Language selection

Search

Patent 2836085 Summary

Third-party information liability

Some of the information on this Web page has been provided by external sources. The Government of Canada is not responsible for the accuracy, reliability or currency of the information supplied by external sources. Users wishing to rely upon this information should consult directly with the source of the information. Content provided by external sources is not subject to official languages, privacy and accessibility requirements.

Claims and Abstract availability

Any discrepancies in the text and image of the Claims and Abstract are due to differing posting times. Text of the Claims and Abstract are posted:

  • At the time the application is open to public inspection;
  • At the time of issue of the patent (grant).
(12) Patent Application: (11) CA 2836085
(54) English Title: SUBSTITUTED 2-BENZYLIDENE-2H-BENZO[B][1,4] THIAZIN-3(4H)-ONES, DERIVATIVES THEREOF, AND THERAPEUTIC USES THEREOF
(54) French Title: 2-BENZYLIDENE-2H-BENZO[B][1,4] THIAZIN-3(4H)-ONES SUBSTITUEES, DERIVES ET USAGE THERAPEUTIQUE DE CELLES-CI
Status: Deemed Abandoned and Beyond the Period of Reinstatement - Pending Response to Notice of Disregarded Communication
Bibliographic Data
(51) International Patent Classification (IPC):
  • C07D 279/16 (2006.01)
  • A61K 31/5415 (2006.01)
  • A61K 47/68 (2017.01)
  • A61P 35/00 (2006.01)
  • A61P 35/02 (2006.01)
  • C07D 417/06 (2006.01)
  • C07D 417/10 (2006.01)
  • C07K 14/025 (2006.01)
  • C07K 16/00 (2006.01)
  • C07K 16/28 (2006.01)
  • C07K 16/40 (2006.01)
  • C12N 9/02 (2006.01)
  • C12N 15/37 (2006.01)
  • C12N 15/53 (2006.01)
(72) Inventors :
  • REDDY, E. PREMKUMAR (United States of America)
  • REDDY, M. V. RAMANA (United States of America)
(73) Owners :
  • TEMPLE UNIVERSITY - OF THE COMMONWEALTH SYSTEM OF HIGHER EDUCATION
  • ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
(71) Applicants :
  • TEMPLE UNIVERSITY - OF THE COMMONWEALTH SYSTEM OF HIGHER EDUCATION (United States of America)
  • ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI (United States of America)
(74) Agent: BORDEN LADNER GERVAIS LLP
(74) Associate agent:
(45) Issued:
(22) Filed Date: 2013-12-05
(41) Open to Public Inspection: 2015-06-05
Availability of licence: N/A
Dedicated to the Public: N/A
(25) Language of filing: English

Patent Cooperation Treaty (PCT): No

(30) Application Priority Data: None

Abstracts

English Abstract


The present invention relates to compounds according to Formula I:
(see formula I)
and salts thereof, wherein R1, R2, R3, R4, Ar, and n are as defined herein.
Methods for
preparing compounds of Formula I are also provided. The present invention
further includes
methods of treating cellular proliferative disorders, such as cancer, with the
compounds of
Formula I.


Claims

Note: Claims are shown in the official language in which they were submitted.


CLAIMS:
1. A compound of Formula I or a salt thereof:
<IMG>
wherein
n is 0, 1, or 2;
R1 is selected from the group consisting of -H, -(C1-C6)alkyl, -(C2-
C6)alkenyl,
-(C2-C6)alkynyl, optionally substituted aryl, optionally substituted
heteroaryl, optionally
substituted aryl-(C1-C6)alkyl, optionally substituted heteroaryl-(C1-C6)alkyl,
-C(=O)(C1-
C6)alkyl, -C(=O)(C2-C6)alkenyl, -C(=O)-optionally substituted aryl, -
C(=O)(CH2)m-optionally
substituted aryl, and -C(=O)(CH2)p-optionally substituted heteroaryl;
R2, R3, and R4 are independently selected from the group consisting of -H,
halogen,
-CN, -NR10R11, -OH, -OR13, -(C1-C6)alkoxy, -NO2, -(C1-C6)alkyl, -(C1-
C6)perfluoroalkyl,
-(C1-C6)perfluoroalkoxy,
-C(=O)NR17R18, -SH, -S(C1-C6)alkyl, -SR13, -S(=O)R13, -S(=O)2R3, -OS(=O)2R13,
-S(=O)q R15, -OS(=O)q R15, -S(=O)2NR17R18, -S(=O)NR17R18, optionally
substituted aryl,
optionally substituted aryl-(C1-C6)alkyl, optionally substituted heteroaryl,
optionally
substituted heteroaryl-(C1-C6)alkyl, optionally substituted (C2-
C9)heterocyclyl, optionally
substituted (C2-C9)heterocyclyl-(C1-C6)alkyl, -NH(CH2)m C(=O)OR14, -
C(=NR14)NR14 2,
-C(=N-OR14)NR14 2, -P(=O)(OR14)2, and -OP(=O)(OR14)2;
104

Ar is optionally substituted heteroaryl, optionally substituted (C10-C14)aryl,
or
<IMG>
wherein
R5, R6, R7, R8, and R9 are independently selected from the group consisting of
-H,
-OH, -OR13, -NO2, halogen, -CN, -NR10R11, -(CH2)m NR10R11, -O(CH2)m NR10R11, -
(C1-
C6)alkyl, -(CH2)m O(C1-C6)alkyl, -(C1-C6)alkoxy, -(C1-C6)perfluoroalkyl, -(C1-
C6)perfluoroalkoxy, -SH, -S(C1-C6)alkyl, -SR13, -S(=O)R15, -S(=O)2R15,
-C(=O)OR15, -C(=O)NR17R18, -OC(=O)R16, -OC(=O)OR12, -OC(=O)NR17R18,
heterocyclyl,
optionally substituted heteroaryl, -NH(CH2)m C(=O)OR14, -OS(=O)2R16,
-C(=NR14)NR14 2, -C(=N-OR14)NR14 2, -P(=O)(OR14)2, and -OP(=O)(OR14)2;
each R10 and R11 is independently selected from the group consisting of -H,
-(C1-C6)alkyl, -(C1-C6)alkoxy, -C(=O)R12, -C(=O)NR17R18, -C(=O)OR12, -
C(=NR14)NR17R18,
R13, optionally substituted aryl, optionally substituted heteroaryl, and -
C(=NR14)R15; or R10
and R11, together with the nitrogen to which they are bound, form an
optionally substituted
(C2-C5)heterocycle;
each R12 is independently selected from the group consisting of -(C1-C6)alkyl,
and
optionally substituted aryl;
each R13 is independently selected from the group consisting of optionally
substituted
aryl and -(CH2)m R16;
105

each R14 is independently selected from the group consisting of -H and -(C1-
C6)alkyl;
or two occurrences of R14 bound to the same nitrogen form a (C2-
C6)heterocycle, together
with the nitrogen atom to which they are bound;
each R15 is independently selected from the group consisting of -H, -(C1-
C6)alkyl,
optionally substituted aryl, and NR14 2;
each R16 is independently selected from the group consisting of -(C1-C6)alkyl,
-NR14 2,
and Ar1;
each R17 and R18 is independently selected from the group consisting of -H,
-(C1-C6)alkyl, -(C1-C6)alkoxy, R13, optionally substituted aryl, and
optionally substituted
heteroaryl; or R17 and R18, together with the nitrogen to which they are
bound, form an
optionally substituted (C2-C5)heterocycle;
m is independently at each occurrence 1 , 2, 3, 4, or 5;
p is independently at each occurrence 0, 1, 2, or 3;
q is independently at each occurrence 0, 1, or 2;
each optionally substituted aryl, optionally substituted (C10-C14)aryl,
optionally
substituted heteroaryl, optionally substituted aryl-(C1-C6)alkyl, optionally
substituted
heteroaryl-(C1-C6)alkyl, optionally substituted (C2-C9)heterocyclyl,
optionally substituted (C2-
C9)heterocyclyl-(C1-C6)alkyl, and optionally substituted (C2-C5)heterocycle is
optionally
substituted with one or more substituents independently selected from the
group consisting of
halogen, -CN, -NR14 2, -(CH2)NR14 2, -O(CH2).NR14 2, -NR14C(=O)(C1-C6)alkyl,
-NR14C(=O)NR14 2, -NR14C(=NR14)NR14 2,
-NH(CH2)m C(=O)OR14, -OH, -NO2, -(C1-C6)alkyl, -(CH2)m O(C1-C6)alkyl, -(C1-
C6)alkoxy,
-(C2-C6)alkenyl, -(C2-C6)alkynyl, -SR14, -S(=O)R15, -S(=O)2R15, -
NR14S(=O)2R15, -(C1-
C6)perfluoroalkyl, -(C1-C6)perfluoroalkoxy, -C(=O)R14, -C(=O)OR14, -C(=O)NR14
2,
-OC(=O)R14, -OC(=O)NR14 2, -OC(=O)O(C1-C6)alkyl, -P(=O)(OR14)2, -
OP(=O)(OR14)2,
heterocyclyl, and heteroaryl;
Ar1 is a radical according to Formula II:
106

<IMG>
wherein R19,R20,R21,R22, and R23 are independently selected from the group
consisting of -H, -OH, -NO2, halogen, -CN,-NR10R11, -(CH2)m NR10R11, -O(CH2)m
NR10R11,
-(C1-C6)alkyl, -(CH2)m O(C1-C6)alkyl, -(C1-C6)alkoxy, -(C1-C6)perfluoroalkyl, -
(C1-
C6)perfluoroalkoxy, -SH, -SR12, -S(=O)R15, -S(=O)2R15, -C(=O)R15, -C(=O)OR15,
-C(=O)NR17R18, -OC(=O)R16, -OC(=O)OR12, -OC(=O)NR17R18, heterocyclyl,
optionally
substituted heteroaryl, -NH(CH2)m C(=O)OR14, - OS (=O)2R16,
-C(=NR14)NR14 2, -C(=N-OR14)NR14 2, -P(=O)(OR14)2, and -OP(=O)(OR14)2;
provided that:
i) at least one of R2, R3, or R4 is other than hydrogen;
ii) when none of R2, R3, and R4 are -OR13, -NHR13, -SR13, -S(=O)R13, or
-S(=O)2R13, and Ar is
<IMG>
then at least one of R6 and R8 is -NO2 and at least R7 is other than hydrogen
or halogen; and
iii) when Ar is optionally substituted heteroaryl and none of R2, R3, or R4
are -OR13,
-NHR13, -SR13, -S(=O)R13, or -S(=O)2R13, then R1 is other than hydrogen.
107

2. A compound according to claim 1, or a salt thereof, wherein n is 0.
3. A compound according to claim 2, or a salt thereof, wherein Ar is
optionally
substituted heteroaryl or
<IMG>
4. A compound according to claim 3, or a salt thereof, wherein at least one
of R2, R3, or
R4 is -OR13, -NHR13, -S(=O)R13, or -S(=O)2R13
5. A compound according to claim 4, or a salt thereof, wherein at least one
of R2, R3, or
R4 is -S(=O)2R13.
6. A compound according to claim 5, or a salt thereof, wherein R13 is -
(CH2)m R16.
7. A compound according to claim 5, or a salt thereof, wherein R2 is -
S(=O)2R13 and R13
is -(CH2)m R16.
8. A compound according to claim 7, or a salt thereof, wherein R16 is Ar1.
108

9. A compound according to claim 8, or a salt thereof, wherein Ar1 is 2,3-
dichlorophenyl,
2,4-dichlorophenyl, 2,5-dichlorophenyl, 2,6-dichlorophenyl, 3,4-
dichlorophenyl, 3,5-
dichlorophenyl, 2,3-dibromophenyl, 2,4-dibromophenyl, 2,5-dibromophenyl, 2,6-
dibromophenyl, 3,4-dibromophenyl, 3,5-dibromophenyl, 2,3-difluorophenyl, 2,4-
difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 3,4-difluorophenyl,
3,5-
difluorophenyl, 2,3-diiodophenyl, 2,4-diiodophenyl, 2,5-diiodophenyl, 2,6-
diiodophenyl, 3,4-
diiodophenyl, 3,5-diiodophenyl, 2-chloro-3-bromophenyl, 2-chloro-4-
bromophenyl, 2-chloro-
5-bromophenyl, 2-chloro-6-bromophenyl, 3-chloro-4-bromophenyl, 3-chloro-5-
bromophenyl,
4-chloro-5-bromophenyl, 2-bromo-3-chlorophenyl, 2-bromo-4-chlorophenyl, 2-
bromo-5-
chlorophenyl, 3-bromo-4-chlorophenyl, 2-chloro-3-fluorophenyl, 2-chloro-4-
fluorophenyl, 2-
chloro-5-fluorophenyl, 2-chloro-6-fluorophenyl, 3-chloro-4-fluorophenyl, 3-
chloro-5-
fluorophenyl, 4-chloro-5-fluorophenyl, 2-fluoro-3-chlorophenyl, 2-fluoro-4-
chlorophenyl, 2-
fluoro-5-chlorophenyl, 3-fluoro-4-chlorophenyl, 2-chloro-3-iodophenyl, 2-
chloro-4-
iodophenyl, 2-chloro-5-iodophenyl, 2-chloro-6-iodophenyl, 3-chloro-4-
iodophenyl, 3-chloro-
5-iodophenyl, 4-chloro-5-iodophenyl, 2-iodo-3-chlorophenyl, 2-iodo-4-
chlorophenyl, 2-iodo-
5-chlorophenyl, 3-iodo-4-chlorophenyl, 2-bromo-3-fluorophenyl, 2-bromo-4-
fluorophenyl, 2-
bromo-5-fluorophenyl, 2-bromo-6-fluorophenyl, 3-bromo-4-fluorophenyl, 3-bromo-
5-
fluorophenyl, 4-bromo-5-fluorophenyl, 2-fluoro-3-bromophenyl, 2-fluoro-4-
bromophenyl, 2-
fluoro-5-bromophenyl, 3-fluoro-4-bromophenyl, 2-bromo-3-iodophenyl, 2-bromo-4-
iodophenyl, 2-bromo-5-iodophenyl, 2-bromo-6-iodophenyl, 3-bromo-4-iodophenyl,
3-bromo-
5-iodophenyl, 4-bromo-5-iodophenyl, 2-iodo-3-bromophenyl, 2-iodo-4-
bromophenyl, 2-iodo-
5-bromophenyl, 3-iodo-4-bromophenyl, 2-fluoro-3-iodophenyl, 2-fluoro-4-
iodophenyl, 2-
fluoro-5-iodophenyl, 2-fluoro-6-iodophenyl, 3-fluoro-4-iodophenyl, 3-fluoro-5-
iodophenyl, 4-
fluoro-5-iodophenyl, 2-iodo-3-fluorophenyl, 2-iodo-4-fluorophenyl, 2-iodo-5-
fluorophenyl, or
3-iodo-4-fluorophenyl.
10. A compound according to claim 9, or a salt thereof, wherein Ar1 is 2,6-
dichlorophenyl.
11. A compound according to claim 10, or a salt thereof, wherein Ar is
109

<IMG>
12. A compound according to claim 11, or a salt thereof, wherein said
compound is
selected from the group consisting of (Z)-4-((6-((2,6-dichlorobenzyl)sulfonyl)-
3-oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-
((6-((2,6-
dichlorobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-6-((2,6-
dichlorobenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dichlorobenzyl)sulfonyl)-2-(4-
methoxy-3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(3-amino-4-
methoxybenzylidene)-6-((2,6-dichlorobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-6-((2,6-dichlorobenzyl)sulfonyl)-2-(4-(4-methylpiperazin-1-yl)benzylidene)-
2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dichlorobenzyl)sulfonyl)-2-(4-
nitrobenzylidene)-
2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-aminobenzylidene)-6-((2,6-
dichlorobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dichlorobenzyl)sulfonyl)-2-(4-fluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-
(4-chlorobenzylidene)-6-((2,6-dichlorobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-(4-bromobenzylidene)-6-((2,6-dichlorobenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-6-((2,6-dichlorobenzyl)sulfonyl)-2-(4-methoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dichlorobenzyl)sulfonyl)-2-(4-
methylbenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dichlorobenzyl)sulfonyl)-2-(2,4,6-trimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-6-((2,6-dichlorobenzyl)sulfonyl)-2-(2,4-dichlorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dichlorobenzyl)sulfonyl)-2-(2,4,6-
110

trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-methyl 2-((4-((6-
((2,6-
dichlorobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl)amino)acetate; (Z)-2-((4-((6-((2,6-
dichlorobenzyl)sulfonyl)-3-oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetic acid; (Z)-
6-((2,6-
dichlorobenzyl)sulfonyl)-2-(4-hydroxy-2,6-dimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-2-(4-chloro-3-nitrobenzylidene)-6-((2,6-
dichlorobenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dichlorobenzyl)sulfonyl)-2-(2,4-
difluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dichlorobenzyl)sulfonyl)-2-(2,4,6-trifluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-methyl 4-((6-((2,6-dichlorobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-
2-ylidene)methyl)benzoate; (Z)-methyl 4-((6-((2,6-dichlorobenzyl)sulfonyl)-3-
oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-3-nitrobenzoate; (Z)-4-((6-
((2,6-
dichlorobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-
nitrophenyl 4-methylbenzenesulfonate; (Z)-2-(4-(2H-tetrazol-5-yl)benzylidene)-
6-((2,6-
dichlorobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-
(benzyloxy)-3-
nitrobenzylidene)-6-((2,6-dichlorobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-
3(4H)-one; and
(Z)-6-((2,6-dichlorobenzyl)sulfonyl)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one.
13. The compound according to claim 12 which is (Z)-6-((2,6-
dichlorobenzyl)sulfonyl)-2-
(4-hydroxy-3-nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one, or salt
thereof.
14. A compound according to claim 10, or a salt thereof, wherein Ar is
optionally
substituted heteroaryl.
15. A compound according to claim 14, or a salt thereof, wherein said
compound is
selected from the group consisting of (Z)-6-((2,6-dichlorobenzyl)sulfonyl)-2-
((2-
(methylthio)pyrimidin-4-yl)methylene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-
((2,6-
111

dichlorobenzyl)sulfonyl)-2-((2-morpholinopyrimidin-4-yl)methylene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-((1H-indol-3-yl)methylene)-6-((2,6-
dichlorobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dichlorobenzyl)sulfonyl)-2-(thiophen-2-ylmethylene)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-((1H-pyrrol-3-yl)methylene)-6-((2,6-dichlorobenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; and (Z)-2-((1-acetyl-1H-indol-3-yl)methylene)-6-((2,6-
dichlorobenzyl)sulfonyl)-
2H-benzo[b][1,4]thiazin-3(4H)-one.
16. A compound according to claim 3, or a salt thereof, wherein at least
one of R6 and R8
is ¨NO2 and at least R7 is other than hydrogen or halogen.
17. A compound according to claim 16, or a salt thereof, wherein said
compound is
selected from the group consisting of (Z)-4-((6-chloro-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-((7-
methoxy-3-oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-
((7-bromo-3-
oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl
acetate; (Z)-4-((7-
methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-2-
nitrophenyl acetate;
(Z)-2-(4-acetoxy-3-nitrobenzylidene)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazine-7-
carboxylic acid; (Z)-4-((7-fluoro-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-((6-fluoro-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-((4-acetyl-
7-methoxy-3-
oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl
acetate; (Z)-2-(4-
(benzyloxy)-3-nitrobenzylidene)-7-fluoro-2H-benzo[b][1,4]thiazin-3(4H)-one;
and (Z)-7-
fluoro-2-(4-hydroxy-3-nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one.
18. A compound according to claim 9, or a salt thereof, selected from the
group consisting
of (Z)-4-((6-((2,6-dibromobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-((6-((2,6-dibromobenzyl)sulfonyl)-
3-oxo-3,4-
112

dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl acetate; (Z)-2-
benzylidene-6-((2,6-
dibromobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dibromobenzyl)sulfonyl)-2-(4-methoxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-(3-amino-4-methoxybenzylidene)-6-((2,6-dibromobenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dibromobenzyl)sulfonyl)-2-(4-(4-
methylpiperazin-1-yl)benzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-
((2,6-
dibromobenzyl)sulfonyl)-2-(4-nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-
(4-aminobenzylidene)-6-((2,6-dibromobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-6-((2,6-dibromobenzyl)sulfonyl)-2-(4-fluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-(4-chlorobenzylidene)-6-((2,6-dibromobenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-2-(4-bromobenzylidene)-6-((2,6-dibromobenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dibromobenzyl)sulfonyl)-2-(4-
methoxybenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dibromobenzyl)sulfonyl)-2-(4-methylbenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-
6-((2,6-dibromobenzyl)sulfonyl)-2-(2,4,6-trimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-6-((2,6-dibromobenzyl)sulfonyl)-2-(2,4-dibromobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dibromobenzyl)sulfonyl)-2-(2,4,6-
trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-methyl 2-((4-((6-
((2,6-
dibromobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl)amino)acetate; (Z)-2-((4-((6-((2,6-
dibromobenzyl)sulfonyl)-3-oxo-
3,4-dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetic acid;
(Z)-6-((2,6-
dibromobenzyl)sulfonyl)-2-(4-hydroxy-2,6-dimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-2-(4-chloro-3-nitrobenzylidene)-6-((2,6-dibromobenzyl)sulfonyl)-
2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dibromobenzyl)sulfonyl)-2-(2,4-
difluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dibromobenzyl)sulfonyl)-2-(2,4,6-trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-methyl 4-((6-((2,6-dibromobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-
2-ylidene)methyl)benzoate; (Z)-methyl 4-((6-((2,6-dibromobenzyl)sulfonyl)-3-
oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-3-nitrobenzoate; (Z)-4-((6-
((2,6-
dibromobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-
113

nitrophenyl 4-methylbenzenesulfonate; (Z)-2-(4-(2H-tetrazol-5-yl)benzylidene)-
6-((2,6-
dibromobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-
(benzyloxy)-3-
nitrobenzylidene)-6-((2,6-dibromobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-
3(4H)-one; (Z)-
6-((2,6-dibromobenzyl)sulfonyl)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-4-((6-((2,6-difluorobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-((6-((2,6-
difluorobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-6-((2,6-
difluorobenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-difluorobenzyl)sulfonyl)-2-(4-
methoxy-3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(3-amino-4-
methoxybenzylidene)-6-((2,6-difluorobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-6-((2,6-difluorobenzyl)sulfonyl)-2-(4-(4-methylpiperazin-1-yl)benzylidene)-
2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-difluorobenzyl)sulfonyl)-2-(4-
nitrobenzylidene)-
2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-aminobenzylidene)-6-((2,6-
difluorobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
difluorobenzyl)sulfonyl)-2-(4-fluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-
(4-chlorobenzylidene)-6-((2,6-difluorobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-(4-bromobenzylidene)-6-((2,6-difluorobenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-6-((2,6-difluorobenzyl)sulfonyl)-2-(4-methoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-difluorobenzyl)sulfonyl)-2-(4-
methylbenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
difluorobenzyl)sulfonyl)-2-(2,4,6-trimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-6-((2,6-difluorobenzyl)sulfonyl)-2-(2,4-difluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-difluorobenzyl)sulfonyl)-2-(2,4,6-
trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-methyl 2-((4-((6-
((2,6-
difluorobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl)amino)acetate; (Z)-2-((4-((6-((2,6-
difluorobenzyl)sulfonyl)-3-oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetic acid; (Z)-
6-((2,6-
difluorobenzyl)sulfonyl)-2-(4-hydroxy-2,6-dimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-2-(4-chloro-3-nitrobenzylidene)-6-((2,6-
difluorobenzyl)sulfonyl)-2H-
114

benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-difluorobenzyl)sulfonyl)-2-(2,4-
difluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-(2,6-
difluorobenzyl)sulfonyl)-2-(2,4,6-trifluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-methyl 4-((6-((2,6-difluorobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-
2-ylidene)methyl)benzoate; (Z)-methyl 4-((6-((2,6-difluorobenzyl)sulfonyl)-3-
oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-3-nitrobenzoate; (Z)-4-((6-
((2,6-
difluorobenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-
nitrophenyl 4-methylbenzenesulfonate; (Z)-2-(4-(2H-tetrazol-5-yl)benzylidene)-
6-((2,6-
difluorobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-
(benzyloxy)-3-
nitrobenzylidene)-6-((2,6-difluorobenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-
3(4H)-one; (Z)-
6-((2,6-difluorobenzyl)sulfonyl)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-4-((6-((2,6-dimethylbenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-((6-((2,6-
dimethylbenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-6-((2,6-
dimethylbenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethylbenzyl)sulfonyl)-2-(4-
methoxy-3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(3-amino-4-
methoxybenzylidene)-6-((2,6-dimethylbenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-6-(2,6-dimethylbenzyl)sulfonyl)-2-(4-(4-methylpiperazin-1-yl)benzylidene)-
2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethylbenzyl)sulfonyl)-2-(4-
nitrobenzylidene)-
2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-aminobenzylidene)-6-((2,6-
dimethylbenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dimethylbenzyl)sulfonyl)-2-(4-fluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-
(4-chlorobenzylidene)-6-((2,6-dimethylbenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-(4-bromobenzylidene)-6-((2,6-dimethylbenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-6-((2,6-dimethylbenzyl)sulfonyl)-2-(4-methoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethylbenzyl)sulfonyl)-2-(4-
methylbenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dimethylbenzyl)sulfonyl)-2-(2,4,6-trimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-6-(2,6-dimethylbenzyl)sulfonyl)-2-(2,4-dimethylbenzylidene)-2H-
115

benzo [b] [1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethylbenzyl)sulfonyl)-2-
(2,4,6-
trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-methyl 2-((4-((6-
((2,6-
dimethylbenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl)amino)acetate; (Z)-2-((4-((6-((2,6-
dimethylbenzyl)sulfonyl)-3-oxo-
3,4-dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetic acid;
(Z)-6-((2,6-
dimethylbenzyl)sulfonyl)-2-(4-hydroxy-2,6-dimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-2-(4-chloro-3-nitrobenzylidene)-6-((2,6-
dimethylbenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethylbenzyl)sulfonyl)-2-(2,4-
difluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dimethylbenzyl)sulfonyl)-2-(2,4,6-trifluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-methyl 4-((6-(2,6-dimethylbenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-
2-ylidene)methyl)benzoate; (Z)-methyl 4-((6-(2,6-dimethylbenzyl)sulfonyl)-3-
oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-3-nitrobenzoate; (Z)-4-((6-
((2,6-
dimethylbenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-
nitrophenyl 4-methylbenzenesulfonate; (Z)-2-(4-(2H-tetrazol-5-yl)benzylidene)-
6-(2,6-
dimethylbenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-
(benzyloxy)-3-
nitrobenzylidene)-6-((2,6-dimethylbenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-
3(4H)-one; and
(Z)-6-((2,6-dimethylbenzyl)sulfonyl)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-4-((6-((2,6-dimethoxybenzyl)sulfonyl)-3-
oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-
((6-((2,6-
dimethoxybenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-6-((2,6-
dimethoxybenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethoxybenzyl)sulfonyl)-2-(4-
methoxy-3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(3-amino-4-
methoxybenzylidene)-6-((2,6-dimethoxybenzyl)sulfonyl)-2H-benzo[b] [1
,4]thiazin-3 (4H)-
one; (Z)-6-((2,6-dimethoxybenzyl)sulfonyl)-2-(4-(4-methylpiperazin-1-
yl)benzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethoxybenzyl)sulfonyl)-2-(4-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-
aminobenzylidene)-6-(2,6-
dimethoxybenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dimethoxybenzyl)sulfonyl)-2-(4-fluorobenzylidene)-2H-benzo[b][1,4]thiazin-
3(4H)-one; (Z)-
116

2-(4-chlorobenzylidene)-6-((2,6-dimethoxybenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-(4-bromobenzylidene)-6-((2,6-dimethoxybenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethoxybenzyl)sulfonyl)-2-(4-
methoxybenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dimethoxybenzyl)sulfonyl)-2-(4-methylbenzylidene)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-6-(2,6-dimethoxybenzyl)sulfonyl)-2-(2,4,6-trimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-(2,6-dimethoxybenzyl)sulfonyl)-2-(2,4-
dimethoxybenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dimethoxybenzyl)sulfonyl)-2-(2,4,6-trifluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-methyl 2-((4-((6-(2,6-dimethoxybenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetate; (Z)-2-((4-((6-(2,6-
dimethoxybenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl)amino)acetic acid; (Z)-6-((2,6-dimethoxybenzyl)sulfonyl)-
2-(4-
hydroxy-2,6-dimethoxybenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-
chloro-3-
nitrobenzylidene)-6-((2,6-dimethoxybenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-6-(2,6-dimethoxybenzyl)sulfonyl)-2-(2,4-difluorobenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-6-((2,6-dimethoxybenzyl)sulfonyl)-2-(2,4,6-
trifluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-methyl 4-((6-(2,6-
dimethoxybenzyl)sulfonyl)-3-oxo-
3,4-dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)benzoate; (Z)-methyl 4-
((6-(2,6-
dimethoxybenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-3-
nitrobenzoate; (Z)-4-((6-((2,6-dimethoxybenzyl)sulfonyl)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl 4-methylbenzenesulfonate;
(Z)-2-(4-
(2H-tetrazol-5-yl)benzylidene)-6-(2,6-dimethoxybenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-2-(4-(benzyloxy)-3-nitrobenzylidene)-6-(2,6-
dimethoxybenzyl)sulfonyl)-2H-
benzo[b][1,4]thiazin-3(4H)-one; and (Z)-6-(2,6-dimethoxybenzyl)sulfonyl)-2-(4-
hydroxy-3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one.
19. A
process for preparing a compound of Formula I according to claim 1, said
process
comprising:
117

condensing an aldehyde of Formula III
<IMG>
with a compound according to Formula IV:
<IMG>
in a reaction mixture, wherein Ar, R1, R2, R3, R4, and n, are as defined in
claim 1; and
isolating from the reaction mixture the compound of Formula I according to
claim 1, or a salt
thereof.
20. The process according to claim 19, wherein said condensation takes
place in the
presence of one or more of an anhydride and a base.
21. The process according to claim 20, wherein said anhydride is acetic
anhydride and
said base is triethylamine.
22. The process according to claim 21, wherein said reaction mixture is
heated to reflux.
118

23. The process according to claim 19, wherein said compound according to
Formula IV
is prepared by:
a) functionalizing 2H-1,4-Benzothiazin-3(4H)-one with chlorosulfonic acid to
give 3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazine-6-sulfonyl chloride, or a
derivative thereof;
b) reacting said 3-oxo-3,4-dihydro-2H-benzo[b] [1 ,4]thiazine-6-sulfonyl
chloride
with sodium sulfite to give a reactive intermediate; and
c) reacting said reactive intermediate with a compound of Formula V:
<IMG>
to give a compound of Formula IV, wherein R19, R20, R21, R22, and R23 are as
defined in claim 1, and X is a leaving group.
24. The process according to claim 19, wherein said compound according to
Formula IV
is prepared by:
a) functionalizing 2H-1,4-Benzothiazin-3(4H)-one, or a derivative thereof,
with
chlorosulfonic acid to give 3-oxo-3,4-dihydro-2H-benzo[b][1,4jthiazine-6-
sulfonyl chloride;
b) reacting said 3-oxo-3,4-dihydro-2H-benzo [b][l ,4]thiazine-6-sulfonyl
chloride
with Zn to form a reactive Zn complex; and
c) reacting said reactive Zn complex with a compound of Formula V:
119

<IMG>
to give a compound of Formula IV, wherein R19, R20, R21, R22, and R23 are as
defined in claim
1, and X is a leaving group.
25. The
process according to claim 19, wherein said compound according to Formula IV
is prepared by:
a) reacting compound A:
<IMG>
with HSCH2CO2R, wherein R is H or (C1-C6)alkyl and R2, R3, and R4 are as
defined in claim 1, to form compound B:
<IMG>
120

b) reducing the nitro group of compound B to give said compound according to
Formula IV.
26. The process according to claim 25, wherein reducing the nitro group of
compound B
comprises treating compound B with Zn and acetic acid.
27. The process according to claim 26, wherein reducing the nitro group of
compound B
comprises treating compound B with Na2S2O4.
28. An antibody conjugate of the formula:
I-L-Ab,
or a salt thereof, wherein
I is a compound according to Formula I as defined in claim 1,
Ab is an antibody; and
-L- is a single bond or a linking group covalently linking said compound of
Formula I
to said antibody.
29. The antibody conjugate of claim 28, or a salt thereof, wherein the
antibody is a
monoclonal antibody or a monospecific polyclonal antibody.
30. The antibody conjugate of claim 29, or a salt thereof, wherein the
antibody is a tumor
specific antibody.
31. A pharmaceutical composition comprising a pharmaceutically acceptable
carrier and a
compound of Formula I, or a pharmaceutically acceptable salt thereof,
according to claim 1.
121

32. A pharmaceutical composition comprising a pharmaceutically acceptable
carrier and
an antibody conjugate according to claim 28, or a pharmaceutically acceptable
salt thereof.
33. A method of treating an individual suffering from a cellular
proliferative disorder,
comprising administering to the individual an effective amount of at least one
compound of
Formula I, or a pharmaceutically acceptable salt thereof, according to claim
1.
34. A method according to claim 33, wherein the cellular proliferative
disorder is selected
from the group consisting of hemangiomatosis in newborn, secondary progressive
multiple
sclerosis, atherosclerosis, chronic progressive myelodegenerative disease,
neurofibromatosis,
ganglioneuromatosis, keloid formation, Paget's disease of the bone,
fibrocystic disease of the
breast, uterine fibroids, Peyronie's disease, Dupuytren's disease, restenosis,
benign
proliferative breast disease, benign prostatic hyperplasia, X linked
lymphocellular
proliferative disorder, post transplantation lymphocellular proliferative
disorder, macular
degeneration, retinopathies, proliferative vitreoretinopathy, non cancerous
lymphocellular
proliferative disorders, and cancer.
35. The method of claim 34 wherein the cellular proliferative disorder is
cancer.
36. The method of claim 35 wherein the compound is (Z)-64(2,6-
dichlorobenzyl)sulfonyl)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one, or pharmaceutically acceptable salt thereof.
37. The method according to claim 36 wherein the activity of one or more
kinases selected
from the group consisting of casein kinase 2, cyclin-dependent kinase 9 and
PIM1 is inhibited
in cancer cells of the individual.
122

38. The method according to claim 35, wherein the cancer is selected from
the group
consisting of ovarian cancer; cervical cancer; breast cancer; prostate cancer;
testicular cancer,
lung cancer, renal cancer; colorectal cancer; skin cancer; brain cancer;
leukemia, including
acute myeloid leukemia, chronic myeloid leukemia, acute lymphoid leukemia, and
chronic
lymphoid leukemia.
39. A method of inducing apoptosis of cancer cells in an individual
afflicted with cancer,
comprising administering to the individual an effective amount of at least one
compound of
Formula I, or a pharmaceutically acceptable salt thereof, according to claim
1.
40. The method of claim 39 wherein the compound is (Z)-6-((2,6-
dichlorobenzyl)sulfonyl)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one, or pharmaceutically acceptable salt thereof.
41. The method according to claim 40 wherein the activity of one or more
kinases selected
from the group consisting of casein kinase 2, cyclin-dependent kinase 9 and
PIM1 is inhibited
in cancer cells of the individual.
42. The method of claim 39, wherein the cancer cells are tumor cells.
43. The method according to claim 42, wherein the tumor cells are selected
from the
group consisting of ovarian, cervical, breast, prostate, testicular, lung,
renal, colorectal, skin
and brain tumor cells.
123

44. A method of treating an individual suffering from a cellular
proliferative disorder,
comprising administering to the individual an effective amount of at least one
antibody
conjugate, or pharmaceutically acceptable salt thereof, according to claim 28.
45. A method of inducing apoptosis of cancer cells in an individual
afflicted with cancer,
comprising administering to the individual an effective amount of at least one
antibody
conjugate, or a pharmaceutically acceptable salt thereof, according to claim
28.
124

Description

Note: Descriptions are shown in the official language in which they were submitted.


CA 02836085 2013-12-05
=
SUBSTITUTED 2-BENZYLIDENE-2H-BENZO[b][1,41THIAZIN-3(4H)-ONES,
DERIVATIVES THEREOF, AND THERAPEUTIC USES THEREOF
Field of the Invention
The invention relates to compounds, methods for their preparation,
compositions
including them. The invention further provides methods for the treatment of
cellular
proliferative disorders, including, but not limited to, cancer.
Background of the Invention
Cellular proliferative disorders such as cancer are among the most common
causes of
death in developed countries. That said, many cellular proliferative disorders
have no
available cures or few, if any, treatment options to slow the progression of
the disease. For
cellular proliferative diseases for which treatments exist, undesirable side
effects and limited
efficacy often call into question the utility of a given treatment. This is
particularly true when
the available treatment option(s) may not appreciably prolong life, but have a
definitive
adverse effect on the quality of time remaining. Thus, identifying new
effective drugs for
cellular proliferative disorders, and in particular cancer, is a continuing
focus of medical
research.
Pim-1
Pim-1 is a proto-oncogene which encodes for a serine/threonine. Pim-1 over-
expression has been implicated in multiple human cancers, including prostate
cancer, acute
myeloid leukemia and other hematopoietic malignancies. Pim-1 has also been
found to be
highly expressed in cell cultures isolated from human tumors and is mainly
involved in cell
cycle progression, apoptosis and transcriptional activation, as well as more
general signal
transduction pathways (Bachmann and Moroy, Int. J. Biochem. Cell Biol.
37(4):726-30
(2005)). Compounds that inhibit the activity of Pim-1 have potential as
anticancer agents.
1

CA 02836085 2013-12-05
CK2 and CDK9
Casein Kinase 2 (CK2) is a ubiquitously expressed serine threonine protein
kinase,
whose activity and expression levels are elevated in a variety of tumor types
(St-Denis and
Litchfield, Cell. Mol. Life Sci., 66:1817-1829 (2009); Trembley et al., Cell.
MoL Life Sci.,
66:1858-1867 (2009)). CK2 is unusual among protein kinases in that it can use
both ATP and
GTP as phosphate donors (St-Denis and Litchfield, supra). CK2 is also unique
in that it is
constitutively active and does not appear to require phosphorylation or other
post-translational
alterations to activate its kinase activity. As a result, the catalytic
activity of CK2 is roughly
proportional to its cellular concentration (Guerra and Issinger, Curr. Med.
Chemistry, 15:
1870-1886 (2008)). CK2 is a tetrameric complexes consisting of two catalytic
(a and/or a')
subunits and two regulatory 13 subunits (Litchfield, Biochem. J. (2003) 369:1-
15).
CK2 phosphorylates a large number of substrates, many of which regulate signal
transduction pathways that in turn mediate cell growth, cell death, DNA
replication and
transcription (St-Denis and Litchfield, supra; Trembley et al., supra; Guerra
and Issinger,
supra). CK2 has also been shown to regulate the activity of tumor suppressors,
cell cycle
regulatory proteins, apoptotic proteins and oncogenes thus making it a key
player in the
development and maintenance of cancer progression (Ahmed et al., Cellular and
Molecular
Life Sciences, 66: 1858-1867 (2009)).
Elevated CK2 activity has been associated with the malignant transformation of
several tissues and higher levels of CK2 are found to correlate with
aggressive behavior of
head and neck cancer and poor prognosis of prostate cancer (Gapany et al., MoL
Med. 1: 659-
666 (1995); Laramas et al., Eur. i Cancer. 43(5):928-34 (2007)). The ability
of CK2 to
phosphorylate (and activate) PTEN and AKT, two key components of tumor cell
survival
pathway, suggests an important role of CK2 in suppressing apoptosis in cancer
cells (Di
Maira et al., Cell Death Differ. 12: 668-677 (2005)). Downregulation of CK2 by
various
approaches results in induction of apoptosis in cultured cell and xenograft
cancer models,
further suggesting its potential as a therapeutic target (Guerra and Issinger,
supra; Ruzzene
and Pinna, Biochim Biophys Acta, 1804(3): 499-504 (2010)).
2

CA 02836085 2013-12-05
Normal cells, which express low levels of CK2, do not appear to have such
undue
reliance on the activity of this kinase and hence are unaffected by reduction
in the levels of
this kinase activity, suggesting that development of potent and selective CK2
inhibitors may
not exhibit undesirable side effects seen with traditional chemotherapeutic
agents.
CDK9 is a cyclin-dependent kinase, which forms a complex with and is regulated
by
two different regulatory cyclins known as cyclin T and cyclin K (Wang and
Fischer, Trends
Pharmacol. Sci. 29(6):302-13 (2008)). CDK9 accumulates on chromatin and limits
the
amount of single-stranded DNA in response to replication stress. CDK9/Cyclin K
complex
was found to play direct role in maintaining genome integrity (Yu and Cortez,
Cell Cycle
10:1, 28-32 (2011)).
Depletion of CDK9 or its cyclin K (but not cyclin T) was found to impair cell
cycle
recovery in response to replication stress, resulting in spontaneous DNA
damage in
replicating cells (Wang et al., supra; Yu et al., EMBO Rep. 11(11):876-82
(2010)).
CDK9 is over-expressed in several cancers, including leukemias and lymphomas
suggesting that inhibitors might be useful in cancers involving high levels of
replication
stress, or in combination with replication stress-inducing chemotherapies
(Wang and Fischer,
supra).
Summary of the Invention
It has been found that certain compounds and compositions are useful for the
treatment of cancer and other cellular proliferative disorders. The
biologically active
compounds of the invention are substituted 2-benzylidene-2H-
benzo[b][1,4]thiazin-3(411)-
ones and related derivatives thereof.
3

CA 02836085 2013-12-05
, =
In certain embodiments, the invention is a compound of Formula I or a salt
thereof:
R4 A r
(0)n
R3
R2 N 0
H
Formula I
wherein
n is 0, 1, or 2;
R1 is selected from the group consisting of -H, -(Ci-C6)alkyl, -(C2-
C6)alkenyl,
-(C2-C6)alkynyl, optionally substituted aryl, optionally substituted
heteroaryl, optionally
substituted aryl-(CI-C6)alkyl, optionally substituted heteroaryl-(CI-C6)alkyl,
-C(=0)(Ci-
C6)alkyl, -C(-0)(C2-C6)alkenyl, -C(-0)-optionally substituted aryl, -C(-
0)(CH2),,,-optionally
substituted aryl, and -C(=0)(CH2)p-optiona11y substituted heteroaryl;
R2, R3, and R4 are independently selected from the group consisting of -H,
halogen,
-CN, -NR1 R 11, -OH, -OW 3, -(C1 -C6)aik0Xy, -NO2, -(Ci-C6)alkyl, -(CI-
C6)perfluoroalkyl,
-(C -C6)perfluoroalkoxy, -C(=0)R15, -C(=0)0R15,
-0C(-0)R12, -0C(-0)0R12,
-C(-0)NR17R18, _ SH, -S(C -C6)alkyl,
-S(=0)R13, -S(-0)2R13, -0S(=0)2R13,
-S(=0),A15, -0S(-0),A15, -S(=0)2NR17R18, -S(=0)NR17R18, optionally substituted
aryl,
optionally substituted aryl-(Ci-C6)alkyl, optionally substituted heteroaryl,
optionally
substituted heteroaryl-(CI-C6)alkyl, optionally substituted (C2-
C9)heterocyclyl, optionally
substituted (C2-C9)heterocycly1-(C -C6)alkyl, -NH(CH2),,C(-0)0R14, -
C(=NR14)NRI42,
-C(=-N-OR14)NR142, -P(=0)(0R14)2, and -0P(=0)(0R14)2;
4

CA 02836085 2013-12-05
Ar is optionally substituted heteroaryl, optionally substituted (Cio-Cia)aryl,
or
R7
R8 R8
R9 R5
../VVV=
wherein
R5, R6, R7, R8, and R9 are independently selected from the group consisting of
-H,
ioRi
-OH, -0R13, -NO2, halogen, -CN, _NR-(CH2)mNRI R11, -0(CH2),,,NR ofti _(C _
C6)alkyl, -(CH2),,,O(C -C6)alkyl,
-(C -C6)alkoxy, -(CI-C6)perfluoroalkyl, -(C 1-
C6)perfluoroalkoxy, -SH, -S(C -C6)alkyl, -SRI 3, -S(-0)e, -S(-0)2R' 5, -
C(=0)R15,
-C(=0)0R15, -C(=0)NRI7R18, -0C(=0)R16, -0Q=0)0R12, -0C(=0)NRI7R18,
heterocyclyl,
optionally substituted heteroaryl, -NH(CH2)mC(=0)0R14,
-0S(=0)2R16,
-C(=NR14)NRI42, -C(=N-ORI4)NR142, -P(-0)(0R14)2, and -0P(=0)(0R14)2;
each RI and R" is independently selected from the group consisting of -H,
-(C1-C6)alkyl, -(CI-C6)alkoxy, -C(=0)RI2, -C(=0)NRI7R18, -C(=0)0R12, -
C(=NRI4)NRI7R18,
R13, optionally substituted aryl, optionally substituted heteroaryl, and -
C(=NR14)R15; or R1
and R", together with the nitrogen to which they are bound, form an optionally
substituted
(C2-05)heterocycle;
each RI2 is independently selected from the group consisting of -(C1-C6)alkyl,
and
optionally substituted aryl;
each RI3 is independently selected from the group consisting of optionally
substituted
aryl and -(CH2),õR16;
each RI4 is independently selected from the group consisting of -H and -(Ci-
C6)alkyl;
or two occurrences of R" bound to the same nitrogen form a (C2-C6)heterocycle,
together
with the nitrogen atom to which they are bound;
5

CA 02836085 2013-12-05
each R15 is independently selected from the group consisting of -H, -(Ci-
C6)alkyl,
optionally substituted aryl, and NR142;
each R16 is independently selected from the group consisting of -(Ci-C6)alkyl,
-NRI42,
and Arl;
each R17 and R18 is independently selected from the group consisting of -H,
-(Ci-C6)alkyl, -(Ci-C6)alkoxy, R13, optionally substituted aryl, and
optionally substituted
heteroaryl; or R17 and R18, together with the nitrogen to which they are
bound, form an
optionally substituted (C2-05)heterocycle;
m is independently at each occurrence 1, 2, 3, 4, or 5;
p is independently at each occurrence 0, 1, 2, or 3;
q is independently at each occurrence 0, 1, or 2;
each optionally substituted aryl, optionally substituted (Cio-Cia)aryl,
optionally
substituted heteroaryl, optionally substituted ary1-(C1-C6)alkyl, optionally
substituted
heteroaryl-(CI-C6)alkyl, optionally substituted (C2-C9)heterocyclyl,
optionally substituted (C2-
C9)heterocycly1-(CI-C6)alkyl, and optionally substituted (C2-05)heterocycle is
optionally
substituted with one or more substituents independently selected from the
group consisting of
halogen, -CN, -NR' 2, -(CH2)õ,NR142, _0(CH2)rnNR142, "NRI4C(=0)(C1-C6)alkyl,
-NR"C(=0)0(C -C6)alkyl, -NR'4C(=0)NR142,
-NR14C(=NR14)NR142.5
-NH(CH2)TIC(=0)0R14, -OH, -NO2, -(C 1-C6)alkyl, -(CH2),T,O(C -C6)alkyl, -(C -
C6)alkoxY,
-(C2-C6)alkenyl, -(C2-C6)alkynyl, -SRI4, -S(=0)R15, -S(=0)2RI5, -NRI4S(=-
0)2R15, -(C1-
C6)perfluoroalkyl, -(C -C6)perfluoroalkoxy, -C(=0)R14, -C(-0)0R14, -
C(=0)NR142,
-0C(=0)R14, -0C(-0)NR142, -0C(-0)0(CI-C6)alkyl, -P(----0)(0R14)2, -
0P(=0)(0R14)2,
heterocyclyl, and heteroaryl;
6

CA 02836085 2013-12-05
Arl is a radical according to Formula II:
R21
R22 0 R19 Rzo
R23
1
Formula II
wherein R19, R20, R21, R22, and R23 are independently selected from the group
consisting of -H, -OH, -NO2, halogen, -CN, -Nee, -(CH2).NeR11, -0(CH2)õ,NR1
R11,
-(C1 -C6)alkyl, -(CH2),n0 (C i-C6)alkyl, - (C 1 -C6)alkoxy,
-(C, -C6)p erfluoro alkyl, -(C i -
C6)perfluoroalkoxy, -SH, -Se, -S(=0)R15, -S(=0)2R15, -C(-0)R15, -C(=-0)0R15,
-Q=0)NRI7R18, -OC(=-0)R16, -0C(=0)0R12, -0C(=0)NRI7R18, heterocyclyl,
optionally
substituted heteroaryl, -NH(CH2),,,C(=0)0R14,
-0S(=0)2R16,
-C(=\TR14)NR142, -C(=-1\1-0R14)NR142, -13(=0)(0R14)2, and -0P(=0)(0R14)2;
provided that:
i) at least one of R2, R3, or R4 is other than hydrogen;
ii) when none of R2, R3, and R4 are -0R13, -NHR13, -SR13, -S(---0)R13, or
-S(=0)2R13, and Ar is
R7
R8 R6
R9 10 R5
I ,
then at least one of R6 and Rg is -NO2 and at least R7 is other than hydrogen
or halogen; and
7

CA 02836085 2013-12-05
=
iii) when Ar is optionally substituted heteroaryl and none of R2, R3, or R4
are
-0R13, -NHR13, -SR13, -S(=0)R13, or -S(=0)2R13, then R1 is other than
hydrogen.
In certain embodiments of a compound of Formula I, or a salt thereof, n is O.
In particular embodiments of a compound of Formula I, or a salt thereof, Ar is
optionally substituted heteroaryl or
R7
R8 R6
R9 116 R5
avvv,
In other embodiments of a compound of Formula I, or a salt thereof, at least
one of R2,
R3, or R4 is -0R13, -NHR13, -SR13, -S(=0)R13, or -S(=0)2R13.
In still other embodiments of a compound of Formula I, or a salt thereof, at
least one
of R2, R3, or R4 is -S(=0)2R13.
In certain embodiments of a compound of Formula I, or a salt thereof, R13 is
-(CH2)mR16.
In some embodiments of a compound of Formula I, or a salt thereof, R2 is
-S(=0)2R13 and R13 is -(CH2)mR16.
In a particular embodiment of a compound of Formula I, or a salt thereof, R16
is Ar'.
In certain embodiments of a compound of Formula I, or a salt thereof, Arl is
2,3-
dichlorophenyl, 2,4-di chl orophenyl, 2,5-
dichlorophenyl, 2,6-dichlorophenyl, 3 ,4-
dichlorophenyl, 3 ,5 -di chlorophenyl, 2,3 -
dibromophenyl, 2,4-dibromophenyl, 2,5 -
dibromophenyl, 2,6-dibromophenyl, 3,4-dibromophenyl, 3
,5-dibromophenyl, 2,3 -
difluorophenyl, 2,4-difluorophenyl, 2,5 -difluorophenyl,
2,6-difluorophenyl, 3,4-
difluorophenyl, 3,5-difluorophenyl, 2,3-diiodophenyl, 2,4-diiodophenyl, 2,5-
diiodophenyl,
8

CA 02836085 2013-12-05
2,6-diiodophenyl, 3,4-diiodophenyl, 3,5-diiodophenyl, 2-chloro-3-bromophenyl,
2-chloro-4-
bromophenyl, 2-chloro-5-bromophenyl, 2-chloro-6-bromophenyl, 3-chloro-4-
bromophenyl,
3-chloro-5-bromophenyl, 4-chloro-5-bromophenyl, 2-bromo-3 -chlorophenyl, 2-
bromo-4-
chlorophenyl, 2-bromo-5-chlorophenyl, 3-bromo-4-chlorophenyl, 2-chloro-3-
fluorophenyl, 2-
chloro-4-fluorophenyl, 2-chloro-5 -fluorophenyl, 2-chloro-6-
fluorophenyl, 3 -chl oro-4 -
fluorophenyl, 3-chloro-5-fluorophenyl, 4-chloro-5-fluorophenyl, 2-fluoro-3-
chlorophenyl, 2-
fluoro-4-chlorophenyl, 2-fluoro-5-chlorophenyl, 3-fluoro-4-chlorophenyl, 2-
chloro-3-
iodophenyl, 2-chloro-4-iodophenyl, 2-chloro-5-iodophenyl, 2-chloro-6-
iodophenyl, 3-chloro-
4-iodophenyl, 3 -chloro-5-iodophenyl, 4-chloro-5-iodophenyl, 2-iodo-3 -
chlorophenyl, 2-iodo-
4-chlorophenyl, 2-iodo-5-chlorophenyl, 3 -iodo-4-chlorophenyl, 2-bromo-3 -
fluorophenyl, 2-
bromo-4-fluorophenyl, 2-bromo-5-fluorophenyl, 2-bromo-6-fluorophenyl, 3-bromo-
4-
fluorophenyl, 3-bromo-5-fluorophenyl, 4-bromo-5-fluorophenyl, 2-fluoro-3-
bromophenyl, 2-
fluoro-4-bromophenyl, 2-fluoro-5-bromophenyl, 3-fluoro-4-bromophenyl, 2-bromo-
3-
iodophenyl, 2-bromo-4-iodophenyl, 2-bromo-5-iodophenyl, 2-bromo-6-iodophenyl,
3-bromo-
1 5 4-iodophenyl, 3 -bromo-5-iodophenyl, 4-bromo-5-iodophenyl, 2-iodo-3-
bromophenyl, 2-iodo-
4-bromophenyl, 2-iodo-5-bromophenyl, 3-iodo-4-bromophenyl, 2-fluoro-3-
iodophenyl, 2-
fluoro-4-iodophenyl, 2-fluoro-5-iodophenyl, 2-fluoro-6-iodophenyl, 3-fluoro-4-
iodophenyl, 3-
fluoro-5-iodophenyl, 4-fluoro-5-iodophenyl, 2-iodo-3-fluorophenyl, 2-iodo-4-
fluorophenyl, 2-
iodo-5-fluorophenyl, or 3-iodo-4-fluorophenyl.
In particular embodiments, Arl is 2,6-dichlorophenyl.
In certain embodiments of a compound of Formula I, or a salt thereof, Ar is
R7
R8 R6
R9 IP R5
%MAP
=
9

CA 02836085 2013-12-05
. = , =
In other embodiments, a compound of Formula I, or a salt thereof, is selected
from the
group consisting of (Z)-44(64(2,6-dichlorobenzyl)sulfony1)-3-oxo-3,4-dihydro-
2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-((6-((2,6-
dichlorobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-642,6-dichlorobenzyesulfony1)-
2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dichlorobenzyl)sulfony1)-2-(4-
methoxy-3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(3-amino-4-
methoxybenzylidene)-64(2,6-dichlorobenzypsulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-64(2,6-dichlorobenzypsulfony1)-2-(4-(4-methylpiperazin-1-y1)benzylidene)-
2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dichlorobenzypsulfony1)-2-(4-
nitrobenzylidene)-
2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-aminobenzylidene)-6-((2,6-
dichlorobenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-64(2,6-
dichlorobenzypsulfony1)-2-(4-fluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-
(4-chlorobenzylidene)-642,6-dichlorobenzypsulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-(4-bromobenzylidene)-64(2,6-dichlorobenzypsulfony1)-2H-benzo
[b][1,4]thiazin-
3(4H)-one; (Z)-6-((2,6-dichlorobenzyl)sulfony1)-2-(4-methoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dichlorobenzyl)sulfony1)-2-(4-
methylbenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dichlorobenzyl)sulfony1)-2-(2,4,6-trimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-6-((2,6-dichlorobenzyl)sulfony1)-2-(2,4-dichlorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dichlorobenzypsulfony1)-2-(2,4,6-
trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-methyl 2-((4-((6-
((2,6-
dichlorobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,41thiazin-2-
ylidene)methyl)phenyl)amino)acetate; (Z)-2-((4-((6-((2,6-
dichlorobenzyl)sulfony1)-3-oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetic acid; (Z)-
6-((2,6-
dichlorobenzyl)sulfony1)-2-(4-hydroxy-2,6-dimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-2-(4-chloro-3-nitrobenzylidene)-64(2,6-dichlorobenzyl)sulfony1)-
2H-
benzo[b][1,41thiazin-3(4H)-one; (Z)-642,6-dichlorobenzyl)sulfony1)-2-(2,4-
difluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dichlorobenzyl)sulfony1)-2-(2,4,6-trifluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one;

CA 02836085 2013-12-05
õ =
(Z)-methyl 4-46-((2,6-dichlorobenzypsulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-
2-ylidene)methyl)benzoate; (Z)-methyl 4-((6-((2,6-dichlorobenzyl)sulfony1)-3-
oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-3-nitrobenzoate; (Z)-4-((6-
((2,6-
dichlorobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-
nitrophenyl 4-methylbenzenesulfonate; (Z)-2-(4-(2H-tetrazol-5-yl)benzylidene)-
642,6-
dichlorobenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-
(benzyloxy)-3-
nitrobenzylidene)-64(2,6-dichlorobenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one; and
(Z)-64(2,6-dichlorobenzypsulfony1)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one.
In certain embodiments of a compound of Formula I, or a salt thereof, Ar is
optionally
substituted heteroaryl.
In a particular embodiment a compound of Formula I, or a salt thereof, is
selected
from the group consisting of (Z)-64(2,6-dichlorobenzypsulfony1)-24(2-
(methylthio)pyrimidin-4-yl)methylene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-
((2,6-
1 5 dichlorobenzyl)sulfony1)-2-((2-morpholinopyrimidin-4-yl)methylene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-((1H-indo1-3-yl)methylene)-6-((2,6-
dichlorobenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-642,6-
dichlorobenzypsulfony1)-2-(thiophen-2-ylmethylene)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-((1H-pyrrol-3-yl)methylene)-6-((2,6-dichlorobenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; and (Z)-2-((1-acety1-1H-indo1-3-y1)methylene)-6-((2,6-
dichlorobenzyl)sulfonyl)-
2H-benzo[b][1,4]thiazin-3(4H)-one.
In certain embodiments of a compound of Formula I, at least one of R6 and R8
is -NO2
and at least re is other than hydrogen or halogen.
In some embodiments, a compound of Formula I, or a salt thereof, is selected
from the
group consisting of (Z)-4-((6-chloro-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-
2-
ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-((7-methoxy-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-((7-bromo-
3-oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-
((7-methy1-
3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl
acetate; (Z)-2-
11

CA 02836085 2013-12-05
'
(4-acetoxy-3-nitrobenzylidene)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazine-7-
carboxylic
acid; (Z)-447-fluoro-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-
nitrophenyl acetate; (Z)-44(6-fluoro-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-
2-
ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-((4-acety1-7-methoxy-3-oxo-3,4-
dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-2-(4-
(benzyloxy)-3-
nitrobenzylidene)-7-fluoro-2H-benzo[b][1,4]thiazin-3(4H)-one; and (Z)-7-fluoro-
2-(4-
hydroxy-3-nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one.
In other embodiments, a compound of Formula I, or a salt thereof, is selected
from the
group consisting of (Z)-44(64(2,6-dibromobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-44(642,6-
dibromobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-6-((2,6-
dibromobenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dibromobenzyl)sulfony1)-2-(4-
methoxy-3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(3-amino-4-
methoxybenzylidene)-6((2,6-dibromobenzypsulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-6-((2,6-dibromobenzyl)sulfony1)-2-(4-(4-methylpiperazin-1-y1)benzylidene)-
2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dibromobenzyl)sulfony1)-2-(4-
nitrobenzylidene)-
2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-aminobenzylidene)-6-((2,6-
dibromobenzypsulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dibromobenzypsulfony1)-2-(4-fluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-
(4-chlorobenzylidene)-64(2,6-dibromobenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-(4-bromobenzylidene)-6-((2,6-dibromobenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-64(2,6-dibromobenzyl)sulfony1)-2-(4-methoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dibromobenzyl)sulfony1)-2-(4-
methylbenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dibromobenzyl)sulfony1)-2-(2,4,6-trimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-6-((2,6-dibromobenzyl)sulfony1)-2-(2,4-dibromobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dibromobenzypsulfony1)-2-(2,4,6-
trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-methyl 2-((4-((6-
((2,6-
dibromobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
12

CA 02836085 2013-12-05
ylidene)methyl)phenyl)amino)acetate; (Z)-2-((4-((6-((2,6-
dibromobenzyl)sulfony1)-3-oxo-
3,4-dihydro-2H-be.nzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetic
acid; (Z)-64(2,6-
dibromobenzyl)sulfony1)-2-(4-hydroxy-2,6-dimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-2-(4-chloro-3-nitrobenzylidene)-6 -((2,6-
dibromobenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dibromobenzyl)sulfony1)-2-(2,4-
difluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-64(2,6-
dibromobenzyl)sulfony1)-2-(2,4,6-trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-methyl 4-((64(2,6-dibromobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-
2-ylidene)methyl)benzoate; (Z)-methyl 4-((6-((2,6-dibromobenzyl)sulfony1)-3-
oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-3-nitrobenzoate; (Z)-
4464(2,6-
dibromobenzypsulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-
nitrophenyl 4-methylbenzenesulfonate; (Z)-2-(4-(2H-tetrazol-5-yl)benzylidene)-
6-((2,6-
dibromobenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-
(benzyloxy)-3-
nitrobenzylidene)-642,6-dibromobenzypsulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-
642,6-dibromobenzyl)sulfony1)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-4-((6-((2,6-difluorobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-46-((2,6-
difluorobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-6-((2,6-
difluorobenzyl)sulfony1)-2H-
benzo[b][1,41thiazin-3(4H)-one; (Z)-642,6-difluorobenzypsulfony1)-2-(4-methoxy-
3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(3-amino-4-
methoxybenzylidene)-642,6-difluorobenzyl)sulfony1)-21-1-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-6-((2,6-difluorobenzyl)sulfony1)-2-(4-(4-methylpiperazin-1-y1)benzylidene)-
2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-642,6-difluorobenzypsulfony1)-2-(4-
nitrobenzylidene)-
2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-aminobenzylidene)-642,6-
difluorobenzypsulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-642,6-
difluorobenzypsulfony1)-2-(4-fluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-
(4-chlorobenzylidene)-6-((2,6-difluorobenzypsulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-(4-bromobenzylidene)-642,6-difluorobenzypsulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-6-((2,6-difluorobenzyl)sulfony1)-2-(4-methoxybenzylidene)-2H-
13

CA 02836085 2013-12-05
benzo [b] [1,4] thiazin-3 (4H)-one; (Z)-64(2,6-difluorobenzyl)sulfony1)-2-(4-
methylbenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
difluorobenzyl)sulfony1)-2-(2,4,6-trimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-64(2,6-difluorobenzypsulfony1)-2-(2,4-difluorobenzylidene)-2H-
benzo [b] [1,4]thiazin-3(4H)-one; (Z)-642,6-difluorobenzyl)sulfony1)-2-(2,4,6-
trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-methyl 24(44642,6-
difluorobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl)amino)acetate; (Z)-24(44(6-((2,6-
difluorobenzyl)sulfony1)-3-oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetic acid; (Z)-
6-((2,6-
difluorobenzyl)sulfony1)-2-(4-hydroxy-2,6-dimethoxybenzylidene)-2H-benzo[b]
[1,4]thiazin-
3(4H)-one; (Z)-2-(4-chloro-3-nitrobenzylidene)-64(2,6-difluorobenzypsulfony1)-
2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-difluorobenzypsulfony1)-2-(2,4-
difluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-642,6-
difluorobenzypsulfony1)-2-(2,4,6-trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-methyl 4-((6-((2,6-difluorobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-
2-ylidene)methyl)benzoate; (Z)-methyl 44(64(2,6-difluorobenzypsulfony1)-3-oxo-
3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-3-nitrobenzoate; (Z)-4-((6-
((2,6-
difluorobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-
nitrophenyl 4-methylbenzenesulfonate; (Z)-2-(4-(2H-tetrazol-5-yl)benzylidene)-
642,6-
difluorobenzyl)sulfony1)-2H-benzo[b][1,41thiazin-3(4H)-one; (Z)-2-(4-
(benzyloxy)-3-
nitrobenzylidene)-642,6-difluorobenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-
64(2,6-difluorobenzyl)sulfony1)-2-(4-hydroxy-3-nitrobenzylidene)-211-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-4-((6-((2,6-dimethylbenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-((6-((2,6-
dimethylbenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-6-((2,6-
dimethylbenzypsulfony1)-211-
benzo[b][1,41thiazin-3(4H)-one; (Z)-642,6-dimethylbenzypsulfony1)-2-(4-methoxy-
3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(3-amino-4-
methoxybenzylidene)-6-((2,6-dimethylbenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-6-((2,6-dimethylbenzyl)sulfony1)-2-(4-(4-methylpiperazin-1-y1)benzylidene)-
2H-
14

CA 02836085 2013-12-05
, . =
benzo[b][1,4]thiazin-3(4H)-one; (Z)-64(2,6-dimethylbenzypsulfony1)-2-(4-
nitrobenzylidene)-
2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-aminobenzylidene)-64(2,6-
dimethylbenzypsulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dimethylbenzyl)sulfony1)-2-(4-fluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-
(4-chlorobenzylidene)-6((2,6-dimethylbenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-(4-bromobenzylidene)-6-((2,6-dimethylbenzypsulfony1)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-642,6-dimethylbenzypsulfony1)-2-(4-methoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethylbenzyl)sulfony1)-2-(4-
methylbenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dimethylbenzyl)sulfony1)-2-(2,4,6-trimethoxybenzylidene)-2H-benzo [b]
[1,4]thiazin-3(4H)-
one; (Z)-6-((2,6-dimethylbenzyl)sulfony1)-2-(2,4-dimethylbenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-64(2,6-dimethylbenzypsulfony1)-2-(2,4,6-
trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-methyl 244464(2,6-
dimethylbenzypsulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl)amino)acetate; (Z)-24(44(6-((2,6-
dimethylbenzyl)sulfony1)-3-oxo-
3,4-dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetic acid;
(Z)-6-((2,6-
dimethylbenzyl)sulfony1)-2-(4-hydroxy-2,6-dimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-2-(4-chloro-3-nitrobenzylidene)-6-((2,6-
dimethylbenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-64(2,6-dimethylbenzyl)sulfony1)-2-(2,4-
difluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dimethylbenzyl)sulfony1)-2-(2,4,6-trifluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-methyl 446-((2,6-dimethylbenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-
2-ylidene)methyDbenzoate; (Z)-methyl 4-((6-((2,6-dimethylbenzyl)sulfony1)-3-
oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-3-nitrobenzoate; (Z)-4-((6-
((2,6-
dimethylbenzypsulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-
nitrophenyl 4-methylbenzenesulfonate; (Z)-2-(4-(2H-tetrazol-5-yObenzylidene)-6-
((2,6-
dimethylbenzyl)sulfonyl)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-
(benzyloxy)-3-
nitrobenzylidene)-6-((2,6-dimethylbenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one; and
(Z)-6-((2,6-dimethylbenzyl)sulfony1)-2-(4-hydroxy-3 -nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-44(642,6-dimethoxybenzyl)sulfony1)-3-oxo-
3,4-

CA 02836085 2013-12-05
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-4-
((6-((2,6-
dimethoxybenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-6-((2,6-
dimethoxybenzypsulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethoxybenzyl)sulfony1)-2-(4-
methoxy-3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(3-amino-4-
methoxybenzylidene)-6-((2,6-dimethoxybenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-
one; (Z)-6-((2,6-dimethoxybenzyl)sulfony1)-2-(4-(4-methylpiperazin-1-
yl)benzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-64(2,6-dimethoxybenzypsulfony1)-2-(4-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-
aminobenzylidene)-6-((2,6-
dimethoxybenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-64(2,6-
dimethoxybenzypsulfony1)-2-(4-fluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-
2-(4-chlorobenzylidene)-6-((2,6-dimethoxybenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-(4-bromobenzylidene)-6-((2,6-dimethoxybenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethoxybenzyl)sulfony1)-2-(4-
methoxybenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-
dimethoxybenzyl)sulfony1)-2-(4-methylbenzylidene)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-6-((2,6-dimethoxybenzyl)sulfony1)-2-(2,4,6-trimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethoxybenzyl)sulfony1)-2-(2,4-
dimethoxybenzylidene)-2H-benzo[b][1,4]thiazin-3(411)-one; (Z)-6-((2,6-
dimethoxybenzyl)sulfony1)-2-(2,4,6-trifluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one; (Z)-methyl 2-44-464(2,6-dimethoxybenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetate; (Z)-2-((4-((6-
((2,6-
dimethoxybenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl)amino)acetic acid; (Z)-6-((2,6-dimethoxybenzyl)sulfony1)-
2-(4-
hydroxy-2,6-dimethoxybenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-
chloro-3-
nitrobenzylidene)-64(2,6-dimethoxybenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-64(2,6-dimethoxybenzypsulfony1)-2-(2,4-difluorobenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one; (Z)-6-((2,6-dimethoxybenzyl)sulfony1)-2-(2,4,6-
trifluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-methyl 44(642,6-dimethoxybenzyl)sulfony1)-
3-oxo-
3,4-dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)benzoate; (Z)-methyl 4-
((6-((2,6-
16

CA 02836085 2013-12-05
dimethoxybenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-3-
nitrobenzoate; (Z)-44(64(2,6-dimethoxybenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl 4-methylbenzenesulfonate;
(Z)-2-(4-
(2H-tetrazol-5-yl)benzylidene)-6-((2,6-dimethoxybenzyl)sulfony1)-2H-benzo[b][1
,4]thiazin-
__ 3(4H)-one; (Z)-2-(4-(benzyloxy)-3-nitrobenzylidene)-6-((2,6-
dimethoxybenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-one; and (Z)-6-((2,6-dimethoxybenzyl)sulfony1)-2-(4-
hydroxy-3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one.
The present invention further provides a process for preparing a compound of
Formula
I. In certain embodiments, the process for preparing a compound of Formula I
comprises
1 0 __ condensing an aldehyde of Formula III
0
Ar/H
Formula III
with a compound according to Formula IV:
R4
(0)n
R3 S
R2 N 0
I
H R1
Formula IV
in a reaction mixture, wherein Ar, RI, R2, R3, R4, and n, are as defined
elsewhere
herein. The method further provides for isolating from the reaction mixture
the compound of
Formula I, or a salt thereof
17

CA 02836085 2013-12-05
=
In certain embodiments, the condensation takes place in the presence of one or
more
of an anhydride and a base. In particular embodiments, the anhydride is acetic
anhydride and
the base is triethylamine.
In other embodiments of the process described herein, a compound of Formula IV
is
prepared by functionalizing 2H-1,4-Benzothiazin-3(4H)-one, or a derivative
thereof, with
chlorosulfonic acid to give 3-oxo-3,4-dihydro-2H-benzo [b][ 1,4]thiazine-6-
sulfonyl chloride;
reacting said 3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazine-6-sulfonyl chloride
with Zn to form
a reactive Zn complex; and reacting said reactive Zn complex with a compound
of Formula
V:
R"
R20
X
R21 1101 R23
R22
Formula V
to give a compound of Formula IV, wherein R19, R20, R21, R22, and R23
are as defined
elsewhere herein, and X is a leaving group.
In still another embodiment of a process described herein, a compound
according to
Formula IV is prepared by reacting compound A:
R4
R3
R2 NO2
A
with HSCH2CO2R, wherein R is H or (Ci-C6)alkyl and R2, R3, and R4 are as
defined
elsewhere herein, to form compound B:
18

CA 02836085 2013-12-05
=
R4
R3
OR
R2 NO2
and reducing the nitro group of compound B to give said compound according to
Formula IV.
In certain embodiments, reducing the nitro group of compound B comprises
treating
compound B with Zn and acetic acid. In other embodiments, reducing the nitro
group of
compound B comprises treating compound B with Na2S204.
The present disclosure further provides an antibody conjugate of the formula:
I-L-Ab
or a salt thereof, wherein I is a compound according to Formula I as defined
above; Ab is an
antibody; and -L- is a single bond or a linking group covalently linking said
compound of
Formula I to said antibody.
In particular embodiments of the antibody conjugate described herein, the
antibody is
a monoclonal antibody or a monospecific polyclonal antibody. In certain
embodiments, the
antibody is a tumor specific antibody.
The present disclosure further provides a pharmaceutical composition
comprising a
pharmaceutically acceptable carrier and a compound of Formula I, or a
pharmaceutically
acceptable salt thereof.
In certain embodiments, the present disclosure provides a pharmaceutical
composition
comprising a pharmaceutically acceptable carrier and an antibody conjugate I-L-
Ab, or a
pharmaceutically acceptable salt thereof, as set forth previously herein.
The present disclosure further provides a method of treating an individual
suffering
from a cellular proliferative disorder, comprising administering to the
individual an effective
amount of at least one compound of Formula I, or a pharmaceutically acceptable
salt thereof.
19

CA 02836085 2013-12-05
, =
In particular embodiments, the cellular proliferative disorder is selected
from the
group consisting of hemangiomatosis in newborn, secondary progressive multiple
sclerosis,
atherosclerosis, chronic progressive myelodegenerative disease,
neurofibromatosis,
ganglioneuromatosis, keloid formation, Paget's disease of the bone,
fibrocystic disease of the
breast, uterine fibroids, Peyronie's disease, Dupuytren's disease, restenosis,
benign
proliferative breast disease, benign prostatic hyperplasia, X linked
lymphocellular
proliferative disorder, post transplantation lymphocellular proliferative
disorder, macular
degeneration, retinopathies, proliferative vitreoretinopathy, non cancerous
lymphocellular
proliferative disorders, and cancer.
In particular embodiments, the cellular proliferative disorder is cancer. In
certain
embodiments, the cancer is selected from the group consisting of ovarian
cancer; cervical
cancer; breast cancer; prostate cancer; testicular cancer, lung cancer, renal
cancer; colorectal
cancer; skin cancer; brain cancer; leukemia, including acute myeloid leukemia,
chronic
myeloid leukemia, acute lymphoid leukemia, and chronic lymphoid leukemia.
In certain embodiments, the cellular proliferative disorder is cancer, the
compound is
(Z)-6-((2,6-dichlorobenzyl)sulfony1)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one or pharmaceutically acceptable salt thereof,
and the activity
of one or more kinases selected from the group consisting of casein kinase 2
(CK2), cyclin-
dependent kinase 9 (CDK9) and PIM1 is inhibited in cancer cells of the
individual.
The present disclosure further provides a method of inducing apoptosis of
cancer cells
in an individual afflicted with cancer, comprising administering to the
individual an effective
amount of at least one compound of Formula I, or a pharmaceutically acceptable
salt thereof.
In certain embodiments of the method of inducing apoptosis of cancer cells in
an
individual afflicted with cancer, the compound is (Z)-6-((2,6-
dichlorobenzyl)sulfony1)-2-(4-
hydroxy-3-nitrobenzylidene)-2H-benzo [b] [1,4]thiazin-3(4H)-one
or pharmaceutically
acceptable salt thereof, and the activity of one or more kinases selected from
the group
consisting of CK2, CDK9 and PIM1 is inhibited in cancer cells of the
individual.

CA 02836085 2013-12-05
. =
In certain embodiments, the cancer cells are tumor cells. In particular
embodiments,
the tumor cells are selected from the group consisting of ovarian, cervical,
breast, prostate,
testicular, lung, renal, colorectal, skin and brain tumor cells.
The present disclosure further provides a method of treating an individual
suffering
from a cellular proliferative disorder, comprising administering to the
individual an effective
amount of at least one antibody conjugate of the formula I-L-Ab, or
pharmaceutically
acceptable salt thereof, as defined elsewhere herein.
The present disclosure further includes a method of inducing apoptosis of
cancer cells
in an individual afflicted with cancer, comprising administering to the
individual an effective
amount of at least one antibody conjugate of the formula I-L-Ab, or a
pharmaceutically
acceptable salt thereof, as defined elsewhere herein.
In particular embodiments of the aforesaid methods and compositions, the
compound
is
(Z)-6-((2, 6- dichlorobenzyl) sulfony1)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one, or pharmaceutically acceptable salt thereof
In certain embodiments, the invention is a compound of Formula I, or salt
thereof, for
use in medicine. In other embodiments, the invention is a compound of Formula
I, or a salt
thereof, for treatment of a cellular proliferative disorder. In other
embodiments, the invention
provides a use of a compound according to Formula I, or a salt thereof, for
preparation of a
medicament for treatment of a cellular proliferative disorder. The present
invention further
provides a medicament for treatment of a cellular proliferative disorder,
containing a
compound of Formula I.
In some embodiments, the invention is an antibody conjugate of a compound of
Formula I, or salt thereof, for use in medicine. In other embodiments, the
invention is an
antibody conjugate of a compound of Formula I, or a salt thereof, for
treatment of a cellular
proliferative disorder. In other embodiments, the invention provides a use of
an antibody
conjugate of a compound according to Formula I, or a salt thereof, for
preparation of a
medicament for treatment of a cellular proliferative disorder. The present
invention further
provides a medicament for treatment of a cellular proliferative disorder,
containing an
antibody conjugate of a compound of Formula I.
21

CA 02836085 2013-12-05
Brief Description Of The Drawings
The foregoing summary, as well as the following detailed description the
embodiments, will be better understood when read in conjunction with the
appended
drawings. For the purpose of illustration, there are shown in the drawings
some embodiments
which may be preferable. It should be understood, however, that the
embodiments depicted
are not limited to the precise arrangements and instrumentalities shown.
Figure 1 is a synthetic scheme for preparing an intermediate useful for the
preparation
of a compound according to Formula I.
Detailed Description of the Invention
The compounds and compositions of the invention are believed to selectively
inhibit
proliferation of cancer cells, and kill various tumor cell types without
killing (or with reduced
killing of) normal cells. Cancer cells are killed at concentrations where
normal cells may be
temporarily growth-arrested but not killed.
The compounds of the invention are believed to inhibit the proliferation of
tumor cells,
and for some compounds, induce cell death. Cell death results from the
induction of
apoptosis. The compounds are believed effective against a broad range of tumor
types,
including but not limited to the following: ovarian cancer, breast cancer,
prostate cancer, lung
cancer, renal cancer, colorectal cancer, brain cancer and leukemia.
The compounds are also believed useful in the treatment of non-cancer cellular
proliferative disorders, including but not limited to the following:
hemangiomatosis in
newborn, secondary progressive multiple sclerosis, chronic progressive
myelodegenerative
disease, neurofibromatosis, ganglioneuromatosis, keloid formation, Paget's
disease of the
bone, fibrocystic disease of the breast, uterine fibroids, Peyronie's disease,
Dupuytren's
disease, restenosis and cirrhosis.
At least one compound of the invention, (Z)-6-((2,6-dichlorobenzyl)sulfony1)-2-
(4-
hydroxy-3-nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one (Compound 42),
inhibits the
activity of at least three kinases ¨ casein kinase 2 (CK2), cyclin-dependent
kinase 9 (CDK9)
and PIM1 ¨ that have been implicated in tumorigenesis and cancer progression.
The
22

CA 02836085 2013-12-05
. '
aforesaid compound is to believed to possess particular utility in inhibiting
the oncogenic
activity of PIM1, in inhibiting the ability of CK2 to induce malignant
transformation and in
enhancing apoptosis of cancer cells otherwise suppressed by CK2, and
inhibiting the pro-
cancer growth activity of CDK9, particularly in cancers involving high levels
of replication
stress.
At least Compound 42 also induces cell cycle arrest in cancer cells. MDA.MB-
231
breast cancer cells treated with increasing concentrations Compound 42
Without wishing to be bound by any mechanism of action, it is believed that
Compound 42 competes with the ATP/GTP phosphate donor in the ATP pocket in the
structure of CK2. The crystal structure of Compound 42 bound to CK2
demonstrates that the
drug mimics not only the shape and electrostatics of GTP/ATP, but also the
hydration patterns
of GTP/ATP (data not shown).
I. Definitions
A. General
As used in the specification and the appended claims, the singular forms "a,"
"an" and
"the" include plural referents unless the context clearly dictates otherwise.
As used herein, the terms "treat" and "treatment" are used interchangeably and
are
meant to indicate a postponement of development of a disorder and/or a
reduction in the
severity of symptoms that will or are expected to develop. The terms further
include
ameliorating existing symptoms, preventing additional symptoms, and
ameliorating or
preventing the underlying metabolic causes of symptoms.
As used herein, "individual" (as in the subject of the treatment) means both
mammals
and non-mammals. Mammals include, for example, humans; non-human primates,
e.g. apes
and monkeys; cattle; horses; sheep; and goats. Non-mammals include, for
example, fish and
birds.
The expression "effective amount", when used to describe therapy to an
individual
suffering from a cancer or other cellular proliferative disorder, refers to
the amount of a
compound according to Formula I that inhibits the abnormal growth or
proliferation, or
23

CA 02836085 2013-12-05
alternatively induces apoptosis of cancer cells, preferably tumor cells,
resulting in a
therapeutically useful and selective cytotoxic effect on proliferative cells.
The term "cellular proliferative disorder" means a disorder wherein unwanted
cell
proliferation of one or more subsets of cells in a multicellular organism
occurs. In some such
disorders, cells are made by the organism at an atypically accelerated rate.
B. Chemical
In the following paragraphs some of the definitions include examples. The
examples
are intended to be illustrative, and not limiting.
The term "alkyl", by itself or as part of another substituent means, unless
otherwise
stated, a straight, branched (chiral or achiral) or cyclic chain hydrocarbon
having the number
of carbon atoms designated (e.g. Ci-C6 means one to six carbons) and includes
straight,
branched chain or cyclic groups. Examples include: methyl, ethyl, propyl,
isopropyl, butyl,
isobutyl, tert-butyl, pentyl, neopentyl, hexyl, cyclohexyl and
cyclopropylmethyl. Most
preferred is (Ci-C3)alkyl, particularly ethyl, methyl and isopropyl.
The term "alkenyl" employed alone or in combination with other terms, means,
unless
otherwise stated, a stable mono-unsaturated or di-unsaturated straight chain,
branched chain
or cyclic hydrocarbon group having the stated number of carbon atoms. Examples
include
vinyl, propenyl (allyl), crotyl, isopentenyl, butadienyl, 1,3-pentadienyl, 1,4-
pentadienyl,
cyclopentenyl, cyclopentadienyl and the higher homologs and isomers. A
functional group
representing an alkene is exemplified by -CH=CH-CH2-.
The term "alkoxy" employed alone or in combination with other terms means,
unless
otherwise stated, an alkyl group having the designated number of carbon atoms,
as defined
above, connected to the rest of the molecule via an oxygen atom, such as, for
example,
methoxy, ethoxy, 1-propoxy, 2-propoxy (isopropoxy) and the higher homologs and
isomers.
Preferred are (Ci-C3)alkoxy, particularly ethoxy and methoxy.
The terms "halo" or "halogen" by themselves or as part of another substituent
mean,
unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom,
preferably, fluorine,
chlorine, or bromine, more preferably, fluorine or chlorine.
24

CA 02836085 2013-12-05
. =
The term "(Cx-Cy)perfluoroalkyl," wherein x < y, means an alkyl group with a
minimum of x carbon atoms and a maximum of y carbon atoms, wherein all
hydrogen atoms
are replaced by fluorine atoms. Preferred is -(CI-C6)perfluoroalkyl, more
preferred is
-(CI-C3)perfluoroalkyl, most preferred is ¨CF3.
The term "(Cx-Cy)perfluoroalkoxy," wherein x < y, means an alkoxy group with a
minimum of x carbon atoms and a maximum of y carbon atoms, wherein all
hydrogen atoms
are replaced by fluorine atoms. Preferred is -(Ci-C6)perfluoroalkoxy, more
preferred is
-(CI-C3)perfluoroalkoxy, most preferred is ¨0CF3.
The term "aromatic" refers to a carbocycle or heterocycle having one or more
polyunsaturated rings having aromatic character (i.e. having (4n + 2)
delocalized ic (pi)
electrons where n is an integer).
The term "aryl" employed alone or in combination with other terms, means,
unless
otherwise stated, a carbocyclic aromatic system containing one or more rings
(typically one,
two or three rings) wherein such rings may be attached together in a pendent
manner, such as
a biphenyl, or may be fused, such as naphthalene. Examples include phenyl;
anthracyl; and
naphthyl. Preferred are phenyl and naphthyl, most preferred is phenyl.
The term "optionally substituted aryl-(Ci-C3)alkyl" means a functional group
wherein
a one to three carbon alkylene chain is attached to an optionally substituted
aryl group, e.g.,
-CH2CH2-phenyl. Similarly, the term "optionally substituted heteroaryl(Ci-
C3)alkyl" means a
functional group wherein a one to three carbon alkylene chain is attached to
an optionally
substituted heteroaryl group, e.g., -CH2CH2-pyridyl.
The term "heterocycle" or "heterocycly1" or "heterocyclic" by itself or as
part of
another substituent means, unless otherwise stated, an unsubstituted or
substituted, stable,
non-aromatic mono- or multi-cyclic ring system which consists of carbon atoms
and at least
one heteroatom selected from the group consisting of N, 0, and S, and wherein
the nitrogen
and sulfur heteroatoms may be optionally oxidized, and the nitrogen atom may
be optionally
quaternized. The heterocyclic system may be attached, unless otherwise stated,
at any
heteroatom or carbon atom which affords a stable structure.

CA 02836085 2013-12-05
The term "heteroaryl" or "heteroaromatic" refers to an unsubstituted or
substituted,
stable, mono- or multi-cyclic ring system having aromatic character which
consists of carbon
atoms and at least one heteroatom selected from the group consisting of N, 0,
and S, and
wherein the nitrogen and sulfur heteroatoms may be optionally oxidized, and
the nitrogen
atom may be optionally quaternized. The heteroaryl or heteroaromatic system
may be
attached, unless otherwise stated, at any heteroatom or carbon atom which
affords a stable
structure.
A polycyclic heteroaryl may include one or more rings which are partially
saturated.
Examples include tetrahydroquinoline and 2,3-dihydrobenzofuryl.
Examples of heterocycles include monocyclic groups such as: aziridine,
oxirane,
thiirane, azetidine, oxetane, thietane, pyrrolidine, pyrroline, imidazoline,
pyrazolidine,
dioxolane, sulfolane, 2,3-dihydrofuran, 2,5-dihydrofuran, tetrahydrofuran,
thiophane,
piperidine, 1,2,3,6-tetrahydropyridine, 1,4-dihydropyridine, piperazine,
morpholine,
thiomorpholine, pyran, 2,3-dihydropyran, tetrahydropyran, 1,4-dioxane, 1,3-
dioxane,
homopiperazine, homopiperidine, 1,3 -dioxepane,
4,7-dihydro-1,3-dioxepin and
hexamethyleneoxide.
Examples of heteroaryl groups include: pyridyl, pyrazinyl, pyrimidinyl,
particularly
2- and 4-pyrimidinyl, pyridazinyl, thienyl, furyl (furanyl), pyrrolyl,
particularly 2-pyrrolyl,
imidazolyl, thiazolyl, oxazolyl, pyrazolyl, particularly 3- and 5-pyrazolyl,
isothiazolyl,
1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4-triazolyl, tetrazolyl, 1,2,3-
thiadiazolyl, 1,2,3-oxadiazolyl,
1,3,4-thiadiazoly1 and 1,3,4-oxadiazo1y1.
Examples of polycyclic heteroaryls include: indolyl, particularly 3-, 4-, 5-,
6- and
7-indolyl, indolinyl, quinolyl, tetrahydroquinolyl, isoquinolyl, particularly
1- and
5-isoquinolyl, 1,2,3,4-tetrahydroisoquinolyl, cinnolinyl, quinoxalinyl,
particularly 2- and
5-quinoxalinyl, quinazolinyl, phthalazinyl, 1,8-naphthyridinyl, 1,4-
benzodioxanyl,
coumarinyl, dihydrocoumarinyl, benzofuryl, 1,5-naphthyridinyl, 2,3-
dihydrobenzofuryl,
1,2-benzisoxazolyl, benzothienyl, particularly 3-, 4-, 5-, 6-, and 7-
benzothienyl, benzoxazolyl,
benzthiazolyl, particularly 2-benzothiazoly1 and 5-benzothiazolyl, purinyl,
benzimidazolyl,
26

CA 02836085 2013-12-05
=
particularly 2-benzimidazolyl, benztriazolyl, thioxanthinyl, carbazolyl,
carbolinyl, acridinyl,
pyrrolizidinyl, and quinolizidinyl.
The aforementioned listing of heterocyclyl and heteroaryl moieties is intended
to be
representative and not limiting.
For compounds of the present invention, when an aromatic or heteroaromatic
ring is
attached to a position and the ring comprises a polycyclic ring which is
partially saturated, the
attachment point on the aromatic or heteroaromatic ring is on a ring atom of
an aromatic ring
component of the polycyclic ring. For example on the partially saturated
heteroaromatic ring,
1,2,3,4-tetrahydroisoquinoline, attachment points would be ring atoms at the 5-
, 6-, 7- and 8-
positions.
The phrase "optionally substituted" means that an atom or group of atoms has
optionally replaced hydrogen as the substituent attached to another group. For
aryl and
heteroaryl groups, the term "optionally substituted" refers to any level of
optional
substitution, namely mono-, di-, tri-, tetra-, or penta-substitution, where
such substitution(s)
are permitted. When present, the substituents are independently selected, and
each optional
substitution may be at any chemically accessible position.
The term "antibody" is intended to encompass not only intact antigen-binding
immunoglobulin molecules, but also to include antigen-binding fragments
thereof such as
Fab, Fab' and F(ab')2 fragments, or any other fragment retaining the antigen-
binding ability of
an intact antibody.
The term "monospecific polyclonal antibody" means an antibody preparation
comprising multiple antibody species having specificity for a single antigen.
The term "peptidyl group" refers to a peptide functional group. Such a
functional
group has a chemical structure that varies from the structure of the
corresponding peptide in
that the structural component of the peptide, i.e., an alpha amino group, a
side chain amino
group, an alpha carboxyl group or a side chain carboxyl group, will form a
different
functionality when bonded to the molecule of which it is to be a substituent.
For example, for
a peptide as shown below:
27

CA 02836085 2013-12-05
=
H2N-Val-Pro-Ala-C(----0)0H
which is a substituent on a compound of Formula I, the peptide is coupled to
the compound of
Formula I such that a carboxyl moiety of said peptide is coupled to a free
amine moiety on the
Formula I compound. Elimination of water results in the formation of an amide
bond. As a
practical result, the corresponding monovalent peptidyl substituent is shown
to the left of the
dotted line in the depiction below of the aforementioned peptide bonded to a
compound of
Formula I:
0
H2N-Val-Pro-Ala-C-- - -NH =
The monovalent peptide group may be attached via either an alpha- or a side
chain
amino group, or an alpha or side chain carboxyl group. The attachment point on
the peptide
group will depend on the functionality at the terminus of the group by which
the peptide
group is connected to the compound of Formula I or an antibody.
Specifically, the peptidyl group may be coupled to a connecting group via an
alpha
amino or a side chain amino group when a connecting group terminates in, for
example:
-C(=S)-, -S(=0)-, or S02.
Likewise, the peptidyl group may be coupled to a connecting group via an alpha
carboxy or a side chain carboxy group when the connecting group terminates in:
-C(=0)NR5-, -SO2NR5-, -NR5-, -S- or ¨0-.
II. Compounds of the Invention
In one aspect, the invention is a compound of Formula I, or a salt thereof:
28

CA 02836085 2013-12-05
R4 Ar
(0)n
R3
R2 1 N 0
RI
Formula I
wherein
n is 0, 1, or 2;
RI is selected from the group consisting of -H, -(Ci-C6)alkyl, -(C2-
C6)alkenyl,
-(C2-C6)alkynyl, optionally substituted aryl, optionally substituted
heteroaryl, optionally
substituted aryl-(C -C6)alkyl, optionally substituted heteroaryl-(Ci-C6)alkyl,
-C(=0)(C t-
C6)alkyl, -C(=0)(C2-C6)alkenyl, -C(=0)-optionally substituted aryl, -
C(=0)(CH2)nroptionally
substituted aryl, and -C(=0)(CH2)p-optiona11y substituted heteroaryl;
1 0
R2, R3, and R4 are independently selected from the group consisting of -H,
halogen,
-CN, -NR1 R11, -OH, -0R13, -(CI-C6)alkoxy, -NO2, -(CI-C6)alkyl, 4C1-
C6)perfluoroalkyl,
-(C -C6)perfluoroalkoxy, -C(=0)R15, -C(=0)0R15, -0C(-0)R12,
-0C(=0)0R12,
-C(=0)NR17R18, -SH, -S(C -C6)alkyl, -SR13, -S(=0)R13, -S(=0)2R13, -0S(=0)2R13,
-S(=0),A15, -0S(=0),A15, -S(=0)2NRI7R18, -S(=0)NR17R18, optionally substituted
aryl,
optionally substituted aryl-(CI-C6)alkyl, optionally substituted heteroaryl,
optionally
substituted heteroaryl-(Ci-C6)alkyl, optionally substituted (C2-
C9)heterocyclyl, optionally
substituted (C2-C9)heterocycly1-(C -C6)alkyl, -NH(CH2),õC(=0)0R14, -
C(=NR14)NRI42,
-C(=N-ORI4)NR142, -P(=0)(0R14)2, and -0P(=0)(0R14)2;
Ar is optionally substituted heteroaryl, optionally substituted (Cio-Ci4)aryl,
or
29

CA 02836085 2013-12-05
R7
R8 R6
R9 R5
aVVV's
=
R5, R6, R7, R8, and R9 are independently selected from the group consisting of
-H,
-OH, -0R13, -NO2, halogen, -CN, -NR10K -(C112),,NRioRii, -0(CH2)n,NR19R11,
-(C -C6)alkyl, -(CH2),õ0(Ci-C6)alkyl, -(CI-C6)alkoxy,
-(C -C6)perfluoroalkyl,
-(C1-C6)perfluoroalkoxy, -SH, -S(Ci-C6)alkyl, -SR13, -S(=0)R15, -S(=0)2R15, -
C(=0)R15,
-C(=0)0R15, -C(=0)NR17R18, -OC(=0)R16, -0C(=0)0R12, -0C(=0)NR17R18,
heterocyclyl,
optionally substituted heteroaryl, -NH(CH2)n,C(=0)0R14, -0S(=0)2R16, -
C(=NR14)NR142,
_Q_N_oRi4)NR142, _
P(=0)(0R14)2, and -0P(=0)(0R14)2;
each R1 and RH is independently selected from the group consisting of -H,
-(Ci-C6)alkyl, -(C1-C6)alkoxy, -C(=0)R12, -C(=0)NR17R18, ..q=0)0R12, -
Q_NR14)NR17R185
R13, optionally substituted aryl, optionally substituted heteroaryl, and -
C(=NR14)R15; or R19
and RH, together with the nitrogen to which they are bound, form an optionally
substituted
(C2-05)heterocycle;
each R12 is independently selected from the group consisting of -(Ci-C6)alkyl,
and
optionally substituted aryl;
each R13 is independently selected from the group consisting of optionally
substituted
aryl and -(CH2),õR16;
each R14 is independently selected from the group consisting of -H and -(Ci-
C6)alkyl;
or two occurrences of Ri4 bound to the same nitrogen form a (C2-
C6)heterocycle, together
with the nitrogen atom to which they are bound;
each R15 is independently selected from the group consisting of -H, -(C1-
C6)alkyl,
optionally substituted aryl, and NRI42;

CA 02836085 2013-12-05
. =
each R16 is independently selected from the group consisting of -(CI-C6)alkyl,
-NR142,
and Arl;
each R17 and R18 is independently selected from the group consisting of -H,
-(Ci-C6)alkyl, -(Ci-C6)alkoxy, R13, optionally substituted aryl, and
optionally substituted
heteroaryl; or R17 and R18, together with the nitrogen to which they are
bound, form an
optionally substituted (C2-05)heterocycle;
m is independently at each occurrence 1, 2, 3, 4, or 5;
p is independently at each occurrence 0, 1, 2, or 3;
q is independently at each occurrence 0, 1, or 2;
1 0
each optionally substituted aryl, optionally substituted (C10-C14)aryl,
optionally
substituted heteroaryl, optionally substituted aryl-(C1-C6)alkyl, optionally
substituted
heteroaryl-(Ci-C6)alkyl, optionally substituted (C2-C9)heterocyclyl,
optionally substituted (C2-
C9)heterocycly1-(CI-C6)alkyl, and optionally substituted (C2-05)heterocycle is
optionally
substituted with one or more substituents independently selected from the
group consisting of
1 5 halogen, -CN, -NR142, -(CH2)mNR142, -0(CH2).NR142, -NRI4C(----0)(C -
C6)alkyl,
-NR'4C(=0)0(C -C6)alkyl, -NR14C(=0)NR142, -NR' 4 -
C( NRI4)NR142,
-NH(CH2),X(=0)0R14, -OH, -NO2, -(CI-C6)alkyl, -(CH2)m0(Ci-C6)alkyl, -(Ci-
C6)alkoxY,
-(C2-C6)alkenyl, -(C2-C6)alkynyl, -SR14, -S(=0)R15, -S(=0)2R15, -
NR14S(=0)2R15, -(Ci-
C6)perfluoro alkyl, -(Ci-C6)p erfluoro alkoxy,
-C(=0)0R14, -C(=0)NR1425
20 -0C(=0)R14, -0C(=0)NR142, -0C(-0)0(Ci-C6)alkyl, -P(-0)(0R14)2, -01)(-
0)(0R14)2,
heterocyclyl, and heteroaryl;
Arl is a radical according to Formula II
R21
R22 R2o
R23 R19
Formula II
31

CA 02836085 2013-12-05
wherein R19, R20, R21, R22, and ic .-.23
are independently selected from the group
consisting of -H, -OH, -NO2, halogen, -CN, R'1,_NR1 -(CH2),õNR K11,
0(CH2)õ,NR10R11,
-(C -C6)alkyl, -(CH2),1O(C -C6)alkyl, -(C -C6)alkoxy,
-(CI-COperfluoroalkyl, -(C 1-
C6)perfluoroalkoxy, -SH, -SR12, -S(=0)R15, -S(=0)2R15, -C(=0)R15, -C(=0)0R15,
-C(=0)NRt7R18,
OC(=0)R16, _OC(=0)0RI 2, -0C(=0)NR17R 8, heterocyclyl, optionally
substituted heteroaryl, -NH(CH2).C(=0)0R14,
-0S(=0)2R16,
_c(=NR14)N---K2 14, _
C(=N-OR14)NR142, -P(=0)(0R14)2, and -0P(=0)(0R14)2;
provided that:
i) at least one of R2, R3, or R4 is other than hydrogen;
ii) when none of R2, R3, and R4 are -0R13, -NHR13, -SR13, -S(=0)R13, or
-S(=0)2R13, and Ar is
R7
R8 R6
R9 R5
%NW
then at least one of R6 and R8 is -NO2 and at least R7 is other than hydrogen
or halogen; and
iii) when Ar is optionally substituted heteroaryl and none of R2, R3, or R4
are
-0R13, -NHR13, -SR13, -S(=0)R13, or -S(=0)2R13, then R1 is other than
hydrogen.
In preferred embodiments of a compound of Formula I, n is 0. In other
embodiments,
however, n is 1 or 2.
The wavy bond in the structure of Formula I indicates either (E), (Z), or a
mixture of
configurations of the double bond to the carbon atom to which Ar and H are
attached.
32

CA 02836085 2013-12-05
. =
In certain embodiments of the invention, the double bond in the compounds of
Formula I can be in the E configuration. In preferred embodiments, the double
bond is in the
Z configuration:
R4 Ar
(0)ri
R3 SH
R2 NO
R1
In certain instances, the double bond in the compound of Formula I is in the Z
configuration, n is 0, and Ar is optionally substituted heteroaryl or
R7
Rs R6
R9 R5
avvvs
In certain embodiments, the optionally substituted heteroaryl group can be
thiophene-
2-y1 (thiene-2-y1), thiophene-3-y1 (thiene-3-y1), indo1-2-yl, indo1-3-yl,
indo1-4-yl, indo1-5-yl,
indo1-6-yl, indo1-7-yl, pyrrol-2-yl, pyrrol-3-yl, pyrimidin-4-yl, pyrimidin-5-
yl, or pyrimidin-6-
y1, any of which can be optionally substituted. In certain embodiments, the
thiophene-2-y1
(thiene-2-y1), thiophene-3-y1 (thiene-3-y1), indo1-2-yl, indo1-3-yl, indo1-4-
yl, indo1-5-yl, indo1-
6-y1, indo1-7-yl, pyrrol-2-yl, pyrrol-3-yl, pyrimidin-4-yl, pyrimidin-5-yl,
and pyrimidin-6-y1
radicals can be optionally substituted with a halogen, an alkyl group, such as
methyl or ethyl,
or an acyl group such as an acetyl group. In particular embodiments, the
acetyl group can be
present on the nitrogen of any of the pyrrolyl or indolyl radicals. In other
instances, the
pyrimidinyl radicals can be substituted with a thioether, such as -SCH3, or a
morpholino
33

CA 02836085 2013-12-05
=
,
group at any of the substitutable position, and in particular embodiments, at
the 2 position of
the pyriminidyl radical.
In certain embodiments of the invention, any of R2, R3, or R4 can be -
S(=0)2R13
wherein R13 is -(CH2).R16, m is 1, and R16 is
R21
R23
R22 IP R R2oio
sfl/VVs
I .
In particular embodiments, R2 is -S(=0)2R13 wherein R13 is -(CH2)mR16, III is
1, and
R16 is
R21
R22 R20
R23 R19
alf \ AP
I
=
In certain embodiments, at least two of R19, R20, R21, R22,
and R23 are halogen atoms
and can be the same or different halogen atoms. In particular embodiments, R19
and R23 are
both halogen atoms (either the same or different), while R20, R21, and R22 are
other than
halogen. In a further embodiment, le9 and R23 are the same halogen atom, while
R20, R21, and
R22 are other than halogen. In certain embodiments, R19 and R23 are both
chlorine atoms and
R20, It -.,21,
and R22 are each hydrogen.
1 5 Particular examples of compounds according to the invention, and
salts thereof, are set
forth in Table 1:
34

CA 02836085 2013-12-05
. .
,
Table 1
Cp Name Structure
d.
#
1 (Z)-4-((6-chloro-3-oxo- 0
3,4-dihydro-2H-
0 CH3
benzo[b][1,4]thiazin-2-
ylidene)methyl)-2- 0 NO2
nitrophenyl acetate
0 S
Cl N 0
H
2 (Z)-4-((7-methoxy-3-oxo- 0
3,4-dihydro-2H- ,,.
0 CH3
benzo[b][1,4]thiazin-2-
ylidene)methyl)-2- lei
NO2
nitrophenyl acetate
CH3
I
010 s
N 0
H

CA 02836085 2013-12-05
,
, .
,
3 (Z)-4-((7-bromo-3-oxo- 0
3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-
0 CH3
ylidene)methyl)-2- 10
NO2
nitrophenyl acetate
Br40 S
N 0
H
4 (Z)-4((7-methy1-3-oxo- 0
3,4-dihydro-2H- ,7.--
,s
0 CH3
benzo[b][1,4]thiazin-2-
op No2
ylidene)methyl)-2-
nitrophenyl acetate
H3C10 S ,/
N 0
H
_
(Z)-2-(4-acetoxy-3- 0
nitrobenzylidene)-3-oxo- ...."--
-,...
0 CH3
3,4-dihydro-2H-
benzo[b][1,4]thiazine-7-
NO2
carboxylic acid
0
ii
7C 0 S /
HO
N 0
H
36

CA 02836085 2013-12-05
. . ,
6 (Z)-4((7-fluoro-3-oxo-3,4- 0
dihydro-2H- .,/...,,.
benzo[b][1,4]thiazin-2- 0 CH3
ylidene)methyl)-2-10 NO2
nitrophenyl acetate
F 10 S ,v-
N 0
H
7 (Z)-4-((6-fluoro-3-oxo-3,4- ' 0
dihydro-2H- ,,-....,
benzo[b][1,4]thiazin-2- 0 CH3
ylidene)methyl)-2-01 NO2
nitrophenyl acetate
0 S
F N 0
H
37

CA 02836085 2013-12-05
8 (Z)-4-((6-((2,6- 9
dichlorobenzyl)sulfony1)- õ,----.....õ
0 0 CH3
3-oxo-3,4-dihydro-2H- II
benzo[b][1,4]thiazin-2- -e N+ 0
ylidene)methyl)-2-
nitrophenyl acetate
Cl
0 S
410 0_
N 0
% H
Cl.
9 (Z)-4-((6-((2,6- 0
dichlorobenzyl)sulfony1)- ./=.,,
0 CH3
3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl 01
acetate
0 S ,,.-
CI
01 0
\ N 0
H
CI
1 0 (Z)-2-benzylidene-6-((2,6-
dichlorobenzyl)sulfony1)-
1110
2H-benzo[b][1,4]thiazin-
3(4H)-onelift S .'
Cl
C1/4
S IgF N 0
H
0
C I
38

CA 02836085 2013-12-05
11 (Z)-6-((2,6- 0 OCH3
dichlorobenzyl)sulfony1)- W
2-(4-methoxy-3- -00
nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-
S
CI =
3(4H)-one
C% = N 0
0
CI
12 (Z)-2-(3-amino-4- OCH3
methoxybenzylidene)-6- H2N
((2,6 =
-
dichlorobenzyl)sulfony1)-
2H-benzo[b][1,4]thiazin-
S
CI
3(4H)-one
140 Cs%S = N 0
0
Cl
39

CA 02836085 2013-12-05
13 (Z)-6-((2,6- CH3
dichlorobenzy1)su1fony1)-
2-(4-(4-methylpiperazin-1-
yl)benzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one
101
CI
0%
S
=
S N 0
0
CI
14 (Z)-6-((2,6- 0
dichlorobenzyl)sulfony1)-
2-(4-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one
S
(01 1 Cl 1.F 0
0
CI

CA 02836085 2013-12-05
. =
15 (Z)-2-(4-
NH2
aminobenzylidene)-6-
((2,6- =
dichlorobenzyl)sulfony1)-
2H-benzo[b][1,4]thiazin-
CI
S
3(4H)-one
o
=
11 1 N 0
0
C I
16 (Z)-6-((2,6-
dichlorobenzyl)sulfony1)-
2-(4-fluorobenzylidene)-
110
2H-benzo [b] [1,4]thiazin-
3 (4H)-one
S
CI
1101 oS N 0
0
C I
17 (Z)-2-(4-
Cl
chlorobenzylidene)-6-
((2,6-
1101
dichlorobenzyl)sulfony1)-
2H-benzo [b] [1 ,4]thiazin-
S
3(4H)-one C I
I o%, siWIN 0
0
Cl
41

CA 02836085 2013-12-05
,
18 (Z)-2-(4- Br
bromobenzylidene)-6- .
((2,6-
dichlorobenzyl)sulfony1)-
2H-benzo[b][1,4]thiazin-
S
3(4H)-one Cl
O101
0%
S N 0
% H
0
C I
19 (Z)-6-((2,6-
OCH3
dichlorobenzyl)sulfony1)-
2-(4-
0
methoxybenzylidene)-2H-
benzo[b][1,4]thiazin-
S
CI
3(4H)-one
1110 12 S $ N 0
% H
0
Cl
20 (Z)-6-((2,6- C H 3
dichlorobenzyl)sulfony1)-
2-(4-methylbenzylidene)-
2H-benzo[b][1,4]thiazin-
3(4H)-one
S
C I
10 ()%s,\ I N 0
\\ H
0
C I
42

CA 02836085 2013-12-05
. .
21 (Z)-6-((2,6- OCH3
dichlorobenzypsulfony1)-
242,4,6-
*
H3C0 OCH3
trimethoxybenzylidene)-
2H-benzo[b][1,4]thiazin- Cl S
3(4H)-one
N 0
% H
0
CI
22 (Z)-6-((2,6- Cl
dichlorobenzyl)sulfony1)-
2-(2,4-
diehlorobenzylidene)-2H-
CI
benzo[b][1,4]thiazin-
1, S
3(4H)-one CI
10 0%
S N 0
% H
0
CI
23 (number intentionally -
skipped)
24 (Z)-methyl 2-((4-((6-((2,6-
y'()CH3
dichlorobenzypsulfony1)-
HN/
3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-
1401
ylidene)methyl)phenyl)ami
no)acetate
Cl S
10 0%
S $ N 0
H
0
Cl
43

CA 02836085 2013-12-05
25 (Z)-2-((4-((6-((2,6-
/
dichlorobenzyl)sulfony1)-
3-oxo-3,4-dihydro-2H-
HN/
benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl)ami
no)acetic acid
CI S
%s N 0
0
CI
26 (Z)-2-((1-acety1-1H-indol- 0
3-yl)methylene)-6-((2,6-
dichlorobenzyl)sulfony1)-
= N
2H-benzo[b][1,4]thiazin-
3(4H)-one S
CI
0\\
\'S N 0
0
CI
27 (Z)-2-((1H-pyrrol-3 NH
-
yl)methylene)-6-((2,6-
dichlorobenzyl)sulfony1)-
2H-benzo[b][1,4]thiazin- CI =
3(4H)-one Cl =
%s
0
CI
44

CA 02836085 2013-12-05
. . =
..
28 (Z)-6-((2,6- OH
dichlorobenzyl)sulfony1)-
2-(4-hydroxy-2,6-
0
H OCH3
3CO
dimethoxybenzylidene)-
ii&
2H-benzo[b][1,4]thiazin- CI S
3(4H)-one 10 Co%
S I N 0
% H
0
CI
29 (Z)-2-(4-chloro-3- 0 Cl
II
nitrobenzylidene)-6-((2,6- N 4 '
dichlorobenzyl)sulfony1)- -o '
0
2H-benzo[b][1,4]thiazin-
3(4H)-one
Cl S
I 0S $ N 0
%
H
0
C I
30 ' (Z)-64(2,6- F
dichlorobenzyl)sulfony1)-
2-(2,4-
F
0
difluorobenzylidene)-2H-
benzo[b][1,4]thiazin-
0 s
3(4H)-one CI
0 0µ\
\ \ S N 0
% H
0
C I

CA 02836085 2013-12-05
31 (Z)-6-((2,6-
dichlorobenzypsulfony1)-
2-(2,4,6 =
-
trifluorobenzylidene)-2H-
benzo [b] [ 1 ,4]thiazin- S
CI
3(4H)-one 01 0%
N 0
0
CI
32 (Z)-methyl 4-((6-((2,6- 0 OCH3
dichlorobenzyl)sulfony1)-
3 -oxo-3 ,4-dihydro-2H-
benzo [b][1 ,4]thiazin-2- 101
ylidene)methyl)benzoate
S
CI
O\
\\S N 0
0
Cl
33 (Z)-6-((2,6-
dichlorobenzyl)sulfony1)- C\S
2-(thiophen-2-
ylmethylene)-2H- CI
S.
benzo [b] [1 ,4]thiazin- 10 0%s 10 No
3(4H)-one
0
CI
46

CA 02836085 2013-12-05
34 4 (Z)-2-((1H-indo1-3-
yl)methylene)-6-((2,6-
NH
dichlorobenzyl)sulfony1)-
2H-benzo[b][1,4]thiazin-
3(4H)-one Cl = =
CZ\
S N 0
0
Cl
35 (Z)-methyl 4-((6-((2,6- 0 OCH3
dichlorobenzyl)sulfony1)-
3-oxo-3,4-dihydro-2H-
,
benzo[b][1,4]thiazin-2- N0-
+
ylidene)methyl)-3- S 0
CI
nitrobenzoate 0,\
N 0
0
CI
36 (Z)-6-((2,6-
-CH3
dichlorobenzyl)sulfony1)- 1
2-((2-
(methylthio)pyrimidin-4- CI
yl)methylene)-2H- c1/4
benzo[b][1,4]thiazin-
0
CI
3(4H)-one
47

CA 02836085 2013-12-05
, .
37 (Z)-6-((2,6- 0
dichlorobenzypsulfony1)-_.-
-' .7' ...-
2-((2-
1 N
morpholinopyrimidin-4-
yl)methylene)-2H- Ci s,
0
benzo[b][1,4]thiazin- 0 sOs
3(4H)-one %
0 H
CI
38 (Z)-4-((6-((2,6- 40
cH3
dichlorobenzyl)sulfony1)- 0
o o's%
3-oxo-3,4-dihydro-2H- II 0
benzo[b][1,4]thiazin-2- -0'N+ 0
ylidene)methyl)-2-
nitrophenyl 4- 10 s
ci
methylbenzenesulfonate 0 0%
N 0
S H
%
0
CI
39 (Z)-2-(4-(2H-tetrazol-5- //
N-----NH
\
yl)benzylidene)-6-((2,6- N N
/
dichlorobenzypsulfony1)-
2H-benzo[b][1,4]thiazin-
0
3(411)-one
0 CI 0 S
0\\
\\S N 0
% H
0
CI
48

CA 02836085 2013-12-05
,
= .
40 (Z)-4-((4-acetyl-7- CH3
methoxy-3-oxo-3,4-
dihydro-2H-
0 0
benzo[b][1,4]thiazin-2- 0
NO2
ylidene)methyl)-2-
nitrophenyl acetate
H3C0 0 S
N 0
H3C 0
41 (Z)-2-(4-(benzyloxy)-3-
0
nitrobenzylidene)-6-((2,6-
dichlorobenzyl)sulfony1)-
2H-benzo[b][1,4]thiazin-
0 0
3(4H)-one
W
-00 0
0 40 S
CI
0 0\
\ \ S N
% H
0
C I
49

CA 02836085 2013-12-05
=
42 (Z)-6-((2,6- OH
dichlorobenzyl)sulfony1)- NO2
2-(4-hydroxy-3-
nitrobenzylidene)-2H-
benzo[b][1,4]thiazin- 0 S
CI
3(4H)-one sCo
N 0
0
CI
43 (Z)-2-(4-(benzyloxy)-3-
nitrobenzylidene)-7-
fluoro-2H-
benzo[b][1,4]thiazin-
3(41-1)-one
0
=
NO2
S
0

CA 02836085 2013-12-05
44 (Z)-7-fluoro-2-(4-hydroxy- OH
3 -nitrobenzylidene)-2H-
NO2
benzo [b] [1,4]thiazin-
3(4H)-one
F S
0
Other exemplary compounds include, but are not limited to the following, and
salts
thereof:
(Z)-44(64(2,6-dibromobenzyl)sulfony1)-3 -oxo-3,4-dihydro -2H-
benzo [b] [1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate;
(Z)-4-((6-((2,6-
dibromobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo [b] [1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-6-((2,6-
dibromobenzypsulfony1)-2H-
benzo [b] [1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-dibromobenzyl)sulfony1)-2-(4-methoxy-3-
nitrobenzylidene)-2H-benzo[b] [1,4]thiazin-3(4H)-one;
(Z)-2-(3 -amino-4-
methoxybenzylidene)-642,6-dibromobenzypsulfony1)-2H-benzo [b] [1,4]thiazin-
3(4H)-one;
(Z)-6-((2,6-dibromobenzyl)sulfony1)-2-(4-(4-methylpiperazin-1-y1)benzylidene)-
2H-
benzo [b] [1,4]thiazin-3(4H)-one; (Z)-64(2,6-dibromobenzypsulfony1)-2-(4-
nitrobenzylidene)-
2H-benzo [b] [1,4]thiazin-3(4H)-one;
(Z)-2-(4-aminobenzylidene)-6-((2,6-
dibromobenzyl)sulfony1)-2H-benzo [b] [1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-
dibromobenzyl)sulfony1)-2-(4-fluorobenzylidene)-2H-benzo [b] [1,4]thiazin-
3(4H)-one; (Z)-2-
(4-ehlorobenzylidene)-6-((2,6-dibromobenzyl)sulfony1)-2H-benzo [b]
[1,4]thiazin-3(4H)-one;
(Z)-2-(4-bromobenzylidene)-64(2,6-dibromobenzypsulfony1)-2H-benzo[b]
[1,4]thiazin-
3 (4H)-one;
(Z)-6-((2,6-dibromobenzyl)sulfony1)-2-(4-methoxybenzylidene)-2H-
benzo [b] [1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-dibromobenzyl)sulfony1)-2-(4-
methylbenzylidene)-2H-benzo [b] [1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-
dibromobenzyl)sulfony1)-2-(2,4,6-trimethoxybenzylidene)-2H-benzo [b]
[1,4]thiazin-3(4H)-
51

CA 02836085 2013-12-05
,
,
one;
(Z)-64(2,6-dibromobenzypsulfony1)-2-(2,4-dibromobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-dibromobenzyl)sulfony1)-2-(2,4,6-
trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-methyl
2-((4-((6-((2,6-
dibromobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl)amino)acetate; (Z)-24(44642,6-dibromobenzyl)sulfony1)-3-
oxo-
3,4-dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetic acid;
(Z)-64(2,6-
dibromobenzypsulfony1)-2-(4-hydroxy-2,6-dimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-(4-chloro-3-nitrobenzylidene)-6-((2,6-dibromobenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-dibromobenzyl)sulfony1)-2-(2,4-
difluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-
dibromobenzypsulfony1)-2-(2,4,6-trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-methyl 4-((6-((2,6-dibromobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-
2-ylidene)methyl)benzoate; (Z)-methyl 4-46-((2,6-dibromobenzypsulfony1)-3-oxo-
3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-3-nitrobenzoate;
(Z)-4-((6-((2,6-
dibromobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-
nitrophenyl 4-methylbenzenesulfonate; (Z)-2-(4-(2H-tetrazol-5-yl)benzylidene)-
6-((2,6-
dibromobenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-2-(4-(benzyloxy)-3-
nitrobenzylidene)-6-((2,6-dibromobenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one; (Z)-
642,6-dibromobenzyl)sulfony1)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-4-46-((2,6-difluorobenzypsulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate;
(Z)-4-((6-((2,6-
difluorobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-6-((2,6-
difluorobenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-642,6-difluorobenzypsulfony1)-2-(4-methoxy-3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-2-(3-amino-4-
methoxybenzylidene)-64(2,6-difluorobenzypsulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-642,6-difluorobenzyl)sulfony1)-2-(4-(4-methylpiperazin-1-yl)benzylidene)-
2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-difluorobenzypsulfony1)-2-(4-
nitrobenzylidene)-
2H-benzo[b][1,41thiazin-3(4H)-one;
(Z)-2-(4-aminobenzylidene)-6-((2,6-
difluorobenzypsulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-
52

CA 02836085 2013-12-05
t =
=
difluorobenzyl)sulfony1)-2-(4-fluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-
(4-chlorobenzylidene)-6-((2,6-difluorobenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-(4-bromobenzylidene)-64(2,6-difluorobenzypsulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-
one;
(Z)-6-((2,6-difluorobenzyl)sulfony1)-2-(4-methoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-difluorobenzyl)sulfony1)-2-(4-
methylbenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-
difluorobenzyl)sulfony1)-2-(2,4,6-trimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one;
(Z)-6-((2,6-difluorobenzyl)sulfony1)-2-(2,4-difluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-difluorobenzyl)sulfony1)-2-(2,4,6-
trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-methyl
24(4464(2,6-
difluorobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl)amino)acetate; (Z)-244-46-((2,6-difluorobenzypsulfony1)-
3-oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetic acid; (Z)-
6-((2,6-
difluorobenzyl)sulfony1)-2-(4-hydroxy-2,6-dimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-(4-chloro-3-nitrobenzylidene)-6-((2,6-difluorobenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-difluorobenzyl)sulfony1)-2-(2,4-
difluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-
difluorobenzyl)sulfony1)-2-(2,4,6-trifluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-methyl 4-((6-((2,6-difluorobenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-
2-ylidene)methyl)benzoate; (Z)-methyl 44(64(2,6-difluorobenzypsulfonyl)-3-oxo-
3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-3-nitrobenzoate;
(Z)-4-((6-((2,6-
difluorobenzyl)sulfony1)-3-0xo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)-2-
nitrophenyl 4-methylbenzenesulfonate; (Z)-2-(4-(2H-tetrazol-5-yl)benzylidene)-
642,6-
difluorobenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-2-(4-(benzyloxy)-3-
nitrobenzylidene)-6-((2,6-difluorobenzypsulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one; (Z)-
642,6-difluorobenzypsulfony1)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-44(64(2,6-dimethylbenzypsulfony1)-3-oxo-3,4-dihydro-211-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate;
(Z)-4-((6-((2,6-
dimethylbenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-6-((2,6-
dimethylbenzyl)sulfony1)-2H-
53

CA 02836085 2013-12-05
= =
benzo[b][1,4[thiazin-3(4H)-one;
(Z)-6-((2,6-dimethylbenzyl)sulfony1)-2-(4-methoxy-3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-2-(3-amino-4-
methoxybenzylidene)-6-((2,6-dimethylbenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-6-((2,6-dimethylbenzyl)sulfony1)-2-(4-(4-methylpiperazin-1-yl)benzylidene)-
2H-
benzo[b][1,4]thiazin-3(4H)-one; (Z)-6-((2,6-dimethylbenzypsulfony1)-2-(4-
nitrobenzylidene)-
2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-2-(4-aminobenzylidene)-6-((2,6-
dimethylbenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-
dimethylbenzyl)sulfony1)-2-(4-fluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-
one; (Z)-2-
(4-chlorobenzylidene)-6-((2,6-dimethylbenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-(4-bromobenzylidene)-6-((2,6-dimethylbenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-6-((2,6-dimethylbenzyl)sulfony1)-2-(4-methoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-dimethylbenzyl)sulfony1)-2-(4-
methylbenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-
dimethylbenzyl)sulfony1)-2-(2,4,6-trimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-
one;
(Z)-64(2,6-dimethylbenzyl)sulfony1)-2-(2,4-dimethylbenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-dimethylbenzyl)sulfony1)-2-(2,4,6-
trifluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(411)-one; (Z)-methyl
2-((4-((6-((2,6-
dimethylbenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo [b] [1,4]thiazin-2-
ylidene)methyl)phenyl)amino)acetate; (Z)-2-((4-((6-((2,6-
dimethylbenzyl)sulfony1)-3-oxo-
3,4-dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)phenyl)amino)acetic acid;
(Z)-6-((2,6-
dimethylbenzyl)sulfony1)-2-(4-hydroxy-2,6-dimethoxybenzylidene)-2H-
benzo[b][1,4]thiazin-
3(4H)-one;
(Z)-2-(4-ehloro-3-nitrobenzylidene)-64(2,6-dimethylbenzyl)sulfony1)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-dimethylbenzyl)sulfony1)-2-(2,4-
difluorobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-
dimethylbenzyl)sulfony1)-2-(2,4,6-trifluorobenzylidene)-2H-
benzo[b][1,4]thiazin-3(4H)-one;
(Z)-methyl 4-((6-((2,6-dimethylbenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo [b]
[1,4]thiazin-
2-ylidene)methyl)benzoate; (Z)-methyl
4-464(2 ,6-dimethylbenzyl)sulfony1)-3 -oxo-3,4-
dihydro-2H-benzo[b][1,4]thiazin-2-ylidene)methyl)-3-nitrobenzoate;
(Z)-4-((6-((2,6-
dimethylbenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo [b] [1,4]thiazin-2-
ylidene)methyl)-2 -
nitrophenyl 4-methylbenzenesulfonate; (Z)-2-(4-(2H-tetrazol-5-yl)benzylidene)-
6-((2,6-
54

CA 02836085 2013-12-05
. . =
dimethylbenzypsulfony1)-2H-benzo[b] [1,4]thiazin-3(4H)-one;
(Z)-2-(4-(benzyloxy)-3-
nitrobenzylidene)-6-((2,6-dimethylbenzyl)sulfony1)-2H-benzo [b] [1,4]thiazin-
3(4H)-one; and
(Z)-6-((2,6-dimethylbenzyl)sulfony1)-2-(4-hydroxy-3-nitrobenzylidene)-2H-
benzo [b] [1,4]thiazin-3(4H)-one;
(Z)-4-((6-((2,6-dimethoxybenzyl)sulfony1)-3-oxo-3,4-
dihydro-2H-benzo[b] [1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl acetate; (Z)-
4-((6-((2,6-
dimethoxybenzyl)sulfony1)-3-oxo-3,4-dihydro-2H-benzo [b] [1,4]thiazin-2-
ylidene)methyl)phenyl acetate; (Z)-2-benzylidene-64(2,6-
dimethoxybenzyl)sulfony1)-2H-
benzo [b] [1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-dimethoxybenzyl)sulfony1)-2-(4-methoxy-3-
nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one;
(Z)-2-(3-amino-4-
methoxybenzylidene)-6-((2,6-dimethoxybenzyl)sulfony1)-2H-benzo [b]
[1,4]thiazin-3 (4H)-
one; (Z)-6-((2,6-dimethoxybenzyl)sulfony1)-2-(4-(4-methylpiperazin-1-
y1)benzylidene)-2H-
benzo[b] [1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-dimethoxybenzyl)sulfony1)-2-(4-
nitrobenzylidene)-2H-benzo [b] [1,4]thiazin-3(4H)-one; (Z)-2-(4-
aminobenzylidene)-6-((2,6-
dimethoxybenzyl)sulfony1)-2H-benzo [b] [1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-
dimethoxybenzyl)sulfony1)-2-(4-fluorobenzylidene)-2H-benzo[b][1,4]thiazin-
3(4H)-one; (Z)-
2-(4-chlorobenzylidene)-6-((2,6-dimethoxybenzyl)sulfony1)-2H-benzo[b]
[1,4]thiazin-3(4H)-
one ;
(Z)-2-(4-bromobenzylidene)-64(2,6-dimethoxybenzypsulfony1)-2H-
benzo [b] [1,4]thiazin-3(4H)-one;
(Z)-64(2,6-dimethoxybenzyl)sulfony1)-2-(4-
methoxybenzylidene)-2H-benzo [b] [1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-
dimethoxybenzyl)sulfony1)-2-(4-methylbenzylidene)-2H-benzo [b] [1,4]thiazin-
3(4H)-one;
(Z)-64(2,6-dimethoxybenzypsulfony1)-2-(2,4,6-trimethoxybenzylidene)-214-
benzo[b] [1,4]thiazin-3(4H)-one;
(Z)-64(2,6-dimethoxybenzypsulfony1)-2-(2,4-
dimethoxybenzylidene)-2H-benzo [b] [1,4]thiazin-3(4H)-one;
(Z)-6-((2,6-
dimethoxybenzyl)sulfony1)-2-(2,4,6-trifluorobenzylidene)-2H-benzo [b]
[1,4]thiazin-3 (4H)-
one; (Z)-methyl 2-((4-
((6-((2,6-dimethoxybenzyl)sulfony1)-3 -oxo-3,4-dihydro-2H-
benzo [b] [1,4]thiazin-2-ylidene)methyl)phenypamino)acetate;
(Z)-2-((4-((6-((2,6-
dimethoxybenzyl)sulfony1)-3 -oxo-3,4-dihydro-2H-benzo [b] [1,4]thiazin-2-
ylidene)methyl)phenyl)amino)acetic acid; (Z)-64(2,6-dimethoxybenzyl)sulfony1)-
2-(4-
hydroxy-2,6-dimethoxybenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one; (Z)-2-(4-
chloro-3 -
nitrobenzylidene)-6-((2,6-dimethoxybenzyl)sulfony1)-2H-benzo [b] [1,4]thiazin-
3(4H)-one;

CA 02836085 2013-12-05
. . =
(Z)-64(2,6-dimethoxybenzyl)sulfony1)-2-(2,4-difluorobenzylidene)-2H-benzo [b]
[1,4]thiazi n-
3 (4H)-one;
(Z)-642,6-dimethoxybenzypsulfony1)-2-(2,4,6-trifluorobenzylidene)-2H-
benzo[b] [1 ,4] thiazin-3 (4H)-one; (Z)-methyl 4 - ((6- ((2,6-
dimethoxybenzyl)sulfony1)-3 -oxo -
3 ,4-dihydro -2H-benzo [b] [1,4]thiazin-2-ylidene)methyl)benzoate;
(Z)-methyl 4-((6-((2,6-
dimethoxybenzyl)sulfony1)-3 -oxo -3 ,4-dihydro-2H-benzo [b] [1,4]thiazin-2-
ylidene)methyl)-3-
nitrobenzoate;
(Z)-44(64(2,6-dimethoxybenzypsulfony1)-3-oxo-3,4-dihydro-2H-
benzo[b][1,4]thiazin-2-ylidene)methyl)-2-nitrophenyl 4-methylbenzenesulfonate;
(Z)-2-(4-
(2H-tetrazol-5-yl)benzylidene)-642,6-dimethoxybenzyl)sulfony1)-2H-benzo [b]
[1,4]thi azin-
3 (411)-one; (Z)-2 -(4-(b enzyl oxy)-3 -nitrob enzylidene)-6 -((2,6-
dimethoxybenzyl)sul fony1)-2H-
benzo [b] [1,4]thiazin-3(4H)-one; and (Z)-6-((2,6-dimethoxybenzyl)sulfony1)-2-
(4-hydroxy-3-
nitrobenzylidene)-2H-benzo [b] [1,4]thiazin-3(4H)-one.
III. Methods for Preparing Compounds of the Invention and Intermediates Useful
in
the Synthesis of Compounds of the Invention
There are provided processes for preparing compounds according to Formula I,
intermediates that are useful in the preparation of such compounds, and
processes for
preparing such intermediates.
The compounds of Formula I can be prepared by condensing an appropriately
substituted aldehyde according to Formula III
0
ArH
Formula III
wherein Ar is as defined elsewhere herein, with a compound according to
Formula IV:
56

CA 02836085 2013-12-05
R4
R3 (0)n
R2 II NO
R1
Formula IV
wherein RI, R2, R3, R4, and n are as defined elsewhere herein.
The aldehyde according to Formula III may be purchased from a commercial
supplier
such as Sigma Aldrich, or the aldehyde of Formula III may be prepared using
procedures
known in the art. The benzothiazinone of Formula IV can be prepared in at
least two ways.
For example, in one embodiment, a benzothiazinone of Formula IV can be
prepared according
to Scheme 1:
Scheme 1
R4 R4 0
R HSCH2CO2R R
R2 NO2 3 0
3 OR
Na0H, Et0H
C --> Reflux, 40 min R2 NO2
A
1. Zn, AcOH, 0 C --> RT, 2hours R4
-or-
R3
2. Na2S204, Et3N; 3N HCI, 50-60 C, 4h .\
_________________________________________ =
R2 N 0
wherein, R2, R3, and R4 are as defined previously herein.
57

CA 02836085 2013-12-05
According to Scheme 1, an appropriately substituted nitro, fluorobenzene
derivative,
A, is reacted with thioglycolic acid (R = H) or an ester of thioglycolic acid
such as methy or
ethyl thioglycolate (R = alkly) to give thioether B. The nitro group in
thioether B can then be
reduced. In particular embodiments, the reduction conditions employ Zn and
acetic acid. In
an alternative embodiment, the reduction protocol employs sodium dithionite
(sodium
hydrosulfite). Other nitro group reduction protocols may also be suitable. The
resulting
amine then undergoes in situ cyclization to give compound C, which is a
compound according
to Formula IV.
Optionally, the amide functionality in compound C can be alkylated or acylated
via
deprotonation of compound C with an appropriate base, examples of which
include, but are
not limited to K2CO3, NaH, potassium bis(trimethylsilyl)amide (KHMDS), sodium
bis(trimethylsilyl)amide (NaHMDS), and lithium tetramethylpiperazine (LiTMP).
The
resulting anion can then be treated with a reactive electrophile, such as, but
not limited to, an
alkyl or alkenyl halide(e.g. CH3I, CH2=CHCH2Br, etc.), or equivalent thereof.
Alternatively,
the anion can be treated with a reactive acyl species, such as, but not
limited to, an acyl
chloride (e.g. acetyl chloride). The resultant product is a compound of
Formula IV wherein
RI is other than hydrogen.
In other embodiments, the amide functionality in compound C can be arylated or
heteroarylated by reacting compound C with CuI, K3PO4, an appropriately
functionalized aryl
or heteroaryl iodide, and trans-1,2-cyclohexanediamine (10 mol% based on the
equivalents of
aryl iodide) all in an appropriate solvent, examples of which include, but are
not limited to,
acetonitrile, toluene, dioxane, and 1,2-dimethoxyethane. Typically the
reaction is warmed,
and in certain embodiments, warmed to reflux. The reaction is typically
carried out under an
inert atmosphere. Upon completion, the reaction can be filtered through a
silica gel or celite
pad to remove solid impurities. The pad can be washed with additional reaction
solvent to
ensure that all product has been removed from the filter pad. The solvent of
the resultant
filtrate can then be evaporated under reduced pressure and the resultant
residue purified by
flash chromatography. As above, the resultant product is a compound of Formula
IV wherein
RI is other than hydrogen.
58

CA 02836085 2013-12-05
A compound according to Formula IV can also be prepared by functionalizing the
commercially available compound 2H-1,4-benzothiazin-3(4H)-one. For example,
according
to Scheme 2, shown in Figure 1, 2H-1,4-benzothiazin-3(4H)-one is reacted with
chlorosulfonic acid to give 3-oxo-3,4-dihydro-2H-benzo [b][1,4]thiazine-6-
sulfonyl chloride
(compound "D"). Compound D can then be treated with sodium sulfite (Na2S03)
and sodium
hydrogen phosphate (Na2HPO4) to give intermediate E. This intermediate can
then be reacted
with an appropriately substituted benzyl group to give compound F, wherein
R19, R20, R21,
R22, and R23
are as defined previously herein and X is leaving group such as a halide,
tosylate,
mesylate, benzensulfonate, methansulfonate, or other equivalent group suitable
for use in SN2
type chemistry. Compound F is a compound according to Formula IV.
Alternatively,
compound D can be reacted with Zn metal to give a reactive Zn complex, G,
which can
subsequently be reacted with the above described benzyl group, to give
compound F. As set
forth previously, compound F is a compound of Formula IV.
Optionally, the amide functionality in compound F can be alkylated or acylated
via
deprotonation of compound F with an appropriate base, examples of which
include, but are
not limited to K2CO3, NaH, potassium bis(trimethylsilyl)amide (KHMDS), sodium
bis(trimethylsilyl)amide (NaHMDS), and lithium tetramethylpiperazine (LiTMP).
The
resulting anion can then be treated with a reactive electrophile, such as, but
not limited to, an
alkyl or alkenyl halide (e.g. CH3I, CH2=CHCH2Br, etc.), or equivalent thereof.
Alternatively,
the anion can be treated with a reactive acyl species, such as, but not
limited to, an acyl
chloride (e.g. acetyl chloride). The resultant product is a compound of
Formula IV wherein
RI is other than hydrogen.
In other embodiments, the amide functionality in compound F can be arylated or
heteroarylated by reacting compound F with CuI, K3PO4, an appropriately
functionalized aryl
or heteroaryl iodide, and trans-1,2-cyclohexanediamine (10 mol% based on the
equivalents of
aryl iodide) all in an appropriate solvent, examples of which include, but are
not limited to,
acetonitrile, toluene, dioxane, and 1,2-dimethoxyethane. Typically the
reaction is warmed,
and in certain embodiments, warmed to reflux. The reaction is typically
carried out under an
59

CA 02836085 2013-12-05
inert atmosphere. Upon completion, the reaction can be filtered through a
silica gel or celite
pad to remove solid impurities. The pad can be washed with additional reaction
solvent to
ensure that all product has been removed from the filter pad. The solvent of
the resultant
filtrate can then be evaporated under reduced pressure and the resultant
residue purified by
flash chromatography. As above, the resultant product is a compound of Formula
IV wherein
RI is other than hydrogen.
The aldehyde of Formula III can then be condensed with the compound of Formula
IV
to give a compound of Formula I. In certain embodiments, the condensation
reaction takes
place in the presence of a suitable base and an anhydride, wherein the
anhydride acts as the
solvent for the reaction. In other embodiments, the reaction can be run in a
solvent other than
the anhydride, examples of which include, but are not limited to toluene and
acetic acid. In
particular embodiments, the base is triethylamine (Et3N) and the anhydride is
acetic anhydride
(Ac20). Other suitable bases, and in particular amine bases, can be
substituted for Et3N.
Example of other suitable bases include, but are not limited to, di-
isopropylethy amine.
Likewise, other suitable anhydrides can be used. Examples include, but are not
limited to
propionoic anhydride and butyric anhydride. The resulting mixture can be then
be warmed,
and in certain embodiments, heated to reflux to give a compound of Formula I.
An exemplary
reaction scheme is provided in Scheme 3:
Scheme 3
R4 R4 Ar
(0)õ (0)õ
R3 R3
0 Et3N, Ac20 SH
Ar/\ HReflux, 2h
R2 NO R2 NO
R1 H R1
Formula III Formula IV Formula I

CA 02836085 2013-12-05
In certain embodiments, the above-described reaction may result in a compound
of
Formula I wherein RI is hydrogen. The nitrogen bound to R1 can be further
functionalized via
selective arylation, heteroarylation, alkylation, or acylation according to
the procedures
described elsewhere herein for compounds C and F.
Compounds of the invention can be prepared and used as their benzothiazinone-1-
oxides, i.e. compounds of Formula I wherein n is 1. These compounds can be
prepared by
oxidizing a compound of Formula I (or appropriately functionalized
intermediate useful for
the preparation of a compound of Formula I) with an oxidizing agent such as
meta-
chloroperoxybenzoic acid (MCPBA) in a solvent such as, but not limited to,
dichloromethane.
After appropriate basic aqueous workup, the oxidized compound of Formula I can
typically
be isolated in high yield and purity after recrystallization or flash
chromatography.
Compounds of the invention can also be prepared and used as their
benzothiazinone-
1,1-dioxides, i.e. compounds of Formula I wherein n = 2. These compounds can
be prepared
by oxidizing a compound of Formula I (or appropriately functionalized
intermediate useful
for the preparation of a compound of Formula I) with an oxidizing agent such
as hydrogen
peroxide in a solvent such as, but not limited to, acetic acid. After
appropriate aqueous
workup, the desired compound can typically be isolated in high yield after
recrystallization or
flash chromatography.
Although compounds of Formula I are generally produced as their Z isomers, the
compounds of the invention can be converted to their E isomeric forms via a
photo-induced
isomerization (either sunlight or UV/VIS radiation) according to the procedure
described in
Kamila, et al., "Synthesis of (Z)-hetarylmethylene and (Z)-substituted
benzylidene derivatives
of 4H-benzo[1,4]thiazine-3-thiones and their subsequent conversion to
benzothiopyrano-[3,2-
b][1,4]benzothiazines" ARKIVOC 2006 (ii) 1-14, the entirety of which is hereby
incorporated
by reference.
61

CA 02836085 2013-12-05
IV. Antibody Conjugates
Another aspect of the invention relates to antibody conjugates of compounds of
Formula I, wherein said antibody conjugates have the Formula I-L-Ab. The
present invention
further includes salts of the antibody conjugates. In the Formula I-L-Ab, "I"
is a compound
of Formula I; Ab is an antibody; and -L- is a single bond or a linking group
covalently
linking said compound of Formula I to said antibody.
In a preferred sub-embodiment of the aforesaid conjugates of the Formula I-L-
Ab, said
antibody (Ab) is a monoclonal antibody or a monospecific polyclonal antibody.
In a more preferred sub-embodiment of the aforesaid conjugates of the formulae
I-L-Ab, the aforesaid antibody (Ab) is a tumor-specific antibody.
Antibodies, preferably monoclonal antibodies and monospecific polyclonal
antibodies,
and most preferably tumor-specific antibodies, may be covalently linked to
compounds of the
present invention. A "tumor-specific antibody" is an antibody which
specifically binds to a
tumor antigen, e.g., an antigen on a tumor cell.
The covalent linker between a compound of Formula I and an antibody may, in
its
simplest form, comprise a single covalent bond connecting the compound of
Formula I to the
antibody. More commonly the compound of Formula I is attached to the antibody
using a
suitable bifunctional linking reagent. The term "bifunctional linking reagent"
refers generally
to a molecule that comprises two reactive moieties which are connected by a
spacer element.
The term "reactive moieties", in this context, refers to chemical functional
groups capable of
coupling with an antibody or a compound of Formula I by reacting with
functional groups on
the antibody and the compound of Formula I.
An example of a covalent bond formed as a linker between a compound of Formula
I
and an antibody is a disulfide bond formed by the oxidation of an antibody and
a compound
of Formula I wherein n is 0 and wherein a linking group is used that contains
one or more
cysteine amino acids. The cysteine residues can be oxidized to form disulfide
links by
dissolving 1 mg of the a suitable compound of Formula I and 0.5 equivalents of
the desired
antibody in 1.5 ml of 0.1% (v/v) 17.5 mM acetic acid, pH 8.4, followed by
flushing with
62

CA 02836085 2013-12-05
, =
nitrogen and then 0.01 M K2Fe(CN)6 After incubation for one hour at room
temperature, the
adduct peptide is purified by HPLC.
Another example of a suitable covalent bond formed as a linker between a
compound
of Formula I and an antibody is an amide bond formed by reacting an amino
group on a
compound of the invention with a carboxylic acid group which forms part of the
primary
structure of the antibody (Ab) (such as, for example a glutamic or aspartic
amino acid
residue). Alternately, an amide bond could be formed if the reacting moieties
were reversed,
i.e., the compound of Formula I could contain a carboxylic acid functionality
and react with
an amino functionality within the Ab structure.
Alternatively, a compound of Formula I and an antibody Ab may be covalently
linked
using a bifunctional linking reagent.
For example, adducts can be prepared by first preparing
S-(-N-hexylsuccinimido)-modified derivatives of an antibody and of a compound
of Formula
I, according to the method of Cheronis et al., J. Med. Chem. 37: 348 (1994)
(the entire
disclosure of which is incorporated herein by reference). N-hexylmaleimide, a
precursor for
the modified antibody and compound of Formula I, is prepared from
N-(methoxycarbonyl)maleimide and N-hexylamine by mixing the two compounds in
saturated NaHCO3 at 0 C according to the procedure of Bodanszky and
Bodanszky, The
Practice of Peptide Synthesis; Springer-Verlag, New York, pp. 29-31 (1984)(the
entire
disclosure of which is incorporated herein by reference). The product of the
resulting reaction
mixture is isolated by extraction into ethyl acetate, followed by washing with
water, dried
over Na2SO4, and is then concentrated in vacuo to produce N-hexylmaleimide as
a light
yellow oil. S-(N-Hexylsuccinimido)-modified antibody and Formula I compound
are then
prepared from a cysteine-containing peptide and N-hexylmaleimide by mixing one
part
peptide with 1.5 parts N-hexylmaleimide in N,N-dimethylformamide (3.3 mL/mM
peptide)
followed by addition to 30 volumes of 0.1 ivt ammonium bicarbonate, pH 7.5.
The
S-alkylation reaction carried out in this manner is complete in 30 minutes.
The resulting
S-(N-hexylsuccinimido)-modified peptide monomer is purified by preparative
reverse-phase
HPLC, followed by lyophilization as a fluffy, white powder.
63

CA 02836085 2013-12-05
,
Bis-succinimidohexane peptide heterodimers (wherein one peptide is the
antibody and
the other peptide is attached to the Formula I compound) may be prepared
according to the
method of Cheronis et al., supra from cysteine-substituted peptides. A mixture
of one part
bismaleimidohexane is made with two parts peptide monomer in N,N-
dimethylformamide
(3.3mL/mM peptide) followed by addition to 0.1 ammonium bicarbonate, pH 7.5.
The
reaction mixture is stirred at room temperature and is usually completed
within 30 minutes.
The resulting bis-succinimidohexane peptide dimer is purified by preparative
reverse-phase
HPLC. After lyophilization the material is a fluffy, white powder.
Covalently linked adducts of the general Formula I¨L¨Ab of the present
invention
may be prepared by utilizing homo-bifunctional linking reagents (wherein the
two reactive
moieties are the same), such as, for example, disuccinimidyl tartrate,
disuccinimidyl suberate,
ethylene glycolbis-(succinimidyl succinate), 1,5-difluoro-2,4-dinitrobenzene
("DFNB"),
4,4'-diisothiocyano-2,2'-disulfonic acid stilbene ("DIDS"), and bis-
maleimidohexane
("BMH").
Alternatively, hetero-bifunctional linking reagents may be employed. Such
agents
include, for example, N-succinimidy1-3-(2-pyridyldithio)propionate ("SPDP"),
sulfo succinimidy1-2-(p-azido sal icylamido)ethy1-1 -3 ' -dithiopropionate
("SASD", Pierce
Chemical Company, Rockford, IL), N-maleimidobenzoyl-N-hydroxy-succinimidyl
ester
("MB S"), m-maleimidobenzoylsulfosuccinimide ester
("sulfo-MBS"),
N-succinimi dyl (4 -io do ac etypaminob enzo ate
("SIAB"), succinimidyl
4-(N-maleimidomethyl)-cyclohexane-1 -carboxylate
("SMCC"),
succinimidy1-4-(p-maleimidophenyl)butyrate
("SMPB"),
sulfosuccinimidy1(4-iodoacetypamino-benzoate ("sulfo-SIAB"),
sulfosuccinimidyl
4- (N-mal eimidomethyl)cyclohexane-1 -c arboxyl ate
(" sulfo-S MCC"), sulfosuccinimidyl
4-(p-ma1eimidopheny1)-butyrate
("sulfo-SMPB"),
bromoacetyl-p-aminobenzoyl-N-hydroxy-succinimidyl ester,
iodoacetyl-N-
hydroxysuccinimidyl ester, and the like.
For hetero-bifunctional linking, a compound of Formula I is derivatized with,
for
example, the N-hydroxysuccinimidyl portion of the bifunctional reagent, and
the resulting
64

CA 02836085 2013-12-05
=
derivatized compound is purified by chromatography. Next, a suitable tumor-
specific Mab is
reacted with the second functional group of the bifunctional linking reagent,
assuring a
directed sequence of binding between components of the desired adduct.
Typical hetero-bifunctional linking agents for forming protein-protein
conjugates have
an amino-reactive N-hydroxysuccinimide ester (NHS-ester) as one functional
group and a
sulfhydryl reactive group as the other functional group. First, epsilon-amino
groups of
surface lysine residues of either the Mab or the Formula I compound are
acylated with the
NHS-ester group of the cross-linking agent. The remaining component,
possessing free
sulfhydryl groups, is reacted with the sulfhydryl reactive group of the cross-
linking agent to
form a covalently cross-linked dimer. Common thiol reactive groups include,
for example,
maleimides, pyridyl disulfides, and active halogens. For example, MBS contains
a NHS-ester
as the amino reactive group, and a maleimide moiety as the sulfhydryl reactive
group.
Photoactive hetero-bifunctional linking reagents, e.g., photoreactive phenyl
azides,
may also be employed. One such reagent, SASD, may be linked to either a Mab or
to a
Formula I compound wherein with an attached peptidyl group, via its NHS-ester
group. The
conjugation reaction is carried out at pH 7 at room temperature for about 10
minutes. Molar
ratios between about 1 and about 20 of the cross-linking agent to the
compounds to be linked
may be used.
Numerous bifunctional linkers, useful as linkers (-L-), exist which have been
used
specifically for coupling small molecules to monoclonal antibodies, and many
of these are
commercially available. Examples include N-succinimidy1-3-(2-pyridyldithio)-
propionate
(SPDP), 2-iminothiolane (2-IT), 3-(4-
carboxamidophenyldithio)propionthioimidate (CDPT),
N-succinimidyl-acetylthioacetate (SATA), ethyl-S-acetyl-propionthioimidate
(AMPT) and
N-succinimidy1-3-(4-carboxamidophenyldithio)propionate (SCDP). Procedures
for
preparation of immunoconjugates using these linkers is detailed in Cattel, et
al,
"Toxin-Targeted Design for Anticancer Therapy II: Preparation and Biological
Comparison
of Different Chemically Linked Gelonin-Antibody Conjugates", 1 Pharm. Sci.,
1993, 82,
699-704, the entire disclosure of which is incorporated herein by reference.

CA 02836085 2013-12-05
=
V. Treatment of Cellular Proliferative Disorders Using Compounds of the
Invention
According to another embodiment of the invention, a method of treating an
individual
suffering from a cellular proliferative disorder, particularly cancer, is
provided, comprising
administering to said individual an effective amount of at least one compound
according to
Formula I, or a pharmaceutically acceptable salt thereof, either alone, or in
combination with
a pharmaceutically acceptable carrier.
According to another embodiment of the invention, a method of inducing
apoptosis of
cancer cells, preferably tumor cells, in an individual afflicted with cancer
is provided,
comprising administering to said individual an effective amount of at least
one compound
according to Formula I, or a pharmaceutically acceptable salt thereof, either
alone, or in
combination with a pharmaceutically acceptable carrier.
According to another embodiment of the invention, a method of treating an
individual
suffering from a cellular proliferative disorder, particularly cancer, is
provided, comprising
administering to said individual an effective amount of at least one conjugate
of the Formula
I-L-Ab, either alone, or in combination with a pharmaceutically acceptable
carrier.
The invention is also directed to the use in medicine of a compound according
to
Formula I, or a pharmaceutically acceptable salt thereof, or a conjugate
according to Formula
I-L-Ab, or a pharmaceutically acceptable salt thereof
The invention is also directed to the use of a compound according to Formula
I, or a
pharmaceutically acceptable salt thereof, or a conjugate according to Formula
I-L-Ab, or a
pharmaceutically acceptable salt thereof, in the preparation of a medicament
for treatment of a
cellular proliferative disorder, particularly cancer, or for inducing
apoptosis of tumor cells in
an individual affected with cancer.
Particular and preferred embodiments of this aspect of the invention are those
wherein
the compound of Formula I used in the method of treatment, either alone or as
part of a
composition, or as a component of the antibody conjugate, is a particular or
preferred
embodiment of the compound of Formula I in the description of the compounds
and
compositions of the invention as provided herein.
66

CA 02836085 2013-12-05
The compounds according to the invention may be administered to individuals
(mammals, including animals and humans) afflicted with a cellular
proliferative disorder such
as cancer, malignant and benign tumors, blood vessel proliferative disorders,
autoimmune
disorders, and fibrotic disorders. In a particular embodiment of the
invention, the individual
treated is a human.
The compounds are believed effective against a broad range of tumor types,
including
but not limited to the following: ovarian cancer; cervical cancer; breast
cancer; prostate
cancer; testicular cancer, lung cancer, renal cancer; colorectal cancer; skin
cancer; brain
cancer; leukemia, including acute myeloid leukemia, chronic myeloid leukemia,
acute
lymphoid leukemia, and chronic lymphoid leukemia.
More particularly, cancers that may be treated by the compounds, compositions
and
methods of the invention include, but are not limited to, the following:
cardiac cancers, including, for example sarcoma, e.g., angiosarcoma,
fibrosarcoma, rhabdomyosarcoma, and liposarcoma; myxoma; rhabdomyoma;
fibroma; lipoma and teratoma;
lung cancers, including, for example, bronchogenic carcinoma, e.g., squamous
cell, undifferentiated small cell, undifferentiated large cell, and
adenocarcinoma;
alveolar and bronchiolar carcinoma; bronchial adenoma; sarcoma; lymphoma;
chondromatous hamartoma; and mesothelioma;
gastrointestinal cancer, including, for example, cancers of the esophagus,
e.g.,
squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, and lymphoma; cancers
of the stomach, e.g., carcinoma, lymphoma, and leiomyosarcoma; cancers of the
pancreas, e.g., ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma,
carcinoid tumors, and vipoma; cancers of the small bowel, e.g.,
adenocarcinoma,
lymphoma, carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma,
neurofibroma, and fibroma; cancers of the large bowel, e.g., adenocarcinoma,
tubular
adenoma, villous adenoma, hamartoma, and leiomyoma;
67

CA 02836085 2013-12-05
genitourinary tract cancers, including, for example, cancers of the kidney,
e.g.,
adenocarcinoma, Wilm's tumor (nephroblastoma), lymphoma, and leukemia; cancers
of the bladder and urethra, e.g., squamous cell carcinoma, transitional cell
carcinoma,
and adenocarcinoma; cancers of the prostate, e.g., adenocarcinoma, and
sarcoma;
cancer of the testis, e.g., seminoma, teratoma, embryonal carcinoma,
teratocarcinoma,
choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma,
adenomatoid tumors, and lipoma;
liver cancers, including, for example, hepatoma, e.g., hepatocellular
carcinoma; cholangiocarcinoma; hepatoblastoma; angiosarcoma; hepatocellular
adenoma; and hemangioma;
bone cancers, including, for example, osteogenic sarcoma (osteosarcoma),
fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma,
malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant
cell
tumor chordoma, osteochrondroma (osteocartilaginous exostoses), benign
chondroma,
chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors;
nervous system cancers, including, for example, cancers of the skull, e.g.,
osteoma, hemangioma, granuloma, xanthoma, and osteitis deformans; cancers of
the
meninges, e.g., meningioma, meningiosarcoma, and gliomatosis; cancers of the
brain,
e.g., astrocytoma, medulloblastoma, glioma, ependymoma, germinoma (pinealoma),
glioblastoma multiform, oligodendroglioma, schwannoma, retinoblastoma, and
congenital tumors; and cancers of the spinal cord, e.g., neurofibroma,
meningioma,
glioma, and sarcoma;
gynecological cancers, including, for example, cancers of the uterus, e.g.,
endometrial carcinoma; cancers of the cervix, e.g., cervical carcinoma, and
pre-tumor
cervical dysplasia; cancers of the ovaries, e.g., ovarian carcinoma, including
serous
cystadenocarcinoma, mucinous cystadeno carcinoma, unclassified carcinoma,
granulosa-thecal cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, and
malignant teratoma; cancers of the vulva, e.g., squamous cell carcinoma,
intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, and melanoma; cancers
of
68

CA 02836085 2013-12-05
. . =
the vagina, e.g., clear cell carcinoma, squamous cell carcinoma, botryoid
sarcoma, and
embryonal rhabdomyosarcoma; and cancers of the fallopian tubes, e.g.,
carcinoma;
hematologic cancers, including, for example, cancers of the blood, e.g., acute
myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia,
chronic
lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, and
myelodysplastic syndrome, Hodgkin's lymphoma, non-Hodgkin's lymphoma
(malignant lymphoma) and Waldenstrom's macroglobulinemia;
skin cancers, including, for example, malignant melanoma, basal cell
carcinoma, squamous cell carcinoma, Kaposi's sarcoma, moles dysplastic nevi,
lipoma, angioma, dermatofibroma, keloids, psoriasis; and
adrenal gland cancers, including, for example, neuroblastoma.
Cancers may be solid tumors that may or may not be metastatic. Cancers may
also
occur, as in leukemia, as a diffuse tissue. Thus, the term "tumor cell", as
provided herein,
includes a cell afflicted by any one of the above identified disorders.
The compounds are also believed useful in the treatment of non-cancer cellular
proliferative disorders, that is, cellular proliferative disorders which are
characterized by
benign indications.
Such disorders may also be known as "cytoproliferative" or
"hyperproliferative" in that cells are made by the body at an atypically
elevated rate.
Non-cancer cellular proliferative disorders believed treatable by compounds
according to the
invention include, for example: hemangiomatosis in newborn, secondary
progressive
multiple sclerosis, atherosclerosis, chronic progressive myelodegenerative
disease,
neurofibromatosis, ganglioneuromatosis, keloid formation, Paget's disease of
the bone,
fibrocystic disease of the breast, uterine fibroids, Peyronie's disease,
Dupuytren's disease,
restenosis, benign proliferative breast disease, benign prostatic hyperplasia,
X-linked
lymphocellular proliferative disorder (Duncan disease), post-transplantation
lymphocellular
proliferative disorder (PTLD), macular degeneration, and retinopathies, such
as diabetic
retinopathies and proliferative vitreoretinopathy (PVR)
69

CA 02836085 2013-12-05
=
,
Other non-cancer cellular proliferative disorders believed treatable by
compounds
according to the invention include the presence of pre-cancerous
lymphoproliferative cells
associated with an elevated risk of progression to a cancerous disorder. Many
non-cancerous
lymphocellular proliferative disorders are associated with latent viral
infections such as
Epstein-Barr virus (EBV) and Hepatitis C. These disorders often begin as a
benign pathology
and progress into lymphoid neoplasia as a function of time.
VI. Salts of Compounds According to the Invention
The compounds of the present invention may take the form of salts when
appropriately
substituted with groups or atoms capable of forming salts. Such groups and
atoms are well
known to those of ordinary skill in the art of organic chemistry. The term
"salts" embraces
addition salts of free acids or free bases which are compounds of the
invention. The term
"pharmaceutically-acceptable salt" refers to salts which possess toxicity
profiles within a
range that affords utility in pharmaceutical applications. Pharmaceutically
unacceptable salts
may nonetheless possess properties such as high crystallinity, which have
utility in the
practice of the present invention, such as for example utility in process of
synthesis,
purification or formulation of compounds of the invention.
Suitable pharmaceutically-acceptable acid addition salts may be prepared from
an
inorganic acid or from an organic acid. Examples of inorganic acids include
hydrochloric,
hydrobromic, hydriodic, nitric, carbonic, sulfuric, and phosphoric acids.
Appropriate organic
acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic,
heterocyclic,
carboxylic and sulfonic classes of organic acids, examples of which include
formic, acetic,
propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric,
ascorbic, glucuronic,
maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, 4-
hydroxybenzoic,
phenylacetic, mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic,
benzenesulfonic,
pantothenic, trifluoromethanesulfonic, 2-hydroxyethanesulfonic, p-
toluenesulfonic, sulfanilic,
cyclohexylaminosulfonic, stearic, alginic, 13-hydroxybutyric, salicylic,
galactaric and
galacturonic acid. Examples of pharmaceutically unacceptable acid addition
salts include, for
example, perchlorates and tetrafluoroborates.

CA 02836085 2013-12-05
Suitable pharmaceutically acceptable base addition salts of compounds of the
invention include, for example, metallic salts including alkali metal,
alkaline earth metal and
transition metal salts such as, for example, calcium, magnesium, potassium,
sodium and zinc
salts. Pharmaceutically acceptable base addition salts also include organic
salts made from
basic amines such as, for example, N,N-dibenzylethylenediamine,
chloroprocaine, choline,
diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine.
Examples
of pharmaceutically unacceptable base addition salts include lithium salts and
cyanate salts.
All of these salts may be prepared by conventional means from the
corresponding
compound according to Formula I by reacting, for example, the appropriate acid
or base with
the compound according to Formula I. Preferably the salts are in crystalline
form, and
preferably prepared by crystallization of the salt from a suitable solvent.
The person skilled in
the art will know how to prepare and select suitable salt forms for example,
as described in
Handbook of Pharmaceutical Salts: Properties, Selection, and Use By P. H.
Stahl and C. G.
Wermuth (Wiley-VCH 2002).
VII. Pharmaceutical Compositions
The compounds of the invention may be administered in the form of a
pharmaceutical
composition, in combination with a pharmaceutically acceptable carrier. The
active
ingredient or agent in such formulations (i.e. a compound of Formula I) may
comprise from
0.1 to 99.99 weight percent of the formulation. "Pharmaceutically acceptable
carrier" means
any carrier, diluent or excipient which is compatible with the other
ingredients of the
formulation and not deleterious to the recipient.
The active agent is preferably administered with a pharmaceutically acceptable
carrier
selected on the basis of the selected route of administration and standard
pharmaceutical
practice. The active agent may be formulated into dosage forms according to
standard
practices in the field of pharmaceutical preparations. See Alphonso Gennaro,
ed., Remington 's
Pharmaceutical Sciences, 18th Edition (1990), Mack Publishing Co., Easton, PA.
Suitable
dosage forms may comprise, for example, tablets, capsules, solutions,
parenteral solutions,
troches, suppositories, or suspensions.
71

CA 02836085 2013-12-05
For parenteral administration, the active agent may be mixed with a suitable
carrier or
diluent such as water, an oil (particularly a vegetable oil), ethanol, saline
solution, aqueous
dextrose (glucose) and related sugar solutions, glycerol, or a glycol such as
propylene glycol
or polyethylene glycol. Solutions for parenteral administration preferably
contain a water
soluble salt of the active agent. Stabilizing agents, antioxidant agents and
preservatives may
also be added. Suitable antioxidant agents include sulfite, ascorbic acid,
citric acid and its
salts, and sodium EDTA. Suitable preservatives include benzalkonium chloride,
methyl- or
propyl-paraben, and chlorbutanol. The composition for parenteral
administration may take
the form of an aqueous or non-aqueous solution, dispersion, suspension or
emulsion.
For oral administration, the active agent may be combined with one or more
solid
inactive ingredients for the preparation of tablets, capsules, pills, powders,
granules or other
suitable oral dosage forms. For example, the active agent may be combined with
at least one
excipient such as fillers, binders, humectants, disintegrating agents,
solution retarders,
absorption accelerators, wetting agents absorbents or lubricating agents.
According to one
tablet embodiment, the active agent may be combined with
carboxymethylcellulose calcium,
magnesium stearate, mannitol and starch, and then formed into tablets by
conventional
tableting methods.
The specific dose of a compound according to the invention to obtain
therapeutic
benefit for treatment of a cellular proliferative disorder will, of course, be
determined by the
particular circumstances of the individual patient including the size, weight,
age and sex of
the patient, the nature and stage of the cellular proliferative disorder, the
aggressiveness of the
cellular proliferative disorder, and the route of administration of the
compound.
For example, a daily dosage from about 0.05 to about 50 mg/kg/day may be
utilized,
more preferably from about 0.1 to about 10 mg/kg/day. Higher or lower doses
are also
contemplated as it may be necessary to use dosages outside these ranges in
some cases. The
daily dosage may be divided, such as being divided equally into two to four
times per day
daily dosing. The compositions are preferably formulated in a unit dosage
form, each dosage
containing from about 1 to about 500mg, more typically, about 10 to about
100mg of active
agent per unit dosage. The term "unit dosage form" refers to physically
discrete units suitable
72

CA 02836085 2013-12-05
as a unitary dosage for human subjects and other mammals, each unit containing
a
predetermined quantity of active material calculated to produce the desired
therapeutic effect,
in association with a suitable pharmaceutical excipient.
The pharmaceutical compositions of the present invention may also be
formulated so
as to provide slow or controlled release of the active ingredient therein
using, for example,
hydropropylmethyl cellulose in varying proportions to provide the desired
release profile,
other polymer matrices, gels, permeable membranes, osmotic systems, multilayer
coatings,
microparticles, liposomes and/or microspheres.
In general, a controlled-release preparation is a pharmaceutical composition
capable of
releasing the active ingredient at the required rate to maintain constant
pharmacological
activity for a desirable period of time. Such dosage forms provide a supply of
a drug to the
body during a predetermined period of time and thus maintain drug levels in
the therapeutic
range for longer periods of time than conventional non-controlled
formulations.
U.S. Patent No. 5,674,533 discloses controlled-release pharmaceutical
compositions in
liquid dosage forms for the administration of moguisteine, a potent peripheral
antitussive.
U.S. Patent No. 5,059,595 describes the controlled-release of active agents by
the use of a
gastro-resistant tablet for the therapy of organic mental disturbances. U.S.
Patent No.
5,591,767 describes a liquid reservoir transdermal patch for the controlled
administration of
ketorolac, a non-steroidal anti-inflammatory agent with potent analgesic
properties. U.S.
Patent No. 5,120,548 discloses a controlled-release drug delivery device
comprised of
swellable polymers. U.S. Patent No. 5,073,543 describes controlled-release
formulations
containing a trophic factor entrapped by a ganglioside-liposome vehicle. U.S.
Patent No.
5,639,476 discloses a stable solid controlled-release formulation having a
coating derived
from an aqueous dispersion of a hydrophobic acrylic polymer. Biodegradable
microparticles
are known for use in controlled-release formulations. U.S. Patent No.
5,733,566 describes the
use of polymeric microparticles that release antiparasitic compositions.
The controlled-release of the active ingredient may be stimulated by various
inducers,
for example pH, temperature, enzymes, water, or other physiological conditions
or
compounds. Various mechanisms of drug release exist. For example, in one
embodiment, the
73

CA 02836085 2013-12-05
controlled-release component may swell and form porous openings large enough
to release
the active ingredient after administration to a patient. The term "controlled-
release
component" in the context of the present invention is defined herein as a
compound or
compounds, such as polymers, polymer matrices, gels, permeable membranes,
liposomes
and/or microspheres, that facilitate the controlled-release of the active
ingredient in the
pharmaceutical composition. In another embodiment, the controlled-release
component is
biodegradable, induced by exposure to the aqueous environment, pH,
temperature, or
enzymes in the body. In another embodiment, sol-gels may be used, wherein the
active
ingredient is incorporated into a sol-gel matrix that is a solid at room
temperature. This
matrix is implanted into a patient, preferably a mammal, having a body
temperature high
enough to induce gel formation of the sol-gel matrix, thereby releasing the
active ingredient
into the patient.
The components used to formulate the pharmaceutical compositions are of high
purity
and are substantially free of potentially harmful contaminants (e.g., at least
National Food
grade, generally at least analytical grade, and more typically at least
pharmaceutical grade).
Particularly for human consumption, the composition is preferably manufactured
or
formulated under Good Manufacturing Practice standards as defined in the
applicable
regulations of the U.S. Food and Drug Administration. For example, suitable
formulations
may be sterile and/or substantially isotonic and/or in full compliance with
all Good
Manufacturing Practice regulations of the U.S. Food and Drug Administration.
VIII. Routes of Administration of Compounds and Compositions of the Invention
The compounds of Formula I may be administered by any route, including oral,
rectal,
sublingual, and parenteral administration. Parenteral administration includes,
for example,
intravenous, intramuscular, intraarterial, intraperitoneal, intranasal,
intravaginal, intravesical
(e.g., to the bladder), intradermal, transdermal, topical or subcutaneous
administration. Also
contemplated within the scope of the invention is the instillation of a drug
in the body of the
patient in a controlled formulation, with systemic or local release of the
drug to occur at a
later time. For example, the drug may be localized in a depot for controlled
release to the
circulation, or for release to a local site of tumor growth.
74

CA 02836085 2013-12-05
One or more compounds useful in the practice of the present inventions may be
administered simultaneously, by the same or different routes, or at different
times during
treatment. The compounds may be administered before, along with, or after
other
medications, including other antiproliferative compounds.
The treatment may be carried out for as long a period as necessary, either in
a single,
uninterrupted session, or in discrete sessions. The treating physician will
know how to
increase, decrease, or interrupt treatment based on patient response.
According to one
emobiment, treatment is carried out for from about four to about sixteen
weeks. The
treatment schedule may be repeated as required.
IX. Isomerism in Compounds of the Invention
A. Geometrical Isomerism
The compounds of Formula I possess an olefinic double bond. The
stereochemistry of
compounds possessing an olefinic double bond is designated using the
nomenclature using E
and Z designations. The compounds are named according to the Cahn-Ingold-
Prelog system,
described in the IUPAC 1974 Recommendations, Section E:
Stereochemistry, in
Nomenclature of Organic Chemistry, John Wiley & Sons, Inc., New York, NY, 4th
ed., 1992,
pp. 127-38, the entire contents of which is incorporated herein by reference.
Using this
system of nomenclature, the four groups about a double bond are prioritized
according to a
series of rules wherein various functional groups are ranked. The isomer with
the two higher
ranking groups on the same side of the double bond is designated Z and the
other isomer, in
which the two higher ranking groups are on opposite sides of the double bond,
is designated
E. This is illustrated schematically, below, where the Cahn-Ingold-Prelog
system priority of
the double bond substituents A is greater than that of B, and the priority of
A' is greater than
that of B'.
A B' A
A'
(E) isomer (Z) isomer

CA 02836085 2013-12-05
= =
The compounds of Formula I prepared according to the methodology described
herein
are believed to be in their Z isomeric forms, but may be converted to their E
isomeric forms as
previously described herein. Thus, both E and Z isomeric forms of each
compound described
herein are contemplated and may be generically described in a compound
including a "wavy"
bond pendant to a double bond, such as:
A
This generic structure indicates that a given compound may have either the E
or Z
configuration, such that the compounds
A
and A
.. would be understood to be described by the single structure provided above.
As used herein,
the presence of a wavy bond pendant to a double bond is not a per se
indication of an impure
mixture of compounds having both E and Z configurations. Rather, as set forth
elsewhere
herein, the presence of a wavy bond indicates indicates either (E), (Z), or a
mixture of
configurations of the double bond to the carbon atom to which the wavy bond is
attached.
.. B. Optical Isomerism
It will be understood that when or if compounds of the present invention
contain one
or more chiral centers, the compounds may exist in, and may be isolated as
pure enantiomeric
or diastereomeric forms or as racemic mixtures. The present invention
therefore includes any
possible enantiomers, diastereomers, racemates or mixtures thereof of the
compounds of the
invention.
76

CA 02836085 2013-12-05
The isomers resulting from the presence of a chiral center comprise a pair of
non-superimposable isomers that are called "enantiomers." Single enantiomers
of a pure
compound are optically active, i.e., they are capable of rotating the plane of
plane polarized
light. Single enantiomers are designated according to the Cahn-Ingold-Prelog
system. Once
the priority ranking of the four groups is determined, the molecule is
oriented so that the
lowest ranking group is pointed away from the viewer. Then, if the descending
rank order of
the other groups proceeds clockwise, the molecule is designated (R) and if the
descending
rank of the other groups proceeds counterclockwise, the molecule is designated
(S). In the
example below, the Cahn-Ingold-Prelog ranking is A> B> C > D. The lowest
ranking atom,
D is oriented away from the viewer.
A A
õoµaD
(R) configuration (S) configuration
The present invention is meant to encompass diastereomers as well as their
racemic
and resolved, diastereomerically and enantiomerically pure forms and salts
thereof.
Diastereomeric pairs may be resolved by known separation techniques including
normal and
reverse phase chromatography, and crystallization.
"Isolated optical isomer" means a compound which has been substantially
purified
from the corresponding optical isomer(s) of the same formula. Preferably, the
isolated isomer
is at least about 80%, more preferably at least 90% pure, even more preferably
at least 98%
pure, most preferably at least about 99% pure, by weight.
Isolated optical isomers may be purified from racemic mixtures by well-known
chiral
separation techniques. According to one such method, a racemic mixture of a
compound
having the structure of Formula I, or a chiral intermediate thereof, is
separated into 99% wt.%
pure optical isomers by HPLC using a suitable chiral column, such as a member
of the series
of DAICEL CHIRALPAK family of columns (Daicel Chemical Industries, Ltd.,
Tokyo,
Japan). The column is operated according to the manufacturer's instructions.
77

CA 02836085 2013-12-05
=
C. Rotational Isomerism
It is understood that due to chemical properties (i.e., resonance lending some
double
bond character to the C-N bond) of restricted rotation about the amide bond
linkage (as
illustrated below) it is possible to observe separate rotamer species and
even, under some
circumstances, to isolate such species. It is further understood that certain
structural elements,
including steric bulk or substituents on the amide nitrogen, may enhance the
stability of a
rotamer to the extent that a compound may be isolated as, and exist
indefinitely, as a single
stable rotamer. The present invention therefore includes any possible stable
rotamers of
Formula I.
0 A0
hindered rotation
)N' ) NI\
A
Examples
The following non-limiting examples are provided to illustrate the invention.
The
synthetic procedures described as "general methods" describe what it is
believed will be
typically effective to perform the synthesis indicated. However, the person
skilled in the art
will appreciate that it may be necessary to vary the procedures for any given
embodiment of
the invention. For example, reaction monitoring, such as by using thin layer
chromatography,
or HPLC may be used to determine the optimum reaction time. Products may be
purified by
conventional techniques that will vary, for example, according to the amount
of side products
produced and the physical properties of the compounds.
On a laboratory scale,
recrystallisation from a suitable solvent, column chromatography, normal or
reverse phase
HPLC, or distillation are all techniques which may be useful. The person
skilled in the art
will appreciate how to vary the reaction conditions to synthesize any given
compound within
the scope of the invention without undue experimentation. See, e.g., Vogel 's
Textbook of
78

CA 02836085 2013-12-05
Practical Organic Chemistry, by A. I. Vogel, et al, Experimental Organic
Chemistry:
Standard and Microscale, by L. M. Harwood et al. (2nd Ed., Blackwell
Scientific Publications,
1998), and Advanced Practical Organic Chemistry, by J. Leonard, et al. (2nd
Edition, CRC
Press 1994).
Synthetic Examples
Example 1: General Preparation of Compound B According to Scheme 1
Thioglycolic acid or methyl thioglycolate (1 mole) was added to a stirred
solution of
sodium hydroxide (2 mole) in ethanol (30 ml) and stirred for 5 min at room
temperature.
Compound A (1 mole) was added to the above solution and the reaction was
refluxed for 1 h.
Progress of the reaction was monitored by TLC. Upon completion, the reaction
mixture was
allowed to cool to room temperature, poured into crushed ice, and subsequently
neutralized
with 10N hydrochloric acid. The resulting solid was collected via filtration,
washed with
water and hexane, and dried to give compound B.
Exemplary compounds produced according to this methodology, along with their
relevant 11-1 NMR characteristics, and liquid chromatography mass spectrum
(LCMS), are
shown in Table 2.
79

CA 02836085 2013-12-05
Table 2
Cpd Cpd. Name Structure 1H NMR LCMS
# (DMSO-do
.
600 MHz)
45 2-((4-Fluoro-2- 0 5 8.14 (d, 1H, (M+Na):
nitrophenyl)thio)ae F N Ar-H, J ---- 8.1 253.98
etic acid
Hz), 7.73-
7.66 (m, 2H,
Ar-H), 4.04
0
(s, 2H, S-
CH2-00)
46 4((2-Methoxy-2-
0O 5 13.64 (brs, 1\1/1D
oxoethyl)thio)-3- -0N+ OH 111, COOH),
nitrobenzoic acid 8.64 (s, 1H,
Ar-H), 8.16
0
(d, 1H, Ar-H,
J = 6.0 Hz),
7.68 (d, 1H,
Ar-H, J = 6.4
Hz) 4.24 (s,
2H, S-CH2-
CO), 3.69 (s,
3H, OCH3)

CA 02836085 2013-12-05
= =
Example 2: General Preparation of Compound C According to Scheme 1
Method 1: Reductive cyclization with Zn/AcOH:
Compound B (8.6 mmole) was dissolved in 90 ml of acetic acid in a round bottom
flask with stirring for 5-10 minutes. The reaction was cooled to 0 C using an
ice bath. Zn
dust (14.4g) was slowly added to the stirring reaction, all the while
maintaining the
temperature at 0 C. Upon complete addition the reaction mixture was
maintained at 0 C for
an additional 60 minutes. The reaction was subsequently warmed to room
temperature and
allowed to stir for an additional two hours. The reaction was subsequently
filtered through a
ceilite pad. The resultant filtrate was then evaporated to give compound C.
Method 2: Reductive cyclization with Na2S204:
Sodium hydrosulfite (10 mmol) was added portion wise (in 4 portions at 5 min
intervals) to a stirred solution of compound B (5 mmol) in 2:1 ethanol:water
(20-30 ml) and
triethylamine (20 mmol). The temperature of the reaction was kept at 50 C
during the addition
of the hydrosulfite. Upon complete addition, the reaction was maintained at 50
C for 1.5
hours and then cooled to room temperature. Subsequently, 3N HC1 (20 ml) was
added and
the reaction was warmed to 60 C. After 1 hour, the reaction mixture was
poured into
crushed ice (300g) with vigorous stirring. A solid precipitate that had formed
was collected
via filtration. The solid was subsequently washed with 1:1 ether/hexane and
dried under
vacuum o provide pure C.
Exemplary compounds produced according to the procedures of Example 2, along
with their relevant IHNMR characteristics, and LCMS peaks, are shown in Table
3.
81

CA 02836085 2013-12-05
. =
Table 3
Cpd Cmpd. Name Structure 11-1 NMR
LCMS
.#
47 6-fluoro-2H- H (CDC13, 600 MHz): 6
(M+1):
benzo[b][1,4]thiazin- 8.70 (brs, 1H, NH),
184.02
3(4H)-one 7.31 (d, 1H, Ar-H, J
8.3 Hz), 6.80 (s, 1H,
Ar-H), 6.69 (d, 1H,
Ar-H, J = 8.6 Hz),
3.47 (s, 2H, C2-H)
48 7-Bromo-2H- Br(DMSO-d6, 300 N/D
benzo[b][1,4]thiazin-= MHz): 6 10.67 (brs,
3(4H)-one N-'/..co 1H, NH), 7.55 (d, 1H,
Ar-H, J = 1.4 Hz),
7.37-7.34 (dd, 1H, Ar-
H, J = 1.5 & 6.4 Hz),
6.91 (d, 1H, Ar-H, J =
6.4 Hz), 3.49 (s, 2H,
C2-H)
49 3-0xo-3,4-dihydro-2H- S (DMSO-d6, 300 N/D
benzo[b][1,4]thiazine-
HO MHz): 6 13.02 (brs,
6-carboxylic acid N-0 1H, COOH), 10.79
o
(brs, 1H, NH), 7.60 (s,
1H, Ar-H), 7.55 (d,
1H, Ar-H, J = 8.4 Hz),
7.47 (d, 1H, Ar-H, J =
8.4 Hz), 3.58 (s, 2H,
C2-H)
82

CA 02836085 2013-12-05
50 6-Amino-2H- NMR (DMSO-d6, N/D
benzo[b][1,4]thiazin- 300 MHz): 10.34
3(4H)-one (3d) H2N
N'O (brs, 1H, NH), 6.96 (d,
1H, Ar-H, J = 6.0 Hz),
6.29-6.26 (m, 2H, Ar-
H), 5.25 (brs, 2H,
NH2), 3.37 (s, 2H, C2-
H)
Example 3: Preparation of 3-oxo-3,4-dihydro-2H-benzo[1,41thiazine-6-sulfonyl
chloride
(Compound D, Scheme 2)
Chlorosulfonic acid (55.93g, 0.48 mole) was added to a 250 mL round bottom
flask
under nitrogen at 10 C. To this, 4H-benzo[1,4]thiazin-3-one, (20 g, 0.12
mole) was added
slowly so that the reaction temperature was maintained below 20 C during the
addition. After
addition, the reaction mixture was stirred at room temperature for 1.5 h and
then heated to 65
C for an additional 1 hour. The reaction was subsequently cooled and poured
into ice-cold
water. A solid precipitate formed and was collected via filtration. The solid
was then washed
with water and dried to provide 23.6 g of the title compound (75 % yield). 11-
1 NMR (CDC13,
300 MHz): 5 9.08 (br s, NH), 7.72-7.88 (m, 31-1, Ar-H), 3.68 (s, 2H, Ar-CH2);
m.p.: 165-167
C; LC-MS (M+1): 263.92
Example 4: Preparation of 6-((2,6-dichlorobenzypsulfony1)-2H-
benzo[b][1,41thiazin-
3(41/)-one
CI
0 N
S,
0
C I 11101
83

CA 02836085 2013-12-05
6((2,6-dichlorobenzyl)sulfony1)-2H-benzo [b] [1,4]thiazin-3(4H)-one.
Method 1: Zn-Mediated Sulfonylation:
Zn dust (1.3 g, 17 mmol), was added to THF/water (2:1) 150 ml and stirred for
15
min. Sold compound D (5 g, 19 mmol) was added to the Zn THF/water mixture over
30 min,
so that the addition exotherm could be controlled. The heterogeneous solution
was then stirred
for 24 h. The reaction was subsequently filtered through celite and the
filtrate collected. The
celite pad was subsequently washed with ethyl acetate. The organic layers were
then
combined and dried over anhydrous Na2SO4. The drying agent was filtered away
and the
filtrate was evaporated under reduced pressure to give Compound G, Scheme 2,
which was
used without further characterization or purification.
2,6-Dichlorobenzyl bromide (3.0 g, 39.6 mmol) in THF/ water (2:1) (70 ml) was
added to the above Zn complex and stirred at 70 C for 5 h. Upon completion of
the reaction,
the reaction mixture was diluted with ice-cold water, quenched with 2N HC1
(5mL), and
stirred for 10 min. A white solid precipitated. The precipitate was collected
via filtration,
washed with cold water, cold methanol (30 ml), and subsequently dried to give
3.5 g of 6-
((2,6-dichlorobenzyl)sulfony1)-2H-benzo[b][1,4]thiazin-3(41/)-one (50 %
yield). NMR
(DMSO-d6, 300 MHz): 8 10.95 (br s, 1H, NH), 7.16-7.80 (m, 6H, Ar-H), 4.88 (s,
2H, Ar-
CH2), 3.46 (s, 2H, C2-H); m.p.: 244-246 C; LC-MS (M+1): 387.96.
Method 2: Sulfite-Mediated Sulfonylation:
Na2HPO4 (2.3 g, 20 mmol) and Na2S03 (4.8 g, 38 mmol) were dissolved in 100 ml
of
water at 30 C. This solution was then added to Compound D (5 g, 19 mmol)
resulting in the
formation of a creamy suspension. The resultant suspension was heated to 60 C
and became
a clear solution. The reaction was stirred at 60 C for 16 h. Subsequently,
2,6-dichlorobenzyl
bromide (4.5 g, 19 mmol) in 10 ml of acetone was added to the reaction over 15
minutes. The
reaction was then stirred for 2 hours at 60 C. Upon completion of the
reaction, the reaction
vessel was cooled and reaction was poured into ice-cold water and stirred for
1 h. A white
precipitate formed and was collected via filtration. The collected solid was
washed with cold
84

CA 02836085 2013-12-05
water and a cold 1:1 water/acetone mixture. The resultant solid was
subsequently dried under
vacuum at 60 C overnight to give 6.8g (93% yield) of white 64(2,6-
dichlorobenzypsulfony1)-2H-benzo [b][1,4]thiazin-3(4H)-one.
11-1 NMR (DMSO-d6, 300
MHz): 5 10.98 (br s, 1H, NH), 7.27-7.65 (m, 6H, Ar-H), 4.95 (s, 2H, Ar-CH2),
3.58 (s, 2H,
C2-H); m.p.: 244-246 C; LC-MS (M+1): 387.96.
Example 5: General Procedure for the Preparation of Compounds of Formula I
According to Scheme 3
A compound according to Formula IV (1 mmol), an aldehyde of Formula III (1
mmol),
triethylamine (4 mmol), and acetic anhydride (10m1) were refluxed under
nitrogen
atmosphere for 2h. The reaction mixture was cooled to room temperature.
Solvent was
removed under reduced pressure and the resulting residue was purified by
column
chromatography to give a compound of Formula I. In certain instances, a solid
precipitate
formed upon completion and/or cooling the reaction. In either case, the
precipitate was
collected via filtration. Subsequently, the filtered solid was washed with an
appropriate
solvent and dried under vacuum to give a compound of Formula I.
Example 6: Additional Data for Compounds of the Invention
The compounds of the invention were prepared using the general procedures
described
above. Table 4, below, identifies compounds of the invention by number as well
as the
starting benzothiazinone and aldehyde used to prepare the compound. Table 4
further
includes the 'H NMR and LCMS peak of the identified compound of the invention.
11-1 NMR
spectra were recorded in either CDC13 or in DMSO-d6, as set forth in the
table.
85

CA 02836085 2013-12-05
Table 4
Cpd. Starting Benzothiazinone Starting Aldehye m.p. 1H NMR (6
LC-MS
( C) ppm)
(M+1)
1H NO2 268- (DMSO-d6; 391.02
Cl NO O 270 300 MHz):
41 0 11.30 (brs,
)r¨CH3 1H, NH),
0 8.42 (d, 1H,
Ar-H, J=
1.5Hz),
8.10-8.07
(dd, 1H, Ar-
H, J= 1.5 &
6.4 Hz),
7.88 (s, 1H,
=CH), 7.62
(d, 1H, Ar-
J= 6.4
Hz), 7.55 (d,
1H, Ar-H, J
= 1.6 Hz),
7.30-7.27
(dd, 1H, Ar-
H, J = 1.7 &
6.5 Hz),
7.08 (d, 1H,
Ar-H, J =
6.5 Hz),
2.37 (s, 3H,
COCH3)
2 H3C0 S.. NO2 238- (DMSO-d6; 387.07
0 240 300 MHz):
NO
11.06 (brs,
CH3 1H, NH),
8.41 (s, 1H,
0 Ar-H), 8.09
(d, 1H, Ar-
H, J= 6.4
Hz), 7.86 (s,
1H, =CH),
7.61 (d, 11-1,
Ar-H, J --
6.3 Hz),
7.03 (d, 1H,
Ar-H, J --
6.7 Hz),
6.99 (s, 1H,
86

CA 02836085 2013-12-05
Ar-H), 6.83
(d, 1H, Ar-
H, J = 6.4
Hz), 3.72 (s,
3H, OCH3),
2.37 (s, 3H,
COCH3)
3 Br 10NO2
270- (DMSO-d6; 436.28
0 272 300 MHz):
0 11.34
(brs,
NO H CH3 1H, NH),
0 8.46 (d,
1H,
Ar-H, J=
1.1Hz),
8.14-8.11
(dd, 1H, Ar-
H, J= 1.2 &
6.4 Hz),
7.92 (s, 1H,
=CH), 7.70
(d, 1H, Ar-
H, J= 1.2
Hz), 7.66 (d,
1H, Ar-H, J
= 6.4 Hz),
7.46-7.43
(dd, 1H, Ar-
H, J = 1.3 &
6.4 Hz),
7.06 (d, 1H,
Ar-H, J =
6.4 Hz),
2.41 (s, 3H,
COCH3)
4 H3C NO2
236- (DMSO-d6; 371.35
0 238 300 MHz):
0 0 11.00
(brs,
-CH3 1H, NH),
0 8.31 (d,
1H,
Ar-H, J =
1.5 Hz),
7.97-7.95
(dd, 1H, Ar-
H, J= 1.5 &
6.4 Hz),
7.73 (s, 1H,
=CH), 7.48
87

CA 02836085 2013-12-05
(d, 1H, Ar-
H, J = 6.3
Hz), 7.06 (s,
1H, Ar-H),
6.91 (d, 1H,
Ar-H, J =
6.2 Hz),
6.86 (d, 1H,
Ar-H, J =
6.1 Hz),
2.24 (s, 3H,
COCH3),
2.10 (s, 3H,
Ar-CH3)
0 NO2 >300 OD1vISO-d6; 401.38
0 300 N1144
0 11.01
(brs,
HO
CH3 1H, NH),
N0 0 8.22 (d,
1H,
Ar-H, J =
1.5 Hz),
7.90-7.88
(dd, 1H, Ar-
H, J= 1.5 &
6.4 Hz),
7.65 (s, 1H,
=CH), 7.48
(s, 1H, Ar-
H), 7.39 (d,
1H, Ar-H, J
= 6.3 Hz),
7.35 (d, 1H,
Ar-H, J =
5.7 Hz),
7.06 (d, 1H,
Ar-H, J =
6.0 Hz),
2.15 (s, 3H,
COCH3)
6 H NO2
238- (DMSO-d6; 375.05
0
0 CH3 240 300
MHz):
N 0 0
11.22 (brs,
)
1H, NH),
8.41 (d, 1H,
Ar-H, J =
1.5 Hz),
8.09-8.06
(dd, 1H, Ar-
88

CA 02836085 2013-12-05
= =
H, J= 1.6 &
6.4 Hz),
7.88 (s, 1H,
=CH), 7.62
(d, 1H, Ar-
H, J = 6.4
Hz), 7.38 (d,
1H, Ar-H, J
= 6.6 Hz),
7.13 (d, 2H,
Ar-H, J =
8.0 Hz),
2.37 (s, 3H,
COCH3)
7 F S NO2
236- (DMSO-d6; 375.09
0
N 0 0 238 300 MHz):
11.25 (brs,
1H, NH),
CH 8.39
(s, 1H,
0 Ar-
H), 8.06
(d, 1H, Ar-
H, J= 6.2
Hz), 7.83 (s,
1H, =CH),
7.58 (d, 1H,
Ar-H, J =
6.3 Hz),
7.38 (t, 1H,
Ar-H, J =
5.0 & 5.2
Hz), 6.91-
6.87 (m, 2H,
Ar-H), 2.37
(s, 3H,
COCH3)
8 6-((2,6- NO2
286- (DMSO-d6; 580.40
dichlorobenzyl)sulfony1)-2H- 0ik 288
300 MHz):
benzo [b] [1,4]thiazin-3(4H)-one 0 11.42
(brs,
(Example 4) CH3 1H, NH),
8.43 (d, 1H,
0
Ar-H, J =
2.4 Hz),
8.11-8.08
89

CA 02836085 2013-12-05
(dd, 1H, Ar-
H, J = 2.1 &
8.5 Hz),
7.91 (s, 1H,
=CH), 7.62
(d, 1H, Ar-
H, J = 1.0
Hz), 7.53 (d,
1H, Ar-H, J
= 1.2 Hz),
7.50 (s, 1H,
Ar-H), 7.48
(d, 1H, Ar-
H, J = 1.8
Hz), 7.42 (d,
1H, Ar-H, J
= 2.1 Hz),
7.38 (t, 1H,
Ar-H, J =-
2.0 & 2.1
Hz), 7.35 (d,
1H, Ar-H, J
= 1.8 Hz),
4.89 (s, 2H,
Ar-CH2),
2.37 (s, 3H,
COCH3)
9 6-((2,6- O 228- NMR not
535.98
dichlorobenzyl)sulfony1)-2H-
411 0 230 recorded
benzo [b] [1,4]thiazin-3 (411)-one H CH3
(Example 4) 0
6-((2,6- Benzaldehyde 210- (DMSO-d6; 477.08
dichlorobenzyl)sulfony1)-2H- 211 300 MHz):
benzo[b] [1,4]thiazin-3 (411)-one 11.28 (brs,
(Example 4) 1H, NH),
7.86 (s, 1H,
=CH), 7.69
(d, 2H, Ar-
H, J = 7.8
Hz), 7.61 (d,
1H, Ar-H, J
= 8.4 Hz),
7.54-7.52
(m, 3H, Ar-
H), 7.47 (s,
1H, Ar-H),
7.45 (s, 1H,
Ar-H), 7.43

CA 02836085 2013-12-05
. =
(d, 2H, Ar-
H, J = 7.8
Hz), 7.36 (d,
1H, Ar-H, J
= 8.4 Hz),
4.88 (s, 2H,
Ar-CH2)
11 6-((2,6- 0 >320 (DMSO-d6;
552.43
dichlorobenzypsulfony1)-2H- 300 MHz):
benzo[b][1,4]thiazin-3(4H)-one H
NO2 11.31 (brs,
(Example 4) 8114.24,
(s,N1HH):
OCH3 Ar-H), 8.00
(d, 1H, Ar-
H, J = 8.4
Hz), 7.84 (s,
1H, =CH),
7.63 (d, 1H,
Ar-H, J =
8.4 Hz),
7.54-7.51
(m, 3H, Ar-
H), 7.47 (s,
1H, Ar-H),
7.43 (d, 1H,
Ar-H, J =
7.2 Hz),
7.37 (d, 1H,
Ar-H, J =
8.4 Hz),
4.89 (s, 2H,
Ar-CH2),
4.00 (s, 3H,
OCH3)
12 6-((2,6- 0 248- NMR not
522.01
dichlorobenzyl)sulfony1)-2H- 250 recorded
benzo[b][1,4]thiazin-3(4H)-one H NH2
(Example 4)
CH3
13 6-((2,6-/ \
140- (DMSO-d6; 575.08
dichlorobenzyl)sulfony1)-2H-
NN¨CH3 142 300 MHz):
benzo[b][1,4]thiazin-3(411)-one H / 9.89 (brs,
(Example 4) 1H, NH),
8.25 (s, 1H,
Ar-H), 7.90
(d, 1H, Ar-
H, J = 7.8
Hz), 7.76 (s,
1H, =CH),
91

CA 02836085 2013-12-05
=
7.45 (d, 2H,
Ar-H, J
8.4 Hz),
7.39 (d, 1H,
Ar-H, J =
7.8 Hz),
7.34 (s, 1H,
Ar-H), 7.27
(d, 1H, Ar-
H, J = 8.4
Hz), 7.09 (d,
1H, Ar-H, J
= 7.8 Hz),
6.94 (d, 2H,
Ar-H, J =
7.2 Hz),
4.83 (s, 2H,
Ar-CH2),
3.55-
3.53(m, 4H,
Pip-H),
2.42-2.39
(m, 4H, Pip-
H), 2.19 (s,
3H, N-CH3)
14 6-((2,6- 4-nitrobenzaldehyde >320 NMR not
522.28
dichlorobenzyl)sulfony1)-2H- recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4)
15 6-((2,6- 4-aminobenzaldehyde >320 NMR not
492.98
dichlorobenzyl)sulfony1)-2H- recorded
benzo[b][1,4]thiazin-3(411)-one
(Example 4)
16 6-((2,6- 4-fluorobenzaldehyde 308 NMR not
495.97
dichlorobenzyl)sulfony1)-2H- recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4)
17 6-((2,6- 4-chlorobenzaldehyde >320 NMR not
511.94
dichlorobenzyl)sulfony1)-2H- recorded
benzo[b][1,4]thiazin-3(41/)-one
(Example 4)
18 6-((2,6- 4-bromobenzaldehyde >310 NMR not
556.89
dichlorobenzyl)sulfony1)-2H- recorded
benzo[b][1,4]thiazin-3(41/)-one
(Example 4)
19 6-((2,6- 4-methoxybenzaldehyde NMR not
507.33
dichlorobenzyl)sulfony1)-2H- recorded
92

CA 02836085 2013-12-05
benzo[b][1,4]thiazin-3(4H)-one
(Example 4)
20 6-((2,6- 4-methylbenzaldehyde NMR not
491,52
dichlorobenzypsulfony1)-2H- recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4)
21 6-((2,6- 2,4,6-trimethoxybenzaldehyde NMR not
567.13
dichlorobenzyl)sulfony1)-2H- recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4)
22 6-((2,6- 2,4-dichlorobenzaldehyde
>310 NMR not 546.30
dichlorobenzyl)sulfony1)-2H- recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4)
23 (number intentionally skipped)
24 6-((2,6- O NMR not 564.36
dichlorobenzyl)sulfony1)-2H- recorded
benzo[b][1,4]thiazin-3(4H)-one H3C0.7
(Example 4)
HN
H
O
25 6-((2,6- O NMR not
550.80
dichlorobenzyl)sulfony1)-2H- recorded
benzo[b][1,4]thiazin-3(4H)-one Ficy)'\
(Example 4)
HN
o
1010
26 6-((2,6- 0 >300 NMR not
558.03
dichlorobenzyl)sulfony1)-2H- recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4)
41111
0
93

CA 02836085 2013-12-05
. =
27 6-((2,6- H NMR
not 466.24
dichlorobenzyl)sulfony1)-2H- N
recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4)
0
28 6-((2,6- 4-hydroxy-2,6- NMR
not 553.85
dichlorobenzypsulfony1)-2H- dimethoxybenzaldehyde
recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4)
29 6-((2,6- 4-
chloro-3-nitrobenzaldehyde >310 NMR not 556.78
dichlorobenzypsulfony1)-2H-
recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4)
30 6-((2,6- 2,4-difluorobenzaldehyde 310-
NMR not 513.69
dichlorobenzyl)sulfony1)-2H- 312
recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4)
31 6-((2,6- 2,4,6-trifluorobenzaldehyde 254-
NMR not 531.53
dichlorobenzyl)sulfony1)-2H- 256
recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4)
32 6-((2,6- H
>300 NMR not 535.98
dichlorobenzyl)sulfony1)-2H-
recorded
benzo[b][1,4]thiazin-3(4H)-one 0
(Example 4)
110 OCH3
o
33 6-((2,6- S H 310-
NMR not 483.93
dichlorobenzyl)sulfony1)-2H- 312
recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4) 0
34 6-((2,6- H NMR
not 515.43
dichlorobenzyl)sulfony1)-2H- N
recorded
benzo[b][1,4]thiazin-3(4H)-one
(Example 4) /
0
94

CA 02836085 2013-12-05
35 6-((2,6- H NO2 260- NMR
not 580.98
dichlorobenzyl)sulfony1)-2H- 264 recorded
benzo[b][1,4]thiazin-3(411)-one 0
(Example 4)
OCH3
O
36 6-((2,6- O >300 NMR
not 525.96
dichlorobenzyl)sulfony1)-2H- recorded
benzo[b][1,4]thiazin-3(41/)-one H
(Example 4)
N
H3C
37 6-((2,6- H >300 NMR
not 564.02
dichlorobenzyl)sulfony1)-2H- 0 recorded
benzo[b][1,43thiazin-3(4H)-one
(Example 4) ¨N\
Ni-N\
38 6-((2,6- 0 0 292- NMR
not 692.44
dichlorobenzyl)sulfonyI)-2H- 294
recorded
benzo[b][1,4]thiazin-3(411)-one =Is14:õ
0"
(Example 4)
0
0=-S=0
CH3
39 6-((2,6- 0 N 250- NMR
not 545.23
dichlorobenzyl)sulfonyI)-2H- 258
recorded
benzo[b][1,4]thiazin-3(4H)-one ,NH
H N
(Example 4)
40 H3C0 S 02N 238-
(CDCI3; 600 429.13
o 240 MHz):
8.50 (s, 1H,
0 0 Ar-H),
7.94
H3C (s, 2H,
<
H3C 0 ID =CH& Ar-
H), 7.53 (d,
1H, Ar-H,
= 8.4 Hz),

CA 02836085 2013-12-05
7.38 (d, 1H,
Ar-H, J
7.4 Hz),
6.93 (s, 1H,
Ar-H), 6.90
(d, 1H, Ar-
H, J = 7.3
Hz), 3.84 (s,
3H, OCH3),
2.67 (s, 3H,
COCH3),
2.45 (s, 3H,
COCH3)
41 6-((2,6- NO2 306- (DMSO-d6; 628.61
dichlorobenzyl)sulfony1)-2H- O= 308 600
MHz):
benzo[b][1,4]thiazin-3(411)-one o 11.31
(brs,
(Example 4) 1H, NH),
8.27 (s, 1H,
Ar-H), 8.00
(d, 1H, Ar-
H, J= 8.8
Hz), 7.86 (s,
1H, =CH),
7.65 (d, 1H,
Ar-H, J
8.3 Hz),
7.62 (d, 1H,
Ar-H, J =
8.8 Hz),
7.55 (d, 2H,
Ar-H, J =
8.0 Hz),
7.51-7.50
(m, 3H, Ar-
H), 7.46-
7.44 (m, 3H,
Ar-H), 7,39
(d, 2H, Ar-
H, J = 7.8
Hz), 5.43 (s,
2H, Ar-
CH2), 4.91
(s, 2H, Ar-
CH2)
42 6-((2,6- 4-hydroxy-3- 312- (DMSO-d6; 538.05
dichlorobenzyl)sulfony1)-2H- nitrobenzaldehyde 314 600
MHz):
benzo[b][1,4]thiazin-3(4H)-one 11.63
(brs,
(Example 4) 1H, OH),
96

CA 02836085 2013-12-05
=
11.27 (brs,
1H,
NH),
8.26 (s, 1H,
Ar-H), 7.90
(d, 1H, Ar-
H, J= 8.3
Hz), 7.82 (s,
1H, =CH),
7.64 (d, 1H,
Ar-H, J =
8.2
Hz),
7.55 (d, 2H,
Ar-H, J =
7.9
Hz),
7.49 (s, 1H,
Ar-H), 7.45
(t, 1H, Ar-
H, J = 7.9 &
8.0
Hz),
7.38 (d, 1H,
Ar-H, J =
8.0
Hz),
7.28 (d, 1H,
Ar-H, J =
8.5
Hz),
4.90 (s, 2H,
__________________________________________________________________ Ar-CH2)
43 F S, NO2 225568- NM
Rdndot 423.45
0
II 0
0
44
4-hydroxy-3-
262- (DMSO-d6; 333.23
nitrobenzaldehyde
264 600 MHz):
11.63 (brs,
N 0 11-1, OH),
11.13 (brs,
1H, NH),
8.24 (d, 1H,
Ar-H, J =
2.0
Hz),
7.89-7.87
(dd, 1H, Ar-
H, J= 2.2 &
8.7
Hz),
7.76 (s, 1H,
=CH), 7.42-
7.40
(dd,
1H, Ar-H, J
= 5.8 & 8.5
97

CA 02836085 2013-12-05
, . . .
Hz), 7.27 (d,
1H, Ar-H, J
= 8.8 Hz),
6.93-6.89
(m, 2H, Ar-
H)
98

CA 02836085 2013-12-05
Example 7: Leukemia and Prostate Cancer Cell Cytotoxicity Assays
The effect of the compounds of the invention on tumor cells was determined by
the
assay described by Latham et al., Oncogene 12:827-837 (1996). Tumor cells K562
(chronic
myelogenous leukemia; leukemia cell line +ve for Bcr-Abl) or DU145 (prostate
cancer) were
plated in 12-well dishes at a cell density of 2.5 x 104 cells well. The plated
cells were treated
24 hours later with a compound of the invention dissolved in DMSO at multiple
concentrations ranging from 0.011..LM to 100 M. The plates were examined 96
hours later
under an inverted microscope, Olympus CK-2 using a 10x objective, and compound
activity
was noted by physical observation. When necessary, the total number of viable
cells was
determined by trypsinizing the wells and counting the number of viable cells,
as determined
by trypan blue exclusion, using a hemacytometer. The results of these assays
are provided
below in Table 5. Where "Lower" and "Upper" value are provided in Table 5, the
IC50 was
determined to be within that range. When only a "Lower" value is provided and
no "Upper"
value is listed, the 1050 was that value, or less than or greater than that
value, as set forth in the
Table.
Table 5
Cpd. # K562 1050 ( m) DU145 1050 (pm)
Lower (pm) Upper (pm) Lower (pm) Upper (pm)
1 10 25 1 10
2 1 10 1 10
3 1 10 10 25
4 1 10 1 10
50 100 >100 N/A
6 1 10 N/A 1.0
7 1 10 1 10
8 0.075 N/A 0.1 N/A
9 <1 N/A <1 N/A
1 10 <1 N/A
11 0.25 0.5 0.25 0.5
12 0.1 0.25 0.25 0.5
13 0.5 1 1 10
14 1 10 1 10
99

CA 02836085 2013-12-05
õ =
15 0.03 N/A 0.04 N/A
16 1 10 1 10
17 1 10 1 10
18 1 10 1 10
22 1 10 1 10
29 1 10 0.5 1.0
30 1 10 1 10
31 1 10 1 10
32 1 10 0.5 1.0
33 0.5 1.0 0.5 1.0
26 0.5 1.0 0.5 1.0
35 1 10 >10 N/A
36 >2.5 N/A 2 N/A
37 1 10 1 10
38 0.1 N/A 0.09 N/A
39 1 10 1 10
40 1.5 N/A 2 N/A
41 0.75 N/A 1.5 N/A
42 0.1 N/A 0.15 N/A
43 >10 N/A >10 N/A
44 0.25 N/A 0.75 N/A
Example 8: Cancer Cell Cytotoxicity Assays: (Z)-64(2,6-
Dichlorobenzyl)sulfony1)-2-(4-
hydroxy-3-nitrobenzylidene)-2H-benzo[b][1,41thiazin-3(4H)-one
The cytotoxicity of Compound 42, (Z)-6-((2,6-diehlorobenzyl)sulfony1)-2-(4-
hydroxy-
3-nitrobenzylidene)-2H-benzo[b][1,4]thiazin-3(4H)-one, was determined
according the assay
method of Example 7 on the prostate (androgen dependent or independent),
breast and
pancreatic cancer cell lines listed in Table 6A, below, and the additional
cell lines listed in
Table 6B, below. The human mesenchymal stem cell line HMSC.hTERT and normal
human
fibroblast cell line HFL was included in the test. The results indicate that
while Compound 42
was toxic to the cancer cell lines, it did not cause cell death in the normal
human stem cell
line, or in the normal human fibroblast cell line, demonstrating the
selectivity of the
compound for cancer cells.
100

CA 02836085 2013-12-05
Table 6A
CELL LINE TUMOR TYPE Gi5o(11M)
DU-145 Prostate Cancer (Androgen Independent) 0.16
LnCap Prostate Cancer (Androgen Dependent) 0.25
MDA.MB-231 Breast Cancer 0.12
MDA.MB-468 Breast Cancer 0.2
BT-474 Breast Cancer 0.2
MCF-7 Breast Cancer 1.0
MiaPaca 2 Pancreatic Cancer 0.09
HMSC.hTERT Human Mesenchymal Stem cells
>10
Table 6B
CELL LINE TMUOR TYPE G150(111\1)
PC3 Prostate Cancer 0.16
22Rv1 Prostate Cancer 0.50
VCAP Prostate Cancer 0.18
RPWE-1 Prostate Cancer 0.10
MCF10A Normal Breast Cancer 2.50
A549 Non-small cell lung cancer 0.16
H23 Non-small cell lung cancer 0.065
HCT-116 Colon cancer 0.15
K562 Chronic myelogenous leukemia
0.16
Z138C Mantle cell lymphoma 0.07
U266B1 Multiple Myeloma 0.10
GRANTA Mantle cell lymphoma 0.08
HFL Normal Fibroblast > 10
Example 9: Casein Kinase 2a Inhibition by (Z)-64(2,6-Dichlorobenzyl)sulfony1)-
2-(4-
hydroxy-3-nitrobenzylidene)-2H-benzo[b][1,41thiazin-3(4H)-one
The ability of Compound 42 to inhibit the activity of casein kinase 2 (CK2)
was
determined as follows using the catalytic CK2 subunits CK2a and CK2a2. Two and
one half
nanograms of CK2a (human full length protein; Invitrogen PV3248), 2.5 ng of a
CK2a form
containing amino acids 3-339) or 10 ng of CK2a2 (human full length protein;
Invitrogen
PV3624) was diluted into kinase buffer (20 mM Tris pH 7.5, 10 mM MgC12, 0.01%
NP-40, 1
mM EGTA, 2 mM DTT) and incubated with varying concentrations of Compound 42 at
room
101

CA 02836085 2013-12-05
temperature for 30 minutes. Kinase reactions were initiated by the addition of
1 us of CK2
substrate a-synuclein or 40 p.M CK2 synthetic peptide substrate (AnaSpec, San
Jose, CA), 20
1.1M cold ATP, and 20 [.tCi y-32P-ATP. The reactions were incubated at 30 C
for 20 minutes,
terminated by the addition of 2x Laemmle sample buffer, boiled for 2 minutes,
resolved by
12% acrylamide SDS-PAGE, and subjected to autoradiography. The IC50 for
Compound 42
inhibition of CK2a was 361.9 nM. The IC50 for Compound 42 inhibition of CK2a2
was
83 .1nM.
Example 10: CDK9 Inhibition by (Z)-64(2,6-Diehlorobenzyl)sulfony1)-2-(4-
hydroxy-3-
nitrobenzylidene)-2H-benzo[b] [1,41thiazin-3(4H)-one
The ability of Compound 42 to inhibit the activity of the kinase CDK9 was
determined
as follows. Twenty-five nanograms of CDK9 (human full length protein;
Invitrogen PV4335)
was diluted into kinase buffer (20 mM Tris pH 7.5, 10 mM MgC12, 0.01% NP-40, 1
mM
EGTA, 2 mM DTT) and incubated with varying concentrations of Compound 42 at
room
temperature for 30 minutes. The kinase reactions were initiated by the
addition of 20 1.11µ4 of
CDK9/Cyclin K specific synthetic peptide substrate (PDKtide, P10-58,
SignalChem), 20 11M
cold ATP, and 20 [xCi 7-32P-ATP. The reactions were incubated at 30 C for 20
minutes and
terminated by the addition of 0.5M o-Phosphoric acid. 10 pi of the reaction
mixture was
spotted on a P-30 Filtermat (PerkinElmer) and developed using autoradiography
or
phosphorimaging. The IC50 for Compound 42 inhibition of CDK9 was determined as
93.68
nM.
Example 11: PIM1 Inhibition by (Z)-64(2,6-Diehlorobenzypsulfony1)-2-(4-hydroxy-
3-
nitrobenzylidene)-2H-benzo[b] [1,4] thiazin-3(4H)-one
The ability of Compound 42 to inhibit the activity of the kinase proto-ocogene
pim-
was determined as follows. Ten nanograms of PIM1 (human full length protein;
Invitrogen
PV3503) was diluted into kinase buffer (20 mM Tris pH 7.5, 10 mM MgC12, 0.01%
NP-40, 1
mM EGTA, 2 mM DTT) and incubated with varying concentrations of Compound 42 at
room
102

CA 02836085 2013-12-05
. .
temperature for 30 minutes. The kinase reactions were initiated by the
addition of 20 M of
PIM1 specific synthetic peptide substrate (K9 Peptide; p70S6 Kinase Substrate;
AnaSpec, San
Jose, CA), 20 p,M cold ATP, and 20 Ci y-32P-ATP. The reactions were incubated
at 30 C
for 20 minutes and terminated by the addition of 0.5M o-Phosphoric acid. 10 I
of the
reaction mixture was spotted on a P-30 Filtermat (PerkinElmer) and developed
using
autoradiography or phosphorimaging. The IC50 for Compound 42 inhibition of
PIM1 was
determined as 226.1 nM.
All references cited herein are incorporated by reference. The present
invention may
be embodied in other specific forms without departing from the spirit or
essential attributes
thereof and, accordingly, reference should be made to the appended claims,
rather than to the
foregoing specification, as indicating the scope of the invention.
1 03

Representative Drawing

Sorry, the representative drawing for patent document number 2836085 was not found.

Administrative Status

2024-08-01:As part of the Next Generation Patents (NGP) transition, the Canadian Patents Database (CPD) now contains a more detailed Event History, which replicates the Event Log of our new back-office solution.

Please note that "Inactive:" events refers to events no longer in use in our new back-office solution.

For a clearer understanding of the status of the application/patent presented on this page, the site Disclaimer , as well as the definitions for Patent , Event History , Maintenance Fee  and Payment History  should be consulted.

Event History

Description Date
Application Not Reinstated by Deadline 2018-12-05
Time Limit for Reversal Expired 2018-12-05
Deemed Abandoned - Failure to Respond to Maintenance Fee Notice 2017-12-05
Inactive: IPC deactivated 2017-09-16
Inactive: IPC expired 2017-01-01
Inactive: IPC assigned 2017-01-01
Inactive: Cover page published 2015-06-22
Application Published (Open to Public Inspection) 2015-06-05
Inactive: IPC assigned 2014-03-13
Inactive: IPC assigned 2014-03-13
Inactive: IPC assigned 2014-03-13
Inactive: IPC assigned 2014-03-13
Inactive: IPC assigned 2014-03-13
Inactive: IPC assigned 2014-03-13
Inactive: IPC assigned 2014-03-13
Inactive: IPC removed 2014-03-13
Inactive: IPC assigned 2014-03-13
Inactive: IPC removed 2014-03-13
Inactive: IPC assigned 2014-03-13
Inactive: First IPC assigned 2014-03-13
Inactive: IPC assigned 2014-03-13
Inactive: IPC assigned 2014-03-13
Inactive: IPC assigned 2014-03-13
Inactive: IPC assigned 2014-03-13
Inactive: IPC assigned 2014-03-11
Inactive: IPC assigned 2014-03-11
Inactive: IPC assigned 2014-03-11
Letter Sent 2014-02-07
Inactive: Single transfer 2014-01-16
Inactive: Filing certificate - No RFE (English) 2013-12-30
Filing Requirements Determined Compliant 2013-12-30
Application Received - Regular National 2013-12-17
Inactive: Pre-classification 2013-12-05

Abandonment History

Abandonment Date Reason Reinstatement Date
2017-12-05

Maintenance Fee

The last payment was received on 2016-11-18

Note : If the full payment has not been received on or before the date indicated, a further fee may be required which may be one of the following

  • the reinstatement fee;
  • the late payment fee; or
  • additional fee to reverse deemed expiry.

Please refer to the CIPO Patent Fees web page to see all current fee amounts.

Fee History

Fee Type Anniversary Year Due Date Paid Date
Application fee - standard 2013-12-05
Registration of a document 2014-01-16
MF (application, 2nd anniv.) - standard 02 2015-12-07 2015-05-21
MF (application, 3rd anniv.) - standard 03 2016-12-05 2016-11-18
Owners on Record

Note: Records showing the ownership history in alphabetical order.

Current Owners on Record
TEMPLE UNIVERSITY - OF THE COMMONWEALTH SYSTEM OF HIGHER EDUCATION
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
Past Owners on Record
None
Past Owners that do not appear in the "Owners on Record" listing will appear in other documentation within the application.
Documents

To view images, click a link in the Document Description column. To download the documents, select one or more checkboxes in the first column and then click the "Download Selected in PDF format (Zip Archive)" or the "Download Selected as Single PDF" button.

List of published and non-published patent-specific documents on the CPD .

If you have any difficulty accessing content, you can call the Client Service Centre at 1-866-997-1936 or send them an e-mail at CIPO Client Service Centre.