-D: R.sub.13.40 min; m/z 583.0
[M+H].sup.+.
b)
N((2,4-Dimethoxybenzyl)oxy)-5-ethyl-2-methoxy-N-(4-methylbenzo[d]isoxaz-
ol-3-yl)benzenesulfonamide A35
[0851] A mixture of
N-(4-bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-m-
ethoxybenzenesulfonamide A34 (200 mg, 0.36 mmol), methylboronic
acid (43 mg, 0.71 mmol), K.sub.2CO.sub.3 (148 mg, 1.07 mmol) and
Pd(dppf)Cl.sub.2 (26 mg, 0.036 mmol) in 1,4-dioxane (15 mL) and
water (3 mL) was heated at 90.degree. C. under N.sub.2 for 4 h. The
mixture was adjusted to pH 2-3 and most of the solvent was removed
under reduced pressure. The residue was diluted with water (10 mL)
and extracted with DCM (25 mL.times.3). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EA=5/1) to give the title
compound (125 mg, 71%) as a white solid, which was used directly in
the next step.
c)
5-Ethyl-2-methoxy-N-(4-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide
198
[0852] A mixture of N-((2,4-di
methoxybenzyl)oxy)-5-ethyl-2-methoxy-N-(4-methyl
benzo[d]isoxazol-3-yl)benzenesulfonamide A35 (125 mg, 0.26 mmol)
and TFA (3 mL) was stirred at RT under for 3 h then diluted with
DCM (100 mL), washed with water, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=3/1) to give the title compound (85 mg, 92%) as a white
solid. LCMS-D: R.sub.t 2.69 min m/z 347.1 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 10.6 (s, 1H), 7.58-7.56 (m, 1H),
7.52-7.46 (m, 3H), 7.18-7.14 (m, 2H), 3.73 (s, 3H), 2.62-2.56 (m,
5H), 1.14 (t, J=7.6 Hz, 3H)
Example 199:
5-Ethyl-N-(4-ethylbenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide
199
##STR00316##
[0853] a)
N-(2,4-Dimethoxybenzyl)-5-ethyl-N-(4-ethylbenzo[d]isoxazol-3-yl)-
-2-methoxybenzenesulfonamide A36
[0854] A mixture of
N-(4-bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-m-
ethoxybenzenesulfonamide A34 (200 mg, 0.36 mmol), ethylboronic acid
(53 mg, 0.71 mmol), K.sub.2CO.sub.3 (148 mg, 1.07 mmol) and
Pd(dppf)Cl.sub.2 (26 mg, 0.036 mmol) in 1,4-dioxane (15 mL) and
H.sub.2O (3 mL) was heated at 90.degree. C. under N.sub.2 for 4 h.
The mixture was adjusted to pH 2-3 and most of the solvent was
removed under reduced pressure. The residue was diluted with water
(15 mL) and extracted with DCM (20 mL.times.3). The combined
organic layers were dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure. The residue was purified
by column chromatography (Pet. ether/EtOAc=5/1) to give the title
compound (120 mg, 66%) as a white solid, which was used directly in
the next step.
b)
5-Ethyl-N-(4-ethylbenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide
199
[0855] A mixture of
N-(2,4-dimethoxybenzyl)-5-ethyl-N-(4-ethylbenzo[d]isoxazol-3-yl)-2-methox-
ybenzenesulfonamide A36 (120 mg, 0.23 mmol) and TFA (3 mL) was
stirred at room temperature under N.sub.2 for 3 h then diluted with
DCM (100 mL), washed with water, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=3/1) to give the title compound (75 mg, 89%) as a white
solid. LCMS-D: R.sub.t 2.80 min 361.1 [M+H].sup.+. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 10.5 (s, 1H), 7.60-7.49 (m, 4H),
7.22-7.18 (m, 2H), 3.81 (s, 3H), 8.09 (q, J=7.6 Hz, 2H), 2.59 (q,
J=7.6 Hz, 2H), 1.25 (t, J=7.6 Hz, 3H), 1.14 (t, J=7.6 Hz, 3H).
Example 200:
N-(4-Cyclopropylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamid-
e 200
##STR00317##
[0856] a)
N-(4-Cyclopropylbenzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-
-ethyl-2-methoxybenzenesulfonamide A37
[0857] A mixture of
N-(4-bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-m-
ethoxybenzenesulfonamide A34 (200 mg, 0.36 mmol),
cyclopropylboronic acid (61 mg, 0.71 mmol), K.sub.2CO.sub.3 (148
mg, 1.07 mmol) and Pd(dppf)Cl.sub.2 (26 mg, 0.036 mmol) in
1,4-dioxane (15 mL) and H.sub.2O (3 mL) was heated at 90.degree. C.
under N.sub.2 for 4 h. The mixture was adjusted to pH 2-3 and most
of the solvent was removed under reduced pressure. The residue was
diluted with water (15 mL) and extracted with DCM (20 mL.times.3).
The combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=5/1) to give the title compound (120 mg, 66%) as a
yellow solid, which was used directly in the next step.
b)
N-(4-Cyclopropylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonam-
ide 200
[0858] A mixture of
N-(4-cyclopropylbenzo[d]isoxazol-3-yl)-N-(2,4-di
methoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A37 (130 mg,
0.25 mmol) and TFA (3 mL) was stirred at RT under N.sub.2 for 3 h
then concentrated under reduced pressure. The residue was purified
by column chromatography (Pet. ether/EtOAc=3/1) to give the title
compound (85 mg, 92%) as a yellow solid. LCMS-D: R.sub.t 2.81 min,
373.1 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.6
(s, 1H), 7.58-7.57 (d, J=2.0 Hz, 1H), 7.53-7.42 (m, 3H), 7.18-7.16
(m, 1H), 6.86-6.84 (m, 1H), 3.74 (s, 3H), 2.72 (m, 1H), 2.60 (q,
J=7.6 Hz, 2H), 1.14 (t, J=7.6 Hz, 3H), 1.02-0.98 (m, 2H), 0.82-0.78
(m, 2H).
Example 201:
N-(4-cyclohexylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
201
##STR00318##
[0859] a)
N-(4-(Cyclohex-1-en-1-yl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxy-
benzyl)-5-ethyl-2-methoxybenzenesulfonamide A38
[0860] A mixture of
N-(4-bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-m-
ethoxybenzenesulfonamide A34 (200 mg, 0.36 mmol),
cyclohex-1-en-1-ylboronic acid (90 mg, 0.71 mmol), K.sub.2CO.sub.3
(148 mg, 1.07 mmol) and Pd(dppf)Cl.sub.2 (26 mg, 0.036 mmol) in
1,4-dioxane (15 mL) and H.sub.2O (3 mL) was heated at 90.degree. C.
under N.sub.2 for 4 h. Most of the solvent was removed under
reduced pressure, the residue was diluted with water (30 mL),
adjusted to pH 1-2 and extracted with DCM (35 mL.times.3). The
combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (DCM/MeOH=100/1)
to give the title compound (150 mg, 75%) as a white solid, which
was used directly in the next step.
b)
N-(4-(Cyclohex-1-en-1-yl)benzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzen-
esulfonamide A39
[0861] A mixture of
N-(4-(cyclohex-1-en-1-yl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-
-ethyl-2-methoxybenzenesulfonamide A38 (150 mg, 0.27 mmol) and TFA
(3 mL) was stirred at RT under N.sub.2 for 3 h then concentrated
under reduced pressure. The residue was purified by column
chromatography (Pet. ether/EtOAc=2/1) to give the title compound
(80 mg, 73%) as a white solid, which was used directly in the next
step.
c)
N-(4-Cyclohexylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonami-
de 201
[0862] A mixture of
N-(4-(cyclohex-1-en-1-yl)benzo[d]isoxazol-3-yl)-5-ethyl-2-methoxy
benzenesulfonamide A39 (80 mg, 0.19 mmol) and 10% Pd/C (16 mg) in
EtOAc (10 mL) was stirred at RT under a H2 atmosphere for 3 h. The
mixture was filtered and the filtrate was concentrated under
reduced pressure to give the title compound (75 mg, 94%) as a white
solid. LCMS-D: R.sub.t 3.24 min, 415.2 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 10.5 (s, 1H), 7.58-7.48 (m, 4H),
7.26 (d, J=7.2 Hz, 1H), 7.19 (d, J=8.8 Hz, 1H), 3.80 (s, 3H),
3.62-3.56 (m, 1H), 2.59 (q, J=7.6 Hz, 2H), 1.91-1.71 (m, 5H),
1.45-1.23 (m, 5H), 1.15 (t, J=7.6 Hz, 3H).
Example 202:
5-Ethyl-2-methoxy-N-(4-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-yl)be-
nzenesulfonamide 202
##STR00319##
[0863] a)
N-(2,4-Dimethoxybenzyl)-5-ethyl-2-methoxy-N-(4-(1-methyl-1H-pyra-
zol-4-yl)benzo[d]isoxazol-3-yl)benzenesulfonamide A40
[0864] A mixture of
N-(4-bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-m-
ethoxybenzenesulfonamide A34 (200 mg, 0.36 mmol),
(1-methyl-1H-pyrazol-4-yl)boronic acid (90 mg, 0.71 mmol),
K.sub.2CO.sub.3 (148 mg, 1.07 mmol) and Pd(dppf)Cl.sub.2 (26 mg,
0.036 mmol) in 1,4-dioxane (15 mL) and H.sub.2O (3 mL) was heated
at 90.degree. C. under N.sub.2 for 4 h. Most of the solvent was
removed under reduced pressure, the residue was diluted with water
(30 mL), adjusted to pH 1-2 and extracted with DCM (35 mL.times.3).
The combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (DCM/MeOH=100/1)
to give the title compound (105 mg, 53%) as a yellow solid, which
was used directly in the next step.
b)
5-Ethyl-2-methoxy-N-(4-(1-methyl-1H-pyrazol-4-34)benzo[d]isoxazol-3-yl)-
benzenesulfonamide 202
[0865] A mixture of
N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxy-N-(4-(1-methyl-1H-pyrazol-4-yl)
benzo[d]isoxazol-3-yl)benzenesulfonamide A40 (105 mg, 0.19 mmol)
and TFA (2 mL) was stirred at RT under N.sub.2 for 3 h then
concentrated under reduced pressure. The residue was purified by
column chromatography (DCM/MeOH=80/1) to give the title compound
(60 mg, 79%) as a yellow solid. LCMS-D: R.sub.t 2.587 min, m/z
413.1 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.2
(s, 1H), 8.12 (s, 1H), 7.90 (s, 1H), 7.66-7.45 (m, 4H), 7.39 (d,
J=7.2 Hz, 1H), 7.13 (d, J=8.4 Hz, 1H), 3.88 (s, 3H), 3.75 (s, 3H),
2.58 (q, J=7.6 Hz, 2H), 1.15 (t, J=7.6 Hz, 3H).
Example 203:
5-Ethyl-2-methoxy-N-(4-(pyrimidin-5-yl)benzo[d]isoxazol-3-yl)benzenesulfo-
namide 203
##STR00320##
[0866] a)
N-(2,4-Dimethoxybenzyl)-5-ethyl-2-methoxy-N-(4-(pyrimidin-5-yl)b-
enzo[d]isoxazol-3-yl)benzenesulfonamide A41
[0867] A mixture of
N-(4-bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-m-
ethoxybenzenesulfonamide A34 (200 mg, 0.36 mmol),
pyrimidin-5-ylboronic acid (43 mg, 0.71 mmol), K.sub.2CO.sub.3 (148
mg, 1.07 mmol) and Pd(dppf)Cl.sub.2 (26 mg, 0.036 mmol) in
1,4-dioxane (15 mL) and H.sub.2O (3 mL) was heated at 90.degree. C.
under N.sub.2 for 4 h. Most of the solvent was removed under
reduced pressure, the residue was diluted with water (30 mL),
adjusted to pH 1-2 and extracted with DCM (35 mL.times.3). The
combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (DCM/MeOH=60/1)
to give the title compound (95 mg, 47%) as a yellow solid, which
was used directly in the next step.
b)
5-Ethyl-2-methoxy-N-(4-(pyrimidin-5-yl)benzo[d]isoxazol-3-yl)benzenesul-
fonamide 203
[0868] A mixture of
N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxy-N-(4-(pyrimidin-5-yl)
benzo[d]isoxazol-3-yl)benzenesulfonamide A41 (95 mg, 0.17 mmol) and
TFA (2 mL) was stirred at RT under N.sub.2 for 3 h then
concentrated under reduced pressure. The residue was purified by
column chromatography (DCM/MeOH=40/1) to give the title compound
(45 mg, 65%) as a red solid. LCMS-D: R.sub.t 2.50 min, m/z 411.1
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.5 (s,
1H), 9.24 (s, 1H), 8.97 (s, 2H), 7.86-7.79 (m, 2H), 7.53-7.38 (m,
3H), 7.13-7.11 (m, 1H), 3.75 (s, 3H), 2.55 (t, J=7.6 Hz, 2H), 1.11
(t, J=7.6 Hz, 3H).
Example 204:
N-(5-Ethylbenzo[d]isoxazol-3-yl)-3-methoxy-5,6,7,8-tetrahydronaphthalene--
2-sulfonamide 204
##STR00321##
[0870] A mixture of
N-(5-bromobenzo[d]isoxazol-3-yl)-3-methoxy-5,6,7,8-tetrahydronaphthalene--
2-sulfonamide 113 (219 mg, 0.5 mmol), ethylboronic acid (185 mg,
2.5 mmol), Pd(dppf)Cl.sub.2 (73 mg, 0.1 mmol) and K.sub.2CO.sub.3
(345 mg, 2.5 mmol) in 1,4-dioxane (20 mL) and water (20 mL) was
heated at 90.degree. C. under N.sub.2 overnight. The mixture was
then adjusted to pH 1 and most of the solvent was removed under
reduced pressure. The residue was diluted with water (50 mL) and
extracted with DCM (100 mL). The organic extract was washed with
water, brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
silica gel chromatography (Pet. ether/EtOAc=10/1 to 2/1) followed
by preparative TLC (Pet. ether/EtOAc=1/1) to give the title
compound (52 mg, 27%) as a white solid. LCMS-D: R.sub.t 3.06 min,
m/z 387.1 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
11.4 (s, 1H), 7.83 (s, 1H), 7.54 (s, 1H), 7.51-7.43 (m, 2H), 6.84
(s, 1H), 3.71 (s, 3H), 2.77-2.65 (m, 6H), 1.75-1.66 (m, 4H), 1.24
(t, J=7.6 Hz, 3H).
Example 205:
N-(5-Cyclopropylbenzo[d]isoxazol-3-yl)-3-methoxy-5,6,7,8-tetrahydronaphth-
alene-2-sulfonamide 205
##STR00322##
[0872] A mixture of
N-(5-bromobenzo[d]isoxazol-3-yl)-3-methoxy-5,6,7,8-tetrahydronaphthalene--
2-sulfonamide 113 (219 mg, 0.5 mmol), cyclopropylboronic acid (215
mg, 2.5 mmol), Pd(dppf)Cl.sub.2 (73 mg, 0.1 mmol) and
K.sub.2CO.sub.3 (345 mg, 2.5 mmol) in 1,4-dioxane (20 mL) and water
(4 mL) was heated at 90.degree. C. under N.sub.2 overnight. The
mixture was then adjusted to pH 1 with conc. HCl and most of the
solvent was removed under reduced pressure. The residue was diluted
with water (30 mL) and extracted with DCM (100 mL). The organic
extract was washed with water, brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by silica gel chromatography (Pet.
ether/EtOAc=10/1 to 2/1) followed by preparative TLC (Pet.
ether/EtOAc=1/1) to give the title compound (50 mg, 25%) as a white
solid. LCMS-D: R.sub.t 3.07 min, m/z 399.1 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 11.4 (s, 1H), 7.68 (s, 1H), 7.54
(s, 1H), 7.47-7.43 (m, 1H), 7.40-7.35 (m, 1H), 6.84 (s, 1H), 3.71
(s, 3H), 2.77-2.65 (m, 4H), 2.08-2.00 (m, 1H), 1.74-1.66 (m, 4H),
1.04-0.96 (m, 2H), 0.68-0.61 (m, 2H).
Example 206:
N-(4-Chloro-5-ethylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfona-
mide 206
##STR00323##
[0874] A mixture of
N-(5-bromo-4-chlorobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxy
benzenesulfonamide 128 (200 mg, 0.449 mmol), ethylboronic acid (166
mg, 2.244 mmol), Pd(dppf)Cl.sub.2 (66 mg, 0.09 mmol) and
K.sub.2CO.sub.3 (310 mg, 2.244 mmol) in 1,4-dioxane (20 mL) and
water (4 mL) was heated at 90.degree. C. under N.sub.2 overnight.
The mixture was then adjusted to pH 2 with 1 M aq. HCl and most of
the solvent was removed under reduced pressure. The residue was
diluted with water and extracted with DCM. The organic extract was
washed with water, brine, dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by silica gel chromatography (Pet. ether/EtOAc=50/1 to
2/1) followed by preparative TLC (DCM/MeOH=10/1) to give the title
compound (15 mg, 9%) as a white solid. LCMS-D: R.sub.t 2.89 min,
m/z 395.1 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
10.4 (s, 1H), 7.64 (s, 2H), 7.61-7.57 (m, 1H), 7.49-7.44 (m, 1H),
7.15-7.10 (m, 1H), 3.66 (s, 3H), 2.80 (q, J=7.6 Hz, 2H), 2.60 (q,
J=7.6 Hz, 2H), 1.21-1.11 (m, 6H).
Example 207:
N-(4-Chloro-5-cyclopropylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenes-
ulfonamide 207
##STR00324##
[0876] A mixture of
N-(5-bromo-4-chlorobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxy
benzenesulfonamide 128 (200 mg, 0.449 mmol), cyclopropylboronic
acid (193 mg, 2.244 mmol), Pd(dppf)Cl.sub.2 (66 mg, 0.09 mmol) and
K.sub.2CO.sub.3 (310 mg, 2.244 mmol) in 1,4-dioxane (20 mL) and
water (4 mL) was heated at 90.degree. C. under N.sub.2 overnight.
The mixture was then adjusted to pH 2 with 1 M aq. HCl and most of
the solvent was removed under reduced pressure. The residue was
diluted with water and extracted with DCM. The organic extract was
washed with water, brine, dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by silica gel chromatography (Pet. ether/EtOAc=50/1 to
3/1) followed by preparative TLC (DCM/MeOH=100/1) to give the title
compound (20 mg, 11%) as a white solid. LCMS-D: R.sub.t 2.91 min,
m/z 407.1 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
10.4 (s, 1H), 7.64-7.56 (m, 2H), 7.52-7.44 (m, 1H), 7.34-7.27 (m,
1H), 7.17-7.11 (m, 1H), 3.67 (s, 3H), 2.60 (q, J=7.6 Hz, 2H),
2.27-2.17 (m, 1H), 1.15 (t, J=7.6 Hz, 3H), 1.08-1.00 (m, 2H),
0.78-0.70 (m, 2H).
Example 208:
N-(5-Cyclopropyl-4-methoxybenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzene-
sulfonamide 208
##STR00325##
[0878] A mixture of
N-(5-bromo-4-methoxybenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxy
benzenesulfonamide 108 (100 mg, 0.227 mmol), cyclopropylboronic
acid (97 mg, 1.133 mmol), Pd(dppf)Cl.sub.2 (33 mg, 0.046 mmol) and
K.sub.2CO.sub.3 (156 mg, 1.133 mmol) in 1,4-dioxane (20 mL) and
water (4 mL) was heated at 90.degree. C. under N.sub.2 overnight.
The mixture was then adjusted to pH 2 with 1 M aq. HCl and most of
the solvent was removed under reduced pressure. The residue was
diluted with water and extracted with DCM. The organic extract was
washed with water, brine, dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by silica gel chromatography (Pet. ether/EtOAc=50/1 to
3/1) followed by preparative TLC (DCM/MeOH=100/1) to give the title
compound (30 mg, 33%) as a white solid. LCMS-D: R.sub.t 2.87 min;
m/z 403.1 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
10.4 (s, 1H), 7.63 (d, J=2.3 Hz, 1H), 7.50-7.45 (m, 1H), 7.32-7.26
(m, 1H), 7.19-7.12 (m, 2H), 3.92 (s, 3H), 3.81 (s, 3H), 2.61 (q,
J=7.6 Hz, 2H), 2.22-2.15 (m, 1H), 1.15 (t, J=7.5 Hz, 3H), 1.03-0.94
(m, 2H), 0.76-0.64 (m, 2H).
Examples 209, 210 and 211
##STR00326## ##STR00327##
[0879] a) tert-Butyl((3,5-dimethoxybenzyl)oxy)dimethylsilane
A42
[0880] To a solution of (3,5-dimethoxyphenyl)methanol (10 g, 0.059
mol) and imidazole (6.07 g, 0.089 mol) in DMF (100 mL) at RT was
added TBSCI (9.86 g, 0.065 mol) and the mixture was stirred
overnight. Water was added and the mixture was extracted with
EtOAc. The organic extract was washed with water, concentrated
under reduced pressure and the residue was purified by column
chromatography (Pet. ether/EtOAc=20/1 to 10/1) to give the title
compound (13 g, 77%) as a colorless oil, which was used directly in
the next step.
b) Lithium
4-(((tert-butyldimethylsilyl)oxy)methyl)-2,6-dimethoxybenzenesu-
lfinate A43
[0881] To a solution of
tert-butyl((3,5-dimethoxybenzyl)oxy)dimethylsilane A42 (5 g, 0.018
mol) and TMEDA (2.26 g, 0.020 mol) in n-hexane (100 mL) at
-70.degree. C. was added n-BuLi (2.5 M solution in hexanes, 7.8 mL,
0.020 mol) and the mixture was stirred at 0.degree. C. for 1 h. The
mixture was then re-cooled to -60.degree. C. and bubbled with
SO.sub.2 gas for 30 min. The resulting precipitate was collected by
filtration and washed with n-hexane to give the title compound (6.2
g, 100%), which was used directly in the next step.
c)
4-(((tert-Butyldimethylsilyl)oxy)methyl)-2,6-dimethoxybenzenesulfonyl
chloride A44
[0882] To a mixture of lithium
4-(((tert-butyldimethylsilyl)oxy)methyl)-2,6-dimethoxy
benzenesulfinate A43 (6.2 g, 0.018 mol) in n-hexane (100 mL) at
0.degree. C. was added SO.sub.2Cl.sub.2 (2.39 g, 0.018 mol) and the
mixture was stirred at 0.degree. C. for 1 h. The solids were
collected by filtration, washed with n-hexane then dissolved in
EtOAc and washed with water. The organic layer was concentrated
under reduced pressure and the residue was purified by column
chromatography (Pet. ether/EtOAc=20/1 to 5/1) to give the title
compound (2.4 g, 36%) as a yellow solid, which was used directly in
the next step.
d)
4-(((tert-Butyldimethylsilyl)oxy)methyl)-2,6-dimethoxy-N-(4-methoxybenz-
o[d]isoxazol-3-yl)benzenesulfonamide A45
[0883] To a solution of 4-methoxybenzo[d]isoxazol-3-amine (819 mg,
4.99 mmol) in THF (20 mL) at -78.degree. C. was added LiHMDS (1 M
solution in THF, 5.98 mL, 5.98 mmol) and the mixture was stirred at
0.degree. C. for 30 min, then cooled to -60.degree. C. A solution
of
4-(((tert-butyldimethylsilyl)oxy)methyl)-2,6-dimethoxybenzenesulfonyl
chloride A44 (1.9 g, 4.99 mmol) in THF (20 mL) was added and the
mixture was stirred at RT overnight. The mixture was adjusted to pH
5 with 1 M aq. HCl and extracted with EtOAc. The organic extract
was concentrated under reduced pressure and the residue was
purified by column chromatography (Pet. ether/EtOAc=100/1 to 20/1)
to give the title compound (170 mg, 7%) as a yellow solid. LCMS-D:
R.sub.t 3.33 min, m/z 509.0 [M+H].sup.+.
e)
4-(Hydroxymethyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benze-
nesulfonamide 209
[0884] To a solution of
4-(((tert-butyldimethylsilyl)oxy)methyl)-2,6-dimethoxy-N-(4-methoxybenzo[-
d]isoxazol-3-yl)benzenesulfonamide A45 (170 mg, 0.334 mmol) in THF
(10 mL) was added TBAF (175 mg, 0.668 mmol) and the mixture was
stirred at RT for 3 h. The reaction was quenched with 1 M aq. HCl
and the mixture was extracted with EtOAc. The organic extract was
concentrated under reduced pressure and the residue was purified by
preparative TLC (DCM/MeOH=100/1 to 50/1) to give the title compound
(30 mg, 23%) as a white solid. LCMS-D: R.sub.t 1.93 min, m/z 394.9
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.40 (s,
1H), 7.55 (t, J=8.2 Hz, 1H), 7.18-7.12 (m, 1H), 6.89-6.82 (m, 1H),
6.72 (s, 2H), 5.39 (t, J=5.8 Hz, 1H), 4.49 (d, J=5.8 Hz, 2H), 3.95
(s, 3H), 3.77 (s, 6H).
f)
4-(Bromomethyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzene-
sulfonamide A46
[0885] A solution of
4-(hydroxymethyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)
benzenesulfonamide 209 (1.0 g, 2.53 mmol), CBr.sub.4 (2.52 g, 7.60
mmol) and PPh.sub.3 (1.99 g, 7.60 mmol) in DCM (20 mL) was stirred
at RT overnight. The mixture was concentrated under reduced
pressure and the residue was purified by column chromatography
(DCM/MeOH=50/1 to 10:1) to give the title compound (600 mg, 52%) as
a yellow solid, which was used directly in the next step.
g)
4-(Azidomethyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzene-
sulfonamide A47
[0886] A solution of
4-(bromomethyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)
benzenesulfonamide A46 (200 mg, 0.44 mmol) and NaN.sub.3 (85 mg,
1.31 mmol) in DMF (5 mL) was heated at 50.degree. C. for 3 h. The
mixture was poured into water and the precipitate was collected by
filtration. The filter cake was washed with water and dried to give
the title compound (100 mg, 54%) as gray solid. LCMS-D: R.sub.t
2.45 min, m/z 420.1 [M+H].sup.+.
h)
4-(Aminomethyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzene-
sulfonamide hydrochloride 210
[0887] A solution of
4-(azidomethyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)
benzenesulfonamide A47 (300 mg, 0.716 mmol) and PPh.sub.3 (207 mg,
0.788 mmol) in THF (8 mL) and EtOH (6 mL) was stirred at RT for 3
h. Water (2 mL) and concentrated HCl (2 mL) were added and the
mixture was heated at 55.degree. C. for 3 h. Most of the organic
solvents were removed under reduced pressure and the aqueous
residue was washed with DCM. The aqueous phase was concentrated
under reduced pressure to give the title compound (150 mg, 53%) as
a yellow solid. LCMS-D: R.sub.t 0.38 min, m/z 394.1 [M+H].sup.+.
.sup.1H NMR (400 MHz, methanol-d.sub.4) .delta. 7.54 (t, J=8.2 Hz,
1H), 7.06-7.01 (m, 1H), 6.87 (s, 2H), 6.86-6.82 (m, 1H), 4.12 (s,
2H), 4.07 (s, 3H), 3.89 (s, 6H).
i)
N-(3,5-Dimethoxy-4-(N-(4-methoxybenzo[d]isoxazol-3-yl)sulfamoyl)benzyl)-
acetamide 211
[0888] A mixture of
4-(aminomethyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)
benzenesulfonamide hydrochloride 210 (130 mg, 0.33 mmol),
NaHCO.sub.3 (83 mg, 0.99 mmol) and acetic anhydride (101 mg, 0.99
mmol) in THF (5 mL) was stirred at RT for 3 h then concentrated
under reduced pressure. The residue was purified by preparative TLC
to give the title compound (50 mg, 34%) as a yellow solid. LCMS-D:
R.sub.t 1.93 min, m/z 436.1 [M+H].sup.+. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.47 (s, 1H), 8.36 (t, J=5.8 Hz, 1H), 7.55
(m, 1H), 7.18-7.12 (m, 1H), 6.89-6.81 (m, 1H), 6.66 (s, 2H), 4.24
(d, J=5.9 Hz, 2H), 3.94 (s, 3H), 3.76 (s, 6H), 1.88 (s, 3H).
Example 212:
2-Fluoro-6-methoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide
212
##STR00328##
[0889] a)
2,6-Difluoro-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamid- e
A48
[0890] To a solution of 4-methoxybenzo[d]isoxazol-3-amine (4.0 g,
24.0 mmol) in THF (200 mL) at -78.degree. C. under nitrogen was
added LiHMDS (1 M solution in THF, 31.2 mL, 31.2 mmol) dropwise and
the mixture was stirred at -78.degree. C. for 3 h. A solution of
2,6-difluorophenylsulfonyl chloride (10.2 g, 48 mmol) in THF (20
mL) was then added dropwise and the mixture was allowed to warm
slowly to RT and stirred overnight. Water was added and the mixture
was extracted with EtOAc (200 mL.times.3). The combined organic
extracts were washed with water, brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (Pet.
ether/EtOAc=6/1 to 2/1) to give the title compound (2.9 g, 35%) as
a white solid. LCMS-D: R.sub.t 1.94 min, m/z 341.1 [M+H].sup.+.
b)
2-Fluoro-6-methoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide
212
[0891] A mixture of
2,6-difluoro-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide
A48 (450 mg, 1.32 mmol) and NaOMe (428 mg, 7.92 mmol) in MeOH (6
mL) was heated at 120.degree. C. overnight then allowed to cool to
RT and filtered. The filtrate was concentrated under reduced
pressure and the residue was purified by preparative TLC
(DCM/MeOH=50/1) to give the title compound (100 mg, 21%) as a white
solid. LCMS-D: R.sub.t 2.22 min, m/z 353.0 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 10.6 (s, 1H), 7.62 (m, 1H), 7.56
(t, J=8.3 Hz, 1H), 7.18 (d, J=8.4 Hz, 1H), 7.04 (d, J=8.5 Hz, 1H),
7.00-6.92 (m, 1H), 6.84 (d, J=8.0 Hz, 1H), 3.85 (s, 3H), 3.80 (s,
3H).
Example 213:
2-Hydroxy-6-methoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide
213
##STR00329##
[0893] A mixture of
2-fluoro-6-methoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide
212 (20 mg, 0.06 mmol), (6-methoxypyridin-3-yl)methanol (50.1 mg,
0.36 mmol) and t-BuOK (20.2 mg, 0.18 mmol) in NMP (1 mL) was heated
at 120.degree. C. for 16 h. LCMS analysis indicated that only the F
hydrolysis product had formed. The mixture was adjusted to pH<4
with aq. HCl, diluted with water and extracted with EtOAc. The
organic extract was washed with water, brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The procedure was repeated with another batch of
2-fluoro-6-methoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide
212 (52 mg, 0.15 mmol), the two crude products were combined and
purified by prep. TLC (EtOAc/Pet. ether=1/13 then DCM/MeOH=50/1 to
100/1) to give the title compound (135 mg, 19%) as a white solid.
LCMS-D: R.sub.t 2.39 min; m/z 350.9 [M+H].sup.+. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 10.2 (s, 1H), 7.57 (t, J=8.4 Hz, 1H),
7.41 (t, J=8.4 Hz, 1H), 7.16 (d, J=8.4 Hz, 1H), 6.84 (d, J=8.0 Hz,
1H), 6.60-6.56 (m, 2H), 3.91 (s, 3H), 3.75 (s, 3H).
Example 214:
2-Methoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)-6-(methylamino)benzenesulfon-
amide 214
##STR00330##
[0895] A solution of
2-fluoro-6-methoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide
212 (30 mg, 0.085 mmol), methylamine HCl (23 mg, 0.34 mmol) and
Et.sub.3N (52 mg, 0.51 mmol) in ethanol (3 mL) was heated at
120.degree. C. in a sealed tube overnight. The mixture was
concentrated under reduced pressure and the residue was purified by
prep. TLC (DCM) to give the title compound (4 mg, 13%) as a yellow
solid. LCMS-D: R.sub.t 2.49 min; m/z 364.0 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.56 (t, J=8.0 Hz, 1H), 7.30 (t,
J=8.0 Hz, 1H), 7.14 (d, J=8.0 Hz, 1H), 6.84 (d, J=8.0 Hz, 1H), 6.33
(d, J=8.0 Hz, 1H), 6.24 (d, J=8.0 Hz, 1H), 3.94 (s, 3H), 3.70 (s,
3H), 2.79 (s, 3H).
Example 215:
2-(Cyclopropylmethoxy)-6-methoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzen-
esulfonamide 215
##STR00331##
[0897] To a solution of
2-fluoro-6-methoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide
212 (30 mg, 0.08 mmol) in THF (4 mL) was added t-BuOK (27 mg, 0.24
mmol) followed by cyclopropylmethanol (17 mg, 0.24 mmol) and the
mixture was heated at 50.degree. C. overnight. The mixture was
diluted with water and extracted with EtOAc (20 mL.times.3). The
combined organic extracts were washed with brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by prep. TLC (Pet.
ether/EtOAc=1/1) to give the title compound (4.4 mg, 13%) as a grey
solid. LCMS-D: R.sub.t 2.54 min, m/z 405.1 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.33 (s, 1H), 7.57 (t, J=8.0 Hz,
1H), 7.48 (t, J=8.0 Hz, 1H), 7.15 (d, J=8.4 Hz, 1H), 6.85 (d, J=8.0
Hz, 1H), 6.77-6.75 (m, 2H), 3.91 (s, 3H), 3.89 (d, J=6.8 Hz, 2H),
3.77 (s, 3H), 0.86-0.83 (m, 1H), 0.43-0.24 (m, 4H).
Example 216:
2-Methoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)-6-((2-methoxypyridin-4-yl)me-
thoxy)benzenesulfonamide 216
##STR00332##
[0899] To a solution of
2-fluoro-6-methoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide
212 (30 mg, 0.08 mmol) in NMP (3 mL) was added t-BuOK (48 mg, 0.43
mmol) followed by (2-methoxypyridin-4-yl)methanol (118 mg, 0.85
mmol) and the mixture was stirred at RT for 6 h. The mixture was
diluted with water, adjusted to pH 5-6 with 1 M aq. HCl and
extracted with EtOAc (20 mL.times.3). The combined organic extracts
were washed with brine, dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by prep. TLC (DCM/MeOH=40/1) to give the title compound (5
mg, 12%) as a yellow solid. LCMS-D: R.sub.t 2.43 min, m/z 472.0
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.65 (s,
1H), 8.07 (d, J=5.2 Hz, 1H), 7.51-7.39 (m, 2H), 7.07-6.98 (m, 3H),
6.79-6.75 (m, 3H), 5.22 (s, 2H), 3.84 (s, 3H), 3.83 (s, 3H), 3.81
(s, 3H).
Examples 217-222 (Table G)
TABLE-US-00011 [0900] TABLE G The following examples were
synthesised according to the methods described for 215 or 216.
Varitaions of conditions have been noted in the table. R--OH Name
and structure Analytical Method Note 217 Isopropanol ##STR00333##
LCMS-C: R.sub.t 2.18 min, m/z 393.0 [M + H].sup.+; .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.13 (s, 1H), 7.58 (t, J = 8.0 Hz, 1H),
7.49 (t, J = 8.0 Hz, 1H), 7.17 (d, J = 8.0 Hz, 1H), 6.87 (d, J =
8.0 Hz, 1H), 6.79-6.75 (m, 2H), 4.75-4.65 (m, 1H), 3.95 (s, 3H),
3.80 (s, (3H), 1.17 (d, J = 6.0 Hz, 6H). As for Example 216
120.degree. C., 16 h; Ratio of fluoride/base/ R--OH 1:3:3 218
Ethanol ##STR00334## LCMS-C: R.sub.t 2.03 min, m/z 379.0 [M+
H].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.13 (s 1H)
7.58 (t, J = 8.0 Hz, 1H), 7.50 (t, J = 8.0 Hz, 1H), 7.17 (d, J =
8.0 Hz, 1H), 6.86 (d, J = 8.0 Hz, 1H), 6.78 (d, J = 8.0 Hz, 2H),
4.01 (q, J = 7.2 Hz, 2H), 3.94 (s, 3H), 3.76 (s, 3H), 1.12 (t, J =
7.2 Hz, 3H). As for Example 216 120.degree. C., 16 h; Ratio of
fluoride/base/ R--OH 1:3:3 219 ##STR00335## ##STR00336## LCMS-D:
R.sub.t 0.31 min, m/z 408.0 [M+ H].sup.+; .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 10.0 (s, 1H), 7.61-7.45 (m, 4H), 7.15 (d, J =
8.4 Hz, 1H), 7.83-6.74 (m, 3H), 4.51 (s, 2H), 3.83 (s, 3H), 3.77
(s, 3H). As for Example 216 120.degree. C., 16 h; Ratio of
fluoride/base/ R--OH 1:6:10; Eluent: DCM/MeOH = 80/1 220
##STR00337## ##STR00338## LCMS-D: R.sub.t 2.11 min, m/z 395.0 [M+
H].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.70 (s, 1H),
7.57-7.48 (m, 2H), 7.15 (d, J = 8.4 Hz, 1H), 6.84-6.80 (m, 3H),
5.10 (br s, 1H), 4.06 (t, J = 5.0 Hz, 2H), 3.90 (s, 3H), 3.80 (s,
3H), 3.63 (t, J = 5.0 Hz, 2H). As for Example 216 RT o/n then
80.degree. C., 16 h; ratio of fluoride/base/ R--OH 1:6:10; Eluent:
DCM/MeOH = 60/1 221 ##STR00339## ##STR00340## LCMS-D: R.sub.t 2.25
min, m/z 445.0 [M+ H].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.62 (s, 1H), 7.54 (t, J = 8.4 Hz, 1H), 7.46 (t, J = 8.4
Hz, 1H), 7.40 (s, 1H), 7.12 (d, J = 8.4 Hz, 1H), 6.86 (d, J = 8.4
Hz, 1H), 6.81 (d, J = 8.0 Hz, 1H), 6.76 (d, J = 8.4 Hz, 1H), 5.04
(s, 2H), 3.86 (s, 3H), 3.75 (s, 3H), 3.73 (s, 3H). As for Example
215 50.degree. C., 16 h; Ratio of fluoride/base/ R--OH 1:3:3 222
##STR00341## ##STR00342## LCMS-D: R.sub.t 2.67 min, m/z 441.1 [M+
H].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.51-7.43 (m,
4H), 7.36- 7.30 (m, 3H), 7.05 (d, J = 8.4 Hz, 1H), 6.79-6.72 (m,
3H), 5.25 (s, 2H), 3.89 (s, 3H), 3.75 (s, 3H). As for Example 215
50.degree. C., 16 h; Ratio of fluoride/base/ R--OH 1:3:3
Example 223:
2,6-Dimethoxy-N-(4-nitrobenzo[d]isoxazol-3-yl)benzenesulfonamide
223
##STR00343##
[0902] To a solution of 4-nitrobenzo[d]isoxazol-3-amine I10 (478
mg, 2.63 mmol) in THF (9 mL) at -78.degree. C. under nitrogen was
added LiHMDS (1 M solution in THF, 5.30 mL, 5.30 mmol) and the
mixture was stirred at -78.degree. C. for 3 h. 2,6-Dimethoxysufonyl
chloride 1111 (1.25 g, 5.27 mmol) was then added and the mixture
was allowed to warm to RT and stirred overnight. The reaction was
quenched with water and the mixture was extracted with EtOAc (80
mL.times.3). The combined organic extracts were washed with water
(100 mL.times.3), brine (50 mL.times.3), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by column chromatography (DCM/MeOH=1/0 to
50/1) to give the title compound (108 mg, 11%) as a yellow solid.
LCMS-D: R.sub.t 2.23 min; m/z 380.1 [M+H].sup.+. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 10.2 (s, 1H), 8.19 (d, J=8.4 Hz, 2H),
7.92 (t, J=8.4 Hz, 1H), 7.79 (d, J=8.4 Hz, 1H), 6.79 (d, J=8.4 Hz,
2H), 3.71 (s, 6H).
Example 224:
N-(4-Aminobenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide
224
##STR00344##
[0904] To a solution of
2,6-dimethoxy-N-(4-nitrobenzo[d]isoxazol-3-yl)benzenesulfonamide
223 (70.0 mg, 0.18 mmol) in EtOAc (30 mL) was added 10% Pd/C (40.0
mg) and the mixture was stirred at RT overnight under a hydrogen
atmosphere. The catalyst was removed by filtration, rinsed with
EtOAc and the filtrate was concentrated under reduced pressure. The
procedure was repeated on an additional batch of
2,6-dimethoxy-N-(4-nitrobenzo[d]isoxazol-3-yl)benzenesulfonamide
223 (30 mg, 0.079 mmol) and the crude products were combined and
purified twice by prep. TLC (DCM/MeOH=50/1) and twice by prep. HPLC
to give the title compound (2.5 mg, 3%) as a white solid. LCMS-D:
R.sub.t 2.25 min; m/z 350.1 [M+H].sup.+. .sup.1H NMR (400 MHz,
FDMSO-d.sub.6) .delta. 7.52 (t, J=8.4 Hz, 1H), 7.25 (t, J=8.0 Hz,
1H), 6.80 (d, J=8.4 Hz, 2H), 6.66 (d, J=8.0 Hz, 1H), 6.43 (d, J=8.0
Hz, 1H), 3.78 (s, 6H).
Example 225:
N-(5-((1H-Pyrazol-1-yl)methyl)benzo[d]isoxazol-3-yl)-5-ethyl-2-methoxyben-
zenesulfonamide 225
##STR00345## ##STR00346##
[0905] a)
N-(5-Bromobenzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-
-2-methoxybenzenesulfonamide A49
[0906] To a solution of
N-(5-bromobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
135 (8.2 g, 19.9 mmol), (2,4-dimethoxyphenyl)methanol (5.03 g, 29.9
mmol) and PPh.sub.3 (13.1 g, 49.9 mmol) in THF (400 mL) at
0.degree. C. under nitrogen was added DIAD (8.06 g, 39.9 mmol)
slowly dropwise and the mixture was stirred at RT overnight. The
solvent was removed under reduced pressure and the residue was
purified by column chromatography (Pet. ether/EtOAc=100/1 to 5/1)
to give the title compound (4.7 g, 42%) as a white solid. LCMS-D:
R.sub.t 3.39 min, m/z 583.1 [M+Na].sup.+
b) Methyl
3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)-
benzo[d]isoxazole-5-carboxylate A50
[0907] A mixture of
N-(5-bromobenzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-2-methox-
ybenzenesulfonamide A49 (1.5 g, 2.67 mmol), Et.sub.3N (10 mL) and
Pd(dppf)Cl.sub.2 (732 mg, 1 mmol) in MeOH (150 mL) was heated at
120.degree. C. under a carbon monoxide atmosphere (20 bar) for 7 h.
The reaction was repeated with 1.7 g
N-(5-bromobenzo[d]isoxazol-3-yl)-N-(2,4-di
methoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A49 and 1.5 g
N-(5-bromobenzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-2-methox-
ybenzenesulfonamide A49 and the mixtures were combined and
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=100/1 to 3/1) to give the
title compound (2.2 g, 49%) as a white solid. LCMS-D: R.sub.t 3.17
min, m/z 563.1 [M+Na].sup.+
c)
3-((N-(2,4-Dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d-
]isoxazole-5-carboxylic acid A51
[0908] To a solution of methyl
3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)
sulfonamido)benzo[d]isoxazole-5-carboxylate A50 (2.2 g, 4.07 mmol)
in methanol (30 mL) was added 3 M aq. NaOH (20 mL) and the mixture
was stirred at RT overnight. The mixture was adjusted to pH 1 with
aq. HCl and the solvent was removed under reduced pressure. The
residue was diluted with water (100 mL) and extracted with DCM (200
mL). The organic extract was washed with water, brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure to give the title compound (1.9 g, 89%) as a light yellow
solid. LCMS-D: R.sub.t 2.99 min, m/z 549.1 [M+Na].sup.+.
d)
N-(2,4-Dimethoxybenzyl)-5-ethyl-N-(5-(hydroxymethyl)benzo[d]isoxazol-3--
yl)-2-methoxybenzenesulfonamide A52
[0909] To a solution of
3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)
benzo[d]isoxazole-5-carboxylic acid (800 mg, 1.52 mmol) A51 in THF
(5 mL) at 0.degree. C. under nitrogen was added BH.sub.3.THF (1 M
solution in THF, 10 mL, 10 mmol) slowly dropwise and the mixture
was allowed to warm to RT and stirred for 2 h. The mixture was
cooled to 0.degree. C. and the reaction was quenched by slow
addition of methanol (20 mL). The solvent was removed under reduced
pressure and the residue was diluted with water and extracted with
DCM (200 mL). The organic extract was washed with water, brine,
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by column
chromatography (DCM/MeOH=100/0 to 40/1) to give the title compound
(700 mg, 90%) as a white solid. LCMS-D: R.sub.t 2.99 min, m/z 513.1
[M+H].sup.+.
e)
N-(5-((1H-Pyrazol-1-yl)methyl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybe-
nzyl)-5-ethyl-2-methoxybenzenesulfonamide A53
[0910] To a solution of
N-(2,4-dimethoxybenzyl)-5-ethyl-N-(5-(hydroxymethyl)
benzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide A52 (200 mg,
0.39 mmol) and Et.sub.3N (197 mg, 1.95 mmol) in DCM (10 mL) at
0.degree. C. under nitrogen was added methanesulfonyl chloride (89
mg, 0.78 mmol) and the mixture was stirred at RT for 1.5 h then
used in next step directly without workup and isolation.
[0911] To a solution of pyrazole (133 mg, 1.95 mmol) in DMF (20 mL)
at 0.degree. C. under nitrogen was added t-BuOK (219 mg, 1.95 mmol)
portion-wise and the mixture was stirred at RT for 1.5 h. The
reaction mixture containing
(3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)
benzo[d]isoxazol-5-yl)methyl methanesulfonate (assumed 0.39 mmol)
was then added slowly and the resulting mixture was stirred at RT
for 30 min. The mixture was diluted with water and extracted with
EtOAc. The organic extract was washed with water (.times.3), brine,
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by column
chromatography (DCM/MeOH=40/1) to give the title compound (60 mg,
27%) as a white solid. LCMS-D: R.sub.t 3.01 min, m/z 563.2
[M+H].sup.+.
f)
N-(5-((1H-Pyrazol-1-yl)methyl)benzo[d]isoxazol-3-yl)-5-ethyl-2-methoxyb-
enzenesulfonamide 225
[0912] A mixture of
N-(5-((1H-pyrazol-1-yl)methyl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenz-
yl)-5-ethyl-2-methoxybenzenesulfonamide A53 (60 mg, 0.107 mmol) and
TFA (5 mL) was stirred at RT for 3 h then concentrated under
reduced pressure. The residue was purified by column chromatography
(DCM/MeOH=20/1) to give the title compound (30 mg, 68%) as a white
solid. LCMS-D: R.sub.t 6.03 min, m/z 412.5 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 11.9 (s, 1H), 7.89 (s, 1H), 7.85
(d, J=2.0 Hz, 1H), 7.68 (d, J=2.0 Hz, 1H), 7.58-7.49 (m, 3H), 7.46
(dd, J=8.4, 2.0 Hz, 1H), 7.08 (d, J=8.8 Hz, 1H), 6.31 (t, J=2.0 Hz,
1H), 5.54 (s, 2H), 3.68 (s, 3H), 2.63 (q, J=7.6 Hz, 2H), 1.16 (t,
J=7.6 Hz, 3H).
Example 226:
N-(5-(Aminomethyl)benzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonam-
ide trifluoroacetate 226
##STR00347##
[0913] a)
3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)-
benzo[d]isoxazole-5-carboxamide A54
[0914] A mixture of
3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)
benzo[d]isoxazole-5-carboxylic acid A51 (600 mg, 1.14 mmol) and
thionyl chloride (5 mL) was heated at 85.degree. C. for 1.5 h then
concentrated under reduced pressure. The residue was dissolved in
in DCM (3 mL) and added dropwise to conc. NH.sub.4OH (10 mL) at
0.degree. C. and the mixture was stirred at RT for 30 min. Most of
the solvent was removed under reduced pressure and the residue was
diluted with water (20 mL). The resulting precipitate was collected
by filtration, washed twice with water and dried to give the title
compound (400 mg, 67%) as a light-yellow solid. LCMS-D: R.sub.1
2.86 min, m/z 526.2 [M+H].sup.+.
b)
N-(5-(Aminomethyl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethy-
l-2-methoxybenzenesulfonamide A55
[0915] A mixture of 3-((N-(2,4-di
methoxybenzyl)-5-ethyl-2-methoxyphenyl)
sulfonamido)benzo[d]isoxazole-5-carboxamide A54 (400 mg, 0.76 mmol)
and BH.sub.3.THF (1 M solution in THF, 20 mL) was heated at
75.degree. C. under nitrogen overnight. The mixture was then cooled
to 0.degree. C. and the reaction was quenched by slow addition of
methanol (10 mL). The solvent was removed under reduced pressure
and the residue was diluted with water and extracted with DCM (100
mL). The organic extract was washed with water, brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography
(DCM/MeOH=100/1 to 10/1) to give the title compound (150 mg, 39%)
as a light-yellow solid, which was used directly in the next
step.
c)
N-(5-(Aminomethyl)benzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfon-
amide trifluoroacetate 226
[0916] A mixture of
N-(5-(aminomethyl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl--
2-methoxybenzenesulfonamide A55 (50 mg, 0.098 mmol) and TFA (5 mL)
was stirred at RT for 3 h then concentrated under reduced pressure.
The residue was purified by prep. TLC (DCM/MeOH=10/1) to give the
title compound (30 mg, 86%) as a white solid. LCMS-D: R.sub.t 1.93
min, m/z 362.1[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 11.8 (br s, 1H), 8.53 (br s, 3H), 8.11 (s, 1H), 7.83 (d,
J=8.4 Hz, 1H), 7.70-7.68 (m, 2H), 7.48 (d, J=8.0 Hz, 1H), 7.12 (d,
J=8.8 Hz, 1H), 4.13 (s, 2H), 3.71 (s, 3H), 2.64 (q, J=7.6 Hz, 2H),
1.17 (t, J=7.6 Hz, 3H).
Example 227: Methyl
((3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazol-5-yl)methyl)
carbamate 227
##STR00348##
[0917] a) Methyl
((3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d-
]isoxazol-5-yl)methyl) carbamate A56
[0918] To a solution of
N-(5-(aminomethyl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl--
2-methoxybenzenesulfonamide A55 (70 mg, 0.137 mmol) and Et.sub.3N
(69 mg, 0.685 mmol) in DCM (20 mL) at 0.degree. C. was added methyl
chloroformate (39 mg, 0.411 mmol) dropwise and the mixture was
stirred at RT for 20 min. The solvent was removed under reduced
pressure and the residue was diluted with water and extracted with
DCM. The organic extract was washed with water, brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by column chromatography
(DCM/MeOH=100/0 to 40/1) to give the title compound (30 mg, 43%) as
a white solid, which was used directly in the next step.
b) Methyl
((3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazol-5-yl)-
methyl) carbamate 227
[0919] A mixture of methyl
((3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)
benzo[d]isoxazol-5-yl)methyl)carbamate A56 (30 mg, 0.0558 mmol) and
TFA (3 mL) was stirred at RT for 3 h then concentrated under
reduced pressure. The residue was purified by column chromatography
(DCM/MeOH=40/1) to give the title compound (15 mg, 64%) as a white
solid. LCMS-D: R.sub.t 2.45 min, m/z 420.1 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 11.7 (s, 1H), 7.91 (s, 1H), 7.79
(t, J=5.6 Hz, 1H), 7.69 (d, J=2.0 Hz, 1H), 7.55-7.43 (m, 3H), 7.09
(d, J=8.4 Hz, 1H), 4.29 (d, J=6.0 Hz, 2H), 3.73 (s, 3H), 3.58 (s,
3H), 2.63 (q, J=7.6 Hz, 2H), 1.16 (t, J=7.6 Hz, 3H).
Example 228:
N-((3-((5-Ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazol-5-yl)methyl)-
acetamide 228
##STR00349##
[0920] a)
N-((3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonam-
ido)benzo[d]isoxazol-5-yl)methyl) acetamide A57
[0921] To a solution of
N-(5-(aminomethyl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl--
2-methoxybenzenesulfonamide A55 (100 mg, 0.195 mmol) and Et.sub.3N
(98 mg, 0.975 mmol) in DCM (10 mL) at 0.degree. C. under nitrogen
was added acetyl chloride (31 mg, 0.391 mmol) and the mixture was
stirred at RT for 30 min. The mixture was diluted with water and
extracted with DCM. The organic extract was washed with water,
brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
prep. TLC (DCM/MeOH=30/1) to give the title compound (60 mg, 56%)
as a white solid, which was used directly in the next step.
b)
N-((3-((5-Ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazol-5-yl)methy-
l)acetamide 228
[0922] A mixture of
N-((3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)
benzo[d]isoxazol-5-yl)methyl)acetamide A57 (60 mg, 0.108 mmol) and
TFA (5 mL) was stirred at RT for 3 h then concentrated under
reduced pressure. The residue was purified by column chromatography
(DCM/MeOH=40/1) to give the title compound (30 mg, 68%) as a white
solid. LCMS-D: R.sub.t 2.35 min, m/z 404.1 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 11.7 (br s, 1H), 8.55 (br s, 1H),
7.85 (s, 1H), 7.69 (s, 1H), 7.47-7.39 (m, 3H), 7.06 (d, J=7.2 Hz,
1H), 4.33 (d, J=3.6 Hz, 2H), 3.72 (s, 3H), 2.60 (q, J=6.8 Hz, 2H),
1.91 (s, 3H), 1.17 (t, J=6.8 Hz, 3H).
Example 229:
5-Ethyl-N-(5-(hydroxymethyl)benzo[d]isoxazol-3-yl)-2-methoxybenzenesulfon-
amide 229
##STR00350##
[0923] a) Methyl
3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxylate
A58
[0924] A mixture of methyl
3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)
benzo[d]isoxazole-5-carboxylate A50 (435 mg, 0.8 mmol) and TFA (9
mL) was stirred at RT for 3 h then concentrated under reduced
pressure. The residue was purified by column chromatography (Pet.
ether/EtOAc=2/1) to give the title compound (200 mg, 89%) as a
light yellow solid. LCMS-D: R.sub.1 2.69 min, m/z 391.1
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.9 (br
s, 1H), 8.82 (s, 1H), 8.18 (dd, J=9.2, 1.6 Hz, 1H), 7.72-7.70 (m,
2H), 7.48 (dd, J=8.8, 2.0 Hz, 1H), 7.10 (d, J=8.8 Hz, 1H), 3.90 (s,
3H), 3.71 (s, 3H), 2.64 (q, J=7.6 Hz, 2H), 1.16 (t, J=7.6 Hz,
3H).
b)
3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxylic
acid A59
[0925] To a solution of methyl
3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxylate
A58 (220 mg, 0.56 mmol) in THF/MeOH (10 mL/10 mL) was added 2 M aq.
NaOH (1.4 mL, 2.8 mmol) and the mixture was stirred at RT
overnight. The solvent was removed under reduced pressure and the
residue was dissolved in water (10 mL) and the solution was
adjusted to pH 2-3. The resulting precipitate was collected by
filtration to give the title compound (180 mg, 85%) as a light
yellow solid. LCMS-D: R.sub.t 2.51 min, m/z 377.2 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 13.2 (s, 1H), 11.86 (s,
1H), 8.78 (s, 1H), 8.16 (dd, J=8.8, 1.2 Hz, 1H), 7.71-7.66 (m, 2H),
7.47 (dd, J=8.4, 1.6 Hz, 1H), 7.10 (d, J=8.4 Hz, 1H), 3.70 (s, 3H),
2.63 (q, J=7.6 Hz, 2H), 1.16 (t, J=7.6 Hz, 3H).
c)
5-Ethyl-N-(5-(hydroxymethyl)benzo[d]isoxazol-3-yl)-2-methoxybenzenesulf-
onamide 229
[0926] A mixture of
3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxylic
acid A59 (50 mg, 0.13 mmol) and BH.sub.3.THF (1 M solution in THF,
5 mL, 5 mmol) was stirred at RT for 5 h under nitrogen. The
reaction was quenched with water (10 mL) and most of the THF was
removed under reduced pressure. The aqueous residue was adjusted to
pH 2-3 and extracted with DCM (15 mL.times.3). The combined organic
layers were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
column chromatography (DCM/MeOH=30/1) to give the title compound
(30 mg, 62%) as a light yellow solid. LCMS-D: R.sub.t 2.46 min, m/z
363.1 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.6
(s, 1H), 7.98 (s, 1H), 7.69 (d, J=2.0 Hz, 1H), 7.53 (s, 2H), 7.46
(dd, J=8.4, 2.0 Hz, 1H), 7.10 (d, J=8.4 Hz, 1H), 5.35 (t, J=5.6 Hz,
1H), 4.59 (d, J=5.6 Hz, 2H), 3.72 (s, 3H), 2.63 (q, J=7.6 Hz, 2H),
1.16 (t, J=7.6 Hz, 3H).
Example 230:
3-((5-Ethyl-2-methoxyphenyl)sulfonamido)-N-methylbenzo[d]isoxazole-5-carb-
oxamide 230
##STR00351##
[0928] A mixture of methyl
3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxylate
A58 (50 mg, 0.13 mmol) and methylamine (33% solution in EtOH, 5 mL)
was heated at 90.degree. C. for 80 min in a sealed tube. The
solvent was removed under reduced pressure and the residue was
purified by prep. TLC to give the desired product (18 mg, 36%) as a
light yellow solid. LCMS-D: R.sub.t 2.39 min, m/z 390.2[M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.8 (s, 1H), 8.53-8.51
(m, 2H), 8.03 (d, J=8.8 Hz, 1H), 7.69 (d, J=2.0 Hz, 1H), 7.62 (d,
J=8.8 Hz, 1H), 7.42 (d, J=8.0 Hz, 1H), 7.06 (d, J=8.4 Hz, 1H), 3.69
(s, 3H), 2.81 (d, J=4.4 Hz, 3H), 2.62 (q, J=7.6 Hz, 2H), 1.16 (t,
J=7.6 Hz, 3H).
Example 231:
3-((5-Ethyl-2-methoxyphenyl)sulfonamido)-N,N-dimethylbenzo[d]isoxazole-5--
carboxamide 231
##STR00352##
[0930] A solution of
3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxylic
acid A59 (70 mg, 0.19 mmol) in thionyl chloride (10 mL) was heated
at 85.degree. C. for 3 h under nitrogen. The mixture was then
concentrated under reduced pressure to give
3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carbonyl
chloride, which was dissolved in THF (2.5 mL) and treated with
dimethylamine (40% solution in water, 5 mL). The mixture was
stirred at RT overnight. Most of the THF was removed under reduced
pressure and the aqueous residue was adjusted to pH 2-3. The
precipitate was collected by filtration to give the title compound
(60 mg, 80%) as a light-yellow solid. LCMS-D: R.sub.t 2.40 min, m/z
404.2 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.13
(s, 1H), 7.71-7.63 (m, 3H), 7.48 (dd, J=8.8, 2.0 Hz, 1H), 7.10 (d,
J=8.8 Hz, 1H), 3.70 (s, 3H), 3.01 (s, 3H), 2.96 (s, 3H), 2.63 (q,
J=7.6 Hz, 2H), 1.16 (t, J=7.6 Hz, 3H).
Example 232:
3-((5-Ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxamide
232
##STR00353##
[0932] To a solution of
3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carbonyl
chloride (63 mg, 0.16 mmol), prepared according to the procedure
described for
3-((5-ethyl-2-methoxyphenyl)sulfonamido)-N,N-dimethylbenzo[d]isoxazole-5--
carboxamide 231, in THF (2.5 mL) was added conc. NH.sub.4OH (5 mL)
and the mixture was stirred at RT over a weekend. Most of the THF
was removed under reduced pressure and the aqueous residue was
adjusted to pH 2-3. The resulting precipitate was collected by
filtration to give the title compound (37 mg, 62%) as a
light-yellow solid. LCMS-D: R.sub.t 2.33 min, m/z 376.1
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.8 (s,
1H), 8.49 (s, 1H), 8.04 (d, J=7.6 Hz, 2H), 7.69 (d, J=2.4 Hz, 1H),
7.54 (d, J=8.4 Hz, 2H), 7.36 (d, J=6.8 Hz, 1H), 7.02 (d, J=8.4 Hz,
1H), 3.68 (s, 3H), 2.61 (q, J=7.6 Hz, 2H), 1.16 (t, J=7.6 Hz,
3H).
Example 233:
N-(5-Cyanobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide
233
##STR00354##
[0934] A solution of
3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxamide
232 (22 mg, 0.058 mmol) in POCl.sub.3 was heated at 110.degree. C.
overnight under nitrogen. The reaction was quenched with ice-water
(25 mL) and the mixture was extracted with DCM (35 mL.times.2). The
combined organic extracts were washed with water (50 mL.times.3),
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give the title compound (19.8 mg, 90%) as
a light-yellow solid. LCMS-D: R.sub.t 2.61 min, m/z 358.1
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.0 (s,
1H), 8.54 (s, 1H), 8.07 (d, J=8.8, 1.2 Hz, 1H), 7.86 (d, J=8.8 Hz,
1H), 7.73 (d, J=2.0 Hz, 1H), 7.49 (dd, J=8.4, 1.6 Hz, 1H), 7.11 (d,
J=8.4 Hz, 1H), 3.70 (s, 3H), 2.65 (q, J=7.6 Hz, 2H), 1.17 (t, J=7.6
Hz, 3H).
Example 234:
5-Ethyl-2-methoxy-N-(5-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide
234
##STR00355##
[0935] a)
N-(2,4-Dimethoxybenzyl)-5-ethyl-2-methoxy-N-(5-methylbenzo[d]iso-
xazol-3-yl)benzenesulfonamide A60
[0936] A mixture of
N-(5-bromobenzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-2-methox-
ybenzenesulfonamide A49 (112 mg, 0.2 mmol), methyl boronic acid (60
mg, 1 mmol), Pd(dppf)Cl.sub.2 (29 mg, 0.04 mmol) and
K.sub.2CO.sub.3 (138 mg, 1 mmol) in 1,4-dioxane (10 mL) and water
(2 mL) was heated at 90.degree. C. under nitrogen overnight. The
solvent was removed under reduced pressure and the residue was
diluted with water (30 mL) and extracted with EtOAc (100 mL). The
organic extract was washed with water, brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by prep. TLC (Pet. ether/EtOAc=3/1) to
give the title compound (60 mg, 61%) as a white solid. LCMS-D:
R.sub.t 2.72 min, m/z 347.2[M-DMB].sup.+.
b)
5-Ethyl-2-methoxy-N-(5-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide
234
[0937] A mixture of
N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxy-N-(5-methylbenzo[d]isoxazol-3-y-
l) benzenesulfonamide A60 (60 mg, 0.12 mmol) and TFA (5 mL) was
stirred at RT for 4 h then concentrated under reduced pressure. The
residue was purified by prep. TLC (Pet. ether/EtOAc=2/1) to give
the title compound (30 mg, 71%) as a white solid. LCMS-D: R.sub.t
2.72 min, m/z 347.2 [M+H].sup.+. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.78 (s, 1H), 7.69 (d, J=2.0 Hz, 1H),
7.49-7.44 (m, 3H), 7.10 (d, J=8.4 Hz, 1H), 3.72 (s, 3H), 2.61 (q,
J=7.6 Hz, 2H), 2.40 (s, 3H), 1.16 (t, J=7.6 Hz, 3H).
Examples 235-238 (Table H)
TABLE-US-00012 [0938] TABLE H The following targets were prepared
according to the procedure described for
5-ethyl-2-methoxy-N-(5-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide
234 Starting Example materials Name and structure LCMS .sup.1H NMR
235 ##STR00356## ##STR00357## LCMS-D: R.sub.t 2.84 min, m/z 361.1
[M + H].sup.+ .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.6 (s,
1H), 7.83 (s, 1H), 7.69 (s, 1H), 7.51-7.44 (m, 3H), 7.10 (d, J =
8.4 Hz, 1H), 3.74 (s, 3H), 2.74 (q, J = 7.6 Hz, 2H), 2.63 (q, J =
7.6 Hz, 2H), 1.24 (t, J = 7.6 Hz, 3H), 1.16 (t, J = 7.6 Hz, 3H).
236 ##STR00358## ##STR00359## LCMS-D: R.sub.t 2.87 min, m/z 373.1
[M + H].sup.+ .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.5 (s,
1H), 7.70 (d, J = 2.8 Hz, 1H), 7.69 (s, 1H), 7.47 (d, J = 8.8 Hz,
2H), 7.38- 7.36 (m, 1H), 7.10 (d, J = 8.8 Hz, 1H), 3.74 (s, 3H),
2.63 (q, J = 7.6 Hz, 2H), 2.06-2.01 (m, 1H), 1.17 (t, J = 7.6 Hz,
3H), 1.02-0.96 (m, 2H), 0.68- 0.64 (m, 2H). 237 ##STR00360##
##STR00361## LCMS-D: R.sub.t 2.57 min, m/z 413.1 [M + H].sup.+
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.6 (s, 1H), 8.17 (s,
1H), 8.10 (s, 1H), 7.82-7.80 (m, 2H), 7.72 (d, J = 1.6 Hz, 1H),
7.60 (d, J = 8.8 Hz, 1H), 7.47 (dd, J = 8.4, 1.6 Hz, 1H), 7.11 (d,
J = 8.8 Hz, 1H), 3.90 (s, 3H), 3.75 (s, 3H), 2.64 (q, J = 7.6 Hz,
2H), 1.16 (t, J = 7.6 Hz, 3H). 238 ##STR00362## ##STR00363##
LCMS-D: R.sub.t 2.52 min, m/z 442.9 [M + H].sup.+ .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 11.8 (s, 1H), 9.24 (s, 1H), 9.14 (s,
2H), 8.44 (s, 1H), 8.09 (d, J = 8.8 Hz, 1H), 7.80-7.74 (m, 2H),
7.48 (d, J = 7.6 Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 3.74 (s, 3H),
2.65 (q, J = 7.6 Hz, 2H), 1.17 (t, J = 7.6 Hz, 3H).
Examples 239-242 (Table I)
[0939] Method IA:
##STR00364##
[0940] To a solution of the amine (0.2 mmol, 1.0 eq.) in pyridine
(2 mL) was added the sulfonyl chloride (1.5 eq.) and the mixture
was heated at 120.degree. C. under microwave irradiation for 2 h.
The mixture was partitioned between water and EtOAc, the layers
were separated and the organic layer was washed with brine, dried
over Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by prep. TLC to give the title
compound. Variations to above conditions have been noted in Table
I.
TABLE-US-00013 TABLE I The following examples were synthesised
according to the method IA. Variations of conditions have been
noted in the table. Intermediates Ex Name and structure Analytical
(if applicable) Method Notes 239 ##STR00365## LCMS-C: R.sub.t 2.45
min; m/z 427.0, 429.0 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO-
d.sub.6) .delta. 11.5 (s, 1H), 7.85 (s, 1H), 7.73 (s, 1H), 7.48 (t,
J = 8.4 Hz, 1H), 6.75 (d, J = 8.4 Hz, 2H), 3.74 (s, 6H), 2.40 (s,
3H). 2,6-Dimethoxy- benzenesulfonyl chloride l111 7-Bromo-5-
methylbenzo[d] isoxazol-3-amine l132 IA 0.2 eq. DMAP used. Organic
layer washed with 0.1 M aq. HCl in workup. Prep. TLC (DCM/MeOH,
20/1) 240 ##STR00366## LCMS-C: R.sub.t 2.33 min; m/z 476.9 [M +
H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. 9.59 (br s,
1H), 7.49 (s, 1H), 7.18-7.06 (m, 1H), 6.86-6.62 (m, 3H), 4.75 (s,
2H), 4.15-4.11 (m, 2H), 3.91 (s, 3H), 3.76 (s, 6H). 2,6-Dimethoxy-
benzenesulfonyl chloride l111 4-Methoxy-6- ((2,2,2-
trifluoroethoxy) methyl)benzo[d] isoxazol-3-amine l118 IA 1 mL
pyridine used. Prep. TLC (Pet. ether/EtOAc = 1/1) 241 ##STR00367##
LCMS-D: R.sub.t 3.32 min; m/z 426.0 [M + H].sup.+; .sup.1H NMR (400
MHz, DMSO- d.sub.6) .delta. 11.4 (br s, 1H), 8.68 (d, J = 3.6 Hz,
1H), 7.96- 7.90 (m, 2H), 7.59- 7.43 (m, 4H), 6.76 (d, J = 8.4 Hz,
2H), 3.76 (s, 6H), 2.34 (s, 3H). 2,6-Dimethoxy- benzenesulfonyl
chloride l111 5-Methyl-6- (pyridin-2- yl)benzo[d] isoxazol-3- amine
l127 IA 5 mL pyridine used; 0.1 eq. DMAP used. Prep. TLC (Pet.
ether/EtOAc = 1/1) 242 ##STR00368## LCMS-C R.sub.t 2.13 min; m/z
430.9 [M + H].sup.+; .sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta.
9.73 (br s, 1H), 7.52 (t, J = 8.4 Hz, 1H), 7.40 (t, J = 73.2 Hz,
1H), 7.04 (d, J = 1.6 Hz, 1H), 6.78 (d, J = 8.4 Hz, 2H), 6.69 (d, J
= 1.6 Hz, 1H), 3.91 (s, 3H), 3.76 (s, 6H). 2,6-Dimethoxy-
benzenesulfonyl chloride l111 6-(Difluoro- methoxy)-4-
methoxybenzo[d] isoxazol-3-amine l121 IA Heated for 1 h. Adjusted
aqueous phase to pH 5 with 1 M aq. HCl in workup. Prep. TLC
(DCM/MeOH = 50/1)
Example 243:
N-(6-(3-Cyanophenyl)-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzene-
sulfonamide 243
##STR00369##
[0942] To a solution of
N-(6-bromo-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamid-
e 89 (50 mg, 0.113 mmol) in 1,4-dioxane (8 mL) and water (2 mL) was
added (3-cyanophenyl)boronic acid (34 mg, 0.226 mmol),
Na.sub.2CO.sub.3 (36 mg, 0.339 mmol) and Pd(PPh.sub.3).sub.4 (14
mg, 0.011 mmol) and the mixture was heated at reflux under a
N.sub.2 atmosphere overnight. The mixture was adjusted to pH 4-5
with 1 M aqueous HCl and extracted with EtOAc. The combined organic
extracts were washed with water, brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by prep. TLC (DCM/MeOH=100/1) to give the
title compound (24 mg, 46%) as a white solid. LCMS-C: R.sub.t 2.44
min, m/z 466.0 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.70 (s, 1H), 8.34 (s, 1H), 8.16 (d, J=7.5 Hz, 1H), 7.90
(d, J=7.7 Hz, 1H), 7.70 (t, J=7.8 Hz, 1H), 7.57 (s, 1H), 7.51 (t,
J=8.4 Hz, 1H), 7.19 (s, 1H), 6.79 (d, J=8.5 Hz, 2H), 4.03 (s, 3H),
3.78 (s, 6H).
Example 244:
2,6-Dimethoxy-N-(4-methoxy-6-methylbenzo[d]isoxazol-3-yl)benzenesulfonami-
de 244
##STR00370##
[0944] To a solution of
N-(6-bromo-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamid-
e 89 (50 mg, 0.113 mmol) in 1,4-dioxane (8 mL) and water (2 mL) was
added methylboronic acid (14 mg, 0.226 mmol), Na.sub.2CO.sub.3 (36
mg, 0.339 mmol) and Pd(PPh.sub.3).sub.4 (14 mg, 0.011 mmol) and the
mixture was heated at reflux under a N.sub.2 atmosphere overnight.
The mixture was adjusted to pH 4-5 with 1 M aqueous HCl and
extracted with EtOAc. The combined organic extracts were washed
with water, brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
prep. HPLC to give the title compound (8 mg, 19%) as a white solid.
LCMS-C: R.sub.t 2.18 min, m/z 379.0 [M+H].sup.+. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.45 (s, 1H), 7.49 (t, J=8.5 Hz, 1H),
6.97 (s, 1H), 6.77 (d, J=8.5 Hz, 2H), 6.70 (s, 1H), 3.91 (s, 3H),
3.77 (s, 6H), 2.42 (s, 3H).
Example 245:
2,6-Dimethoxy-N-(4-methoxy-6-(pyridin-4-yl)benzo[d]isoxazol-3-yl)benzenes-
ulfonamide 245
##STR00371##
[0946] To a solution of
N-(6-bromo-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamid-
e 89 (50 mg, 0.113 mmol) in 1,4-dioxane (8 mL) and water (2 mL) was
added pyridin-4-ylboronic acid (28 mg, 0.226 mmol),
Na.sub.2CO.sub.3 (36 mg, 0.339 mmol) and Pd(PPh.sub.3).sub.4 (14
mg, 0.011 mmol) and the mixture was heated at reflux under a
N.sub.2 atmosphere overnight. The mixture was adjusted to pH 4-5
with 1 M aqueous HCl and extracted with EtOAc. The combined organic
extracts were washed with water, brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by prep. TLC (DCM/MeOH=30/1) to give the
title compound (22 mg, 44%) as a white solid. LCMS-C: R.sub.t 0.58
min, m/z 442.0 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.75 (s, 1H), 8.69 (d, J=5.3 Hz, 2H), 7.84 (d, J=5.3 Hz,
2H), 7.63 (s, 1H), 7.50 (t, J=8.4 Hz, 1H), 7.20 (s, 1H), 6.78 (d,
J=8.5 Hz, 2H), 4.03 (s, 3H), 3.78 (s, 6H).
Example 246:
2,6-Dimethoxy-N-(4-methoxy-6-(pyridin-3-yl)benzo[d]isoxazol-3-yl)benzenes-
ulfonamide 246
##STR00372##
[0948] To a solution of
N-(6-bromo-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamid-
e 89 (50 mg, 0.113 mmol) in 1,4-dioxane (8 mL) and water (2 mL) was
added 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (47
mg, 0.226 mmol), Na.sub.2CO.sub.3 (36 mg, 0.339 mmol) and
Pd(PPh.sub.3).sub.4 (14 mg, 0.011 mmol) and the mixture was heated
at reflux under a N.sub.2 atmosphere overnight. The mixture was
adjusted to pH 4-5 with 1 M aqueous HCl and extracted with EtOAc.
The combined organic extracts were washed with water, brine, dried
over Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by prep. TLC (DCM/MeOH=30/1) to
give the title compound (9 mg, 18%) as a white solid. LCMS-C:
R.sub.t 0.77 min, m/z 442.0 [M+H].sup.+. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.67 (s, 1H), 9.02 (d, J=2.5 Hz, 1H), 8.63
(dd, J=4.8, 1.6 Hz, 1H), 8.21 (dt, J=8.0, 2.0 Hz, 1H), 7.56 (s,
1H), 7.54-7.48 (m, 2H), 7.17 (s, 1H), 6.79 (d, J=8.5 Hz, 2H), 4.03
(s, 3H), 3.79 (s, 6H).
Example 247:
2,6-Dimethoxy-N-(4-methoxy-6-(6-methoxypyridin-3-yl)benzo[d]isoxazol-3-yl-
)benzenesulfonamide 247
##STR00373##
[0950] To a solution of
N-(6-bromo-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamid-
e 89 (50 mg, 0.113 mmol) in 1,4-dioxane (8 mL) and water (2 mL) was
added (6-methoxypyridin-3-yl)boronic acid (35 mg, 0.226 mmol),
Na.sub.2CO.sub.3 (36 mg, 0.339 mmol) and Pd(PPh.sub.3).sub.4 (14
mg, 0.011 mmol) and the mixture was heated at reflux under a
N.sub.2 atmosphere overnight. The mixture was adjusted to pH 4-5
with 1 M aqueous HCl and extracted with EtOAc. The combined organic
extracts were washed with water, brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by prep. TLC (DCM/MeOH=60/1) to give the
title compound (45 mg, 85%) as a white solid. LCMS-C: R.sub.t 2.33
min, m/z 472.0 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.61 (s, 1H), 8.63 (d, J=2.6 Hz, 1H), 8.15 (dd, J=8.7, 2.6
Hz, 1H), 7.51 (t, J=8.5 Hz, 1H), 7.47 (s, 1H), 7.10 (s, 1H), 6.94
(d, J=8.6 Hz, 1H), 6.79 (d, J=8.5 Hz, 2H), 4.02 (s, 3H), 3.91 (s,
3H), 3.79 (s, 6H).
Example 248:
2,6-Dimethoxy-N-(4-methoxy-6-(3-methoxy-5-methylphenyl)benzo[d]isoxazol-3-
-yl)benzenesulfonamide 248
##STR00374##
[0951] a)
2-(3-Methoxy-5-methylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaboro-
lane A61
[0952] A mixture of 1-bromo-3-methoxy-5-methylbenzene (500 mg, 2.49
mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane)
(1.9 g, 7.49 mmol), potassium acetate (977 mg, 9.96 mmol) and
Pd(dppf)Cl.sub.2 (196 mg, 0.25 mmol) in 1,4-dioxane (20 mL) was
heated at reflux under N.sub.2 for 3 h. The mixture was diluted
with EtOAc (300 mL), washed with water (50 mL.times.3) and the
organic layer was dried over Na.sub.2SO.sub.4, filtered,
concentrated under reduced pressure. The residue was purified by
column chromatography (Pet. ether/EtOAc=20/1) to give the title
compound (300 mg, 49%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.08 (s, 1H), 6.94 (d, J=2.6 Hz, 1H), 6.87
(s, 1H), 3.73 (s, 3H), 2.30-2.25 (m, 3H), 1.28 (s, 12H).
b)
2,6-Dimethoxy-N-(4-methoxy-6-(3-methoxy-5-methylphenyl)benzo[d]isoxazol-
-3-yl)benzenesulfonamide 248
[0953] To a solution of
N-(6-bromo-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamid-
e 89 (50 mg, 0.11 mmol) in 1,4-dioxane (8 mL) and water (2 mL) was
added
2-(3-methoxy-5-methylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
A61 (55 mg, 0.22 mmol), K2003 (61 mg, 0.44 mmol) and
Pd(PPh.sub.3).sub.4 (13 mg, 0.011 mmol) and the mixture was heated
at reflux under a N.sub.2 atmosphere overnight. The mixture was
adjusted to pH 4-5 with 1 M aqueous HCl and extracted with EtOAc.
The combined organic extracts were washed with water, brine, dried
over Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by prep. HPLC to give the title
compound (4.3 mg, 8%) as a white solid. LCMS-C: R.sub.t 2.56 min,
m/z 485.3 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
9.62 (s, 1H), 7.51 (t, J=8.5 Hz, 1H), 7.43 (s, 1H), 7.18 (s, 1H),
7.10 (s, 1H), 7.05 (s, 1H), 6.83 (s, 1H), 6.79 (d, J=8.5 Hz, 2H),
4.02 (s, 3H), 3.81 (s, 3H), 3.79 (s, 6H), 2.36 (s, 3H).
Example 249:
2,6-Dimethoxy-N-(4-methoxy-6-(2-methoxypyridin-4-yl)benzo[d]isoxazol-3-yl-
)benzenesulfonamide 249
##STR00375##
[0955] To a solution of
N-(6-bromo-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamid-
e 89 (50 mg, 0.11 mmol) in 1,4-dioxane (4 mL) and water (1 mL) was
added (2-methoxypyridin-4-yl)boronic acid (33 mg, 0.22 mmol),
Na.sub.2CO.sub.3 (35 mg, 0.33 mmol) and Pd(PPh.sub.3).sub.4 (13 mg,
0.011 mmol) and the mixture was heated at reflux under a N.sub.2
atmosphere overnight. The mixture was poured into water and
extracted with DCM. The combined organic extracts were dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by prep. HPLC to give the title compound
(5 mg, 10%) as a white solid. LCMS-C: R.sub.t 2.29 min, m/z 472.0
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.71 (br
s, 1H), 8.26 (d, J=5.4 Hz, 1H), 7.59 (s, 1H), 7.50 (t, J=8.5 Hz,
1H), 7.42 (dd, J=5.4, 1.6 Hz, 1H), 7.25 (s, 1H), 7.16 (s, 1H), 6.78
(d, J=8.5 Hz, 2H), 4.02 (s, 3H), 3.90 (s, 3H), 3.78 (s, 6H).
Example 250:
2,6-Dimethoxy-N-(4-methoxy-6-(5-methoxypyridin-3-yl)benzo[d]isoxazol-3-yl-
)benzenesulfonamide 250
##STR00376##
[0957] To a solution of
N-(6-bromo-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamid-
e 89 (50 mg, 0.11 mmol) in 1,4-dioxane (4 mL) and water (1 mL) was
added
3-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(52 mg, 0.22 mmol), Na.sub.2CO.sub.3 (35 mg, 0.33 mmol) and
Pd(PPh.sub.3).sub.4 (13 mg, 0.011 mmol) and the mixture was heated
at reflux under a N.sub.2 atmosphere overnight. The mixture was
poured into water and extracted with DCM. The combined organic
extracts were dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure.
[0958] The residue was purified by prep. HPLC to give the title
compound (5 mg, 10%) as a white solid. LCMS-C: R.sub.t 2.25 min,
m/z 472.0 [M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
9.67 (s, 1H), 8.63 (s, 1H), 8.38 (d, J=2.7 Hz, 1H), 7.81-7.79 (m,
1H), 7.59 (s, 1H), 7.51 (t, J=8.5 Hz, 1H), 7.18 (s, 1H), 6.79 (d,
J=8.5 Hz, 2H), 4.04 (s, 3H), 3.94 (s, 3H), 3.79 (s, 6H).
Example 251:
N-(6-(3-Cyano-5-methoxyphenyl)-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimeth-
oxybenzenesulfonamide 251
##STR00377##
[0959] a)
3-Methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoni-
trile A62
[0960] A mixture of 3-bromo-5-methoxybenzonitrile (500 mg, 2.35
mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane)
(1.8 g, 7.07 mmol), potassium acetate (923 mg, 9.4 mmol) and
Pd(dppf)Cl.sub.2.DCM (196 mg, 0.24 mmol) was heated at reflux under
N.sub.2 for 4 h. Water was added and the mixture was extracted with
EtOAc (20 mL.times.3). The combined organic extracts were washed
with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by prep. TLC
(EtOAc/Pet. ether=1/5) to give the title compound (600 mg, 98%) as
a yellow oil. LCMS-C: R.sub.t 2.66 min; m/z 260.0 [M+H].sup.+.
b)
N-(6-(3-Cyano-5-methoxyphenyl)-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dime-
thoxybenzenesulfonamide 251
[0961] To a solution of
N-(6-bromo-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamid-
e 89 (415 mg, 0.94 mmol) in 1,4-dioxane (80 mL) and water (20 mL)
was added
3-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitri-
le A62 (500 mg, 2.82 mmol), Na.sub.2CO.sub.3 (399 mg, 3.77 mmol)
and Pd(PPh.sub.3).sub.4 (116 mg, 0.1 mmol) and the mixture was
heated at reflux under a N.sub.2 atmosphere overnight. The mixture
was diluted with water and extracted with EtOAc. The combined
organic extracts were dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
prep. TLC (DCM/MeOH=30/1) to give the title compound (40 mg, 8.6%)
as a white solid. LCMS-C: R.sub.t 2.48 min, m/z 495.9 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.69 (br s, 1H), 7.89
(s, 1H), 7.67 (s, 1H), 7.57 (m, 1H), 7.49 (s, 2H), 7.15 (s, 1H),
6.77 (d, J=8.5 Hz, 2H), 4.02 (s, 3H), 3.90 (s, 3H), 3.77 (s,
6H).
Example 252:
2,6-Dimethoxy-N-(5-methyl-7-phenylbenzo[d]isoxazol-3-yl)benzenesulfonamid-
e 252
##STR00378##
[0963] To a solution of
N-(7-bromo-5-methylbenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide
239 (50 mg, 0.117 mmol) in 1,4-dioxane (8 mL) and water (2 mL) was
added phenylboronic acid (22 mg, 0.176 mmol), Na.sub.2CO.sub.3 (50
mg, 0.468 mmol) and Pd(PPh.sub.3).sub.4 (14 mg, 0.012 mmol) and the
mixture was heated at reflux under a N.sub.2 atmosphere overnight.
The mixture was diluted with water and extracted with EtOAc (30
mL.times.3). The combined organic extracts were washed with brine,
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by prep. TLC (Pet.
ether/EtOAc=3/1) to give the title compound (40 mg, 80%) as a white
solid. LCMS-C: R.sub.t 2.49 min; m/z 425.0 [M+H].sup.+. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.87-7.80 (m, 2H), 7.67-7.55 (m,
2H), 7.48 (t, J=7.7 Hz, 2H), 7.42-7.32 (m, 2H), 6.69 (d, J=8.4 Hz,
2H), 3.70 (s, 6H), 2.45 (s, 3H).
Assays
Protein Preparation
KAT5
[0964] Molecular Biology: A codon optimized DNA sequence (for
expression in Escherichia coli) encoding amino acid residues 2 to
461 (Uniprot Q92993-2) of human KAT5 isoform was synthesised by
GenScript USA Inc (Piscataway, N.J., USA). This was ligated into a
modified pET43a E. coli expression vector designed to encode an
N-terminal hexahistidine tag followed by a tobacco etch virus
protease (TEV) cleavage site and by the KAT5 sequence. The
resulting protein sequence is listed below.
TABLE-US-00014 (SEQ ID NO: 1)
MGHHHHHHGTENLYFQGSAEVGEIIEGCRLPVLRRNQDNEDEWPLAEILS
VKDISGRKLFYVHYIDFNKRLDEWVTHERLDLKKIQFPKKEAKTPTKNGL
PGSRPGSPEREVKRKVEVVSPATPVPSETAPASVFPQNGAARRAVAAQPG
RKRKSNCLGTDEDSQDSSDGIPSAPRMTGSLVSDRSHDDIVTRMKNIECI
ELGRHRLKPWYFSPYPQELTTLPVLYLCEFCLKYGRSLKCLQRHLTKCDL
RHPPGNEIYRKGTISFFEIDGRKNKSYSQNLCLLAKCFLDHKTLYYDTDP
FLFYVMTEYDCKGFHIVGYFSKEKESTEDYNVACILTLPPYQRRGYGKLL
IEFSYELSKVEGKTGTPEKPLSDLGLLSYRSYWSQTILEILMGLKSESGE
RPQITINEISEITSIKKEDVISTLQYLNLINYYKGQYILTLSEDIVDGHE
RAMLKRLLRIDSKCLHFTPKDWSKRGKWAS*
[0965] Protein Expression: To produce recombinant KAT5 protein,
expression plasmid was transformed into E. coli BL21 DE3 strain and
grown with shaking at 37.degree. C. in 1 L volumes of Terrific
broth (TB) supplemented with 100 .mu.g/mL Ampicillin and 50 .mu.M
zinc until an OD600 of 0.8 was reached. Cultures were transferred
to 18.degree. C. and protein expression induced by the addition of
Isopropyl .beta.-D-1-thiogalactopyranoside to a final concentration
of 0.5 mM and the cultures shaken overnight for further 16 hours.
Following expression, cell cultures were centrifuged at
5000.times.g for 20 min and cell pellet stored frozen at
-20.degree. C.
[0966] Protein Purification: Protein purification was initiated by
thawing the cell pellet (25 g wet weight) in Lysis buffer (50 mM
Hepes pH 7.4, 500 mM NaCl, 5 mM imidazole, 5% [v/v] glycerol, 0.1%
[w/v] CHAPS, 2 mM 2-mercaptoethanol, 3 mM MgCl2, 0.5 mg/mL
lysozyme, benzonase endonuclease [EMD Millipore], 1 mM PMSF,
complete protease inhibitor tablets EDTA-free [Roche]) using a
ratio of 6 mL of buffer per 1 g of cells. Cells were further lysed
by sonication using a Misonix Liquid Processor (6.times.30 second
pulses, amplitude 60 [70 watts]) and then centrifuged at
48,000.times.g at 4.degree. C. Supernatant (cell lysate) was mixed
with 20 mL of Q-Sepharose FF resin (GE Healthcare) pre-equilibrated
with Q buffer (20 mM Hepes pH 7.4, 1 M NaCl). The unbound fraction
from Q-Sepharose FF was then incubated with 5 mL of complete
His-Tag Purification Resin (Roche), pre-equilibrated with IMAC Wash
Buffer (20 mM hepes pH 7.4, 500 mM NaCl, 35 mM imidazole). The
resin was washed with IMAC Wash Buffer, and bound KAT5 eluted with
IMAC Elution buffer (20 mM hepes pH 7.4, 500 mM NaCl, 300 mM
imidazole). IMAC-eluted protein was immediately desalted into
Storage buffer (50 mM Na citrate pH 6.5, 500 mM NaCl, 5% [v/v]
glycerol) using 2.times. HiPrep 26/10 desalting columns (GE
Healthcare) in series. Desalted protein was further purified by
passing through a HiLoad 26/60 Superdex 75 column pre-equilibrated
in Storage buffer. Finally, KAT5 protein was concentrated to 1.5
mg/mL using Amicon Ultra centrifugal filter unit (Utra-15 MWCO 10
kDa), flash-frozen in liquid nitrogen and stored in -70.degree. C.
freezer.
[0967] KAT6A
[0968] Molecular Biology: The DNA sequence encoding amino acid
residues 507 to 778 (Uniprot Q92794-1) of human KAT6A was amplified
by PCR and was ligated into a modified pET E. coli expression
vector designed to encode a NusA solubility tag followed by a
hexahistidine tag and a tobacco etch virus protease (TEV) cleavage
site and by the KAT6A sequence. The resulting protein sequence is
listed below.
TABLE-US-00015 (SEQ ID NO: 2)
MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDVRVQID
RKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESV
TFDRITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNI
SLDLGNNAEAVILREDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRS
KPEMLIELFRIEVPEIGEEVIEIKAAARDPGSRAKIAVKTNDKRIDPVGA
CVGMRGARVQAVSTELGGERIDIVLWDDNPAQFVINAMAPADVASIVVDE
DKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMTVDDLQAKHQA
EAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLDE
PTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAA
RGVCTLEDLAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEATSGSG
HHHHHHSAGENLYFQGAMGRCPSVIEFGKYEIHTWYSSPYPQEYSRLPKL
YLCEFCLKYMKSRTILQQHMKKCGWFHPPVNEIYRKNNISVFEVDGNVST
IYCQNLCLLAKLFLDHKTLYYDVEPFLFYVLTQNDVKGCHLVGYFSKEKH
CQQKYNVSCIMILPQYQRKGYGRFLIDFSYLLSKREGQAGSPEKPLSDLG
RLSYMAYWKSVILECLYHQNDKQISIKKLSKLTGICPQDITSTLHHLRML
DFRSDQFVIIRREKLIQDHMAKLQLNLRPVDVDPECLRWTP
[0969] Protein Expression: To produce recombinant KAT6A protein,
expression plasmid was transformed into E. coli BL21 DE3 strain and
grown with shaking at 37.degree. C. in 1 L volumes of Terrific
broth (TB) supplemented with 100 .mu.g/mL Ampicillin until an OD600
of 0.8 was reached. Cultures were transferred to 18.degree. C. and
protein expression induced by the addition of Isopropyl
.beta.-D-1-thiogalactopyranoside to a final concentration of 0.5 mM
and the cultures shaken overnight for further 16 hours. Following
expression, cell cultures were centrifuged at 5000.times.g for 20
min and cell pellet stored frozen at -20.degree. C.
[0970] Protein Purification: Protein purification was initiated by
thawing the cell pellet (40 g wet weight) in Lysis buffer (25 mM
Tris-HCl pH 7.8, 500 mM NaCl, 5 mM DTT, 0.01% [v/v] Triton-X 100,
5% [v/v] glycerol, 2 mM MgCl2, 10 mM Imidazole, 0.5 mg/mL lysozyme,
benzonase endonuclease [EMD Millipore], 1 mM PMSF, complete
protease inhibitor tablets EDTA-free [Roche]) using a ratio of 5 mL
of buffer per 1 g of cells. Cells were further lysed by 3 passes
(at 15000 psi) through an ice cooled Avestin C.sub.5 cell crusher
and then centrifuged at 48,000.times.g at 4.degree. C. Supernatant
(cell lysate) was filtered through a 5 .mu.m filter and applied
onto 5 mL HiTrap IMAC Sepharose FF column (GE Healthcare)
pre-equilibrated with IMAC wash buffer (25 mM Tris-HCl pH 7.8, 500
mM NaCl, 5 mM DTT, 0.01% [v/v] Triton-X 100, 5% [v/v] glycerol, 20
mM Imidazole) using a Profinia Affinity chromatography purification
system (Bio-Rad). The IMAC column was then washed with IMAC Wash
buffer and bound KAT6A protein eluted with IMAC Elution buffer (25
mM Tris-HCl pH 7.8, 500 mM NaCl, 5% [v/v] glycerol, 5 mM DTT, 250
mM Imidazole). IMAC-eluted protein was further purified by passing
through a HiLoad 26/60 Superdex 200 column pre-equilibrated in
Storage buffer (25 mM Tris-HCl pH 7.8, 500 mM NaCl, 5 mM DTT, 5%
[v/v] glycerol). Finally, KAT6A protein was concentrated to 1 mg/mL
using Amicon Ultra centrifugal filter unit (Utra-15 MWCO 10 kDa),
flash-frozen in liquid nitrogen and stored in -70.degree. C.
freezer.
[0971] KAT6B was obtained from SignalChem, catalog ID:
K315-381BG
[0972] KAT7
[0973] Molecular Biology: A codon optimized DNA sequence encoding
amino acid residues 325 to 611 (Uniprot 095251-1) of human KAT7 was
synthesised by GenScript USA Inc (Piscataway, N.J., USA). This was
ligated into a modified pET43a E. coli expression vector designed
to encode an N-terminal hexahistidine tag followed by a tobacco
etch virus protease (TEV) cleavage site and by the KAT7 sequence.
The resulting protein sequence is listed below.
TABLE-US-00016 (SEQ ID NO: 3)
MGHHHHHHGTENLYFQGSRLQGQITEGSNMIKTIAFGRYELDTWYHSPYP
EEYARLGRLYMCEFCLKYMKSQTILRRHMAKCVWKHPPGDEIYRKGSISV
FEVDGKKNKIYCQNLCLLAKLFLDHKTLYYDVEPFLFYVMTEADNTGCHL
IGYFSKEKNSFLNYNVSCILTMPQYMRQGYGKMLIDFSYLLSKVEEKVGS
PERPLSDLGLISYRSYWKEVLLRYLHNFQGKEISIKEISQETAVNPVDIV
STLQALQMLKYWKGKHLVLKRQDLIDEWIAKEAKRSNSNKTMDPSCLKWT PPKGTAS
[0974] Protein Expression: To produce recombinant KAT7 protein,
expression plasmid was transformed into E. coli BL21 DE3 RIL strain
and grown with shaking at 37.degree. C. in 1 L volumes of Terrific
broth (TB) supplemented with 100 .mu.g/mL Ampicillin and 50 .mu.M
zinc until an OD600 of 0.8 was reached. Cultures were transferred
to 18.degree. C. and protein expression induced by the addition of
Isopropyl .beta.-D-1-thiogalactopyranoside to a final concentration
of 0.5 mM and the cultures shaken overnight for further 16 hours.
Following expression, cell cultures were centrifuged at
5000.times.g for 20 min and cell pellet stored frozen at
-20.degree. C.
[0975] Protein Purification: Protein purification was initiated by
thawing the cell pellet (10 g wet weight) in Lysis buffer (50 mM
Hepes pH 7.5, 300 mM NaCl, 5 mM DTT, 5 mM Imidazole, 0.05% [v/v]
Brij 35, 10% [v/v] glycerol, 3 mM MgCl2, 0.5 mg/mL lysozyme,
benzonase endonuclease [EMD Millipore], 1 mM PMSF, complete
protease inhibitor tablets EDTA-free [Roche]) using a ratio of 10
mL of buffer per 1 g of cells. Cells were further lysed by
sonication using a Misonix Liquid Processor (6.times.30 second
pulses, amplitude 60 [70 watts]) and then centrifuged at
48,000.times.g at 4.degree. C. Supernatant (cell lysate) was
incubated with 1 mL of complete His-Tag Purification Resin (Roche),
pre-equilibrated with IMAC Wash Buffer 1 (25 mM Hepes pH 7.5, 800
mM NaCl, 5 mM imidazole, 10% [v/v] glycerol, 5 mM DTT, 0.01% [v/v]
Brij 35, 50 mM arginine, 50 mM glutamic acid). The resin was
sequentially washed with IMAC Wash buffer 1 and IMAC Wash buffer 2
(25 mM hepes pH 7.5, 300 mM NaCl, 20 mM imidazole, 10% [v/v]
glycerol, 5 mM DTT, 0.01% [v/v] Brij 35, 50 mM arginine, 50 mM
glutamic acid). Bound KAT7 protein was eluted with IMAC Elution
buffer (25 mM hepes pH 7.5, 200 mM NaCl, 500 mM imidazole, 10%
[v/v] glycerol, 5 mM DTT 0.01% [v/v] Brij 35, 50 mM arginine, 50 mM
glutamic acid). The eluting protein was collected directly into 4
volumes of Desalt Buffer (50 mM Na citrate pH 6.5, 200 mM NaCl,
0.01% [v/v] Brij 35, 10% [v/v] glycerol, 5 mM DTT) to bring the
final imidazole concentration to 100 mM. IMAC-eluted protein was
immediately desalted into Desalt buffer using 2.times. HiPrep 26/10
desalting columns (GE Healthcare) in series. Desalted protein was
further purified by passing through a HiLoad 26/60 Superdex 75
column pre-equilibrated in Storage Buffer (50 mM Na citrate pH 6.5,
200 mM NaCl, 10% [v/v] glycerol, 5 mM DTT). Finally, KAT7 protein
was concentrated to 3.5 mg/mL using Amicon Ultra centrifugal filter
unit (Utra-15 MWCO 10 kDa), flash-frozen in liquid nitrogen and
stored in -70.degree. C. freezer.
[0976] KAT8
[0977] Molecular Biology: A codon optimized DNA sequence (for
expression in E. coli) encoding amino acid residues 177 to 447
(Uniprot Q9H7Z6-1) of human KAT8 was synthesised by Thermo Fisher
Scientific GENEART GmbH (Regensberg, Germany). This was ligated
into pPROEX Hta E. coli expression vector designed to encode an
N-terminal hexahistidine tag followed by a tobacco etch virus
protease (TEV) cleavage site and by the KAT8 sequence. The
resulting protein sequence is listed below.
TABLE-US-00017 (SEQ ID NO: 4)
MSYYHHHHHHDYDIPTTENLYFQGAKYVDKIHIGNYEIDAWYFSPFPEDY
GKQPKLWLCEYCLKYMKYEKSYRFHLGQCQWRQPPGKEIYRKSNISVYEV
DGKDHKIYCQNLCLLAKLFLDHKTLYFDVEPFVFYILTEVDRQGAHIVGY
FSKEKESPDGNNVACILTLPPYQRRGYGKFLIAFSYELSKLESTVGSPEK
PLSDLGKLSYRSYWSWVLLEILRDFRGTLSIKDLSQMTSITQNDIISTLQ
SLNMVKYWKGQHVICVTPKLVEEHLKSAQYKKPPITVDSVCLKWAP*
[0978] Protein Expression: To produce recombinant KAT8 protein,
expression plasmid was transformed into E. coli BL21 DE3 strain and
grown with shaking at 37.degree. C. in 1 L volumes of Terrific
broth (TB) supplemented with 100 .mu.g/mL Ampicillin until an OD600
of 0.8 was reached. Cultures were transferred to 18.degree. C. and
protein expression induced by the addition of Isopropyl
.beta.-D-1-thiogalactopyranoside to a final concentration of 0.5 mM
and the cultures shaken overnight for further 16 hours. Following
expression, cell cultures were centrifuged at 5000.times.g for 20
min and cell pellet stored frozen at -20.degree. C.
[0979] Protein Purification: Protein purification was initiated by
thawing the cell pellet (34 g wet weight) in Lysis buffer (20 mM
Hepes pH 7.5, 500 mM NaCl, 5 mM Imidazole, 5% [v/v] glycerol, 0.01%
[v/v] Triton-X 100, 5 mM 2-mercaptoethanol, 2 mM MgCl.sub.2, 0.5
mg/mL lysozyme, benzonase endonuclease [EMD Millipore], 1 mM PMSF,
complete protease inhibitor tablets EDTA-free [Roche]) using a
ratio of 3 mL of buffer per 1 g of cells. Cells were further lysed
by 3 passes (at 15000 psi) through an ice cooled Avestin C.sub.5
cell crusher and then centrifuged at 48,000.times.g at 4.degree. C.
Supernatant (cell lysate) was filtered through a 0.2 .mu.m filter
and applied onto 5 mL HiTrap IMAC Sepharose FF column (GE
Healthcare) pre-equilibrated with IMAC wash buffer 1 (20 mM Hepes
pH 7.5, 500 mM NaCl, 0.5 mM TCEP, 5 mM Imidazole) using a Profinia
Affinity chromatography purification system (Bio-Rad). The IMAC
column was then sequentially washed with IMAC Wash buffer 1 and
IMAC Wash buffer 2 (20 mM Hepes pH 7.5, 500 mM NaCl, 0.5 mM TCEP,
10 mM Imidazole) and bound KAT8 protein eluted with IMAC Elution
buffer (20 mM Hepes pH 7.5, 500 mM NaCl, 0.5 mM TCEP, 500 mM
Imidazole). IMAC-eluted protein was further purified by passing
through a HiLoad 26/60 Superdex 200 column pre-equilibrated in
Storage buffer (20 mM Hepes pH 7.5, 500 mM NaCl, 1 mM TCEP).
Finally, KAT8 protein was concentrated to 0.2 mg/mL using Amicon
Ultra centrifugal filter unit (Utra-15 MWCO 10 kDa), flash-frozen
in liquid nitrogen and stored in -70.degree. C. freezer.
[0980] Acetyltransferase Biochemical Assay
[0981] To determine the inhibition of KAT enzymatic activity by
test compounds, assay reactions were conducted in a volume of 8
.mu.L. in 384-well low volume assay plates. The reactions were
performed in assay buffer (100 mM Tris-HCl, pH 7,8, 15 mM NaCl, 1
mM EDTA, 0.01% Tween-20, 1 mM Dithiothreitol, and 0.01% m/v chicken
egg white albumin).
[0982] Reactions were set up with 1 .mu.M Acetyl coenzyme A, 100 nM
of full-length recombinant histone labelled by limited
biotinylation (KAT6A, KAT6B, KAT7: H3,1, KAT5, KAT8: H4),
10/5/8/40/20 nM of KAT5/KAT6A/KAT6B/KAT7/KAT8 enzyme respectively,
and an acetyl-lysine specific antibody (H3.1: Cell Signaling
Technology, H4: Abcam). 11-point dilution series of the test
compounds were prepared in DMSO; a volume of 100 nL was transferred
using a pin tool into assay plates containing substrates, before
adding enzyme to start the reaction. Positive (no compound, DMSO
only) and negative (AcCoA omitted) control reactions were included
on the same plates and received the same amount of DMSO as the
compound treated wells. After adding all reagents, the plates were
sealed with adhesive seals and incubated for 90 min at room
temperature. An additional 4 .mu.L of assay buffer containing
AlphaScreen.RTM. Protein A acceptor beads and Streptavidin donor
beads (PerkinElmer, Waltham, Mass.) to a final concentration of 8
.mu.g/mL was then added. After incubation for 2 hours the plates
were read using an EnVision 2103 multi label plate reader
(PerkinElmer) in HTS AlphaScreen.RTM. mode. IC.sub.50 values were
obtained from the raw readings by calculating percent inhibition (%
I) for each reaction relative to controls on the same plate (%
I=(I-CN)/(CP-CN) where CN/CP are the averages of the
negative/positive reactions, respectively), then fitting the % I
data vs. compound concentration [I] to %
I=(A+((B-A)/(1+((C/[I]){circumflex over ( )}D)))) where A is the
lower asymptote, B is the upper asymptote, C is the IC.sub.50
value, and D is the slope.
[0983] The results are shown in tables 1 to 5 below:
TABLE-US-00018 TABLE 1 (TIP60-KAT5) Example IC50 (.mu.M) 1
>125.000 2 >125.000 3 =65.106 4 =35.221 5 =114.325 6 =94.934
7 >125.000 8 >125.000 9 =40.976 10 =93.664 11 >125.000 12
>125.000 13 =119.896 14 >125.000 15 =7.294 16 =30.179 17
=27.659 18 =118.055 19 =64.983 20 =81.458 21 >125.000 22 =38.877
23 =72.865 24 >125.000 25 =120.445 26 >125.000 27 >125.000
28 >125.000 29 >125.000 30 >125.000 31 >125.000 32
=56.003 33 =90.452 34 >125.000 35 =33.836 36 =38.979 37
>125.000 38 =80.086 39 >125.000 40 =121.024 41 =65.079 42
=11.568 43 =23.002 44 =60.208 45 =34.341 46 >125.000 47 =123.081
48 =92.895 49 =74.577 50 >125.000 51 =87.660 52 >125.000 53
>125.000 54 >125.000 55 >125.000 56 >125.000 57
>125.000 58 >125.000 59 >125.000 60 >125.000 61 =85.401
62 >125.000 63 >125.000 64 >125.000 65 >125.000 66
=123.371 67 =114.876 68 >125.000 69 =124.236 70 =36.766 71
=28.431 72 >125.000 73 =2.161 74 =8.132 75 =33.535 76 =2.578 77
=51.770 78 >125.000 79 =17.451 80 =10.913 81 =31.488 82 =8.247
83 =17.898 84 =50.464 85 =28.466 86 =97.635 88 >125.000 89
=5.785 90 =90.684 91 =2.183 92 =1.387 93 =53.063 94 =124.242 95
=13.967 96 =28.640 97 =6.181 98 =31.286 99 =9.699 100 =51.711 101
=98.204 102 =9.330 103 =69.326 104 =28.112 105 >125.000 106
=14.254 107 =74.084 108 =41.946 109 =69.262 110 >125.000 111
=26.608 112 =16.494 113 =4.836 114 =59.355 115 =23.660 116 =10.463
117 >125.000 118 =37.082 119 =8.726 120 =5.781 121 =29.010 122
=58.591 123 =59.355 124 =27.879 125 >125.000 126 =51.346 127
=49.699 128 =6.301 129 =18.536 130 >125.000 131 =69.740 132
=53.030 133 =45.717 134 =51.523 135 =4.788 136 >125.000 137
=14.209 138 =5.115 139 =34.292 140 =47.841 141 >125.000 142
=32.183 143 >125.000 144 =11.487 145 =20.967 146 =21.951 147
>125.000 148 >125.000 149 >125.000 150 =50.012 151
>125.000 152 =38.945 153 =4.055 154 =25.313 155 =0.672 156
>125.000 157 >125.000 158 >125.000 159 >125.000 160
=46.217 161 >125.000 162 =13.954 163 >125.000 164 >125.000
165 >125.000 166 =3.093 167 =32.157 168 =3.415 169 =63.197 170
=4.826 171 =4.374 172 =9.070 173 =4.050 174 =1.833 175 =43.215 176
=5.670 177 =7.082 178 =7.535 179 =32.129 180 =25.476 181 =23.827
182 =65.635 183 =33.231 184 >125.000 185 >125.000 186 =84.702
187 =65.279 188 =69.912 189 =31.983 190 =19.884 191 =68.651 192
=29.888 193 =53.125 194 =37.844 195 =16.708 196 =95.399 197
>125.000 198 =73.654 199 =14.892 200 =4.733 201 =21.848 202
=110.730 203 =74.251 204 =4.565 205 =3.729 206 =6.361 207 =4.605
208 =28.461 209 =28.674 210 >125.000 211 =7.931 212 =9.302 213
>125.000 214 >125.000 215 >125.000 216 =123.671 217
>125.000 218 >125.000 219 >125.000 220 >125.000 221
>125.000 222 =26.758 223 >125.000 224 >125.000 225
>125.000 226 =80.348 227 >125.000 228 =124.363 229
>125.000 230 >125.000 231 >125.000 232 >125.000 233
>125.000 234 =10.877 235 =9.044 236 =10.055 237 >125.000 238
>125.000 239 =17.373 240 =122.168 241 =26.350 242 =27.314 243
=18.076 244 =30.172 245 =57.554 246 =27.066
247 =96.592 248 =9.331 249 =69.187 250 =12.220 251 =46.983 252
=101.708
TABLE-US-00019 TABLE 2 (MOZ-KAT6A) Example IC50 (.mu.M) 1 =91.387 2
=94.607 3 =23.759 4 =6.223 5 =31.283 6 =30.156 7 =62.687 8 =113.061
9 =26.816 10 =17.503 11 >125.000 12 >125.000 13 =63.854 14
=45.004 15 =1.907 16 =15.105 17 =11.820 18 =74.876 19 =32.241 20
=29.373 21 =24.799 22 =5.206 23 =21.776 24 >125.000 25 =33.179
26 =60.096 27 =71.527 28 =117.346 29 >125.000 30 =89.484 31
=36.075 32 =31.124 33 =6.847 34 >125.000 35 =4.632 36 =17.653 37
=24.848 38 =27.525 39 =38.220 40 =2.128 41 =4.274 42 =5.947 43
=5.971 44 =10.569 45 =2.085 46 =36.202 47 =12.863 48 =7.410 49
=8.133 50 =123.076 51 =15.032 52 =85.314 53 =90.683 54 =63.015 55
=103.246 56 =72.793 57 =56.212 58 =28.364 59 =49.410 60 =116.146 61
=51.918 62 =43.709 63 =2.558 64 =26.746 65 =27.934 66 =14.554 67
=22.711 68 =85.089 69 =42.890 70 =31.339 71 =11.578 72 =46.210 73
=4.547 74 =3.914 75 =23.533 76 =0.688 77 =10.814 78 =93.778 79
=13.890 80 =3.473 81 =43.616 82 =6.128 83 =13.571 84 =18.678 85
=3.866 86 =5.890 88 =41.205 89 =0.285 90 =4.779 91 =0.009 92 =0.006
93 =0.181 94 =6.105 95 =0.430 96 =1.203 97 =0.061 98 =5.602 99
=2.099 100 =0.972 101 =3.798 102 =0.143 103 =0.810 104 =0.786 105
=2.903 106 =0.782 107 =14.870 108 =3.089 109 =1.207 110 =7.890 111
=0.842 112 =1.463 113 =0.775 114 =29.278 115 =28.986 116 =0.560 117
=85.409 118 =10.003 119 =0.570 120 =0.310 121 =1.236 122 =24.400
123 =26.864 124 =11.011 125 =24.458 126 =10.472 127 =9.165 128
=0.250 129 =0.772 130 =2.956 131 =3.106 132 =7.454 133 =4.449 134
=6.449 135 =0.563 136 =34.274 137 =8.579 138 =2.892 139 =2.144 140
=2.256 141 =39.557 142 =3.296 143 =32.391 144 =0.261 145 =0.127 146
=0.153 147 =93.597 148 =23.965 149 =5.272 150 =4.966 151 =27.867
152 =6.276 153 =0.437 154 =0.516 155 =2.116 156 =1.535 157 =23.036
158 =62.560 159 =66.556 160 =1.048 161 =1.871 162 =0.147 163 =0.884
164 =27.173 165 =99.899 166 =0.123 167 =8.666 168 =10.006 169
=24.793 170 =1.504 171 =1.876 172 =0.037 173 =0.015 174 =0.013 175
=0.441 176 =0.030 177 =0.127 178 =1.440 179 =2.864 180 =9.437 181
=1.058 182 =15.217 183 =5.601 184 >125.000 185 =104.789 186
=25.161 187 =5.664 188 =9.228 189 =4.347 190 =9.733 191 =12.582 192
=9.394 193 =1.950 194 =1.507 195 =1.322 196 =13.919 197 =20.970 198
=2.818 199 =0.709 200 =0.364 201 =1.482 202 =18.907 203 =22.648 204
=0.400 205 =0.115 206 =0.302 207 =0.104 208 =1.629 209 =2.029 210
=8.532 211 =2.128 212 =2.117 213 =8.280 214 =36.431 215 =4.469 216
=0.625 217 =10.237 218 =6.594 219 =33.313 220 =3.497 221 =37.464
222 =0.655 223 =25.496 224 =50.368 225 =3.625 226 =10.774 227
=41.520 228 =75.246 229 =85.020 230 =48.075 231 =58.983 232 =46.464
233 =9.950 234 =0.381 235 =0.395 236 =0.318 237 =4.950 238 =12.039
239 =2.132 240 =1.828 241 =0.157 242 =3.232 243 =0.654 244 =2.126
245 =3.901 246 =0.676
247 =3.476 248 =0.139 249 =1.271 250 =0.423 251 =1.156 252
=4.160
TABLE-US-00020 TABLE 3 (HBO-KAT7) Example IC50 (.mu.M) 1 =53.948 2
=15.521 3 =23.243 4 =5.168 5 =6.011 6 =6.277 7 =14.175 8
>125.000 9 =8.418 10 =54.053 11 =60.488 12 =49.922 13 =63.834 14
=15.174 15 =1.456 16 =9.635 17 =14.575 18 =34.064 19 =36.094 20
=41.258 21 =25.506 22 =6.300 23 =23.893 24 =33.854 25 =41.948 26
=34.465 27 =32.121 28 =54.786 29 =94.098 30 =11.481 31 =53.590 32
=14.923 33 =11.409 34 =31.493 35 =8.748 36 =26.267 37 =114.461 38
=6.698 39 =10.116 40 =19.929 41 =11.845 42 =8.384 43 =12.914 44
=10.794 45 =6.833 46 =74.439 47 =29.419 48 =54.767 49 =77.831 50
>125.000 51 =28.277 52 >125.000 53 >125.000 54 >125.000
55 >125.000 56 >125.000 57 >125.000 58 =16.607 59
>125.000 60 >125.000 61 >125.000 62 =70.618 63 =60.060 64
=105.707 65 =59.720 66 =113.991 67 >125.000 68 >125.000 69
=94.149 70 =32.029 71 =21.593 72 =11.413 73 =0.508 74 =1.665 75
=5.748 76 =0.937 77 =12.022 78 >125.000 79 =4.059 80 =1.129 81
=6.726 82 =1.496 83 =3.792 84 =20.038 85 =1.769 86 =1.981 88 =7.509
89 =0.168 90 =18.889 91 =0.079 92 =0.060 93 =2.799 94 =16.059 95
=0.754 96 =4.265 97 =0.444 98 =3.411 99 =5.739 100 =2.240 101
=32.782 102 =0.568 103 =5.928 104 =3.132 105 =63.160 106 =3.977 107
=11.732 108 =2.038 109 =4.067 110 =17.497 111 =4.536 112 =3.014 113
=0.914 114 =41.609 115 =95.520 116 =1.435 117 >125.000 118
=0.852 119 =1.146 120 =0.503 121 =5.211 122 =4.122 123 =4.198 124
=13.017 125 =85.834 126 =28.884 127 =9.487 128 =0.618 129 =1.318
130 =21.712 131 =23.780 132 =11.785 133 =5.342 134 =27.644 135
=1.426 136 =28.316 137 =6.558 138 =3.683 139 =10.043 140 =9.895 141
=24.336 142 =1.165 143 =7.883 144 =0.286 145 =0.259 146 =0.511 147
>125.000 148 =36.783 149 =9.432 150 =8.039 151 =25.571 152
=9.866 153 =0.507 154 =3.128 155 =0.248 156 =1.975 157 =15.514 158
=44.862 159 =37.620 160 =1.577 161 =2.506 162 =0.969 163 =13.103
164 =61.638 165 =30.654 166 =0.328 167 =6.854 168 =36.401 169
=8.646 170 =0.982 171 =0.292 172 =0.426 173 =0.169 174 =0.031 175
=5.949 176 =0.138 177 =0.021 178 =0.482 179 =13.481 180 =1.759 181
=5.137 182 =15.608 183 =4.480 184 =30.983 185 =60.714 186 =47.092
187 =21.961 188 =40.080 189 =8.016 190 =18.459 191 =1.576 192
=24.230 193 =15.518 194 =6.872 195 =4.963 196 =23.280 197 =45.687
198 =6.951 199 =0.632 200 =0.678 201 =0.773 202 =54.375 203 =23.360
204 =3.045 205 =0.936 206 =0.626 207 =0.257 208 =4.654 209 =1.054
210 =6.539 211 =0.917 212 =1.048 213 =20.519 214 >125.000 215
=16.630 216 =3.106 217 =18.406 218 =6.444 219 =60.688 220 =11.008
221 =63.721 222 =1.422 223 =43.526 224 >125.000 225 =31.135 226
=37.985 227 =63.139 228 >125.000 229 =115.218 230 =109.417 231
>125.000 232 >125.000 233 =17.872 234 =3.739 235 =0.820 236
=1.070 237 =18.016 238 =94.180 239 =3.003 240 =10.198 241 =2.510
242 =1.785 243 =0.720 244 =1.113 245 =2.761 246 =0.478
247 =5.897 248 =0.375 249 =0.889 250 =0.050 251 =0.473 252
=1.303
TABLE-US-00021 TABLE 4 (MOF-KAT8) Example IC50 (.mu.M) 11 =35.565
18 =63.474 19 =40.985 20 =61.440 21 =39.655 26 =29.480 73 =3.459 81
=9.425 93 =96.545 94 =60.943 95 =6.447 96 =53.767 100 =62.604 102
=11.485 107 =42.081 109 =9.122 111 =3.221 132 =15.285 151 =34.717
154 =2.570 161 =117.753 162 =10.649 163 >125.000 164 >125.000
165 >125.000 166 =2.920 168 =6.458 170 =9.666 173 =3.522 178
=0.770 204 =3.063 205 =3.441 211 =29.532 214 >125.000 216
=11.172 219 =69.922 221 =27.895 223 =42.389 224 =112.662 236
=3.682
TABLE-US-00022 TABLE 5 (QKF-KAT6B) Example IC50 (.mu.M) 73 =4.047
91 =0.037 92 =0.053 95 =1.640 96 =3.395 97 =0.049 102 =0.282 116
=1.616 143 =50.697 144 =0.057 146 =0.291 147 >125.000 162 =0.180
166 =0.227 172 =0.042 173 =0.040 174 =0.018 175 =0.821 176 =0.121
177 =0.092 178 =2.455 207 =1.454
[0984] Histone H3 Lysine 23 Acetylation Biomarker Assay
[0985] Compounds were tested for their ability to inhibit
acetylation of the histone H3K23 marker in the following assay:
[0986] The cell line U2OS was seeded at a density of 9,000 cells
per well in 96 well optical quality tissue culture plates in RPMI
medium and 10% foetal bovine serum, and allowed to adhere for 24
hours under standard culture conditions (37 degree Celsius, 5%
CO2). At the end of this period the medium was aspirated. Compound
dilutions prepared in DMSO were added to medium, with negative
control wells reserved for treatment with DMSO only and 100%
inhibition positive controls receiving a potent inhibitor compound
(e.g. cas 2055397-28-7, benzoic acid, 3-fluoro-5-(2-pyridinyl)-,
2-[(2-fluorophenyl)sulfonyl]hydrazide) (Baell, J., Nguyen, H. N.,
Leaver, D. J., Cleary, B. L., Lagiakos, H. R., Sheikh, B. N.,
Thomas. T. J., Aryl sulfonohydrazides, WO2016198507A1, 2016) at 10
.mu.M concentration and 200 .mu.L transferred to the cells. After
incubation for 24 hours, the cells were fixed with 3.7%
formaldehyde in PBS for 20 minutes at room temperature, washed
(5.times.5 minutes) with phosphate buffer saline containing 0.1%
Tween 20 and blocked with Odyssey blocking buffer (LI-COR, Lincoln,
Nebr.) containing 0.1% TritonX100. Anti-H3K23ac specific antibody
(Abcam ab177275) in Odyssey blocking buffer containing 0.1% Tween
20 was added and incubated for 16 hours at 4 degree Celsius. After
washing (as above), a secondary antibody labelled with Alexa647 dye
(LifeTechnologies) and Hoechst 33342 (1 .mu.g/mL, SigmaAldrich)
were added for 1 hour incubation. Plates were washed as previously
and read on a PerkinElmer Phenix high content imaging platform.
Using a Columbus image analysis pipeline, individual nuclei were
located by Hoechst 33342 stain and the acetylation level was
calculated from the Alexa647-related intensity in the same area.
The resulting mean intensity per cell was directly converted to
percent inhibition relative to controls on the same plate and the
data fitted against a four-parameter logistic model to determine
the 50% inhibitory concentration (IC50).
[0987] The results are shown in table 6 below:
TABLE-US-00023 TABLE 6 Example IC50 (.mu.M) 73 >10.000 86 =1.579
91 =0.182 92 =0.013 95 >10.000 96 >10.000 97 =0.045 116
=3.455 144 =0.128 145 =0.098 146 =0.045 147 >10.000 155
>10.000 162 =0.144 166 =0.398 172 =0.014 173 =0.020 174 =0.017
176 =0.685 177 =1.486 178 >10.000 191 >10.000 242 =0.672 250
=3.713
[0988] Histone H3 Lysine 14 Acetylation Biomarker Assay
[0989] Compounds were tested for their ability to inhibit
acetylation of the histone H3 Lysine 14 marker in the following
assay:
[0990] The cell line U2OS was seeded at a density of 3,000 cells
per well in 384-well optical quality tissue culture plates in RPMI
medium supplemented with 10% foetal bovine serum and 10 mM Hepes.
The cells were allowed to adhere for 24 hours under standard
culture conditions (37 degree Celsius, 5% CO2). At the end of this
period the cells were washed with serum free medium. Compound
dilutions prepared in DMSO were added to the serum free medium,
with negative control wells reserved for treatment with DMSO only
and 100% inhibition positive controls receiving a potent inhibitor
compound (e.g.
(Z)-4-fluoro-N-((3-hydroxyphenyl)sulfonyl)-5-methyl-[1,1'-biphenyl]-3-car-
bohydrazonic acid) at 10 .mu.M concentration. After incubation for
24 hours, the cells were fixed with 4% formaldehyde in PBS for 15
minutes at room temperature, washed with phosphate buffer saline
and blocked with blocking buffer containing 0.2% TritonX100 and 2%
BSA. Anti-H3K14ac specific antibody (Cell Signalling Technologies)
in blocking buffer was added and incubated overnight at 4 degree
Celsius. After washing, a secondary antibody labelled with
AlexaFluor 488 dye (ThermoFisher) and Hoechst 33342 (1 .mu.g/mL,
Life Technologies) were added for 2 hours incubation at room
temperature. Plates were washed and read on a PerkinElmer Opera HCS
high content imaging platform. Using a Columbus image analysis
pipeline, individual nuclei were located by Hoechst 33342 stain and
the acetylation level was calculated from the AlexaFluor
488-related intensity in the same area. The resulting mean
intensity per cell was converted to percent inhibition relative to
controls on the same plate and the data fitted against a
four-parameter logistic model to determine the 50% inhibitory
concentration (IC50).
[0991] The results are shown in table 7 below:
TABLE-US-00024 TABLE 7 Example IC50 (.mu.M) 73 =12.451 74 =31.675
75 >40.000 80 =11.511 82 =12.600 89 =3.957 91 =0.874 92 =0.703
95 =9.089 97 =1.093 102 =3.686 144 =2.954 145 =3.383 146 =2.304 147
>40.000 155 =26.130 162 =1.527 163 =20.356 166 =3.686 171 =2.574
172 =1.453 173 =0.533 174 =0.068 176 =1.071 177 =0.135 178 =3.068
191 =17.835 211 >40.000 212 =10.347 218 >40.000 236
>40.000 250 =3.482 252 =24.719
[0992] H2A.Z Lysine 7 Acetylation Biomarker Assay
[0993] Compounds were tested for their ability to inhibit the
histone H2A.Z Lysine 7 acetylation marker in the following
assay:
[0994] The cell line U2OS was seeded at a density of 3,000 cells
per well in 384-well optical quality tissue culture plates in RPMI
medium supplemented with 10% foetal bovine serum and 10 mM Hepes.
The cells were allowed to adhere for 24 hours under standard
culture conditions (37 degree Celsius, 5% CO.sub.2). At the end of
this period the cells were washed with serum free medium. Compound
dilutions prepared in DMSO were added to the serum free medium,
with negative control wells reserved for treatment with DMSO only
and 100% inhibition positive controls receiving a potent inhibitor
compound enantiomer 1 of
7-iodo-N-(2-(oxazol-2-yl)-2-phenylethyl)-2H-benzo[e][1,2,4]thiadiazine-3--
carboxamide 1,1-dioxide, which is compound 146 of co-pending
application GB1713962.7, filed on 31 Aug. 2018, at 30 .mu.M
concentration. After incubation for 24 hours, the cells were fixed
with 4% formaldehyde in PBS for 15 minutes at room temperature,
washed with phosphate buffer saline and blocked with blocking
buffer containing 0.2% TritonX100 and 2% BSA. Anti-H2A.ZK7ac
specific antibody (Abcam) in blocking buffer was added and
incubated overnight at 4 degree Celsius. After washing, a secondary
antibody labelled with AlexaFluor 488 dye (ThermoFisher) and
Hoechst 33342 (1 .mu.g/mL, Life Technologies) were added for 2
hours incubation at room temperature. Plates were washed and read
on a PerkinElmer Opera HCS high content imaging platform. Using a
Columbus image analysis pipeline, individual nuclei were located by
Hoechst 33342 stain and the acetylation level was calculated from
the AlexaFluor 488-related intensity in the same area. The
resulting mean intensity per cell was converted to percent
inhibition relative to controls on the same plate and the data
fitted against a four-parameter logistic model to determine the 50%
inhibitory concentration (IC.sub.50).
[0995] The results are shown in table 8 below:
TABLE-US-00025 TABLE 8 Example IC50 (.mu.M) 73 =36.725 89 =19.816
91 =8.022 92 =4.619 95 >40.000 97 =27.988 102 >40.000 144
=18.752 145 >40.000 146 =15.065 147 >40.000 155 >40.000
162 =29.540 166 =33.304 168 >40.000 172 =29.159 173 =26.225 174
=1.544 176 =7.612 177 =1.088 178 =5.589 191 >40.000 234
>40.000 236 >40.000 250 =13.197 252 >40.000
STATEMENTS OF INVENTION
[0996] 1. A compound of formula (I), or a pharmaceutically
acceptable salt thereof, for use in a method of therapy:
##STR00379##
wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are independently
selected from: [0997] (i) H; [0998] (ii) C.sub.1-3 alkyl,
optionally substituted by: [0999] hydroxy, [1000] C.sub.1-2 alkoxy,
optionally substituted by one or more fluoro groups, [1001]
NH.sub.2, [1002] phenyl, [1003] C.sub.5-6 heteroaryl, [1004]
C.sub.1-4 alkyl carbamoyl, [1005] acylamido, or [1006] one or more
fluoro groups; [1007] (iii) C.sub.1-3 alkoxy, optionally
substituted by C.sub.3-6 cycloalkyl or by one or more fluoro
groups; [1008] (iv) C.sub.3-6 cycloalkyl; [1009] (v) halo; [1010]
(vi) COR.sup.C, where R.sup.C is selected from NR.sup.N1R.sup.N2,
where R.sup.N1 and R.sup.N2 are independently selected from H and
methyl; [1011] (vii) cyano, NH.sub.2, or NO.sub.2; and [1012]
(viii) phenyl or C.sub.5-6 heteroaryl, optionally substituted by
methyl, cyano, hydroxy or methoxy; Ar is a phenyl, napthyl or
C.sub.5-10 heteroaryl group, which groups are optionally
substituted by one or more groups selected from: [1013] (i)
C.sub.1-4 alkyl, optionally substituted by hydroxy, C.sub.1-2
alkoxy, NH.sub.2, C.sub.1-4 alkyl carbamoyl, or by one or more
fluoro groups; [1014] (ii) C.sub.3-6 cycloalkyl; [1015] (iii)
hydroxy; cyano; NR.sup.N3R.sup.N4, where R.sup.N3 and R.sup.N4 are
independently selected from H and methyl; or acylamido; [1016] (iv)
halo; [1017] (v) C.sub.1-3 alkoxy, optionally substituted by
hydroxy, C(O)NH.sub.2, C.sub.3-6 cycloalkyl, phenyl, C.sub.5-6
heteroaryl, or by one or more fluoro groups; [1018] (vi) phenoxy,
optionally substituted by fluoro; [1019] (vii) phenyl or C.sub.5-6
heteroaryl; [1020] (viii) SF.sub.5 or SO.sub.2CH.sub.3; [1021] (ix)
--(CH.sub.2).sub.n--Y--, where Y is O or CH.sub.2, and n is 2 or 3;
or [1022] (x) C.sub.1-4 alkyl ester. 2. A compound for use
according to statement 1, wherein at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 is H. 3. A compound for use according to either
statement 1 or statement 2, wherein one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 are H. 4. A compound for use according to
either statement 1 or statement 2, wherein two of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 are H. 5. A compound for use according to
either statement 1 or statement 2, wherein three of R.sup.1,
R.sup.2, R.sup.3 and R.sup.4 are H. 6. A compound for use according
to either statement 1 or statement 2, wherein at least one of
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is not H. 7. A compound for
use according to any one of statements 1 to 6, wherein at least one
of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is C.sub.1-3 alkyl,
optionally substituted by: [1023] hydroxy, [1024] C.sub.1-2 alkoxy,
optionally substituted by one or more fluoro groups, [1025]
NH.sub.2, [1026] phenyl, [1027] C.sub.5-6 heteroaryl, [1028]
C.sub.1-4 alkyl carbamoyl, [1029] acylamido, or [1030] one or more
fluoro groups. 8. A compound for use according to statement 7,
wherein the C.sub.1-3 alkyl group is unsubstituted. 9. A compound
for use according to statement 7, wherein the C.sub.1-3 alkyl group
is perfluorinated. 10. A compound for use according to statement 7,
wherein the C.sub.1-3 alkyl group is substituted, by a group
selected from: [1031] (i) hydroxy; [1032] (ii) C.sub.1-2 alkoxy;
[1033] (iii) NH.sub.2; [1034] (iv) phenyl; [1035] (v) C.sub.5-6
heteroaryl; [1036] (vi) C.sub.1-4 alkyl carbamoyl; and [1037] (vii)
acylamido. 11. A compound for use according to any one of
statements 1 to 10, wherein at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 is C.sub.1-3 alkoxy, optionally substituted by
C.sub.3-6 cycloalkyl or by one of more fluoro groups. 12. A
compound for use according to statement 11, wherein the C.sub.1-3
alkoxy group is unsubstituted. 13. A compound for use according to
statement 11, wherein the C.sub.1-3 alkoxy group is perfluorinated.
14. A compound for use according to statement 11, wherein the
C.sub.1-3 alkoxy group is substituted by C.sub.3-6 cycloalkyl. 15.
A compound for use according to any one of statements 1 to 14,
wherein at least one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is
C.sub.3-6 cycloalkyl. 16. A compound for use according to statement
15, wherein the C.sub.3-6 cycloalkyl group is cyclopropyl. 17. A
compound for use according to any one of statements 1 to 16,
wherein at least one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is
halo. 18. A compound for use according to any one of statements 1
to 17, wherein at least one of R.sup.1, R.sup.2, R.sup.3 and
R.sup.4 is COR.sup.C, where R.sup.C is selected from
NR.sup.N1R.sup.N2, where R.sup.N1 and R.sup.N2 are independently
selected from H and methyl. 19. A compound for use according to any
one of statements 1 to 18, wherein at least one of R.sup.1,
R.sup.2, R.sup.3 and R.sup.4 is selected from cyano, NH.sub.2 and
NO.sub.2. 20. A compound for use according to statement 19, wherein
at least one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is cyano. 21.
A compound for use according to statement 19, wherein at least one
of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is NH.sub.2. 22. A
compound for use according to statement 19, wherein at least one of
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is NO.sub.2. 23. A compound
for use according to any one of statements 1 to 22, wherein at
least one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is phenyl or
C.sub.5-6 heteroaryl, optionally substituted by methyl, cyano,
hydroxy or methoxy. 24. A compound for use according to statement
23, wherein at least one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4
is phenyl, optionally substituted by methyl or methoxy. 25. A
compound for use according to statement 23, wherein at least one of
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is C.sub.5-6 heteroaryl,
optionally substituted by one or more methyl groups. 26. A compound
for use according to statement 1, wherein R.sup.4 is methoxy,
R.sup.2 is CH.sub.2OCH.sub.3 or CH.sub.2OCH.sub.2CH.sub.3 and
R.sup.1 and R.sup.3 are H. 27. A compound for use according to
statement 1, wherein R.sup.4 is methoxy, R.sup.2 is phenyl,
optionally substituted by methyl or methoxy, and R.sup.1 and
R.sup.3 are H. 28. A compound for use according to statement 1,
wherein R.sup.4 is methoxy, R.sup.2 is C.sub.5-6 heteroaryl,
optionally substituted by methyl. 29. A compound for use according
to statement 1, wherein R.sup.4 is methoxy, R.sup.2 is C.sub.6
heteroaryl, optionally substituted by methyl. 30. A compound for
use according to statement 1, wherein R.sup.4 is methoxy and
R.sup.1, R.sup.2 and R.sup.3 are H. 31. A compound for use
according to statement 1, wherein R.sup.4 is chloro, R.sup.2 is
C.sub.1-3 alkyl or bromo, and R.sup.1 and R.sup.3 are H. 32. A
compound for use according to statement 1, wherein R.sup.4 is
chloro and R.sup.1, R.sup.2 and R.sup.4 are H. 33. A compound for
use according to statement 1, wherein R.sup.3 is C.sub.1-3 alkyl
and R.sup.1, R.sup.2 and R.sup.4 are H. 34. A compound for use
according to any one of statements 1 to 33, wherein Ar is phenyl,
which may be unsubstituted or substituted. 35. A compound for use
according to any one of statements 1 to 33, wherein Ar is napthyl,
which may be unsubstituted or substituted. 36. A compound for use
according to any one of statements 1 to 33, wherein Ar is
substituted phenyl. 37. A compound for use according to any one of
statements 1 to 33, wherein Ar is a C.sub.5-10 heteroaryl group,
which may be unsubstituted or substituted. 38. A compound for use
according to statement 37, wherein the C.sub.5-10 heteroaryl group
is selected from: quinolinyl, benzothiazolyl, quinoxalinyl,
benzooxadiazolyl, benzothiadiazolyl, benzofuran and benzotriazolyl.
39. A compound for use according to statement 37, wherein the
C.sub.5-10 heteroaryl group is quinolinyl or benzothiazolyl. 40. A
compound for use according to any one of statements 1 to 33,
wherein Ar is the group:
##STR00380##
[1037] 41. A compound for use according to any one of statements 33
to 39, wherein Ar is substituted by C.sub.1-4 alkyl, optionally
substituted by hydroxy, C.sub.1-2 alkoxy, NH.sub.2, C.sub.1-4 alkyl
carbamoyl, or by one or more fluoro groups. 42. A compound for use
according to statement 41, wherein the C.sub.1-4 alkyl group is
unsubstituted. 43. A compound for use according to statement 41,
wherein the C.sub.1-4 alkyl group is perfluorinated. 44. A compound
for use according to statement 41, wherein the C.sub.1-4 alkyl
group is substituted, by a group selected from: [1038] (i) hydroxy;
[1039] (ii) C.sub.1-2 alkoxy; [1040] (iii) NH.sub.2; and [1041]
(iv) C.sub.1-4 alkyl carbamoyl. 45. A compound for use according to
any one of statements 33 to 39, wherein Ar is substituted by
C.sub.3-6 cycloalkyl. 46. A compound for use according to statement
45, wherein the C.sub.3-6 cycloalkyl group is cyclohexyl. 47. A
compound for use according to any one of statements 33 to 39,
wherein Ar is substituted by hydroxy; cyano; NR.sup.N3R.sup.N4,
where R.sup.N3 and R.sup.N4 are independently selected from H and
methyl; or acylamido. 48. A compound for use according to statement
47, wherein the substituent is hydroxy. 49. A compound for use
according to statement 47, wherein the substituent is cyano. 50. A
compound for use according to statement 47, wherein the substituent
is NR.sup.N3R.sup.N4, where R.sup.N3 and R.sup.N4 are independently
selected from H and methyl. 51. A compound for use according to
statement 47, wherein the substituent acylamido. 52. A compound for
use according to any one of statements 33 to 39, wherein Ar is
substituted by halo. 53. A compound for use according to any one of
statements 33 to 39, wherein Ar is substituted by C.sub.1-3 alkoxy,
optionally substituted by hydroxy, C(O)NH.sub.2, C.sub.3-6
cycloalkyl, phenyl, C.sub.5-6 heteroaryl, or by one of more fluoro
groups. 54. A compound for use according to statement 53, wherein
the C.sub.1-3 alkoxy group is unsubstituted. 55. A compound for use
according to statement 53, wherein the C.sub.1-3 alkoxy group is
perfluorinated. 56. A compound for use according to statement 53,
wherein the C.sub.1-3 alkoxy group is substituted by hydroxy. 57. A
compound for use according to statement 53, wherein the C.sub.1-3
alkoxy group is substituted by C(O)NH.sub.2. 58. A compound for use
according to statement 53, wherein the C.sub.1-3 alkoxy group is
substituted by C.sub.3-6 cycloalkyl. 59. A compound for use
according to statement 53, wherein the C.sub.1-3 alkoxy group is
substituted by phenyl. 60. A compound for use according to
statement 53, wherein the C.sub.1-3 alkoxy group is substituted by
C.sub.5-6 heteroaryl. 61. A compound for use according to any one
of statements 33 to 39, wherein Ar is substituted by phenoxy,
optionally substituted by fluoro. 62. A compound for use according
to statement 61, wherein Ar is substituted by phenoxy. 63. A
compound for use according to statement 61, wherein Ar is
substituted by OC.sub.6H.sub.4F. 64. A compound for use according
to any one of statements 33 to 39, wherein Ar is substituted by
phenyl or C.sub.5-6 heteroaryl. 65. A compound for use according to
statement 64, wherein Ar is substituted by phenyl. 66. A compound
for use according to statement 64, wherein Ar is substituted by
C.sub.5-6 heteroaryl. 67. A compound for use according to any one
of statements 33 to 39, wherein Ar is substituted by SF.sub.5 or
SO.sub.2CH.sub.3. 68. A compound for use according to statement 67,
wherein Ar is substituted by SF.sub.5. 69. A compound for use
according to statement 67, wherein Ar is substituted by
SO.sub.2CH.sub.3. 70. A compound for use according to any one of
statements 33 to 39, wherein Ar is substituted by
--(CH.sub.2).sub.n--Y--, where Y is O or CH.sub.2, and n is 2 or 3.
71. A compound for use according to statement 70, where Ar is
phenyl. 72. A compound for use according to any one of statements
33 to 39, wherein Ar is substituted by C.sub.1-4 alkyl ester. 73. A
compound for use according to statement 72, where Ar is substituted
by C(O)OCH.sub.3. 74. A compound for use according to statement 72,
where Ar is substituted by C(O)OC(CH.sub.3).sub.3. 75. A compound
for use according to any one of statements 1 to 33 and 41 to 74,
wherein Ar is represented by the formula (Ar-1):
##STR00381##
[1041] where Y is either N or C--R.sup.A4, and Z is either N or
C--R.sup.A5; and R.sup.A1, R.sup.A2, R.sup.A3, R.sup.A4 (if
present) and R.sup.A5 (if present) are independently selected from
H and the optional substituents for Ar. 76. A compound for use
according to statement 75, wherein R.sup.A2 is ethyl. 77. A
compound for use according to statement 75, wherein R.sup.A3 is
selected from cycloalkyl; phenoxy; phenyl; C.sub.5-6 heteroaryl;
SF.sub.5; and SO.sub.2CH.sub.3. 78. A compound for use according to
any one of statements 1 to 33, wherein Ar is
5-ethyl-2-methoxyphenyl. 79. A compound for use according to any
one of statements 1 to 33, wherein Ar is
5-CF.sub.3-2-methoxyphenyl. 80. A compound for use according to any
one of statements 1 to 33, wherein Ar is 2,6-dimethoxyphenyl. 81. A
compound for use according to statement 1, with the proviso that
when: [1042] R.sub.1, R.sub.2, R.sup.3 and R.sup.4 are H, [1043] Ar
is not 4-aminophenyl. 82. A pharmaceutical composition comprising a
compound as defined in any one of statements 1 to 81 and a
pharmaceutically acceptable excipient. 83. A method of treatment of
cancer, comprising administering to a patient in need of treatment,
a compound as defined in any one of statements 1 to 81 or a
pharmaceutical composition according to statement 82. 84. A method
according to statement 83, wherein the compound is administered
simultaneously or sequentially with radiotherapy and/or
chemotherapy. 85. The use of a compound as defined in any one of
statements 1 to 81 in the manufacture of a medicament for treating
cancer. 86. A compound as defined in any one of statements 1 to 81
or a pharmaceutical composition according to statement 82 for use
in the treatment of cancer. 87. A compound or pharmaceutical
composition according to statement 86, wherein the treatment is for
simultaneous or sequential administration with radiotherapy and/or
chemotherapy. 88. A compound as defined in any one of statements 1
to 81 or a pharmaceutical salt thereof. 89. A compound according to
statement 88, wherein at least one of R.sup.1, R.sup.2, R.sup.3 and
R.sup.4 is not H. 90. A compound according to statement 89, wherein
R.sup.3 is not CF.sub.3. 91. A compound according to statement 89,
wherein R.sup.3 is not substituted C.sub.1-3 alkyl. 92. A compound
according to statement 89, wherein R.sup.3 is ethyl or propyl. 93.
A compound according to statement 89, wherein R.sup.3 is not
C.sub.1-3 alkyl, optionally substituted by hydroxy, C.sub.1-2
alkoxy, NH.sub.2, phenyl, C.sub.5-6 heteroaryl, C.sub.1-4 alkyl
carbamoyl, acylamido or by one or more fluoro groups. 94. A
compound according to statement 88, wherein R.sup.4 is methoxy. 95.
A compound according to statement 88, wherein R.sup.4 is Cl, and
R.sup.1, R.sup.2 and R.sup.3 are H. 96. A compound according to
statement 88, wherein R.sup.4 is Cl, and R.sup.2 is C.sub.1-3 alkyl
or bromo, and R.sup.1 and R.sup.3 are H. 97. A compound according
to statement 88, wherein R.sup.3 is C.sub.1-3 alkyl and R.sup.1,
R.sup.2 and R.sup.4 are H. 98. A compound according to statement
88, with the proviso that when: [1044] R.sup.1, R.sup.2 and R.sup.3
are H, and R.sup.4 is methoxy, [1045] Ar is not unsubstituted
napthyl. 99. A compound according to statement 88, with the proviso
that when: [1046] R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are H,
[1047] Ar is not 2,4,6-trimethylphenyl. 100. A compound according
to statement 88, with the proviso that when: [1048] R.sup.1,
R.sup.2 and R.sup.4 are H, and R.sup.3 is CF.sub.3, [1049] Ar is
not 2-(difluromethoxy)phenyl. 101. A compound according to
statement 88, with the proviso that when: [1050] R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 are H, [1051] Ar is not
4-fluoro-3-methyl-phenyl. 102. A compound for use according to
statement 88, with the proviso that when: [1052] R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 are H, [1053] Ar is not 4-aminophenyl. 103. A
method of synthesis of a compound as defined in any one of
statements 88 to 102. 104. A compound for use according to
statement 1, wherein: R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are
independently selected from: [1054] (i) H; [1055] (ii) C.sub.1-3
alkyl, optionally substituted by: hydroxy, C.sub.1-2 alkoxy,
NH.sub.2, phenyl, C.sub.5-6 heteroaryl, C.sub.1-4 alkyl carbamoyl,
acylamido, or one or more fluoro groups; [1056] (iii) C.sub.1-3
alkoxy, optionally substituted by C.sub.3-6 cycloalkyl or by one of
more fluoro groups; [1057] (iv) C.sub.3-6 cycloalkyl; [1058] (v)
halo; [1059] (vi) COR.sup.C, where R.sup.C is selected from
NR.sup.N1R.sup.N2, where R.sup.N1 and R.sup.N2 are independently
selected from H and methyl; [1060] (vii) cyano, NH.sub.2, NO.sub.2;
[1061] (viii) phenyl or C.sub.5-6 heteroaryl, optionally
substituted by methyl, hydroxy or methoxy; Ar is a phenyl, napthyl,
or C.sub.5-10 heteroaryl group, which groups are optionally
substituted by one or more groups selected from: [1062] (i)
C.sub.1-4 alkyl, optionally substituted by hydroxy, C.sub.1-2
alkoxy, NH.sub.2, C.sub.1-4alkyl carbamoyl, or by one or more
fluoro groups; [1063] (ii) C.sub.3-6 cycloalkyl; [1064] (iii)
hydroxy; cyano; NR.sup.N3R.sup.N4, where R.sup.N3 and R.sup.N4 are
independently selected from H and methyl; acylamido; [1065] (iv)
halo; [1066] (v) C.sub.1-3 alkoxy, optionally substituted by
hydroxy, C(O)NH.sub.2, C.sub.3-6 cycloalkyl, phenyl, C.sub.5-6
heteroaryl, or by one of more fluoro groups; [1067] (vi) phenoxy,
optionally substituted by fluoro; [1068] (vii) phenyl, C.sub.5-6
heteroaryl [1069] (viii) SF.sub.5, SO.sub.2CH.sub.3; [1070] (ix)
--(CH.sub.2).sub.n--Y--, where Y is O or CH.sub.2, and n is 2 or 3;
105. A compound for use according to statement 104, wherein at
least one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is not H. 106. A
compound for use according to either statement 104 or 105, wherein
at least one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is C.sub.1-3
alkyl, optionally substituted by: hydroxy, C.sub.1-2 alkoxy,
NH.sub.2, phenyl, C.sub.5-6 heteroaryl, C.sub.1-4 alkyl carbamoyl,
acylamido, or one or more fluoro groups. 107. A compound for use
according to any one of statements 104 to 106, wherein at least one
of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is C.sub.1-3 alkoxy,
optionally substituted by C.sub.3-6 cycloalkyl or by one of more
fluoro groups. 108. A compound for use according to any one of
statements 104 to 107, wherein at least one of R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 is C.sub.3-6 cycloalkyl. 109. A compound for
use according to any one of statements 104 to 108, wherein at least
one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is halo. 110. A
compound for use according to any one of statements 104 to 109,
wherein at least one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is
COR.sup.C, where R.sup.C is selected from NR.sup.N1R.sup.N2, where
R.sup.N1 and R.sup.N2 are independently selected from H and methyl.
111. A compound for use according to any one of statements 104 to
110, wherein at least one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4
is selected from cyano, NH.sub.2 and NO.sub.2. 112. A compound for
use according to any one of statements 104 to 111, wherein at least
one of R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is phenyl or C.sub.5-6
heteroaryl, optionally substituted by methyl, hydroxy or methoxy.
113. A compound for use according to statement 104, wherein: (a)
R.sup.4 is methoxy, R.sup.2 is CH.sub.2OCH.sub.3 or
CH.sub.2OCH.sub.2CH.sub.3 and R.sup.1 and R.sup.3 are H; (b)
R.sup.4 is methoxy, R.sup.2 is phenyl, optionally substituted by
methyl or methoxy, and R.sup.1 and R.sup.3 are H; (c) R.sup.4 is
methoxy, R.sup.2 is C.sub.5-6 heteroaryl, optionally substituted by
methyl; (d) R.sup.4 is methoxy and R.sup.1, R.sup.2 and R.sup.3 are
H; (e) R.sup.4 is chloro, R.sup.2 is C.sub.1-3 alkyl or bromo, and
R.sup.1 and R.sup.3 are H; (f) R.sup.4 is chloro and R.sup.1,
R.sup.2 and R.sup.4 are H; or (g) R.sup.3 is C.sub.1-3 alkyl and
R.sup.1, R.sup.2 and R.sup.4 are H. 114. A compound for use
according to any one of statements 104 to 113, wherein Ar is:
[1071] (a) phenyl, which may be unsubstituted or substituted;
[1072] (b) napthyl, which may be unsubstituted or substituted; or
[1073] (c) a C.sub.5-10 heteroaryl group, which may be
unsubstituted or substituted. 115. A compound for use according to
statement 114, wherein Ar is substituted by C.sub.1-4 alkyl,
optionally substituted by hydroxy, C.sub.1-2 alkoxy, NH.sub.2,
C.sub.1-4 alkyl carbamoyl, or by one or more fluoro groups. 116. A
compound for use according to statement 114, wherein Ar is
substituted by C.sub.3-6 cycloalkyl. 117. A compound for use
according to statement 114, wherein Ar is substituted by hydroxy;
cyano; NR.sup.N3R.sup.N4, where R.sup.N3 and R.sup.N4 are
independently selected from H and methyl; or acylamido. 118. A
compound for use according to statement 114, wherein Ar is
substituted by halo. 119. A compound for use according to statement
114, wherein Ar is substituted by C.sub.1-3 alkoxy, optionally
substituted by hydroxy, C(O)NH.sub.2, C.sub.3-6 cycloalkyl, phenyl,
C.sub.5-6 heteroaryl, or by one of more fluoro groups. 120. A
compound for use according to statement 114, wherein Ar is
substituted by phenoxy, optionally substituted by fluoro. 121. A
compound for use according to statement 114, wherein Ar is
substituted by phenyl or C.sub.5-6 heteroaryl. 122. A compound for
use according to statement 114, wherein Ar is substituted by
SF.sub.5 or SO.sub.2CH.sub.3. 123. A compound for use according to
statement 114, wherein Ar is substituted by
--(CH.sub.2).sub.n--Y--, where Y is O or CH.sub.2, and n is 2 or 3.
124. A compound for use according to any one of statements 104 to
113, wherein Ar is represented by the formula (Ar-1):
##STR00382##
[1073] where Y is either N or C--R.sup.A4, and Z is either N or
C--R.sup.A5; and R.sup.A1, R.sup.A2, R.sup.A3, R.sup.A4 (if
present) and R.sup.A5 (if present) are independently selected from
H and the optional substituents for Ar. 125. A compound for use
according to statement 124, wherein: (a) R.sup.A2 is ethyl; or (b)
R.sup.A3 is selected from cycloalkyl; phenoxy; phenyl; C.sub.5-6
heteroaryl; SF.sub.5; and SO.sub.2CH.sub.3. 126. A compound for use
according to any one of statements 104 to 113, wherein Ar is: (a)
5-ethyl-2-methoxyphenyl; (b) 5-CF.sub.3-2-methoxyphenyl; or (c)
2,6-dimethoxyphenyl. 127. A compound as defined in any one of
statements 104 to 126 or a pharmaceutical salt thereof. 128. A
compound according to statement 127, wherein: (a) at least one of
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 is not H; (b) R.sup.3 is not
CF.sub.3; (c) R.sup.3 is not substituted C.sub.1-3 alkyl; (d)
R.sup.3 is ethyl or propyl; (e) R.sup.3 is not C.sub.1-3 alkyl,
optionally substituted by hydroxy, C.sub.1-2 alkoxy, NH.sub.2,
phenyl, C.sub.5-6 heteroaryl, C.sub.1-4 alkyl carbamoyl, acylamido
or by one or more fluoro groups; (f) R.sup.4 is methoxy; (g)
R.sup.4 is Cl, and R.sup.1, R.sup.2 and R.sup.3 are H; (h) R.sup.4
is Cl, and R.sup.2 is C.sub.1-3 alkyl or bromo, and R.sup.1 and
R.sup.3 are H; or (i) R.sup.3 is C.sub.1-3 alkyl and R.sup.1,
R.sup.2 and R.sup.4 are H.
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Sequence CWU 1
1
41480PRTArtificial SequenceHuman KAT5 isoform 1Met Gly His His His
His His His Gly Thr Glu Asn Leu Tyr Phe Gln1 5 10 15Gly Ser Ala Glu
Val Gly Glu Ile Ile Glu Gly Cys Arg Leu Pro Val 20 25 30Leu Arg Arg
Asn Gln Asp Asn Glu Asp Glu Trp Pro Leu Ala Glu Ile 35 40 45Leu Ser
Val Lys Asp Ile Ser Gly Arg Lys Leu Phe Tyr Val His Tyr 50 55 60Ile
Asp Phe Asn Lys Arg Leu Asp Glu Trp Val Thr His Glu Arg Leu65 70 75
80Asp Leu Lys Lys Ile Gln Phe Pro Lys Lys Glu Ala Lys Thr Pro Thr
85 90 95Lys Asn Gly Leu Pro Gly Ser Arg Pro Gly Ser Pro Glu Arg Glu
Val 100 105 110Lys Arg Lys Val Glu Val Val Ser Pro Ala Thr Pro Val
Pro Ser Glu 115 120 125Thr Ala Pro Ala Ser Val Phe Pro Gln Asn Gly
Ala Ala Arg Arg Ala 130 135 140Val Ala Ala Gln Pro Gly Arg Lys Arg
Lys Ser Asn Cys Leu Gly Thr145 150 155 160Asp Glu Asp Ser Gln Asp
Ser Ser Asp Gly Ile Pro Ser Ala Pro Arg 165 170 175Met Thr Gly Ser
Leu Val Ser Asp Arg Ser His Asp Asp Ile Val Thr 180 185 190Arg Met
Lys Asn Ile Glu Cys Ile Glu Leu Gly Arg His Arg Leu Lys 195 200
205Pro Trp Tyr Phe Ser Pro Tyr Pro Gln Glu Leu Thr Thr Leu Pro Val
210 215 220Leu Tyr Leu Cys Glu Phe Cys Leu Lys Tyr Gly Arg Ser Leu
Lys Cys225 230 235 240Leu Gln Arg His Leu Thr Lys Cys Asp Leu Arg
His Pro Pro Gly Asn 245 250 255Glu Ile Tyr Arg Lys Gly Thr Ile Ser
Phe Phe Glu Ile Asp Gly Arg 260 265 270Lys Asn Lys Ser Tyr Ser Gln
Asn Leu Cys Leu Leu Ala Lys Cys Phe 275 280 285Leu Asp His Lys Thr
Leu Tyr Tyr Asp Thr Asp Pro Phe Leu Phe Tyr 290 295 300Val Met Thr
Glu Tyr Asp Cys Lys Gly Phe His Ile Val Gly Tyr Phe305 310 315
320Ser Lys Glu Lys Glu Ser Thr Glu Asp Tyr Asn Val Ala Cys Ile Leu
325 330 335Thr Leu Pro Pro Tyr Gln Arg Arg Gly Tyr Gly Lys Leu Leu
Ile Glu 340 345 350Phe Ser Tyr Glu Leu Ser Lys Val Glu Gly Lys Thr
Gly Thr Pro Glu 355 360 365Lys Pro Leu Ser Asp Leu Gly Leu Leu Ser
Tyr Arg Ser Tyr Trp Ser 370 375 380Gln Thr Ile Leu Glu Ile Leu Met
Gly Leu Lys Ser Glu Ser Gly Glu385 390 395 400Arg Pro Gln Ile Thr
Ile Asn Glu Ile Ser Glu Ile Thr Ser Ile Lys 405 410 415Lys Glu Asp
Val Ile Ser Thr Leu Gln Tyr Leu Asn Leu Ile Asn Tyr 420 425 430Tyr
Lys Gly Gln Tyr Ile Leu Thr Leu Ser Glu Asp Ile Val Asp Gly 435 440
445His Glu Arg Ala Met Leu Lys Arg Leu Leu Arg Ile Asp Ser Lys Cys
450 455 460Leu His Phe Thr Pro Lys Asp Trp Ser Lys Arg Gly Lys Trp
Ala Ser465 470 475 4802791PRTArtificial SequenceHuman KAT6A isoform
2Met Asn Lys Glu Ile Leu Ala Val Val Glu Ala Val Ser Asn Glu Lys1 5
10 15Ala Leu Pro Arg Glu Lys Ile Phe Glu Ala Leu Glu Ser Ala Leu
Ala 20 25 30Thr Ala Thr Lys Lys Lys Tyr Glu Gln Glu Ile Asp Val Arg
Val Gln 35 40 45Ile Asp Arg Lys Ser Gly Asp Phe Asp Thr Phe Arg Arg
Trp Leu Val 50 55 60Val Asp Glu Val Thr Gln Pro Thr Lys Glu Ile Thr
Leu Glu Ala Ala65 70 75 80Arg Tyr Glu Asp Glu Ser Leu Asn Leu Gly
Asp Tyr Val Glu Asp Gln 85 90 95Ile Glu Ser Val Thr Phe Asp Arg Ile
Thr Thr Gln Thr Ala Lys Gln 100 105 110Val Ile Val Gln Lys Val Arg
Glu Ala Glu Arg Ala Met Val Val Asp 115 120 125Gln Phe Arg Glu His
Glu Gly Glu Ile Ile Thr Gly Val Val Lys Lys 130 135 140Val Asn Arg
Asp Asn Ile Ser Leu Asp Leu Gly Asn Asn Ala Glu Ala145 150 155
160Val Ile Leu Arg Glu Asp Met Leu Pro Arg Glu Asn Phe Arg Pro Gly
165 170 175Asp Arg Val Arg Gly Val Leu Tyr Ser Val Arg Pro Glu Ala
Arg Gly 180 185 190Ala Gln Leu Phe Val Thr Arg Ser Lys Pro Glu Met
Leu Ile Glu Leu 195 200 205Phe Arg Ile Glu Val Pro Glu Ile Gly Glu
Glu Val Ile Glu Ile Lys 210 215 220Ala Ala Ala Arg Asp Pro Gly Ser
Arg Ala Lys Ile Ala Val Lys Thr225 230 235 240Asn Asp Lys Arg Ile
Asp Pro Val Gly Ala Cys Val Gly Met Arg Gly 245 250 255Ala Arg Val
Gln Ala Val Ser Thr Glu Leu Gly Gly Glu Arg Ile Asp 260 265 270Ile
Val Leu Trp Asp Asp Asn Pro Ala Gln Phe Val Ile Asn Ala Met 275 280
285Ala Pro Ala Asp Val Ala Ser Ile Val Val Asp Glu Asp Lys His Thr
290 295 300Met Asp Ile Ala Val Glu Ala Gly Asn Leu Ala Gln Ala Ile
Gly Arg305 310 315 320Asn Gly Gln Asn Val Arg Leu Ala Ser Gln Leu
Ser Gly Trp Glu Leu 325 330 335Asn Val Met Thr Val Asp Asp Leu Gln
Ala Lys His Gln Ala Glu Ala 340 345 350His Ala Ala Ile Asp Thr Phe
Thr Lys Tyr Leu Asp Ile Asp Glu Asp 355 360 365Phe Ala Thr Val Leu
Val Glu Glu Gly Phe Ser Thr Leu Glu Glu Leu 370 375 380Ala Tyr Val
Pro Met Lys Glu Leu Leu Glu Ile Glu Gly Leu Asp Glu385 390 395
400Pro Thr Val Glu Ala Leu Arg Glu Arg Ala Lys Asn Ala Leu Ala Thr
405 410 415Ile Ala Gln Ala Gln Glu Glu Ser Leu Gly Asp Asn Lys Pro
Ala Asp 420 425 430Asp Leu Leu Asn Leu Glu Gly Val Asp Arg Asp Leu
Ala Phe Lys Leu 435 440 445Ala Ala Arg Gly Val Cys Thr Leu Glu Asp
Leu Ala Glu Gln Gly Ile 450 455 460Asp Asp Leu Ala Asp Ile Glu Gly
Leu Thr Asp Glu Lys Ala Gly Ala465 470 475 480Leu Ile Met Ala Ala
Arg Asn Ile Cys Trp Phe Gly Asp Glu Ala Thr 485 490 495Ser Gly Ser
Gly His His His His His His Ser Ala Gly Glu Asn Leu 500 505 510Tyr
Phe Gln Gly Ala Met Gly Arg Cys Pro Ser Val Ile Glu Phe Gly 515 520
525Lys Tyr Glu Ile His Thr Trp Tyr Ser Ser Pro Tyr Pro Gln Glu Tyr
530 535 540Ser Arg Leu Pro Lys Leu Tyr Leu Cys Glu Phe Cys Leu Lys
Tyr Met545 550 555 560Lys Ser Arg Thr Ile Leu Gln Gln His Met Lys
Lys Cys Gly Trp Phe 565 570 575His Pro Pro Val Asn Glu Ile Tyr Arg
Lys Asn Asn Ile Ser Val Phe 580 585 590Glu Val Asp Gly Asn Val Ser
Thr Ile Tyr Cys Gln Asn Leu Cys Leu 595 600 605Leu Ala Lys Leu Phe
Leu Asp His Lys Thr Leu Tyr Tyr Asp Val Glu 610 615 620Pro Phe Leu
Phe Tyr Val Leu Thr Gln Asn Asp Val Lys Gly Cys His625 630 635
640Leu Val Gly Tyr Phe Ser Lys Glu Lys His Cys Gln Gln Lys Tyr Asn
645 650 655Val Ser Cys Ile Met Ile Leu Pro Gln Tyr Gln Arg Lys Gly
Tyr Gly 660 665 670Arg Phe Leu Ile Asp Phe Ser Tyr Leu Leu Ser Lys
Arg Glu Gly Gln 675 680 685Ala Gly Ser Pro Glu Lys Pro Leu Ser Asp
Leu Gly Arg Leu Ser Tyr 690 695 700Met Ala Tyr Trp Lys Ser Val Ile
Leu Glu Cys Leu Tyr His Gln Asn705 710 715 720Asp Lys Gln Ile Ser
Ile Lys Lys Leu Ser Lys Leu Thr Gly Ile Cys 725 730 735Pro Gln Asp
Ile Thr Ser Thr Leu His His Leu Arg Met Leu Asp Phe 740 745 750Arg
Ser Asp Gln Phe Val Ile Ile Arg Arg Glu Lys Leu Ile Gln Asp 755 760
765His Met Ala Lys Leu Gln Leu Asn Leu Arg Pro Val Asp Val Asp Pro
770 775 780Glu Cys Leu Arg Trp Thr Pro785 7903307PRTArtificial
SequenceHuman KAT7 isoform 3Met Gly His His His His His His Gly Thr
Glu Asn Leu Tyr Phe Gln1 5 10 15Gly Ser Arg Leu Gln Gly Gln Ile Thr
Glu Gly Ser Asn Met Ile Lys 20 25 30Thr Ile Ala Phe Gly Arg Tyr Glu
Leu Asp Thr Trp Tyr His Ser Pro 35 40 45Tyr Pro Glu Glu Tyr Ala Arg
Leu Gly Arg Leu Tyr Met Cys Glu Phe 50 55 60Cys Leu Lys Tyr Met Lys
Ser Gln Thr Ile Leu Arg Arg His Met Ala65 70 75 80Lys Cys Val Trp
Lys His Pro Pro Gly Asp Glu Ile Tyr Arg Lys Gly 85 90 95Ser Ile Ser
Val Phe Glu Val Asp Gly Lys Lys Asn Lys Ile Tyr Cys 100 105 110Gln
Asn Leu Cys Leu Leu Ala Lys Leu Phe Leu Asp His Lys Thr Leu 115 120
125Tyr Tyr Asp Val Glu Pro Phe Leu Phe Tyr Val Met Thr Glu Ala Asp
130 135 140Asn Thr Gly Cys His Leu Ile Gly Tyr Phe Ser Lys Glu Lys
Asn Ser145 150 155 160Phe Leu Asn Tyr Asn Val Ser Cys Ile Leu Thr
Met Pro Gln Tyr Met 165 170 175Arg Gln Gly Tyr Gly Lys Met Leu Ile
Asp Phe Ser Tyr Leu Leu Ser 180 185 190Lys Val Glu Glu Lys Val Gly
Ser Pro Glu Arg Pro Leu Ser Asp Leu 195 200 205Gly Leu Ile Ser Tyr
Arg Ser Tyr Trp Lys Glu Val Leu Leu Arg Tyr 210 215 220Leu His Asn
Phe Gln Gly Lys Glu Ile Ser Ile Lys Glu Ile Ser Gln225 230 235
240Glu Thr Ala Val Asn Pro Val Asp Ile Val Ser Thr Leu Gln Ala Leu
245 250 255Gln Met Leu Lys Tyr Trp Lys Gly Lys His Leu Val Leu Lys
Arg Gln 260 265 270Asp Leu Ile Asp Glu Trp Ile Ala Lys Glu Ala Lys
Arg Ser Asn Ser 275 280 285Asn Lys Thr Met Asp Pro Ser Cys Leu Lys
Trp Thr Pro Pro Lys Gly 290 295 300Thr Ala Ser3054296PRTArtificial
SequenceHuman KAT8 isoform 4Met Ser Tyr Tyr His His His His His His
Asp Tyr Asp Ile Pro Thr1 5 10 15Thr Glu Asn Leu Tyr Phe Gln Gly Ala
Lys Tyr Val Asp Lys Ile His 20 25 30Ile Gly Asn Tyr Glu Ile Asp Ala
Trp Tyr Phe Ser Pro Phe Pro Glu 35 40 45Asp Tyr Gly Lys Gln Pro Lys
Leu Trp Leu Cys Glu Tyr Cys Leu Lys 50 55 60Tyr Met Lys Tyr Glu Lys
Ser Tyr Arg Phe His Leu Gly Gln Cys Gln65 70 75 80Trp Arg Gln Pro
Pro Gly Lys Glu Ile Tyr Arg Lys Ser Asn Ile Ser 85 90 95Val Tyr Glu
Val Asp Gly Lys Asp His Lys Ile Tyr Cys Gln Asn Leu 100 105 110Cys
Leu Leu Ala Lys Leu Phe Leu Asp His Lys Thr Leu Tyr Phe Asp 115 120
125Val Glu Pro Phe Val Phe Tyr Ile Leu Thr Glu Val Asp Arg Gln Gly
130 135 140Ala His Ile Val Gly Tyr Phe Ser Lys Glu Lys Glu Ser Pro
Asp Gly145 150 155 160Asn Asn Val Ala Cys Ile Leu Thr Leu Pro Pro
Tyr Gln Arg Arg Gly 165 170 175Tyr Gly Lys Phe Leu Ile Ala Phe Ser
Tyr Glu Leu Ser Lys Leu Glu 180 185 190Ser Thr Val Gly Ser Pro Glu
Lys Pro Leu Ser Asp Leu Gly Lys Leu 195 200 205Ser Tyr Arg Ser Tyr
Trp Ser Trp Val Leu Leu Glu Ile Leu Arg Asp 210 215 220Phe Arg Gly
Thr Leu Ser Ile Lys Asp Leu Ser Gln Met Thr Ser Ile225 230 235
240Thr Gln Asn Asp Ile Ile Ser Thr Leu Gln Ser Leu Asn Met Val Lys
245 250 255Tyr Trp Lys Gly Gln His Val Ile Cys Val Thr Pro Lys Leu
Val Glu 260 265 270Glu His Leu Lys Ser Ala Gln Tyr Lys Lys Pro Pro
Ile Thr Val Asp 275 280 285Ser Val Cys Leu Lys Trp Ala Pro 290
295
* * * * *