U.S. patent application number 15/426294 was filed with the patent office on 2017-05-25 for novel compounds.
The applicant listed for this patent is GlaxoSmithKline LLC. Invention is credited to James BAILEY, Romain Luc Marie Gosmini, Olivier Mirguet, Jason Witherington.
Application Number | 20170145021 15/426294 |
Document ID | / |
Family ID | 43969589 |
Filed Date | 2017-05-25 |
United States Patent
Application |
20170145021 |
Kind Code |
A1 |
BAILEY; James ; et
al. |
May 25, 2017 |
NOVEL COMPOUNDS
Abstract
Benzodiazepine compounds of formula (I) ##STR00001## and salts
thereof, pharmaceutical compositions containing such compounds and
their use in therapy.
Inventors: |
BAILEY; James; (Stevenage,
GB) ; Gosmini; Romain Luc Marie; (Les Ulis, FR)
; Mirguet; Olivier; (Les Ulis, FR) ; Witherington;
Jason; (Stevenage, GB) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
GlaxoSmithKline LLC |
Wilmington |
DE |
US |
|
|
Family ID: |
43969589 |
Appl. No.: |
15/426294 |
Filed: |
February 7, 2017 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
14753326 |
Jun 29, 2015 |
9598420 |
|
|
15426294 |
|
|
|
|
13504971 |
Jun 18, 2012 |
9102677 |
|
|
PCT/EP10/66697 |
Nov 3, 2010 |
|
|
|
14753326 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 19/02 20180101;
A61P 37/06 20180101; A61P 37/00 20180101; A61K 9/0019 20130101;
A61P 29/00 20180101; A61P 35/00 20180101; A61P 43/00 20180101; A61K
31/5517 20130101; C07D 487/04 20130101; A61P 37/02 20180101 |
International
Class: |
C07D 487/04 20060101
C07D487/04; A61K 9/00 20060101 A61K009/00; A61K 31/5517 20060101
A61K031/5517 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 5, 2009 |
GB |
0919433.3 |
Jun 22, 2010 |
GB |
1010509.6 |
Aug 6, 2010 |
EP |
PCT/EP2010/061518 |
Aug 25, 2010 |
GB |
1014231.3 |
Claims
1. A compound of formula (I) or a salt thereof ##STR00141## where
R.sup.1 is C.sub.1-3alkyl; R.sup.2 is --NR.sup.2aR.sup.2a' or
--OR.sup.2b; wherein one of R.sup.2a or R.sup.2'a is hydrogen, and
R.sup.2b or the other of R.sup.2a or R.sup.2'a is selected from
C.sub.1-6alkyl, haloC.sub.1-6alkyl,
R.sup.2cR.sup.2'cN--C.sub.1-6alkyl, carbocyclyl,
carbocyclylC.sub.1-4alkyl, heterocyclyl and
heterocyclylC.sub.1-4alkyl, wherein any of the carbocyclyl or
heterocyclyl groups are optionally substituted by one or more
groups selected from halogen, C.sub.1-6alkyl, haloC.sub.1-6alkyl,
C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, carbonyl, --CO-carbocyclyl,
azido, amino, hydroxyl, nitro and cyano, wherein the
--CO-carbocyclyl group may be optionally substituted by one or more
groups selected from halogen, C.sub.1-6alkyl, haloC.sub.1-6alkyl,
C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, azido, nitro and cyano; or
two adjacent groups on any of the carbocyclyl or heterocyclyl
groups together with the interconnecting atoms form a 5- or
6-membered ring which ring may contain 1 or 2 heteroatoms
independently selected from O, S and N; or R.sup.2a and R.sup.2'a
together with the N-atom to which they are attached form a 4-, 5-,
6- or 7-membered ring which optionally contains 1 or 2 heteroatoms
independently selected from O, S and N; wherein the 4-, 5-, 6- or
7-membered ring is optionally substituted by C.sub.1-6alkyl,
hydroxyl or amino; R.sup.2c and R.sup.2'c are independently
hydrogen or C.sub.1-6alkyl; each R.sup.3 is independently selected
from hydrogen, hydroxyl, halo, C.sub.1-6alkyl, halo C.sub.1-6alkyl,
C.sub.1-6alkoxy, halo C.sub.1-6alkoxy, nitro, cyano, CF.sub.3,
--OCF.sub.3, --COOR.sup.5, --C.sub.1-4alkylNR.sup.6R.sup.7 and
--C.sub.1-4alkylOH; R.sup.4 is hydroxyl, halo, C.sub.1-6alkyl,
haloC.sub.1-6alkyl, C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, nitro,
cyano, CF.sub.3, --OCF.sub.3, --COOR.sup.5; or
--OS(O).sub.2C.sub.1-4alkyl; R.sup.5 is C.sub.1-3alkyl; and n is an
integer 1 to 5.
2. A compound or a salt thereof according to claim 1, wherein
R.sup.1 is methyl.
3. A compound or a salt thereof according to claim 1, wherein
R.sup.2 is --OR.sup.2b and R.sup.2b is C.sub.1-6alkyl.
4. A compound or a salt thereof according to claim 1, wherein
R.sup.2 is --NR.sup.2aR.sup.2a'.
5. A compound or a salt thereof according to claim 4, wherein
R.sup.2a is hydrogen and R.sup.2'a is C.sub.1-6alkyl.
6. A compound or a salt thereof according to claim 4, wherein
R.sup.2'a is selected from R.sup.2cR.sup.2'cN--CH.sub.2--,
R.sup.2cR.sup.2'cN--CH.sub.2CH.sub.2--,
R.sup.2cR.sup.2'cN--CH.sub.2CH.sub.2CH.sub.2-- and
R.sup.2cR.sup.2'cN--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--, and wherein
R.sup.2c and R.sup.2'c are selected from hydrogen and methyl.
7. A compound or a salt thereof according to claim 4, wherein
R.sup.2a is hydrogen and R.sup.2'a is selected from:
##STR00142##
8. A compound or a salt thereof according to claim 4, wherein
R.sup.2a is hydrogen and R.sup.2'a is heterocyclylC.sub.1-4alkyl,
wherein the heterocyclyl is selected from: ##STR00143##
9. A compound or a salt thereof according to claim 1, wherein
R.sup.4 is 8-MeO.
10. A compound or a salt thereof according to claim 1 wherein the
chiral carbon atom depicted by * is the S-enantiomer.
11. A compound or a salt thereof according to claim 1 which is any
one of Examples 1-82 or a salt thereof.
12. A compound which is
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide ##STR00144## or a salt
thereof.
13. A pharmaceutical composition which comprises a compound or a
salt thereof according to claim 12 and one or more pharmaceutically
acceptable carriers, diluents or excipients.
14. A combination pharmaceutical product comprising a compound or a
salt thereof according to claim 12 together with one or more other
therapeutically active agents.
15. A method for treatment of a disease or condition, for which a
bromodomain inhibitor is indicated, in a subject in need thereof
which comprises administering a therapeutically effective amount of
compound or a salt thereof according to claim 12.
16. A method for treatment according to claim 15, wherein the
disease or condition is a chronic autoimmune and/or inflammatory
condition.
17. A method for treatment according to claim 15, wherein the
disease or condition is cancer.
18. A method for treatment according to claim 15, wherein the
subject is a human.
19. A method for inhibiting a bromodomain which comprises
contacting the bromodomain with a compound or a salt thereof
according to claim 12.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to benzodiazepine compounds,
pharmaceutical compositions containing such compounds and to their
use in therapy.
BACKGROUND OF THE INVENTION
[0002] The genomes of eukaryotic organisms are highly organised
within the nucleus of the cell. The long strands of duplex DNA are
wrapped around an octomer of histone proteins (most usually
comprising two copies of histones H2A, H2B H3 and H4) to form a
nucleosome. This basic unit is then further compressed by the
aggregation and folding of nucleosomes to form a highly condensed
chromatin structure. A range of different states of condensation
are possible, and the tightness of this structure varies during the
cell cycle, being most compact during the process of cell division.
Chromatin structure plays a critical role in regulating gene
transcription, which cannot occur efficiently from highly condensed
chromatin. The chromatin structure is controlled by a series of
post translational modifications to histone proteins, notably
histones H3 and H4, and most commonly within the histone tails
which extend beyond the core nucleosome structure. These
modifications include acetylation, methylation, phosphorylation,
ubiquitinylation, SUMOylation. These epigenetic marks are written
and erased by specific enzymes, which place the tags on specific
residues within the histone tail, thereby forming an epigenetic
code, which is then interpreted by the cell to allow gene specific
regulation of chromatin structure and thereby transcription.
[0003] Histone acetylation is most usually associated with the
activation of gene transcription, as the modification loosens the
interaction of the DNA and the histone octomer by changing the
electrostatics. In addition to this physical change, specific
proteins bind to acetylated lysine residues within histones to read
the epigenetic code. Bromodomains are small (.about.110 amino acid)
distinct domains within proteins that bind to acetylated lysine
resides commonly but not exclusively in the context of histones.
There is a family of around 50 proteins known to contain
bromodomains, and they have a range of functions within the
cell.
[0004] The BET family of bromodomain containing proteins comprises
4 proteins (BRD2, BRD3, BRD4 and BRD-t) which contain tandem
bromodomains capable of binding to two acetylated lysine residues
in close proximity, increasing the specificity of the interaction.
BRD2 and BRD3 are reported to associate with histones along
actively transcribed genes and may be involved in facilitating
transcriptional elongation (Leroy et al, Mol. Cell. 2008
30(1):51-60), while BRD4 appears to be involved in the recruitment
of the pTEF-.beta. complex to inducible genes, resulting in
phosphorylation of RNA polymerase and increased transcriptional
output (Hargreaves et al, Cell, 2009 138(1): 129-145). It has also
been reported that BRD4 or BRD3 may fuse with NUT (nuclear protein
in testis) forming novel fusion oncogenes, BRD3-NUT or BRD4-NUT, in
a highly malignant form of epithelial neoplasia (French et al.
Cancer Research, 2003, 63, 304-307 and French et al. Journal of
Clinical Oncology, 2004, 22 (20), 4135-4139). Data suggests that
BRD-NUT fusion proteins contribute to carcinogensesis (Oncogene,
2008, 27, 2237-2242). BRD-t is uniquely expressed in the testes and
ovary. All family members have been reported to have some function
in controlling or executing aspects of the cell cycle, and have
been shown to remain in complex with chromosomes during cell
division--suggesting a role in the maintenance of epigenetic
memory. In addition some viruses make use of these proteins to
tether their genomes to the host cell chromatin, as part of the
process of viral replication (You et al Cell, 2004
117(3):349-60).
[0005] Japanese patent application JP2008-156311 discloses a
benzimidazole derivative which is said to be a BRD2 bromodomain
binding agent which has utility with respect to virus
infection/proliferation.
[0006] Patent application WO2009/084693A1 discloses a series of
thienotriazolodiazepiene derivatives that are said to inhibit the
binding between an acetylated histone and a bromodomain containing
protein which are said to be useful as anti-cancer agents.
[0007] A novel class of compounds have been found which inhibit the
binding of bromodomains with its cognate acetylated proteins, more
particularly a class of compounds that inhibit the binding of BET
family bromodomains to acetylated lysine residues. Such compounds
will hereafter be referred to as "bromodomain inhibitors".
SUMMARY OF THE INVENTION
[0008] In a first aspect of the present invention, there is
provided a compound of formula (I) or a salt thereof, more
particularly a compound of formula (I) or a pharmaceutically
acceptable salt thereof.
##STR00002##
[0009] In a second aspect of the present invention, there is
provided a pharmaceutical composition comprising a compound of
formula (I) or a pharmaceutically acceptable salt thereof and one
or more pharmaceutically acceptable carriers, diluents or
excipients.
[0010] In a third aspect of the present invention, there is
provided a compound of formula (I), or a pharmaceutically
acceptable salt thereof for use in therapy, in particular in the
treatment of diseases or conditions for which a bromodomain
inhibitor is indicated.
[0011] In a fourth aspect of the present invention, there is
provided a method of treating diseases or conditions for which a
bromodomain inhibitor is indicated in a subject in need thereof
which comprises administering a therapeutically effective amount of
compound of formula (I) or a pharmaceutically acceptable salt
thereof.
[0012] In a fifth aspect of the present invention, there is
provided the use of a compound of formula (I), or a
pharmaceutically acceptable salt thereof in the manufacture of a
medicament for the treatment of diseases or conditions for which a
bromodomain inhibitor is indicated.
DETAILED DESCRIPTION OF THE INVENTION
[0013] The present invention relates to compounds of formula (I) or
a salt thereof
##STR00003##
where R.sup.1 is C.sub.1-3alkyl; R.sup.2 is --NR.sup.2aR.sup.2a' or
--OR.sup.2b; wherein one of R.sup.2a or R.sup.2'a is hydrogen, and
R.sup.2b or the other of R.sup.2a or R.sup.2'a is selected from
C.sub.1-6alkyl, haloC.sub.1-6alkyl,
R.sup.2cR.sup.2'cN--C.sub.1-6alkyl, carbocyclyl,
carbocyclylC.sub.1-4alkyl, heterocyclyl and
heterocyclylC.sub.1-4alkyl, wherein any of the carbocyclyl or
heterocyclyl groups are optionally substituted by one or more
groups selected from halogen, C.sub.1-6alkyl, haloC.sub.1-6alkyl,
C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, carbonyl, --CO-carbocyclyl,
azido, amino, hydroxyl, nitro and cyano, wherein the
--CO-carbocyclyl group may be optionally substituted by one or more
groups selected from halogen, C.sub.1-6alkyl, haloC.sub.1-6alkyl,
C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, azido, nitro and cyano; or
two adjacent groups on any of the carbocyclyl or heterocyclyl
groups together with the interconnecting atoms form a 5- or
6-membered ring which ring may contain 1 or 2 heteroatoms
independently selected from O, S and N; or R.sup.2a and R.sup.2'a
together with the N-atom to which they are attached form a 4-, 5-,
6- or 7-membered ring which optionally contains 1 or 2 heteroatoms
independently selected from O, S and N; wherein the 4-, 5-, 6- or
7-membered ring is optionally substituted by C.sub.1-6alkyl,
hydroxyl or amino; R.sup.2c and R.sup.2'c are independently
hydrogen or C.sub.1-6alkyl; each R.sup.3 is independently selected
from hydrogen, hydroxyl, halo, C.sub.1-6alkyl, halo C.sub.1-6alkyl,
C.sub.1-6alkoxy, halo C.sub.1-6alkoxy, nitro, cyano, CF.sub.3,
--OCF.sub.3, --COOR.sup.5, --C.sub.1-4alkylNR.sup.6R.sup.7 and
--C.sub.1-4alkylOH; R.sup.4 is hydroxyl, halo, C.sub.1-6alkyl,
haloC.sub.1-6alkyl, C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, nitro,
cyano, CF.sub.3, --OCF.sub.3, --COOR.sup.5; or
--OS(O).sub.2C.sub.1-4alkyl; R.sup.5 is C.sub.1-3alkyl; and n is an
integer 1 to 5.
[0014] In one embodiment the present invention relates to compounds
of formula (Ic) or a salt thereof
##STR00004##
where R.sup.1 is C.sub.1-3alkyl; R.sup.2 is --NR.sup.2aR.sup.2a' or
--OR.sup.2b; wherein one of R.sup.2a or R.sup.2'a is H, and
R.sup.2b or the other of R.sup.2a or R.sup.2'a is selected from
C.sub.1-6alkyl, haloC.sub.1-6alkyl,
R.sup.2cR.sup.2'cN--C.sub.1-6alkyl, carbocyclyl,
carbocyclylC.sub.1-4alkyl, heterocyclyl and
heterocyclylC.sub.1-4alkyl, wherein any of the carbocyclyl or
heterocyclyl groups are optionally substituted by one or more
groups selected from halogen, C.sub.1-6alkyl, haloC.sub.1-6alkyl,
C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, carbonyl, --CO-carbocyclyl,
azido, nitro and cyano, wherein the --CO-carbocyclyl group may be
further optionally substituted by one or more groups selected from
halogen, C.sub.1-6alkyl, haloC.sub.1-6alkyl, C.sub.1-6alkoxy,
haloC.sub.1-6alkoxy, azido, nitro and cyano; or two adjacent groups
on any of the carbocyclyl or heterocyclyl groups together with the
interconnecting atoms form a 5- or 6-membered ring which ring may
contain 1 or 2 heteroatoms independently selected from O, S and N;
or R.sup.2a and R.sup.2'a together with the N-atom to which they
are attached form a 5-, 6- or 7-membered ring which may optionally
contain 1 or 2 heteroatoms independently selected from O, S and N;
wherein the 5-, 6- or 7-membered ring may be further optionally
substituted by C.sub.1-6alkyl; R.sup.2c and R.sup.2'c are
independently H and C.sub.1-6alkyl; each R.sup.3 is independently
selected from H, hydroxy, halo, C.sub.1-6alkyl, halo
C.sub.1-6alkyl, C.sub.1-6alkoxy, halo C.sub.1-6alkoxy, nitro,
cyano, CF.sub.3, --OCF.sub.3 and --COOR.sup.5; R.sup.4 is hydroxy,
halo, C.sub.1-6alkyl, halo C.sub.1-6alkyl, C.sub.1-6alkoxy, halo
C.sub.1-6alkoxy, nitro, cyano, CF.sub.3, --OCF.sub.3; --COOR.sup.5
or --OS(O).sub.2C.sub.1-4alkyl; R.sup.5 is C.sub.1-3alkyl; and n is
an integer 1 to 5.
[0015] In one embodiment of the invention the S-enantiomers are
preferred.
[0016] As used herein, the term "alkyl" refers to straight or
branched hydrocarbon chains containing the specified number of
carbon atoms. For example, C.sub.1-6alkyl means a straight or
branched alkyl containing at least 1, and at most 6, carbon atoms.
Examples of "alkyl" as used herein include, but are not limited to,
methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, isobutyl,
isopropyl, t-butyl and 1,1-dimethylpropyl.
[0017] As used herein, the term "alkoxy" refers to a straight or
branched alkoxy group containing the specified number of carbon
atoms. For example, C.sub.1-6alkoxy means a straight or branched
alkoxy group containing at least 1, and at most 6, carbon atoms.
Examples of "alkoxy" as used herein include, but are not limited
to, methoxy, ethoxy, propoxy, prop-2-oxy, butoxy, but-2-oxy,
2-methylprop-1-oxy, 2-methylprop-2-oxy, pentoxy or hexyloxy.
[0018] As used herein, the term "halogen" or "halo" refers to the
elements fluorine, chlorine, bromine and iodine. Examples of
suitable halogens are fluorine, chlorine and bromine.
[0019] Unless otherwise indicated, any carbocyclyl group contains 3
to 14 ring-atoms for example, 3 to 10 ring-atoms, or in a further
example, 3 to 8 ring-atoms, and may be saturated, unsaturated or
aromatic. Examples of saturated carbocyclyl groups include
cyclopropyl, cyclopentyl or cyclohexyl. Examples of unsaturated
carbocyclyl groups include those which contain up to 3 double
bonds. An example of a suitable aromatic carbocyclyl group is
phenyl. The term carbocylic should be similarly construed. In
addition, the term carbocyclyl includes any fused combination of
carbocyclyl groups, for example naphthyl, anthryl, phenanthryl,
indanyl, indenyl, azulenyl, azulanyl, and fluorenyl.
[0020] Unless otherwise indicated, any heterocyclyl group contains
4 to 7 ring-atoms, for example, 5 to 7 ring-atoms up to 4 of which
may be hetero-atoms such as nitrogen, oxygen and sulfur, and may be
saturated, unsaturated or aromatic. Examples of heterocyclyl groups
are azetidinyl, furyl, thienyl, pyrrolyl, pyrrolinyl, pyrrolidinyl,
imidazolyl, dioxolanyl, oxazolyl, thiazolyl, imidazolyl,
imidazolinyl, imidazolidinyl, pyrazolyl, pyrazolinyl,
pyrazolidinyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl,
thiadiazolyl, pyranyl, pyridyl, piperidinyl, homopiperazinyl,
dioxanyl, morpholino, dithianyl, thiomorpholino,
thiomorpholino-1,1-dioxide, pyridazinyl, pyrimidinyl, pyrazinyl,
piperazinyl, sulfolanyl, tetrazolyl, triazinyl, azepinyl,
oxazepinyl, thiazepinyl, diazepinyl and thiazolinyl. In addition,
the term heterocyclyl includes fused heterocyclyl groups, for
example benzimidazolyl, benzoxazolyl, imidazopyridinyl,
benzoxazinyl, benzothiazinyl, oxazolopyridinyl, benzofuranyl,
quinolinyl, quinazolinyl, quinoxalinyl, dihydroquinazolinyl,
benzothiazolyl, phthalimido, benzofuranyl, benzodiazepinyl, indolyl
and isoindolyl. The term heterocyclic should be similarly
construed.
[0021] As used herein, the term "substituted" refers to
substitution with the named substituent or substituents. With
multiple degrees of substitution (e.g. 1 or 2) the groups can be
the same or different.
[0022] In one embodiment of the invention, R.sup.1 represents
methyl.
[0023] In one embodiment of the invention R.sup.2 represents
--OR.sup.2b.
[0024] Thus there is provided a compound of formula (Ia)
##STR00005##
wherein R.sup.1, R.sup.2b, R.sup.3, R.sup.4 and n are as defined
above for compounds of formula (I).
[0025] In one embodiment of the invention R.sup.2 is
--NR.sup.2aR.sup.2a'.
[0026] In one embodiment of the invention one of R.sup.2a or
R.sup.2'a is hydrogen, and the other of R.sup.2a or R.sup.2'a is
selected from C.sub.1-6alkyl, carbocyclyl,
carbocyclylC.sub.1-4alkyl, heterocyclyl and
heterocyclylC.sub.1-4alkyl, wherein any of the carbocyclyl or
heterocyclyl groups are optionally substituted by one or more
groups selected from halogen, C.sub.1-6alkyl, haloC.sub.1-6alkyl,
C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, carbonyl, --CO-carbocyclyl,
azido, amino, hydroxyl, nitro and cyano, wherein the
--CO-carbocyclyl group is optionally substituted by one or more
groups selected from halogen, C.sub.1-6alkyl, haloC.sub.1-6alkyl,
C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, azido, nitro and cyano. In
another embodiment one of R.sup.2a or R.sup.2'a is hydrogen, and
the other of R.sup.2a or R.sup.2'a is selected from C.sub.1-6alkyl,
phenyl, benzyl, benzofuranyl, pyridinyl, thiazoyl, furanyl, and
morpholinyl, wherein the phenyl, benzyl, benzofuranyl, pyridinyl,
thiazoyl, furanyl, or morpholinyl groups are optionally substituted
by one or more substituents selected from halogen, C.sub.1-6alkyl,
haloC.sub.1-6alkyl, C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, carbonyl,
azido, nitro and cyano. In another embodiment R.sup.2a is hydrogen
and R.sup.2'a is C.sub.1-6alkyl. In a further embodiment R.sup.2'a
is ethyl.
[0027] In one embodiment of the invention R.sup.2a is hydrogen and
R.sup.2'a is R.sup.2cR.sup.2'cN--C.sub.1-6alkyl. In another
embodiment R.sup.2'a is selected from
R.sup.2cR.sup.2'cN--CH.sub.2--,
R.sup.2cR.sup.2'cN--CH.sub.2CH.sub.2--,
R.sup.2cR.sup.2'cN--CH.sub.2CH.sub.2CH.sub.2-- and
R.sup.2cR.sup.2'cN--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--. In a
further embodiment R.sup.2c and R.sup.2'c are selected from
hydrogen and methyl.
[0028] In one embodiment of the invention R.sup.2a is hydrogen and
R.sup.2'a is carbocyclyl. In another embodiment R.sup.2'a is
cyclopentyl or cyclohexyl, wherein each group is optionally
substituted once by amino or hydroxyl.
[0029] In one embodiment of the invention R.sup.2a is hydrogen and
R.sup.2'a is heterocyclyl. In another embodiment R.sup.2'a is
selected from pyrrolidinyl, piperidinyl, tetrahydrofuranyl and
tetrahydropyran, wherein each group is optionally substituted by
C.sub.1-6alkyl. In another embodiment R.sup.2'a is selected
from:
##STR00006##
[0030] In a further embodiment R.sup.2'a is selected from:
##STR00007##
[0031] In one embodiment of the invention R.sup.2a is hydrogen and
R.sup.2'a is heterocyclylC.sub.1-4alkyl, wherein the heterocyclyl
is optionally substituted once by amino, hydroxyl or methyl. In
another embodiment R.sup.2'a is selected from
heterocyclyl-CH.sub.2--, heterocyclyl-CH.sub.2CH.sub.2--,
heterocyclyl-CH.sub.2CH.sub.2CH.sub.2-- and
heterocyclyl-CH.sub.2CH.sub.2CH.sub.2CH.sub.2--, wherein the
heterocyclyl is optionally substituted once by amino, hydroxyl or
methyl. In another embodiment heterocyclyl is selected from
pyrrolidinyl, piperidinyl, tetrahydropyranyl, piperazinyl,
morpholino and thiomorpholinodioxide, wherein each group is
optionally substituted once by amino, hydroxyl or methyl. In
another embodiment heterocyclyl is selected from:
##STR00008##
[0032] In a further embodiment heterocyclyl is selected from:
##STR00009##
[0033] In one embodiment of the invention, R.sup.2a and R.sup.2'a
together with the N-atom to which they are attached form a 5-, 6-
or 7-membered ring which optionally contains 1 or 2 heteroatoms
independently selected from O, S and N; wherein the 5-, 6- or
7-membered ring is optionally substituted by C.sub.1-6alkyl.
[0034] In another embodiment of the invention, R.sup.2a and
R.sup.2'a together with the N-atom to which they are attached form
a 4-, 5-, 6- or 7-membered ring which optionally contains 1 or 2
heteroatoms independently selected from O, S and N; wherein the
4-5-, 6- or 7-membered ring is optionally substituted by
C.sub.1-6alkyl, hydroxyl or amino. In another embodiment the 4-,
5-, 6- or 7-membered ring is selected from:
##STR00010##
[0035] In one embodiment of the invention, R.sup.2b represents
C.sub.1-6alkyl, carbocyclyl or carbocyclylC.sub.1-4alkyl, wherein
the carbocyclyl group is optionally substituted by one or more
groups selected from halogen, C.sub.1-6alkyl, haloC.sub.1-6alkyl,
C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, carbonyl, --CO-carbocyclyl,
azido, nitro and cyano, and wherein the --CO-carbocyclyl group may
be further optionally substituted by one or more groups selected
from halogen, C.sub.1-6alkyl, haloC.sub.1-6alkyl, C.sub.1-6alkoxy,
haloC.sub.1-6alkoxy, azido, nitro and cyano. In another embodiment
of the invention, R.sup.2b represents C.sub.1-6alkyl, phenyl, or
benzyl wherein the phenyl or benzyl group is optionally substituted
by one or more groups selected from halogen, C.sub.1-6alkyl,
haloC.sub.1-6alkyl, C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, carbonyl,
--CO-carbocyclyl, azido, nitro and cyano, and wherein the
--CO-carbocyclyl group may be further optionally substituted by one
or more groups selected from halogen, C.sub.1-6alkyl,
haloC.sub.1-6alkyl, C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, azido,
nitro and cyano. In another embodiment R.sup.2b represents methyl,
ethyl, n-butyl, t-butyl, benzyl, iso-propyl, iso-butyl, or phenyl.
In a further embodiment, R.sup.2b represents ethyl, t-butyl,
benzyl, iso-propyl, iso-butyl, or phenyl;
[0036] In one embodiment of the invention, R.sup.3 represents halo,
C.sub.1-6alkyl, C.sub.1-6alkoxy or CF.sub.3. In another embodiment,
R.sup.3 represents chloro, fluoro, methoxy or CF.sub.3. In another
embodiment, R.sup.3 represents 3-fluoro, 4-chloro, 4-fluoro,
4-methoxy, or 4-CF.sub.3. In a further embodiment R.sup.3
represents 4-chloro.
[0037] In one embodiment of the invention, R.sup.4 represents
hydroxyl or C.sub.1-6alkoxy. In another embodiment, R.sup.4
represents MeO.
[0038] In one embodiment of the invention, R.sup.4 is located in
position 8 such as 8-MeO.
[0039] Thus in one embodiment the invention provides a compound of
formula (Ib)
##STR00011##
wherein: R.sup.1, R.sup.2 and R.sup.3 and n are as defined above
for compounds of formula (I). n may for example represent an
integer in the range 1 to 3 such as 1.
[0040] While the embodiments for each variable have generally been
listed above separately for each variable this invention includes
those compounds in which several or each embodiment in formula (I)
is selected from each of the embodiments listed above. Therefore,
this invention is intended to include all combinations of
embodiments for each variable described hereinabove including salts
thereof.
[0041] In one embodiment, examples of compounds of the present
invention include: [0042]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-(2-phenylethyl)acetamide; [0043]
6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4-[2-oxo-2-(1-piperidinyl)ethyl-
]-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; [0044]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-(2-furanylmethyl)acetamide; [0045]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-cyclohexylacetamide; [0046]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-(2,2,2-trifluoroethyl)acetamide; [0047]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(3-fluorophenyl)acetamide; [0048]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(4-fluorophenyl)acetamide; [0049]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(4-fluorophenyl)acetamide; [0050]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-4-pyridinylacetamide; [0051]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-3-pyridinylacetamide; [0052]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-1,3-thiazol-2-ylacetamide; [0053]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-2-pyridinylacetamide; [0054]
N-(4-azidophenyl)-2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1-
,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamide; [0055]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-(tetrahydro-2-furanylmethyl)acetamide;
[0056]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(9-oxo-9H-fluoren-2-yl)acetamide;
[0057]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[3-(phenylcarbonyl)phenyl]acetamide;
[0058]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-{[4-(phenylcarbonyl)phenyl]methyl}acetamide;
[0059]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazo-
lo[4,3-a][1,4]benzodiazepin-4-yl]-N-[4-iodo-2-(phenylcarbonyl)phenyl]aceta-
mide; [0060]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(4-{[4-(methyloxy)phenyl]carbonyl}phenyl)acet-
amide; [0061]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(9-oxo-9H-fluoren-1-yl)acetamide;
[0062]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-{4-[(4-iodophenyl)carbonyl]phenyl}acetamide;
[0063]
6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4-[2-(4-morpholinyl)-2-o-
xoethyl]-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; [0064]
N-(1,3-benzodioxol-5-ylmethyl)-2-[6-(4-chlorophenyl)-1-methyl-8-(methylox-
y)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamide;
[0065]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[(1S)-1-phenylethyl]acetamide; [0066]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[(1R)-1-phenylethyl]acetamide; [0067]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide; [0068]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(cyclopropylmethyl)acetamide; [0069]
methyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate; [0070]
2-[(4S)-6-(3-fluorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(4-fluorophenyl)acetamide; [0071]
2-[(4S)-6-(3-fluorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide; [0072]
2-[6-(4-chlorophenyl)-8-hydroxy-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]be-
nzodiazepin-4-yl]-N-(4-fluorophenyl)acetamide; [0073] ethyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate; [0074] 1-methylethyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate; [0075] butyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate; [0076]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-[2-(methylamino)ethyl]acetamide; [0077]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-4-piperidinylacetamide; [0078]
1-{[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-
-a][1,4]benzodiazepin-4-yl]acetyl}-N-methyl-4-piperidinamine;
[0079]
(S)-1-(4-aminopiperidin-1-yl)-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-
-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)ethanone; [0080]
(S)-N-(azetidin-3-yl)-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f-
][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)acetamide; [0081]
(S)-1-(3-aminoazetidin-1-yl)-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H--
benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)ethanone; [0082]
(S)-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f][1,2,4]triazolo[4-
,3-a][1,4]diazepin-4-yl)-1-(4-hydroxypiperidin-1-yl)ethanone;
[0083]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[2-(4-morpholinyl)ethyl]acetamide;
[0084]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[2-(1H-imidazol-4-yl)ethyl]acetamide;
[0085]
(S)-tert-butyl-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f][1,2,4-
]triazolo[4,3-a][1,4]diazepin-4-yl)acetate; and [0086]
(S)-piperidin-4-yl-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f][1-
,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)acetate; [0087] or a salt
thereof.
[0088] In another embodiment, examples of compounds of the present
invention include: [0089]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-(2-phenylethyl)acetamide; [0090]
6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4-[2-oxo-2-(1-piperidinyl)ethyl-
]-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; [0091]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-(2-furanylmethyl)acetamide; [0092]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-cyclohexylacetamide; [0093]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-(2,2,2-trifluoroethyl)acetamide; [0094]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(3-fluorophenyl)acetamide; [0095]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(4-fluorophenyl)acetamide; [0096]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(4-fluorophenyl)acetamide; [0097]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-4-pyridinylacetamide; [0098]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-3-pyridinylacetamide; [0099]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-1,3-thiazol-2-ylacetamide; [0100]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-2-pyridinylacetamide; [0101]
N-(4-azidophenyl)-2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1-
,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamide; [0102]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-(tetrahydro-2-furanylmethyl)acetamide;
[0103]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(9-oxo-9H-fluoren-2-yl)acetamide;
[0104]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[3-(phenylcarbonyl)phenyl]acetamide;
[0105]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-{[4-(phenylcarbonyl)phenyl]methyl}acetamide;
[0106]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazo-
lo[4,3-a][1,4]benzodiazepin-4-yl]-N-[4-iodo-2-(phenylcarbonyl)phenyl]aceta-
mide; [0107]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(4-{[4-(methyloxy)phenyl]carbonyl}phenyl)acet-
amide; [0108]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(9-oxo-9H-fluoren-1-yl)acetamide;
[0109]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-{4-[(4-iodophenyl)carbonyl]phenyl}acetamide;
[0110]
6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4-[2-(4-morpholinyl)-2-o-
xoethyl]-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; [0111]
N-(1,3-benzodioxol-5-ylmethyl)-2-[6-(4-chlorophenyl)-1-methyl-8-(methylox-
y)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamide;
[0112]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[(1S)-1-phenylethyl]acetamide; [0113]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[(1R)-1-phenylethyl]acetamide; [0114]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide; [0115]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(cyclopropylmethyl)acetamide; [0116]
methyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate; [0117]
2-[(4S)-6-(3-fluorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(4-fluorophenyl)acetamide; [0118]
2-[(4S)-6-(3-fluorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide; [0119]
2-[6-(4-chlorophenyl)-8-hydroxy-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]be-
nzodiazepin-4-yl]-N-(4-fluorophenyl)acetamide; [0120] ethyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate; [0121] 1-methylethyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate; [0122] butyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate; [0123]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-[2-(methylamino)ethyl]acetamide; and [0124]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-4-piperidinylacetamide; [0125] or a salt
thereof.
[0126] In a further embodiment, examples of compounds of the
present invention include: [0127]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide; [0128]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-(2-phenylethyl)acetamide; [0129]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-(2-furanylmethyl)acetamide; [0130]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(2-fluorophenyl)acetamide; [0131]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(3-fluorophenyl)acetamide; [0132]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(4-fluorophenyl)acetamide; [0133]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-2-pyridinylacetamide; [0134]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-3-pyridinylacetamide; [0135]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-4-pyridinylacetamide; [0136]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-1,3-thiazol-2-ylacetamide; [0137]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(cyclopropylmethyl)acetamide; [0138]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[(1S)-1-phenylethyl]acetamide; [0139]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-{[4-(phenylcarbonyl)phenyl]methyl}acetamide;
[0140]
N-ethyl-2-[(4S)-6-(3-fluorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,-
4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamide; [0141]
N-(4-fluorophenyl)-2-[(4S)-6-(3-fluorophenyl)-1-methyl-8-(methyloxy)-4H-[-
1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamide; [0142]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-[2-(dimethylamino)ethyl]acetamide; [0143]
methyl-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo-
[4,3-a][1,4]benzodiazepin-4-yl]acetate; [0144] 1-methylethyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate; [0145]
butyl-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[-
4,3-a][1,4]benzodiazepin-4-yl]acetate; [0146]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[2-(methylamino)ethyl]acetamide;
[0147]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-4-piperidinylacetamide; [0148]
1-{[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-
-a][1,4]benzodiazepin-4-yl]acetyl}-N-methyl-4-piperidinamine;
[0149]
1-{[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-
-a][1,4]benzodiazepin-4-yl]acetyl}-4-piperidinamine; [0150]
N-3-azetidinyl-2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,-
4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamide; [0151]
1-{[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-
-a][1,4]benzodiazepin-4-yl]acetyl}-3-azetidinamine; [0152]
(S)-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f][1,2,4]triazolo[4-
,3-a][1,4]diazepin-4-yl)-1-(4-hydroxypiperidin-1-yl)ethanone;
[0153]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[2-(4-morpholinyl)ethyl]acetamide;
[0154]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[2-(1H-imidazol-4-yl)ethyl]acetamide;
[0155]
(S)-tert-butyl-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f][1,2,4-
]triazolo[4,3-a][1,4]diazepin-4-yl)acetate; [0156]
(S)-piperidin-4-yl-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f][1-
,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)acetate; [0157]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[2-(dimethylamino)ethyl]acetamide;
[0158]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[3-(dimethylamino)propyl]acetamide;
[0159]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-4-pyrimidinylacetamide; [0160]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(5-methyl-3-isoxazolyl)acetamide;
[0161]
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-(tetrahydro-2H-pyran-4-yl)acetamide;
[0162]
[4-(methyloxy)phenyl]methyl-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methylox-
y)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetate; [0163]
2-methylpropyl-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]-
triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetate; [0164]
tetrahydro-2H-pyran-4-yl-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)--
4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetate; [0165]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-4-pyridinylacetamide; [0166]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-(cyclopropylmethyl)acetamide; [0167]
2-[6-(4-chlorophenyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepi-
n-4-yl]-N-4-pyridinylacetamide; [0168]
2-[6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-1,3-thiazol-2-ylacetamide; and [0169]
methyl-[6-(4-chlorophenyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodi-
azepin-4-yl]acetate; or a salt thereof.
[0170] In one embodiment of the invention, there is provided a
compound which is
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]tria-
zolo[4,3-a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a salt
thereof. In another embodiment there is provided a compound which
is
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a pharmaceutically
acceptable salt thereof. In a further embodiment there is provided
a compound which is
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide.
[0171] It will be appreciated that the present invention covers
compounds of formula (I) as the free base and as salts thereof, for
example as a pharmaceutically acceptable salt thereof. In one
embodiment the invention relates to compounds of formula (I) as the
free base. In another embodiment the invention to compounds of
formula (I) or a pharmaceutically acceptable salt thereof.
[0172] Because of their potential use in medicine, salts of the
compounds of formula (I) are desirably pharmaceutically acceptable.
Suitable pharmaceutically acceptable salts can include acid or base
addition salts. For a review on suitable salts see Berge et al., J.
Pharm. Sci., 66:1-19, (1977). Typically, a pharmaceutically
acceptable salt may be readily prepared by using a desired acid or
base as appropriate. The resultant salt may precipitate from
solution and be collected by filtration or may be recovered by
evaporation of the solvent.
[0173] A pharmaceutically acceptable base addition salt can be
formed by reaction of a compound of formula (I) with a suitable
inorganic or organic base, (e.g. triethylamine, ethanolamine,
triethanolamine, choline, arginine, lysine or histidine),
optionally in a suitable solvent, to give the base addition salt
which is usually isolated, for example, by crystallisation and
filtration. Pharmaceutically acceptable base salts include ammonium
salts, alkali metal salts such as those of sodium and potassium,
alkaline earth metal salts such as those of calcium and magnesium
and salts with organic bases, including salts of primary, secondary
and tertiary amines, such as isopropylamine, diethylamine,
ethanolamine, trimethylamine, dicyclohexyl amine and
N-methyl-D-glucamine.
[0174] A pharmaceutically acceptable acid addition salt can be
formed by reaction of a compound of formula (I) with a suitable
inorganic or organic acid (such as hydrobromic, hydrochloric,
sulphuric, nitric, phosphoric, succinic, maleic, acetic, propionic,
fumaric, citric, tartaric, lactic, benzoic, salicylic, glutamaic,
aspartic, p-toluenesulfonic, benzenesulfonic, methanesulfonic,
ethanesulfonic, naphthalenesulfonic such as 2-naphthalenesulfonic,
or hexanoic acid), optionally in a suitable solvent such as an
organic solvent, to give the salt which is usually isolated, for
example, by crystallisation and filtration. A pharmaceutically
acceptable acid addition salt of a compound of formula (I) can
comprise or be, for example, a hydrobromide, hydrochloride,
sulfate, nitrate, phosphate, succinate, maleate, acetate,
propionate, fumarate, citrate, tartrate, lactate, benzoate,
salicylate, glutamate, aspartate, p-toluenesulfonate,
benzenesulfonate, methanesulfonate, ethanesulfonate,
naphthalenesulfonate (e.g. 2-naphthalenesulfonate) or hexanoate
salt.
[0175] Other non-pharmaceutically acceptable salts, e.g. formates,
oxalates or trifluoroacetates, may be used, for example, in the
isolation of the compounds of formula (I), and are included within
the scope of this invention.
[0176] The invention includes within its scope all possible
stoichiometric and non-stoichiometric forms of the salts of the
compounds of formula (I).
[0177] It will be appreciated that many organic compounds can form
complexes with solvents in which they are reacted or from which
they are precipitated or crystallized. These complexes are known as
"solvates". For example, a complex with water is known as a
"hydrate". Solvents with high boiling points and/or capable of
forming hydrogen bonds such as water, xylene, N-methyl
pyrrolidinone, methanol and ethanol may be used to form solvates.
Methods for identification of solvates include, but are not limited
to, NMR and microanalysis. Solvates of the compounds of formula (I)
are within the scope of the invention.
[0178] The invention includes within its scope all possible
stoichiometric and non-stoichiometric forms of the solvates of the
compounds of formula (I).
[0179] The invention encompasses all prodrugs, of the compounds of
formula (I) and pharmaceutically acceptable salts thereof, which
upon administration to the recipient are capable of providing
(directly or indirectly) a compound of formula (I) or a
pharmaceutically acceptable salt thereof, or an active metabolite
or residue thereof. Such derivatives are recognizable to those
skilled in the art, without undue experimentation. Nevertheless,
reference is made to the teaching of Burger's Medicinal Chemistry
and Drug Discovery, 5.sup.th Edition, Vol 1: Principles and
Practice, which is incorporated herein by reference to the extent
of teaching such derivatives.
[0180] The compounds of formula (I) may be in crystalline or
amorphous form. Furthermore, some of the crystalline forms of the
compounds of formula (I) may exist as polymorphs, which are
included within the scope of the present invention. Polymorphic
forms of compounds of formula (I) may be characterized and
differentiated using a number of conventional analytical
techniques, including, but not limited to, X-ray powder diffraction
(XRPD) patterns, infrared (IR) spectra, Raman spectra, differential
scanning calorimetry (DSC), thermogravimetric analysis (TGA) and
solid state nuclear magnetic resonance (SSNMR).
[0181] Certain of the compounds described herein may contain one or
more chiral atoms so that optical isomers, e.g.--enantiomers or
diastereoisomers may be formed. Accordingly, the present invention
encompasses all isomers of the compounds of formula (I) whether as
individual isomers isolated such as to be substantially free of the
other isomer (i.e. pure) or as mixtures (i.e. racemates and racemic
mixtures).
[0182] Similarly the invention also extends to conformational
isomers of compounds of formula (I) and any geometric (cis and/or
trans) isomers of said compounds.
[0183] An individual isomer isolated such as to be substantially
free of the other isomer (i.e. pure) may be isolated such that less
than 10%, particularly less than about 1%, for example less than
about 0.1% of the other isomer is present.
[0184] Separation of isomers may be achieved by conventional
techniques known to those skilled in the art, e.g. by fractional
crystallisation, chromatography or HPLC.
[0185] Certain compounds of formula (I) may exist in one of several
tautomeric forms. It will be understood that the present invention
encompasses all tautomers of the compounds of formula (I) whether
as individual tautomers or as mixtures thereof.
[0186] It will be appreciated from the foregoing that included
within the scope of the invention are solvates, isomers and
polymorphic forms of the compounds of formula (I) and salts
thereof.
[0187] The compounds of formula (I) and pharmaceutically acceptable
salts thereof may be made by a variety of methods, including
standard chemistry. Any previously defined variable will continue
to have the previously defined meaning unless otherwise indicated.
Illustrative general synthetic methods are set out below and then
specific compounds of formula (I) and pharmaceutically acceptable
salts thereof are prepared in the working Examples. These processes
form further aspects of the present invention.
[0188] Throughout the specification, general formulae are
designated by Roman numerals (I), (II), (III), (IV) etc. Subsets of
these general formulae are defined as (Ia), (Ib), (Ic) etc.
[0189] Compounds of formula (Ic), i.e. compounds of general formula
(I) where R.sup.2 is OR.sup.2b, may be prepared according to
reaction scheme 1 by reacting compounds of formula (Id), i.e.
compounds of general formula (I) where R.sup.2 is OH, with
R.sup.2bOH in the presence of DIC and DMAP at room temperature.
Alternatively compounds of formula (Ic) may be prepared by reacting
compounds of formula (Id) with oxalyl chloride followed by addition
of R.sup.2bOH.
##STR00012##
[0190] Compounds of formula (Ie), i.e. compounds of general formula
(I) where R.sup.2 is NR.sup.2aR.sup.2'a may be prepared according
to reaction scheme 2 by reacting compounds of formula (Id), i.e.
compounds of general formula (I) where R.sup.2 is OH, with
R.sup.2aR.sup.2'aNH, in the presence of HATU or HBTU and DIEA at
room temperature. Alternatively compounds of formula (Ie) may be
prepared by reacting compounds of formula (Id) with oxalyl chloride
followed by addition of R.sup.2aR.sup.2'aNH in the presence of
triethylamine.
##STR00013##
[0191] Alternatively compounds of formula (Ie), i.e. compounds of
general formula (I) where R.sup.2 is NR.sup.2aR.sup.2'a, may be
prepared according to reaction scheme 3 by reacting compounds of
formula (III) with hydrazine followed by reaction of the resulting
hydrazone (II) with R.sup.1C(OR).sub.3. Preferably hydrazone (II)
is used without further purification and is reacted with
R.sup.1C(OR).sub.3.
##STR00014##
wherein R represents C.sub.1-4alkyl such as methyl.
[0192] Compounds of formula (Id), i.e. compounds of general formula
(I) where R.sup.2 is OH, may be prepared according to reaction
scheme 4. Suitable reaction conditions comprise reacting compounds
of formula (Ic), i.e. compounds of general formula (I) where
R.sup.2 is OR.sup.2b, with alkaline hydroxide preferably sodium
hydroxide or lithium hydroxide.
##STR00015##
[0193] Compounds of formula (Ic), i.e. compounds of general formula
(I) where R.sup.2 is OR.sup.2b may be prepared according to
reaction scheme 5 by reacting compounds of formula (IV) with
AcOH.
##STR00016##
[0194] Compounds of formula (IV) may be prepared according to
reaction scheme 6 by reacting compounds of formula (VI) with
hydrazine below 15.degree. C. followed by reaction of the resulting
hydrazone (V) with R.sup.1COCl at 0.degree. C. Generally hydrazone
(V) is used without further purification and is reacted with
R.sup.1COCl at, for example 0.degree. C.
##STR00017##
[0195] Compounds of formula (III) may be prepared according to
reaction scheme 7 by reacting compounds of formula (VII) with
R.sup.2aR.sup.2'aNH in the presence of HATU and DIEA at, for
example room temperature.
##STR00018##
[0196] Compounds of formula (VII) may be prepared according to
reaction scheme 8. Suitable reaction conditions comprise reacting
compounds of formula (VI) with alkaline hydroxide such as sodium
hydroxide.
##STR00019##
[0197] Compounds of formula (VI) may be prepared according to
reaction scheme 9 from compounds of formula (VIII) by treatment
with Lawesson's reagent or P.sub.4S.sub.10. Suitable reaction
conditions comprise reacting compounds of formula (VIII) with
P.sub.4S.sub.10 in 1,2-dichloroethane at, for example 70.degree.
C.
##STR00020##
[0198] Compounds of formula (VIII) may be prepared according to
reaction scheme 10, by reacting compounds of formula (X) with an
organic base such as triethylamine followed by reaction of the
resulting amine (IX) with acetic acid. Generally, amine (IX) is
used without further purification and is reacted with AcOH at, for
example 60.degree. C.
##STR00021##
[0199] Compounds of formula (X) may be prepared according to
reaction scheme 11, by reacting compounds of formula (XI) with the
acylchloride (XII) derived from protected aspartic acid.
##STR00022##
[0200] Compounds of formula (XI) may be prepared according to
procedures described in Synthesis 1980, 677-688.
[0201] Acyl chlorides (XII) may be prepared according to procedures
described in J. Org. Chem., 1990, 55, 3068-3074 and J. Chem. Soc.
Perkin Trans. 1, 2001, 1673-1695
[0202] It will be appreciated by those skilled in the art that it
may be advantageous to protect one or more functional groups of the
compounds described in the above processes. Examples of protecting
groups and the means for their removal can be found in T. W. Greene
`Protective Groups in Organic Synthesis` (4th edition, J. Wiley and
Sons, 2006). Suitable amine protecting groups include acyl (e.g.
acetyl, carbamate (e.g. 2',2',2'-trichloroethoxycarbonyl,
benzyloxycarbonyl or t-butoxycarbonyl) and arylalkyl (e.g. benzyl),
which may be removed by hydrolysis (e.g. using an acid such as
hydrochloric acid in dioxane or trifluoroacetic acid in
dichloromethane) or reductively (e.g. hydrogenolysis of a benzyl or
benzyloxycarbonyl group or reductive removal of a
2',2',2'-trichloroethoxycarbonyl group using zinc in acetic acid)
as appropriate. Other suitable amine protecting groups include
trifluoroacetyl (--COCF.sub.3) which may be removed by base
catalysed hydrolysis.
[0203] It will be appreciated that in any of the routes described
above, the precise order of the synthetic steps by which the
various groups and moieties are introduced into the molecule may be
varied. It will be within the skill of the practitioner in the art
to ensure that groups or moieties introduced at one stage of the
process will not be affected by subsequent transformations and
reactions, and to select the order of synthetic steps
accordingly.
[0204] Certain intermediate compounds described above are believed
to be novel and therefore form a yet further aspect of the
invention.
[0205] The compounds of formula (I) and salts thereof are
bromodomain inhibitors, and thus are believed to have potential
utility in the treatment of diseases or conditions for which a
bromodomain inhibitor is indicated.
[0206] The present invention thus provides a compound of formula
(I) or a pharmaceutically acceptable salt thereof for use in
therapy. In one embodiment there is provided
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a pharmaceutically
acceptable salt thereof for use in therapy. The compounds of
formula (I) or pharmaceutically salts thereof can be for use in the
treatment of diseases or conditions for which a bromodomain
inhibitor indicated.
[0207] In one embodiment there is provided a compound of formula
(I) or a pharmaceutically acceptable salt thereof for use in the
treatment of diseases or conditions for which a bromodomain
inhibitor is indicated. In another embodiment, there is provided a
compound or a pharmaceutically acceptable salt thereof for use in
the treatment of a chronic autoimmune and/or inflammatory
condition. In a further embodiment, there is provided a compound or
a pharmaceutically acceptable salt thereof for use in the treatment
of cancer, such as midline carcinoma.
[0208] In one embodiment there is provided
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a pharmaceutically
acceptable salt thereof for use in the treatment of diseases or
conditions for which a bromodomain inhibitor is indicated. In
another embodiment, there is provided
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a pharmaceutically
acceptable salt thereof for use in the treatment of a chronic
autoimmune and/or inflammatory condition. In a further embodiment,
there is provided
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a pharmaceutically
acceptable salt thereof for use in the treatment of cancer, such as
midline carcinoma.
[0209] In one embodiment there is provided the use of a compound of
formula (I) or a pharmaceutically acceptable salt thereof in the
manufacture of a medicament for the treatment of diseases or
conditions for which a bromodomain inhibitor is indicated. In
another embodiment, there is provided the use of a compound of
formula (I) or a pharmaceutically acceptable salt thereof in the
manufacture of a medicament for the treatment of a chronic
autoimmune and/or inflammatory condition. In a further embodiment,
there is provided the use of a compound of formula (I) or a
pharmaceutically acceptable salt thereof in the manufacture of a
medicament for the treatment of cancer, such as midline
carcinoma.
[0210] In one embodiment there is provided the use of
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a pharmaceutically
acceptable salt thereof in the manufacture of a medicament for the
treatment of diseases or conditions for which a bromodomain
inhibitor is indicated. In another embodiment, there is provided
the use of
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a pharmaceutically
acceptable salt thereof in the manufacture of a medicament for the
treatment of a chronic autoimmune and/or inflammatory condition. In
a further embodiment, there is provided the use of
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a pharmaceutically
acceptable salt thereof in the manufacture of a medicament for the
treatment of cancer, such as midline carcinoma.
[0211] In one embodiment there is provided a method for the
treatment of a disease or condition, for which a bromodomain
inhibitor is indicated, in a subject in need thereof which
comprises administering a therapeutically effective amount of
compound of formula (I) or a pharmaceutically acceptable salt
thereof. In another embodiment there is provided a method for
treatment of a chronic autoimmune and/or inflammatory condition, in
a subject in need thereof which comprises administering a
therapeutically effective amount of compound of formula (I) or a
pharmaceutically acceptable salt thereof. In a further embodiment
there is provided a method for treatment of cancer, such as midline
carcinoma, in a subject in need thereof which comprises
administering a therapeutically effective amount of compound of
formula (I) or a pharmaceutically acceptable salt thereof.
[0212] In one embodiment there is provided a method for treatment
of a disease or condition, for which a bromodomain inhibitor is
indicated, in a subject in need thereof which comprises
administering a therapeutically effective amount of
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a pharmaceutically
acceptable salt thereof. In another embodiment there is provided a
method for treatment of a chronic autoimmune and/or inflammatory
condition, in a subject in need thereof which comprises
administering a therapeutically effective amount of
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a pharmaceutically
acceptable salt thereof. In a further embodiment there is provided
a method for treatment of cancer, such as midline carcinoma, in a
subject in need thereof which comprises administering a
therapeutically effective amount of
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]tri-
azolo[4,3-a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a
pharmaceutically acceptable salt thereof.
[0213] In one embodiment the subject in need thereof is a mammal,
particularly a human.
[0214] As used herein, the term "effective amount" means that
amount of a drug or pharmaceutical agent that will elicit the
biological or medical response of a tissue, system, animal or human
that is being sought, for instance, by a researcher or clinician.
Furthermore, the term "therapeutically effective amount" means any
amount which, as compared to a corresponding subject who has not
received such amount, results in improved treatment, healing,
prevention, or amelioration of a disease, disorder, or side effect,
or a decrease in the rate of advancement of a disease or disorder.
The term also includes within its scope amounts effective to
enhance normal physiological function.
[0215] Bromodomain inhibitors are believed to be useful in the
treatment of a variety of diseases or conditions related to
systemic or tissue inflammation, inflammatory responses to
infection or hypoxia, cellular activation and proliferation, lipid
metabolism, fibrosis and in the prevention and treatment of viral
infections.
[0216] Bromodomain inhibitors may be useful in the treatment of a
wide variety of chronic autoimmune and inflammatory conditions such
as rheumatoid arthritis, osteoarthritis, acute gout, psoriasis,
systemic lupus erythematosus, multiple sclerosis, inflammatory
bowel disease (Crohn's disease and Ulcerative colitis), asthma,
chronic obstructive airways disease, pneumonitis, myocarditis,
pericarditis, myositis, eczema, dermatitis, alopecia, vitiligo,
bullous skin diseases, nephritis, vasculitis, atherosclerosis,
Alzheimer's disease, depression, retinitis, uveitis, scleritis,
hepatitis, pancreatitis, primary biliary cirrhosis, sclerosing
cholangitis, Addison's disease, hypophysitis, thyroiditis, type I
diabetes and acute rejection of transplanted organs.
[0217] Bromodomain inhibitors may be useful in the treatment of a
wide variety of acute inflammatory conditions such as acute gout,
giant cell arteritis, nephritis including lupus nephritis,
vasculitis with organ involvement such as glomerulonephritis,
vasculitis including giant cell arteritis, Wegener's
granulomatosis, Polyarteritis nodosa, Behcet's disease, Kawasaki
disease, Takayasu's Arteritis and acute rejection of transplanted
organs.
[0218] Bromodomain inhibitors may be useful in the prevention or
treatment of diseases or conditions which involve inflammatory
responses to infections with bacteria, viruses, fungi, parasites or
their toxins, such as sepsis, sepsis syndrome, septic shock,
endotoxaemia, systemic inflammatory response syndrome (SIRS),
multi-organ dysfunction syndrome, toxic shock syndrome, acute lung
injury, ARDS (adult respiratory distress syndrome), acute renal
failure, fulminant hepatitis, burns, acute pancreatitis,
post-surgical syndromes, sarcoidosis, Herxheimer reactions,
encephalitis, myelitis, meningitis, malaria, SIRS associated with
viral infections such as influenza, herpes zoster, herpes simplex
and coronavirus.
[0219] Bromodomain inhibitors may be useful in the prevention or
treatment of conditions associated with ischaemia-reperfusion
injury such as myocardial infarction, cerebro-vascular ischaemia
(stroke), acute coronary syndromes, renal reperfusion injury, organ
transplantation, coronary artery bypass grafting, cardio-pulmonary
bypass procedures and pulmonary, renal, hepatic, gastro-intestinal
or peripheral limb embolism.
[0220] Bromodomain inhibitors may be useful in the treatment of
disorders of lipid metabolism via the regulation of APO-A1 such as
hypercholesterolemia, atherosclerosis and Alzheimer's disease.
[0221] Bromodomain inhibitors may be useful in the treatment of
fibrotic conditions such as idiopathic pulmonary fibrosis, renal
fibrosis, post-operative stricture, keloid formation, scleroderma
and cardiac fibrosis.
[0222] Bromodomain inhibitors may be useful in the prevention and
treatment of viral infections such as herpes virus, human papilloma
virus, adenovirus, poxvirus and other DNA viruses.
[0223] Bromodomain inhibitors may be useful in the treatment of
cancer, including hematological, epithelial including lung, breast
and colon carcinomas, midline carcinomas, mesenchymal, hepatic,
renal and neurological tumours.
[0224] In one embodiment the disease or condition for which a
bromodomain inhibitor is indicated is selected from diseases
associated with systemic inflammatory response syndrome, such as
sepsis, burns, pancreatitis, major trauma, haemorrhage and
ischaemia. In this embodiment the bromodomain inhibitor would be
administered at the point of diagnosis to reduce the incidence of:
SIRS, the onset of shock, multi-organ dysfunction syndrome, which
includes the onset of acute lung injury, ARDS, acute renal,
hepatic, cardiac and gastro-intestinal injury and mortality. In
another embodiment the bromodomain inhibitor would be administered
prior to surgical or other procedures associated with a high risk
of sepsis, haemorrhage, extensive tissue damage, SIRS or MODS
(multiple organ dysfunction syndrome). In a particular embodiment
the disease or condition for which a bromodomain inhibitor is
indicated is sepsis, sepsis syndrome, septic shock and
endotoxaemia. In another embodiment, the bromodomain inhibitor is
indicated for the treatment of acute or acute on chronic
pancreatitis. In another embodiment the bromodomain inhibitor is
indicated for the treatment of burns.
[0225] In one embodiment the disease or condition for which a
bromodomain inhibitor is indicated is selected from herpes simplex
infections and reactivations, cold sores, herpes zoster infections
and reactivations, chickenpox, shingles, human papilloma virus,
cervical neoplasia, adenovirus infections, including acute
respiratory disease, and poxvirus infections such as cowpox and
smallpox and African swine fever virus. In one particular
embodiment a bromodomain inhibitor is indicated for the treatment
of Human papilloma virus infections of skin or cervical
epithelia.
[0226] The term "diseases or conditions for which a bromodomain
inhibitor is indicated", is intended to include any of or all of
the above disease states.
[0227] In one embodiment, there is provided a method for inhibiting
a bromodomain which comprises contacting the bromodomain with a
compound of formula (I) or a pharmaceutically acceptable salt
thereof.
[0228] While it is possible that for use in therapy, a compound of
formula (I) as well as pharmaceutically acceptable salts thereof
may be administered as the raw chemical, it is common to present
the active ingredient as a pharmaceutical composition.
[0229] The present invention therefore provides in a further aspect
a pharmaceutical composition comprising a compound of formula (I)
or a pharmaceutically acceptable salt thereof and one or more
pharmaceutically acceptable carriers, diluents or excipients. The
compounds of the formula (I) and pharmaceutically acceptable salts
thereof, are as described above. In one embodiment there is
provided a pharmaceutical composition comprising
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-ethylacetamide or a pharmaceutically
acceptable salt thereof and one or more pharmaceutically acceptable
carriers, diluents or excipients. The carrier(s), diluent(s) or
excipient(s) must be acceptable in the sense of being compatible
with the other ingredients of the composition and not deleterious
to the recipient thereof. In accordance with another aspect of the
invention there is also provided a process for the preparation of a
pharmaceutical composition including admixing a compound of the
formula (I), or a pharmaceutically acceptable salt thereof, with
one or more pharmaceutically acceptable carriers, diluents or
excipients. The pharmaceutical composition can be for use in the
treatment of any of the conditions described herein.
[0230] Since the compounds of formula (I) and pharmaceutically
acceptable salts thereof are intended for use in pharmaceutical
compositions it will be readily understood that they are each
preferably provided in substantially pure form, for example, at
least 60% pure, more suitably at least 75% pure and preferably at
least 85% pure, especially at least 98% pure (% in a weight for
weight basis).
[0231] Pharmaceutical compositions may be presented in unit dose
forms containing a predetermined amount of active ingredient per
unit dose. Preferred unit dosage compositions are those containing
a daily dose or sub-dose, or an appropriate fraction thereof, of an
active ingredient. Such unit doses may therefore be administered
more than once a day. Preferred unit dosage compositions are those
containing a daily dose or sub-dose (for administration more than
once a day), as herein above recited, or an appropriate fraction
thereof, of an active ingredient.
[0232] Pharmaceutical compositions may be adapted for
administration by any appropriate route, for example by the oral
(including buccal or sublingual), rectal, inhaled, intranasal,
topical (including buccal, sublingual or transdermal), vaginal or
parenteral (including subcutaneous, intramuscular, intravenous or
intradermal) route. Such compositions may be prepared by any method
known in the art of pharmacy, for example by bringing into
association the active ingredient with the carrier(s) or
excipient(s).
[0233] In one embodiment there is provided a pharmaceutical
composition comprising a compound of formula (I) or a
pharmaceutically acceptable salt thereof adapted for oral
administration.
[0234] In one embodiment the pharmaceutical composition is adapted
for parenteral administration, particularly intravenous
administration.
[0235] Pharmaceutical compositions adapted for parenteral
administration include aqueous and non-aqueous sterile injection
solutions which may contain anti-oxidants, buffers, bacteriostats
and solutes which render the composition isotonic with the blood of
the intended recipient; and aqueous and non-aqueous sterile
suspensions which may include suspending agents and thickening
agents. The compositions may be presented in unit-dose or
multi-dose containers, for example sealed ampoules and vials, and
may be stored in a freeze-dried (lyophilized) condition requiring
only the addition of the sterile liquid carrier, for example water
for injections, immediately prior to use. Extemporaneous injection
solutions and suspensions may be prepared from sterile powders,
granules and tablets.
[0236] Pharmaceutical compositions adapted for oral administration
may be presented as discrete units such as capsules or tablets;
powders or granules; solutions or suspensions in aqueous or
non-aqueous liquids; edible foams or whips; or oil-in-water liquid
emulsions or water-in-oil liquid emulsions.
[0237] For instance, for oral administration in the form of a
tablet or capsule, the active drug component can be combined with
an oral, non-toxic pharmaceutically acceptable inert carrier such
as ethanol, glycerol, water and the like. Powders suitable for
incorporating into tablets or capsules may be prepared by reducing
the compound to a suitable fine size (e.g. by micronisation) and
mixing with a similarly prepared pharmaceutical carrier such as an
edible carbohydrate, as, for example, starch or mannitol.
Flavoring, preservative, dispersing and coloring agent can also be
present.
[0238] Capsules may be made by preparing a powder mixture, as
described above, and filling formed gelatin sheaths. Glidants and
lubricants such as colloidal silica, talc, magnesium stearate,
calcium stearate or solid polyethylene glycol can be added to the
powder mixture before the filling operation. A disintegrating or
solubilizing agent such as agar-agar, calcium carbonate or sodium
carbonate can also be added to improve the availability of the
medicament when the capsule is ingested.
[0239] Moreover, when desired or necessary, suitable binders,
glidants, lubricants, sweetening agents, flavours, disintegrating
agents and coloring agents can also be incorporated into the
mixture. Suitable binders include starch, gelatin, natural sugars
such as glucose or beta-lactose, corn sweeteners, natural and
synthetic gums such as acacia, tragacanth or sodium alginate,
carboxymethylcellulose, polyethylene glycol, waxes and the like.
Lubricants used in these dosage forms include sodium oleate, sodium
stearate, magnesium stearate, sodium benzoate, sodium acetate,
sodium chloride and the like. Disintegrators include, without
limitation, starch, methyl cellulose, agar, bentonite, xanthan gum
and the like. Tablets are formulated, for example, by preparing a
powder mixture, granulating or slugging, adding a lubricant and
disintegrant and pressing into tablets. A powder mixture is
prepared by mixing the compound, suitably comminuted, with a
diluent or base as described above, and optionally, with a binder
such as carboxymethylcellulose, an aliginate, gelatin, or polyvinyl
pyrrolidone, a solution retardant such as paraffin, a resorption
accelerator such as a quaternary salt and/or an absorption agent
such as bentonite, kaolin or dicalcium phosphate. The powder
mixture can be granulated by wetting with a binder such as syrup,
starch paste, acadia mucilage or solutions of cellulosic or
polymeric materials and forcing through a screen. As an alternative
to granulating, the powder mixture can be run through the tablet
machine and the result is imperfectly formed slugs broken into
granules. The granules can be lubricated to prevent sticking to the
tablet forming dies by means of the addition of stearic acid, a
stearate salt, talc or mineral oil. The lubricated mixture is then
compressed into tablets. The compounds of the present invention can
also be combined with a free flowing inert carrier and compressed
into tablets directly without going through the granulating or
slugging steps. A clear or opaque protective coating consisting of
a sealing coat of shellac, a coating of sugar or polymeric material
and a polish coating of wax can be provided. Dyestuffs can be added
to these coatings to distinguish different unit dosages.
[0240] Oral fluids such as solution, syrups and elixirs can be
prepared in dosage unit form so that a given quantity contains a
predetermined amount of the compound. Syrups can be prepared by
dissolving the compound in a suitably flavored aqueous solution,
while elixirs are prepared through the use of a non-toxic alcoholic
vehicle. Suspensions can be formulated by dispersing the compound
in a non-toxic vehicle. Solubilizers and emulsifiers such as
ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol
ethers, preservatives, flavor additive such as peppermint oil or
natural sweeteners or saccharin or other artificial sweeteners, and
the like can also be added.
[0241] Where appropriate, dosage unit compositions for oral
administration can be microencapsulated. The formulation can also
be prepared to prolong or sustain the release as for example by
coating or embedding particulate material in polymers, wax or the
like.
[0242] The compounds of formula (I) and pharmaceutically acceptable
salts thereof can also be administered in the form of liposome
delivery systems, such as small unilamellar vesicles, large
unilamellar vesicles and multilamellar vesicles. Liposomes can be
formed from a variety of phospholipids, such as cholesterol,
stearylamine or phosphatidylcholines.
[0243] Pharmaceutical compositions adapted for topical
administration may be formulated as ointments, creams, suspensions,
lotions, powders, solutions, pastes, gels, sprays, aerosols or
oils.
[0244] For treatments of the eye or other external tissues, for
example mouth and skin, the compositions are preferably applied as
a topical ointment or cream. When formulated in an ointment, the
active ingredient may be employed with either a paraffinic or a
water-miscible ointment base. Alternatively, the active ingredient
may be formulated in a cream with an oil-in-water cream base or a
water-in-oil base.
[0245] Pharmaceutical compositions adapted for topical
administrations to the eye include eye drops wherein the active
ingredient is dissolved or suspended in a suitable carrier,
especially an aqueous solvent.
[0246] Dosage forms for nasal or inhaled administration may
conveniently be formulated as aerosols, solutions, suspensions,
gels or dry powders.
[0247] For compositions suitable and/or adapted for inhaled
administration, it is preferred that the compound of formula (I) or
pharmaceutically acceptable salt thereof is in a
particle-size-reduced form e.g. obtained by micronisation. The
preferable particle size of the size-reduced (e.g. micronised)
compound or salt is defined by a D50 value of about 0.5 to about 10
microns (for example as measured using laser diffraction).
[0248] Aerosol formulations, e.g. for inhaled administration, can
comprise a solution or fine suspension of the active substance in a
pharmaceutically acceptable aqueous or non-aqueous solvent. Aerosol
formulations can be presented in single or multidose quantities in
sterile form in a sealed container, which can take the form of a
cartridge or refill for use with an atomising device or inhaler.
Alternatively the sealed container may be a unitary dispensing
device such as a single dose nasal inhaler or an aerosol dispenser
fitted with a metering valve (metered dose inhaler) which is
intended for disposal once the contents of the container have been
exhausted.
[0249] Where the dosage form comprises an aerosol dispenser, it
preferably contains a suitable propellant under pressure such as
compressed air, carbon dioxide or an organic propellant such as a
hydrofluorocarbon (HFC). Suitable HFC propellants include
1,1,1,2,3,3,3-heptafluoropropane and 1,1,1,2-tetrafluoroethane. The
aerosol dosage forms can also take the form of a pump-atomiser. The
pressurised aerosol may contain a solution or a suspension of the
active compound. This may require the incorporation of additional
excipients e.g. co-solvents and/or surfactants to improve the
dispersion characteristics and homogeneity of suspension
formulations. Solution formulations may also require the addition
of co-solvents such as ethanol.
[0250] For pharmaceutical compositions suitable and/or adapted for
inhaled administration, the pharmaceutical composition may be a dry
powder inhalable composition. Such a composition can comprise a
powder base such as lactose, glucose, trehalose, mannitol or
starch, the compound of formula (I) or a pharmaceutically
acceptable salt thereof (preferably in particle-size-reduced form,
e.g. in micronised form), and optionally a performance modifier
such as L-leucine or another amino acid and/or metals salts of
stearic acid such as magnesium or calcium stearate. Preferably, the
dry powder inhalable composition comprises a dry powder blend of
lactose e.g. lactose monohydrate and the compound of formula (I) or
a pharmaceutically acceptable salt thereof. Such compositions can
be administered to the patient using a suitable device such as the
DISKUS.RTM. device, marketed by GlaxoSmithKline which is for
example described in GB 2242134 A.
[0251] The compounds of formula (I) and pharmaceutically acceptable
salts thereof may be formulated as a fluid formulation for delivery
from a fluid dispenser, for example a fluid dispenser having a
dispensing nozzle or dispensing orifice through which a metered
dose of the fluid formulation is dispensed upon the application of
a user-applied force to a pump mechanism of the fluid dispenser.
Such fluid dispensers are generally provided with a reservoir of
multiple metered doses of the fluid formulation, the doses being
dispensable upon sequential pump actuations. The dispensing nozzle
or orifice may be configured for insertion into the nostrils of the
user for spray dispensing of the fluid formulation into the nasal
cavity. A fluid dispenser of the aforementioned type is described
and illustrated in WO2005/044354 A1.
[0252] A therapeutically effective amount of a compound of formula
(I) or a pharmaceutically acceptable salt thereof will depend upon
a number of factors including, for example, the age and weight of
the animal, the precise condition requiring treatment and its
severity, the nature of the formulation, and the route of
administration, and will ultimately be at the discretion of the
attendant physician or veterinarian. In the pharmaceutical
composition, each dosage unit for oral or parenteral administration
preferably contains from 0.01 to 3000 mg, more preferably 0.5 to
1000 mg, of a compound of formula (I) or a pharmaceutically
acceptable salt thereof calculated as the free base. Each dosage
unit for nasal or inhaled administration preferably contains from
0.001 to 50 mg, more preferably 0.01 to 5 mg, of a compound of the
formula (I) or a pharmaceutically acceptable salt thereof,
calculated as the free base.
[0253] The compounds of formula (I) and pharmaceutically acceptable
salts thereof can be administered in a daily dose (for an adult
patient) of, for example, an oral or parenteral dose of 0.01 mg to
3000 mg per day or 0.5 to 1000 mg per day, or a nasal or inhaled
dose of 0.001 to 50 mg per day or 0.01 to 5 mg per day, of the
compound of formula (I) or a pharmaceutically acceptable salt
thereof, calculated as the free base. This amount may be given in a
single dose per day or more usually in a number (such as two,
three, four, five or six) of sub-doses per day such that the total
daily dose is the same. An effective amount of a pharmaceutically
acceptable salt thereof, may be determined as a proportion of the
effective amount of the compound of formula (I) per se.
[0254] The compounds of formula (I) and pharmaceutically acceptable
salts thereof may be employed alone or in combination with other
therapeutic agents. Combination therapies according to the present
invention thus comprise the administration of at least one compound
of formula (I) or a pharmaceutically acceptable salt thereof, and
the use of at least one other pharmaceutically active agent.
Preferably, combination therapies according to the present
invention comprise the administration of at least one compound of
formula (I) or a pharmaceutically acceptable salt thereof, and at
least one other pharmaceutically active agent. The compound(s) of
formula (I) and pharmaceutically acceptable salts thereof, and the
other pharmaceutically active agent(s) may be administered together
in a single pharmaceutical composition or separately and, when
administered separately this may occur simultaneously or
sequentially in any order. The amounts of the compound(s) of
formula (I) and pharmaceutically acceptable salts thereof, and the
other pharmaceutically active agent(s) and the relative timings of
administration will be selected in order to achieve the desired
combined therapeutic effect. Thus in a further aspect, there is
provided a combination comprising a compound of formula (I) or
pharmaceutically acceptable salt thereof and at least one other
pharmaceutically active agent. In one embodiment there is provided
a combination pharmaceutical product comprising a compound of
formula (I) or a pharmaceutically acceptable salt thereof together
with one or more other therapeutically active agents.
[0255] Thus in one aspect, the compounds and pharmaceutical
compositions according to the invention may be used in combination
with or include one or more other therapeutic agents, for example
selected from antibiotics, anti-virals, glucocorticosteroids,
muscarinic antagonists and beta-2 agonists.
[0256] It will be appreciated that when the compounds of the
present invention are administered in combination with other
therapeutic agents normally administered by the inhaled,
intravenous, oral or intranasal route, that the resultant
pharmaceutical composition may be administered by the same routes.
Alternatively the individual components of the composition may be
administered by different routes.
[0257] One embodiment of the invention encompasses combinations
comprising one or two other therapeutic agents.
[0258] It will be clear to a person skilled in the art that, where
appropriate, the other therapeutic ingredient(s) may be used in the
form of salts, for example as alkali metal or amine salts or as
acid addition salts, or prodrugs, or as esters, for example lower
alkyl esters, or as solvates, for example hydrates, to optimise the
activity and/or stability and/or physical characteristics, such as
solubility, of the therapeutic ingredient. It will be clear also
that, where appropriate, the therapeutic ingredients may be used in
optically pure form.
[0259] The combinations referred to above may conveniently be
presented for use in the form of a pharmaceutical composition and
thus pharmaceutical compositions comprising a combination as
defined above together with a pharmaceutically acceptable diluent
or carrier represent a further aspect of the invention.
[0260] The compounds of formula (I) and salts thereof may be
prepared by the methods described below or by similar methods. Thus
the following Intermediates and Examples serve to illustrate the
preparation of the compounds of formula (I) and salts thereof, and
are not to be considered as limiting the scope of the invention in
any way.
General Experimental Details
[0261] All temperatures referred to are in .degree. C.
Abbreviations
[0262] TLC--thin layer chromatography [0263] AcOH--acetic acid
[0264] AcCl--acetyl chloride [0265] PPTS--pyridinium
p-toluenesulfonate [0266] DCM--dichloromethane [0267]
1,2-DCE--1,2-dichloroethane [0268] DIC--Diisopropylcarbodiimide
[0269] DIEA--N,N-diisopropylethylamine [0270]
DMF--N,N-dimethylformamide [0271] DMAP--4-dimethylaminopyridine
[0272] Fmoc--9H-fluoren-9-ylmethyl)oxy]carbonyl [0273]
HATU--O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate [0274]
HBTU--O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate [0275] Et.sub.2O--diethyl ether [0276]
EtOAc--ethyl acetate [0277] i-Pr.sub.2O--di-isopropyl ether [0278]
Config.--absolute configuration [0279]
Lawesson's--2,4-bis(4-methoxyphenyl)-1,3-dithia-2,4-diphosphetane-2,4-dis-
ulphide Reagent [0280] MeCN--acetonitrile [0281] MeOH--methanol
[0282] Rt--retention time [0283] THF--tetrahydrofuran [0284]
RT--room temperature [0285] Pd/C--palladium on carbon [0286]
COMU--1-cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-ca-
rbenium hexafluorophosphate
[0287] LC/MS refers to analyses by analytical HPLC which were
conducted on three kinds of apparatus: [0288] a) On a Supelcosil
LCABZ+PLUS column (3 .mu.m, 3.3 cm.times.4.6 mm ID) eluting with
0.1% HCO.sub.2H and 0.01 M ammonium acetate in water (solvent A),
and 95% acetonitrile and 0.05% HCO.sub.2H in water (solvent B),
using the following elution gradient 0-0.7 minutes 0% B, 0.7-4.2
minutes 0.fwdarw.100% B, 4.2-5.3 minutes 100% B, 5.3-5.5 minutes
100.fwdarw.0% B at a flow rate of 3 mL/minute. The mass spectra
(MS) were recorded on a Fisons VG Platform mass spectrometer using
electrospray positive ionisation [(ES+ve to give [M+H].sup.+ and
[M+NH.sub.4].sup.+ molecular ions] or electrospray negative
ionisation [(ES-ve to give [M-H]- molecular ion] modes. Analytical
data from this apparatus are given with the following format:
[M+H].sup.+ or [M-H].sup.-. [0289] b) On a Chromolith Performance
RP 18 column (100.times.4.6 mm id) eluting with 0.01M ammonium
acetate in water (solvent A) and 100% acetonitrile (solvent B),
using the following elution gradient 0-4 minutes 0.fwdarw.100% B,
4-5 minutes 100% B at a flow rate of 5 mL/minute. The mass spectra
(MS) were recorded on a micromass Platform-LC mass spectrometer
using atmospheric pressure chemical positive ionisation [AP+ve to
give MH.sup.+ molecular ions] or atmospheric pressure chemical
negative ionisation [AP-ve to give (M-H).sup.- molecular ions]
modes. Analytical data from this apparatus are given with the
following format: [M+H]+ or [M-H]- preceded by the acronym APCI to
specify between both mass spectrometry analyses sources. [0290] c)
On an Acquity UPLC BEH C18 column (2.1 mm.times.50 mm id, 1.7 .mu.m
packing diameter) at 40.degree. C., eluting with 0.1% v/v solution
of formic acid in water (solvent system A) and 0.1% v/v solution of
formic acid in acetonitrile (solvent system B). The gradient
employed was:
TABLE-US-00001 [0290] Solvent Flow Rate System Time (min) (ml/min)
% A % B 0 1 97 3 1.5 1 0 100 1.9 1 0 100 2.0 1 97 3
[0291] The UV detection system was an averaged signal from
wavelength of 210 nm to 350 nm and mass spectra were recorded on a
mass spectrometer using alternate-scan positive and negative mode
electrospray ionization.
[0292] LC/HRMS: Analytical HPLC was conducted on a Uptisphere-hsc
column (3 .mu.m 33.times.3 mm id) eluting with 0.01M ammonium
acetate in water (solvent A) and 100% acetonitrile (solvent B),
using the following elution gradient 0-0.5 minutes 5% B, 0.5-3.75
minutes 5.fwdarw.100% B, 3.75-4.5 100% B, 4.5-5 100.fwdarw.5% B,
5-5.5 5% B at a flow rate of 1.3 mL/minute. The mass spectra (MS)
were recorded on a micromass LCT mass spectrometer using
electrospray positive ionisation [ES+ve to give MH.sup.+ molecular
ions] or electrospray negative ionisation [ES-ve to give (M-H)-
molecular ions] modes.
[0293] Mass directed auto-prep HPLC refers to the method where the
material was purified by high performance liquid chromatography on
a HPLCABZ+5 .mu.m column (5 cm.times.10 mm i.d.) with 0.1%
HCO.sub.2H in water and 95% MeCN, 5% water (0.5% HCO.sub.2H)
utilising the following gradient elution conditions: 0-1.0 minutes
5% B, 1.0-8.0 minutes 5.fwdarw.30% B, 8.0-8.9 minutes 30% B,
8.9-9.0 minutes 30.fwdarw.95% B, 9.0-9.9 minutes 95% B, 9.9-10
minutes 95.fwdarw.0% B at a flow rate of 8 mL/minute. The Gilson
202-fraction collector was triggered by a VG Platform Mass
Spectrometer on detecting the mass of interest.
[0294] Proton NMR (.sup.1H NMR) spectra were recorded at ambient
temperature on a Bruker Avance 300 DPX spectrometer using solvant
as internal standard and proton chemical shifts are expressed in
ppm in the indicated solvent. The following abbreviations are used
for multiplicity of NMR signals: s=singlet, d=doublet, t=triplet,
q=quadruplet, dd=double doublet, m=multiplet.
[0295] TLC (thin layer chromatography) refers to the use of TLC
plates sold by Merck coated with silica gel 60 F254.
Intermediate 1: Methyl
[(4S)-6-(3-fluorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate
##STR00023##
[0297] The crude Intermediate 5 (assumed 9.3 mmol) was suspended in
a mixture of THF and AcOH (1/1) and stirred overnight before being
concentrated under reduced pressure. The crude mixture was
suspended in DCM and washed with sat. NaHCO.sub.3 solution. The
organic layer was dried over Na.sub.2SO.sub.4, filtered and
concentrated in vacuo and the resulting solid was triturated in
i-Pr.sub.2O and filtered. The crude product was purified by
flash-chromatography on silica gel (DCM/MeOH:95/5) to give the
title compound which will be used in the next step without further
purification. LC/MS: m/z 395 [M+H].sup.+, Rt 2.71 min.
Intermediate 2:
[(4S)-6-(4-Chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetic acid
##STR00024##
[0299] To a solution of Example 34 (28 g, 68 mmol) in THF (450 mL)
at RT was added 1N NaOH (136 mL, 136 mmol). The reaction mixture
was stirred at this temperature for 5 h before being cooled down
and quenched with 1N HCl (136 mL). THF was removed under reduced
pressure and the aqueous layer was extracted with DCM. The combined
organic layers were dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The crude solid was
recrystallised in CH.sub.3CN to give the title compound (23.9 g,
89% yield) as a pale yellow powder. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta.7.55-7.48 (m, 2H), 7.41 (d, J=8.9 Hz, 1H),
7.38-7.31 (m, 2H), 7.22 (dd, J=2.9 and 8.9 Hz, 1H), 6.90 (d, J=2.9
Hz, 1H), 4.59 (dd, J=6.9 and 6.9 Hz, 1H), 3.81 (s, 3H), 3.70 (dd,
J=6.9 and 25.7 Hz, 1H), 3.61 (dd, J=6.9 and 25.7 Hz, 1H), 2.63 (s,
3H). LC/MS: m/z 397 [M(.sup.35Cl)+H].sup.+, Rt 2.11 min.
[0300] Intermediate 3 of formula (Ib), i.e. compounds of general
formula (I) where R.sup.2 is OH was prepared by methods analogous
to that described for Intermediate 2 starting from Intermediate 1
and using dioxane instead of THF as solvant.
Intermediate 3:
[(4S)-6-(3-Fluorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetic acid
##STR00025##
[0302] LC/MS: m/z 381 [M+H].sup.+, Rt 2.08 min.
Intermediate 4: Methyl
[(3S)-2-[(1Z)-2-acetylhydrazino]-5-(4-chlorophenyl)-7-(methyloxy)-3H-1,4--
benzodiazepin-3-yl]acetate
##STR00026##
[0304] To a suspension of Intermediate 6 (30.2 g, 77.7 mmol) in THF
(800 mL) at 0.degree. C. was added hydrazine monohydrate (11.3 mL,
233 mmol) dropwise. The reaction mixture was stirred for 4 h
between 0.degree. C. and 15.degree. C. before being cooled at
0.degree. C. Et.sub.3N (32.4 mL, 230 mmol) was then added slowly
and AcCl (16.3 mL, 230 mmol) was added dropwise. The mixture was
allowed to warm to RT and stir for 1 h then quenched with water and
concentrated under reduced pressure. The resulting aqueous layer
was then extracted with DCM and the organic layer was dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
crude title compound (34 g, 100% yield) which was used without
further purification. LC/MS: m/z 429 [M(.sup.35Cl)+H].sup.+, Rt
2.83 min.
[0305] The method described above could equally be applied to the
racemic form of Intermediate 6 to give the product in racemic
form.
[0306] Intermediate 5 was prepared by methods analogous to that
described for Intermediate 4 starting from Intermediate 7.
Intermediate 5: Methyl
[(3S)-2-[(1Z)-2-acetylhydrazino]-5-(3-fluorophenyl)-7-(methyloxy)-3H-1,4--
benzodiazepin-3-yl]acetate
##STR00027##
[0308] LC/MS: m/z 413 [M+H].sup.+, Rt 2.66 min.
Intermediate 6: Methyl
[(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-thioxo-2,3-dihydro-1H-1,4-benzod-
iazepin-3-yl]acetate
##STR00028##
[0310] A suspension of P.sub.4S.sub.10 (85.8 g, 190 mmol) and
Na.sub.2CO.sub.3 (20.5 g, 190 mmol) in 1,2-DCE (1.5 L) at RT was
stirred for 1 h before Intermediate 9 (40 g, 107 mmol) was added.
The resulting mixture was stirred at 65.degree. C. for 4 h before
being cooled and filtered. The solid was washed with DCM and the
filtrate washed with sat. NaHCO.sub.3. The organic layer was dried
over Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The title compound was precipitated from a
DCM/i-Pr.sub.2O mixture and filtered. The filtrate was then
concentrated and purified by flash chromatography (DCM/MeOH:98/2)
to afford another batch of product. The title compound was obtained
combining the two fractions (30.2 g, 73%) as a yellow powder.
LC/MS: m/z 389 [M(.sup.35Cl)+H].sup.+, Rt 3.29 min.
[0311] Alternatively, Intermediate 9 could be reacted with
Lawesson's reagent (0.6 equiv.) in refluxing toluene to give the
title compound as a racemic mixture.
[0312] Intermediate 7 was prepared by methods analogous to that
described for Intermediate 6 starting from Intermediate 10.
Intermediate 7: Methyl
[(3S)-5-(3-fluorophenyl)-7-(methyloxy)-2-thioxo-2,3-dihydro-1H-1,4-benzod-
iazepin-3-yl]acetate
##STR00029##
[0314] LC/MS: m/z 373 [M+H].sup.+, Rt 3.11 min.
Intermediate 8:
2-[5-(4-Chlorophenyl)-7-(methyloxy)-2-thioxo-2,3-dihydro-1H-1,4-benzodiaz-
epin-3-yl]-N-cyclohexylacetamide
##STR00030##
[0316] To a solution of Intermediate 11 (500 mg, 1.3 mmol) in THF
(50 mL) at RT was added DIEA (0.35 mL, 2.0 mmol) followed by HATU
(760 mg, 2.0 mmol). The reaction mixture was stirred for 5 min
before cyclohexylamine (0.23 mL, 2.0 mmol) was added. The mixture
was stirred for 30 h and a further 0.5 equiv. of HATU, DIEA and
cyclohexylamine were added with stirring. The organic layer was
washed with 1N NaOH. The aqueous layer was extracted with DCM and
the organics were dried over Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude solid was then purified by flash
chromatography on silica gel to give the title compound (420 mg,
62%) as a yellow foam. R.sub.f=0.68 (DCM/MeOH:90/10). LC/MS: m/z
456 [M(.sup.35Cl)+H].sup.+, Rt 3.11 min.
Intermediate 9: Methyl
[(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-oxo-2,3-dihydro-1H-1,4-benzodiaz-
epin-3-yl]acetate
##STR00031##
[0318] To a solution of the crude Intermediate 12 (assumed 0.2 mol)
in DCM (500 mL) was added Et.sub.3N (500 mL, 3.65 mol) and the
resulting mixture was refluxed for 24 h before being concentrated.
The resulting crude amine was dissolved in 1,2-DCE (1.5 L) and AcOH
(104 mL, 1.8 mol) was added carefully. The reaction mixture was
then stirred at 60.degree. C. for 2 h before being concentrated in
vacuo and dissolved in DCM. The organic layer was washed with 1N
HCl and the aqueous layer was extracted with DCM (.times.3). The
combined organic layers were washed twice with water, and brine,
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The crude solid was recrystallised in MeCN
leading to the title compound (51 g) as a pale yellow solid. The
filtrate could be concentrated and recrystallised in MeCN to give
another 10 g of Intermediate 9 (total: 61 g, 69% yield based on
recovered Intermediate 12). R.sub.f=0.34 (DCM/MeOH:95/5). LC/MS m/z
373 [M(.sup.35Cl)+H].sup.+, Rt 2.76 min.
[0319] Intermediate 10 was prepared by methods analogous to that
described for Intermediate 9 starting from Intermediate 13.
Intermediate 10: Methyl
[(3S)-5-(3-fluorophenyl)-7-(methyloxy)-2-oxo-2,3-dihydro-1H-1,4-benzodiaz-
epin-3-yl]acetate
##STR00032##
[0321] LC/MS: m/z 357 [M+H].sup.+, Rt 2.83 min.
Intermediate 11:
[5-(4-Chlorophenyl)-7-(methyloxy)-2-thioxo-2,3-dihydro-1H-1,4-benzodiazep-
in-3-yl]acetic acid
##STR00033##
[0323] To a solution of racemic Intermediate 6 (2.6 g, 6.7 mmol) in
1,4-dioxane (70 mL) at RT was added slowly 1N NaOH (13.5 mL) and
the reaction mixture was stirred overnight before adding another 1N
NaOH (13.5 mL). The mixture was then further stirred for 2 h before
1,4-dioxane was removed in vacuo. The aqueous layer was acidified
with 1N HCl to ca pH 6 and extracted with DCM. The organics were
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give the title compound (1.9 g, 76% yield) as a
yellow solid. LC/MS: m/z 375 [M(.sup.35Cl)+H].sup.+, Rt=1.6
min.
Intermediate 12: Methyl
N'-[2-[(4-chlorophenyl)carbonyl]-4-(methyloxy)phenyl]-N.sup.2-{[(9H-fluor-
en-9-ylmethyl)oxy]carbonyl}-L-.alpha.-asparaginate
##STR00034##
[0325] A mixture of Methyl
N-{[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-.alpha.-aspartyl
chloride (prepared from J. Org. Chem. 1990, 55, 3068-3074 and J.
Chem. Soc. Perkin Trans. 1 2001, 1673-1695) (221 g, 0.57 mol) and
Intermediate 14 (133 g, 0.5 mol) in CHCl.sub.3 (410 mL) was stirred
at 60.degree. C. for 1.5 h before being cooled and concentrated
under reduced pressure and used without further purification.
LC/MS: m/z 613 [M(.sup.35Cl)+H].sup.+, Rt=3.89 min.
[0326] Intermediate 13 was prepared by methods analogous to that
described for Intermediate 12 starting from methyl
N-{[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-.alpha.-aspartyl
chloride (prepared from J. Org. Chem. 1990, 55, 3068-3074 and J.
Chem. Soc. Perkin Trans. 1 2001, 1673-1695) and
(2-Amino-5-methoxy-phenyl)-(3-fluoro-phenyl)-methanone (Synthesis
1980, 677-688).
Intermediate 13: Methyl
N'-[2-[(3-fluorophenyl)carbonyl]-4-(methyloxy)phenyl]-N.sup.2-{[(9H-fluor-
en-9-ylmethyl)oxy]carbonyl}-L-.alpha.-asparaginate
##STR00035##
[0328] LC/MS: m/z 597 [M+H].sup.+, Rt 3.75 min.
Intermediate 14:
[2-amino-5-(methyloxy)phenyl](4-chlorophenyl)methanone
##STR00036##
[0330] To a solution of Intermediate 15 (40.0 g, 0.21 mol) in a
toluene (560 mL)/ether (200 mL) mixture at 0.degree. C. was added
dropwise a solution of 4-chlorophenylmagnesium bromide (170 mL, 1M
in Et.sub.2O, 0.17 mol). The reaction mixture was allowed to warm
to RT and stirred for 1 h before being quenched with 1N HCl. The
aqueous layer was extracted with EtOAc (3.times.) and the combined
organics were washed with brine, dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure. The crude
compound was then dissolved in EtOH (400 mL) and 6N HCl (160 mL)
was added. The reaction mixture was refluxed for 2 h before being
concentrated under reduced pressure. The resulting solid was
filtered and washed twice with ether before being suspended in
EtOAc and neutralised with 1N NaOH. The aqueous layer was extracted
with EtOAc (3.times.) and the combined organics were washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The title compound was obtained as a yellow solid
(39 g, 88% yield) which was used without further purification.
Intermediate 15: 2-methyl-6-(methyloxy)-4H-3,1-benzoxazin-4-one
##STR00037##
[0332] A solution of 5-methoxyanthranilic acid (7.8 g, 46.5 mmol)
was refluxed in acetic anhydride (60 mL) for 2 h15 before being
cooled and concentrated under reduced pressure. The crude residue
was then concentrated twice in the presence of toluene before being
filtered and washed with ether to yield to the title compound (6.8
g, 77% yield) as a beige solid; LC/MS: m/z 192 [M+H].sup.+, Rt 1.69
min.
Intermediate 16 1,1-Dimethylethyl
[2-({[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4-
,3-a][1,4]benzodiazepin-4-yl]acetyl}amino)ethyl]methylcarbamate
##STR00038##
[0334] Intermediate 2 (104 mg, 0.262 mmol), 1,1-dimethylethyl
(2-aminoethyl)methylcarbamate (0.049 mL, 0.275 mmol), HATU (120 mg,
0.314 mmol) and DIPEA (0.110 mL, 0.629 mmol) were dissolved in DMF
(2.5 mL) and stirred at 50.degree. C. under N.sub.2 overnight. The
reaction was allowed to cool next morning, then the DMF evaporated
under vacuum and the crude oil partitioned between EtOAc and
saturated aqueous sodium bicarbonate solution. The organic phase
was washed with saturated aqueous brine, dried over anhydrous
magnesium sulfate and evaporated under vacuum to give a colourless
oil. Purification of the residue by flash chromatography on silica
gel (DCM-methanol 0.5 to 10% gradient) gave the title compound as a
yellow oil (189 mg, 0.342 mmol, 130% yield). LC/MS
(M(.sup.35Cl)+H).sup.+ 553, (M(.sup.37Cl)+H).sup.+ 555; RT 1.07
min.
Intermediate 17 1,1-Dimethylethyl
4-({[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,-
3-a][1,4]benzodiazepin-4-yl]acetyl}amino)-1-piperidinecarboxylate
##STR00039##
[0336] Intermediate 2 (100 mg, 0.252 mmol), 1,1-dimethylethyl
4-amino-1-piperidinecarboxylate (0.054 mL, 0.265 mmol), HATU (115
mg, 0.302 mmol) and DIPEA (0.106 mL, 0.605 mmol) were dissolved in
(DMF) (2.5 mL) and stirred at 50.degree. C. under N.sub.2
overnight. The reaction was allowed to cool next morning, then the
DMF evaporated under vacuum and the crude oil partitioned between
EtOAc and saturated aqueous sodium bicarbonate solution. The
organic phase was washed with saturated aqueous brine, dried over
anhydrous magnesium sulfate and evaporated under vacuum to give a
colourless oil. Purification of the residue by flash chromatography
on silica gel (DCM-methanol 0.5 to 10% gradient) gave the title
compound as a yellow oil (165 mg, 0.285 mmol, 113% yield). LC/MS
(M(.sup.35Cl)+H).sup.+ 579, (M(.sup.37Cl)+H).sup.+ 581; RT 1.11
min.
[0337] Intermediates 18-21 were prepared from Intermediate 2 and
the appropriate amine using the procedure described in Intermediate
17.
TABLE-US-00002 ##STR00040## Inter- Mass Spec me- (M(.sup.35Cl) +
H).sup.+, RT diate A (M(.sup.37Cl) + H).sup.+ (mins) 18
##STR00041## 593, 595 1.18 19 ##STR00042## 579, 581 1.09 20
##STR00043## 551, 553 1.07 21 ##STR00044## 551, 553 1.04
[0338] LC/MS was carried out using apparatus c).
Intermediate 22:
[(4S)-6-(4-chlorophenyl)-8-hydroxy-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4-
]benzodiazepin-4-yl]acetic acid
##STR00045##
[0340] A solution of Intermediate 2 (400 mg, 1 mmol) in DCM (8 mL)
was cooled to -78.degree. C. A solution of boron tribromide (1.26
g, 476 .mu.l, 5 mmol) in dichloromethane (2 mL) was added slowly.
The reaction mixture was a stirred at -78.degree. C. for 30 minutes
under nitrogen then allowed to warm to RT and stirred for 4 hours.
The reaction mixture was poured onto ice/water (.about.100 mL) and
the organic phase was separated. The aqueous phase was extracted
with DCM (2.times.10 mL). The aqueous phase was evaporated to
dryness. The residue was extracted with MeOH (3.times.10 mL) and
the combined organic phase was evaporated and the residue
re-evaporated from toluene (.times.2) to give the title compound as
a yellow solid (383 mg, 1.0 mmol, 99% yield). LC/MS
(M(.sup.35Cl)+H).sup.+ 383, (M(.sup.37Cl)+H).sup.+ 385; RT 0.73
min. (using LC/MS apparatus c)).
EXAMPLES
Example 1:
2-[(4S)-6-(4-Chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]tri-
azolo[4,3-a][1,4]benzodiazepin-4-yl]-N-ethylacetamide
##STR00046##
[0342] To a solution of Intermediate 2 (16.0 g, 40 mmol) in THF at
RT was added DIEA (14 mL, 80 mmol) followed by HATU (30.4 g, 80
mmol). The reaction mixture was stirred for 3 h at this temperature
and ethylamine (40 mL, 2M in THF, 80 mmol) was added. The mixture
was stirred for 48 h before being concentrated under reduced
pressure. The crude material was suspended in water and extracted
with DCM. The organic layer was dried over Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The crude solid was purified by
chromatography on SiO.sub.2 (DCM/MeOH 95/5) and the resulting solid
recrystallised in MeCN. The solid was then dissolved in DCM and
precipited with i-Pr.sub.2O to give the title compound (8 g, 47%
yield) as a white solid.
[0343] R.sub.f=0.48 (DCM/MeOH:90/10). Mp>140.degree. C. (becomes
gummy). .sup.1H RMN (300 MHz, CDCl.sub.3) .delta.7.53-7.47 (m, 2H),
7.39 (d, J=8.9 Hz, 1H), 7.37-7.31 (m, 2H), 7.20 (dd, J=2.9 and 8.9
Hz, 1H), 6.86 (d, J=2.9 Hz, 1H), 6.40 (m, 1H), 4.62 (m, 1H), 3.80
(s, 3H), 3.51 (dd, J=7.3 and 14.1 Hz, 1H), 3.46-3.21 (m, 3H), 2.62
(s, 3H), 1.19 (t, J=7.3 Hz, 3H). LC/MS: m/z 424
[M(.sup.35Cl)+H].sup.+, Rt 2.33 min.
[0344] Examples 2 to 24 i.e. compounds of general formula (I) where
R.sup.2 is NR.sup.2aR.sup.2'a, (see Table 1) were prepared by
methods analogous to that described for Example 1 from Intermediate
2 using the appropriate amine. Intermediate 3 was used in place of
Intermediate 1 for Examples 23 and 24.
##STR00047##
TABLE-US-00003 TABLE 1 Ex R.sup.3 Config. R.sup.2a R.sup.2'a
Physical data 2 4-Cl (R,S) ##STR00048## H LC/MS: m/z 500
[M(.sup.35Cl) + H].sup.+, Rt 2.78 min. 3 4-Cl (R,S)
--(CH.sub.2).sub.5-- LC/HRMS (M + H).sup.+ calc for
C.sub.25H.sub.26.sup.35ClN.sub.5O.sub.2 464.1853; found 464.1846.
Rt 2.68 min. 4 4-Cl (R,S) ##STR00049## H LC/HRMS (M + H).sup.+ calc
for C.sub.25H.sub.22.sup.35ClN.sub.5O.sub.3 476.1489; found
476.1471. Rt 2.56 min. 5 4-Cl (R,S) ##STR00050## H LC/HRMS (M +
H).sup.+ calc for C.sub.22H.sub.18.sup.35ClF.sub.3N.sub.5O.sub.2
478.1258; found 478.1256. Rt 2.60 min. 6 4-Cl (R,S) ##STR00051## H
LC/HRMS (M + H).sup.+ calc for
C.sub.25H.sub.26.sup.35ClN.sub.5O.sub.3 480.1845; found 480.1830.
Rt 2.37 min. 7 4-Cl (R,S) --(CH.sub.2).sub.2--O--(CH.sub.2).sub.2--
LC/MS: m/z 466 [M(.sup.35Cl) + H].sup.+, Rt 2.66 min 8 4-Cl (R,S)
##STR00052## H LC/HRMS (M + H).sup.+ calc for
C.sub.28H.sub.24.sup.35ClN.sub.5O.sub.4 530.1595; found 530.1578.
Rt 2.67 min. Mp 255.degree. C. 9 4-Cl (S) ##STR00053## H LC/MS: m/z
490 [M(.sup.35Cl) + H].sup.+, Rt 3.15 min. Mp 264.degree. C. 10
4-Cl (S) ##STR00054## H LC/MS: m/z 490 [M(.sup.35Cl) + H].sup.+, Rt
3.25 min. Mp 248.degree. C. 11 4-Cl (S) ##STR00055## H LC/HRMS (M +
H).sup.+ calc for C.sub.26H.sub.21.sup.35ClFN.sub.5O.sub.2
490.1446; found 490.1399. Rt 2.81 min. Mp 246.degree. C. 12 4-Cl
(S) ##STR00056## H LC/MS: m/z 473 [M(.sup.35Cl) + H].sup.+, Rt 2.96
min. 13 4-Cl (S) ##STR00057## H LC/MS: m/z 473 [M(.sup.35Cl) +
H].sup.+, Rt 2.75 min. Mp >160.degree. C. 14 4-Cl (S)
##STR00058## H LC/HRMS (M + H).sup.+ calc for
C.sub.25H.sub.21.sup.35ClN.sub.6O.sub.2 473.1493; found 473.1419.
Rt 2.41 .min. Mp 223.degree. C. 15 4-Cl (S) ##STR00059## H LC/HRMS
(M + H).sup.+ calc for C.sub.23H.sub.19.sup.35ClN.sub.6O.sub.2S
479.1057; found 479.1050. Rt 2.58 min. Mp 249.degree. C. 16 4-Cl
(S) ##STR00060## H LC/HRMS (M + H).sup.+ calc for
C.sub.24H.sub.24.sup.35ClN.sub.5O.sub.2 450.1697; found 450.1613.
Rt 2.50 min. Mp 128.degree. C. 17 4-Cl (S) ##STR00061## H LC/MS:
m/z 500 [M(.sup.35Cl) + H].sup.+, Rt 2.73 min. 18 4-Cl (S)
##STR00062## H LC/MS: m/z 500 [M(.sup.35Cl) + H].sup.+, Rt 4.23
min. 19 4-Cl (S) ##STR00063## H LC/MS: m/z 513 [M(.sup.35Cl) +
H].sup.+, Rt 3.33 min. 20 4-Cl (S) ##STR00064## H LC/HRMS (M +
H).sup.+ calc for C.sub.33H.sub.24.sup.35ClN.sub.5O.sub.3 574.1646;
found 574.1616. Rt 3.05 min. 21 4-Cl (S) ##STR00065## H LC/MS: m/z
576 [M(.sup.35Cl) + H].sup.+, Rt 3.42 min 22 4-Cl (S) ##STR00066##
H LC/MS: m/z 590 [M(.sup.35Cl) + H].sup.+, Rt 3.20 min. 23 3-F (S)
##STR00067## H LC/MS: m/z 408 [M(.sup.35Cl) + H].sup.+, Rt 2.50
min. 24 3-F (S) ##STR00068## H LC/MS: m/z 474 [M(.sup.35Cl) +
H].sup.+, Rt 3.01 min.
Example 25:
2-[(4S)-6-(4-Chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[4-(4-methoxybenzoyl)phenyl]acetamide
##STR00069##
[0346] To a solution of Intermediate 2 (1 g, 2.5 mmol) in THF at
0.degree. C. was added dropwise oxalyl chloride (1.26 mL, 2.0 M in
DCM, 2.5 mmol). DMF (1 mL) was then added dropwise and the reaction
mixture was stirred for 1 h before a further 1 equiv of
oxalylchloride was added. The reaction mixture was stirred another
1 h. One tenth of the latter solution was then added to a solution
of (4-aminophenyl)[4-(methyloxy)phenyl]methanone (50 mg, 0.22 mmol)
and Et.sub.3N in THF. The mixture was stirred overnight before
being quenched with 1N NaOH. The aqueous layer was extracted with
EtOAc and organics were washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
product was purified by flash chromatography on silica gel (DCM
then DCM/MeOH:95/5) and the solid was triturated in i-Pr.sub.2O to
give the title compound (38 mg, 26% yield) as a ochre powder.
LC/MS: m/z 606 [M(.sup.35Cl)+H].sup.+, Rt 3.41 min.
[0347] Examples 26 to 29 i.e. compounds of general formula (I)
where R.sup.2 is NR.sup.2aR.sup.2'a, (see Table 2) were prepared by
methods analogous to that described for Example 25 using the
appropriate amine.
##STR00070##
TABLE-US-00004 TABLE 2 Ex Config. R.sup.2a R.sup.2'a Physical data
26 (R,S) ##STR00071## H LC/HRMS (M + H).sup.+ calc for
C.sub.24H.sub.28.sup.35ClN.sub.6O.sub.2 467.1962; found 467.1964.
Rt 2.60 min. 27 (S) ##STR00072## H LC/MS: m/z 574 [M(.sup.35Cl) +
H].sup.+, Rt 3.66 min 28 (S) ##STR00073## H LC/MS: m/z 702
[M(.sup.35Cl) + H].sup.+, Rt = 3.78 min 29 (S) ##STR00074## H
LC/MS: m/z 702 [M(.sup.35Cl) + H].sup.+, Rt = 3.79 min
Example 30:
2-[6-(4-Chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,-
4]benzodiazepin-4-yl]-N-cyclohexylacetamide
##STR00075##
[0349] To a solution of Intermediate 8 (420 mg, 0.9 mmol) in MeOH
(40 mL) at 0.degree. C. was added hydrazine monohydrate (0.22 mL,
4.6 mmol). After 15 min at 0.degree. C., the reaction mixture was
allowed to warm RT for 1 h and a further 2.5 equiv. of hydrazine
hydrate before being heated to 40.degree. C. for 15 min. The
mixture was then concentrated to half volume and water was added.
The aqueous layer was extracted with DCM (.times.2) and the
organics were dried over Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude product was then dissolved in MeOH
(30 mL) and trimethylorthoacetate (0.17 mL, 1.3 mmol) and PPTS (45
mg, 0.18 mmol) were added. The reaction mixture was refluxed for 45
min before being concentrated under reduced pressure. The crude
mixture was dissolved in DCM and washed with water, dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
solid was then purified by flash chromatography on silica gel to
give the title compound (70 mg, 16% yield over two steps) as a
white solid. R.sub.f=0.5 (DCM/MeOH:90/10). Mp>260.degree. C.
LC/HRMS (M+H).sup.+ calculated for
C.sub.26H.sub.28.sup.35ClN.sub.5O.sub.2 478.2010; found 478.1968.
Rt 2.76 min.
Example 31:
(4S)-6-(4-Chlorophenyl)-4-{2-[(4-fluorophenyl)amino]-2-oxoethyl}-1-methyl-
-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-8-yl
methanesulfonate
##STR00076##
[0351] To a solution of Example 32 (20 mg, 42 .mu.mol) and
Et.sub.3N (9 .mu.L, 63 .mu.mol) in THF (1 mL) at RT was added
methanesulfonyl chloride (5 .mu.L, 63 .mu.mol). The reaction
mixture was stirred for 5 h before another Et.sub.3N (63 .mu.mol)
and methanesulfonyl chloride (63 .mu.mol) were added twice. After
concentration, the crude mixture was dissolved in DCM and washed
successively with 1N HCl, 1N NaOH and brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
solid was triturated in a DCM/i-Pr.sub.2O mixture to give the title
compound (9 mg, 39% yield) as a white solid. LC/MS: m/z 554
[M(.sup.35Cl)+H].sup.+, Rt 3.04 min.
Example 32:
2-[(4S)-6-(4-Chlorophenyl)-8-hydroxy-1-methyl-4H-[1,2,4]triazolo[4,3-a][1-
,4]benzodiazepin-4-yl]-N-(4-fluorophenyl)acetamide
##STR00077##
[0353] A mixture of a solution of Example 11 (50 mg, 0.1 mmol) in
DCM (1 mL) and BBr.sub.3 (0.51 mL, 1M in DCM, 0.51 mmol) at
0.degree. C. was allowed to stir and warm to RT for 5 h before
being quenched with water. The organic layer was then washed with a
saturated hydrosulfite solution and brine before being dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
solid was triturated in a DCM/i-Pr.sub.2O mixture to give the title
compound (47 mg, 99% yield) as a pale yellow solid. Mp=222.degree.
C. (decomposition). LC/MS: m/z 476 [M(.sup.35Cl)+H].sup.+, Rt 2.82
min.
[0354] Example 33 was prepared by a method analogous to that
described for Example 32 starting from Example 1.
Example 33:
2-[(4S)-6-(4-Chlorophenyl)-8-hydroxy-1-methyl-4H-[1,2,4]triazolo[4,3-a][1-
,4]benzodiazepin-4-yl]-N-ethylacetamide
##STR00078##
[0356] LC/HRMS (M+H).sup.+ calc for
C.sub.21H.sub.20.sup.35ClN.sub.5O.sub.2 410.1384; found 410.1324.
Rt 2.05 min.
Example 34: Methyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate
##STR00079##
[0358] The crude Intermediate 4 (34 g, 79 mmol) was suspended in
THF (200 mL) and AcOH (200 mL) was added at RT. The reaction
mixture was stirred at this temperature overnight before being
concentrated to dryness. The residue was suspended in saturated
NaHCO.sub.3 and extracted with DCM. The organic layer was dried
over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude solid was purified by chromatography on SiO.sub.2
(DCM/MeOH:90/10) to give the title compound (28 g, 86% yield) as a
yellow powder. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.7.54-7.47
(m, 2H), 7.40 (d, J=8.8 Hz, 1H), 7.37-7.31 (m, 2H), 7.22 (dd, J=2.8
and 8.8 Hz, 1H), 6.89 (d, J=2.8 Hz, 1H), 4.61 (dd, J=6.4 and 7.8
Hz, 1H), 3.82 (s, 3H), 3.78 (s, 3H), 3.66 (dd, J=7.8 and 16.9 Hz,
1H), 3.60 (dd, J=6.4 and 16.9 Hz, 1H), 2.62 (s, 3H). LC/MS m/z 411
[M(.sup.35Cl)+H].sup.+, Rt 2.88 min.
[0359] The method described above could equally be applied to the
racemic form of Intermediate 4 to give the product in racemic
form.
Example 35: Ethyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate
##STR00080##
[0361] To a solution of Intermediate 2 (200 mg, 0.5 mmol) in THF
(10 mL) at 0.degree. C. was added oxalylchloride (0.37 mL, 2M in
DCM, 0.75 mmol). After stirring 3 hours, excess ethanol was added
and the reaction was allowed to warm to room temperature and stir
an additional 24 hours. The mixture was concentrated to dryness and
the residue taken up in water and extracted with DCM to afford a
crude mixture which was purified by Mass Directed Auto Prep to give
the title compound as a white powder (30 mg, 15%).
[0362] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.7.54-7.47 (m, 2H),
7.39 (d, J=9.1 Hz, 1H), 7.37-7.32 (m, 2H), 7.22 (dd, J=2.8 and 9.1
Hz, 1H), 6.89 (d, J=2.8 Hz, 1H), 4.61 (dd, J=6.2 and 7.9 Hz, 1H),
4.23 (q, J=7.2 Hz, 2H), 3.82 (s, 3H), 3.64 (dd, J=7.9 and 17.0 Hz,
1H), 3.58 (dd, J=6.3 and 17.0 Hz, 1H), 2.63 (s, 3H), 1.33 (t, J=7.2
Hz, 3H). LC/HRMS (M+H).sup.+ calc for
C.sub.22H.sub.21.sup.35ClN.sub.4O.sub.3 425.1333; found 425.1328.
Rt 2.80 min.
Example 36: 1-Methylethyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate
##STR00081##
[0364] To a solution of Intermediate 2 (200 mg, 0.5 mmol) in
isopropanol were added at room temperature DIC (80 .mu.L, 0.55
mmol) and DMAP (67 mg, 0.55 mmol). After stirring 48 hours at RT,
the crude mixture was concentrated under reduced pressure. Water
was added and the compound was extracted with DCM and the organic
layer dried over Na.sub.2SO.sub.4. Purification of the compound by
Mass Directed Auto Prep afforded the title compound as white foam
(40 mg, 19%).
[0365] .sup.1H RMN (300 MHz, CDCl.sub.3) .delta.7.49 (d, J=8.3 Hz,
2H), 7.39 (d, J=8.9 Hz, 1H), 7.34 (d, J=8.3 Hz, 2H), 7.21 (dd,
J=2.6 and 8.9 Hz, 1H), 6.88 (d, J=2.6 Hz, 1H), 5.08 (m, 1H), 4.59
(m, 1H), 3.81 (s, 3H), 3.70-3.48 (m, 2H), 2.61 (s, 3H), 1.31 (d,
J=6.4 Hz, 3H), 1.28 (d, J=6.4 Hz, 3H). LC/HRMS (M+H).sup.+ calc for
C.sub.23H.sub.23.sup.35ClN.sub.4O.sub.3 439.1537; found 439.1527;
Rt 2.89 min.
Example 37: Butyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate
##STR00082##
[0367] To a solution of Intermediate 2 (200 mg, 0.5 mmol) in
butanol were added at room temperature DIC (80 .mu.L, 0.55 mmol)
and DMAP (67 mg, 0.55 mmol). After stirring 48 hours at room
temperature the crude mixture is concentrated under reduced
pressure. Water was added and the compound was extracted with DCM
and the organic layer dried over Na.sub.2SO.sub.4. Purification of
the compound by Mass Directed Auto Prep afforded the title compound
as white foam (70 mg, 31%).
[0368] .sup.1H RMN (300 MHz, CDCl.sub.3) .delta.7.51 (d, J=8.4 Hz,
2H), 7.39 (d, J=8.9 Hz, 1H), 7.34 (d, J=8.4 Hz, 2H), 7.22 (dd,
J=2.6 and 8.8 Hz, 1H), 6.88 (d, J=2.6 Hz, 1H), 4.60 (dd, J=5.7 and
8.5 Hz, 1H), 4.18 (t, J=6.5 Hz, 2H), 3.82 (s, 3H), 3.66 (dd, J=8.5
and 16.8 Hz, 1H), 3.56 (dd, J=5.7 and 16.8 Hz, 1H), 2.62 (s, 3H),
1.66 (m, 2H), 1.42 (m, 2H), 0.95 (t, J=7.3 Hz, 3H). LC/HRMS
(M+H).sup.+ calc for C.sub.24H.sub.25.sup.35ClN.sub.4O.sub.3
453.1693; found 453.1704; Rt 3.06 min.
Example 38:
2-[(4S)-6-(4-Chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[2-(methylamino)ethyl]acetamide
trifluoroacetic acid salt
##STR00083##
[0370] Intermediate 16 (189 mg, 0.342 mmol) and trifluoroacetic
acid (0.022 mL, 0.285 mmol) were dissolved in DCM (3 mL) and
stirred at 25.degree. C. under N.sub.2 for 2 h. The solvent was
evaporated under vacuum and the residue obtained dissolved in DCM
and re-evaporated under vacuum to give a yellow oil which was dried
under high vacuum to afford the title compound as a yellow solid
(153 mg, 0.270 mmol, 95% yield). LC/MS (M(.sup.35Cl)+H).sup.+ 453,
(M(.sup.37Cl)+H).sup.+ 455; RT 0.74 min.
Example 39:
2-[(4S)-6-(4-Chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-4-piperidinylacetamide trifluoroacetic
acid salt
##STR00084##
[0372] Intermediate 17 (165 mg, 0.285 mmol) and trifluoroacetic
acid (0.022 mL, 0.285 mmol) were dissolved in DCM (3 mL) and
stirred at 25.degree. C. under N.sub.2 for 2 h. The solvent was
evaporated under vacuum and the residue obtained dissolved in DCM
and re-evaporated under vacuum to give a yellow oil which was dried
under high vacuum to afford the title compound as a yellow solid
(161 mg, 0.272 mmol, 95% yield). LC/MS (M(.sup.35Cl)+H).sup.+ 479,
(M(.sup.37Cl)+H).sup.+ 481; RT 0.73 min.
Example 40:
1-{[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-
-a][1,4]benzodiazepin-4-yl]acetyl}-N-methyl-4-piperidinamine formic
acid salt
##STR00085##
[0374] TFA (0.5 mL, 6.49 mmol) was added to a solution of
Intermediate 18 (142 mg, 0.239 mmol) in DCM (2 mL) at 25.degree. C.
under N.sub.2 and the mixture stirred for 2 hours. The solvent was
evaporated under vacuum to give a yellow residue which was
dissolved in MeOH (1 ml) and purified by Mass Directed AutoPrep to
give the title compound as a cream solid (81 mg, 0.150 mmol, 62.8%
yield) LC/MS (M(.sup.35Cl)+H).sup.+ 493, (M(.sup.37Cl)+H).sup.+
495; RT 0.76 min (using LC/MS apparatus c)).
Examples 41 to 43
[0375] Examples 41 to 43 were prepared by a method analogous to
that described for Example 40 starting from the appropriately
protected Intermediate.
TABLE-US-00005 ##STR00086## Mass Spectrum From (M(.sup.35Cl) +
H).sup.+, RT Example Name Intermediate A (M(.sup.37Cl) + H).sup.+
(mins) 41 (S)-1-(4- aminopiperidin-1- yl)-2-(6-(4- chlorophenyl)-8-
methoxy-1-methyl- 4H- 19 ##STR00087## 479, 481 0.75 benzo[f][1,2,4]
triazolo[4,3- a][1,4]diazepin-4- yl)ethanone 42 (S)-N-(azetidin-3-
yl)-2-(6-(4- chlorophenyl)-8- methoxy-1-methyl- 4H- 20 ##STR00088##
451, 453 0.73 benzo[f][1,2,4] triazolo[4,3- a][1,4]diazepin-4-
yl)acetamide 43 (S)-1-(3- aminoazetidin-1- yl)-2-(6-(4-
chlorophenyl)-8- methoxy-1-methyl- 4H- 21 ##STR00089## 451, 453
0.73 benzo[f][1,2,4] triazolo[4,3- a][1,4]diazepin-4-
yl)ethanone
[0376] LC/MS was carried out using apparatus c).
Example 44:
(S)-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f][1,2,4]triazolo[4-
,3-a][1,4]diazepin-4-yl)-1-(4-hydroxypiperidin-1-yl)ethanone
##STR00090##
[0378] Example 44 was prepared by a method analogous to that
described for Intermediate 17 starting from Intermediate 2 and the
appropriate amine.
[0379] LC/HRMS (M(.sup.35Cl)+H).sup.+, (M(.sup.37Cl)+H).sup.+;
found 480, 482. Rt 0.86 min (using LC/MS apparatus c)).
Example 45:
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[2-(4-morpholinyl)ethyl]acetamide
##STR00091##
[0381] Intermediate 2 (104 mg, 0.262 mmol),
[2-(4-morpholinyl)ethyl]amine (0.036 mL, 0.275 mmol), COMU (135 mg,
0.314 mmol) and DIPEA (0.055 mL, 0.314 mmol) were dissolved in DMF
(3 mL) and stirred at 25.degree. C. overnight. The solvent was
evaporated to give an orange oil which was partitioned between DCM
and saturated aqueous sodium bicarbonate solution. The mixture was
separated on a hydrophobic frit and the chlorinated phase
evaporated under vacuum to give an orange oil which was purified by
flash chromatography on silica gel (DCM-MeOH 5-20% gradient) to
give the title compound as a an orange solid (64 mg, 0.126 mmol,
48.0% yield). LC/MS (M(.sup.35Cl)+H).sup.+ 509,
(M(.sup.37Cl)+H).sup.+ 511; RT 0.77 min (using LC/MS apparatus
c)).
Example 46:
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[2-(1H-imidazol-4-yl)ethyl]acetamide
##STR00092##
[0383] Intermediate 2 (104 mg, 0.262 mmol), histamine (30.6 mg,
0.275 mmol), COMU (135 mg, 0.314 mmol) and DIPEA (0.055 mL, 0.314
mmol) were dissolved in DMF (3 mL) and stirred at 25.degree. C.
overnight. More histamine (30.6 mg, 0.275 mmol) and COMU (135 mg,
0.314 mmol) and DIPEA (0.055 mL, 0.314 mmol) were added and the
reaction stirred for a further 6 hours then allowed to sit at RT
over 56 hours. The solvent was evaporated to give an orange oil
which was partitioned between DCM and saturated aqueous sodium
bicarbonate solution. The mixture was separated on a hydrophobic
frit and the chlorinated phase evaporated under vacuum to give a
yellow oil which was purified by flash chromatography on silica gel
(DCM-MeOH 5-20% gradient) to give the title compound as a white
solid (66 mg, 0.135 mmol, 51.4% yield). LC/MS
(M(.sup.35Cl)+H).sup.+ 490; RT 0.74 min (using LC/MS apparatus
c)).
Example 47:
(S)-tert-butyl-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f][1,2,4-
]triazolo[4,3-a][1,4]diazepin-4-yl)acetate
##STR00093##
[0385] Oxalyl chloride (0.114 mL, 1.298 mmol) was added to a cold
(0.degree. C.) solution of Intermediate 2 (103 mg, 0.260 mmol) in
DCM (2 mL) under N.sub.2. DMF (0.020 mL, 0.260 mmol) was then added
and the reaction allowed to warm to RT over 1 h. The crude acid
chloride mixture was poured into a solution of tert-butanol (0.124
mL, 1.298 mmol) in DCM (2 mL) and Hunig's base (0.045 mL, 0.260
mmol) was added and stirred for 10 mins. Then more tert-butanol (1
mL) was added and the solution stirred for 1 h. The reaction
mixture was evaporated and re-dissolved in tert-butanol (1 mL)
twice more, then the solvent was evaporated a final time to give a
yellow solid. Purification of the solid by flash chromatography on
silica gel (cyclohexane-ethyl acetate 10 to 50% gradient) gave the
title compound as a colourless oil (25 mg, 0.055 mmol, 21.27%
yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.50 (d, J=9 Hz,
2H), 7.39 (d, J=9 Hz, 1H), 7.20 (dd, J=9, 3 Hz, 1H) 7.33 (s, 2H),
6.89 (d, J=3 Hz, 1H), 4.56 (dd, J=8, 6 Hz, 1H), 3.81 (s, 3H),
3.24-3.71 (m, 2H), 2.62 (s, 3H), 1.50 (s, 9H). LC/MS
(M(.sup.35Cl)+H).sup.+ 453, (M(.sup.37Cl)+H).sup.+ 455; RT 1.19 min
(using LC/MS apparatus c)).
Example 48:
(S)-piperidin-4-yl-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f][1-
,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)acetate
##STR00094##
[0387] Intermediate 2 (103 mg, 0.260 mmol), 1,1-dimethylethyl
4-hydroxy-1-piperidinecarboxylate (0.101 mL, 0.389 mmol), EDC (74.6
mg, 0.389 mmol) and DMAP (6.34 mg, 0.052 mmol) were dissolved in
DCM (2 mL) at RT under N.sub.2 and the reaction mixture allowed to
stir overnight. The reaction mixture was quenched with saturated
aqueous sodium bicarbonate solution and the mixture was filtered
through a hydrophobic frit, and the chlorinated phase evaporated
under vacuum to give a yellow residue. Purification of the solid by
flash chromatography on silica gel (DCM-MeOH 5-20% gradient) gave
the title compound as a colourless oil which was used immediately
in the next reaction.
[0388] The colourless oil (157 mg, 0.271 mmol) and trifluoroacetic
acid (0.5 mL, 6.49 mmol) were dissolved in DCM (2 mL) at 0.degree.
C. under N.sub.2 and the mixture allowed to warm to RT over 1 h.
The solvent was evaporated under vacuum and the residue obtained
dissolved in DCM and re-evaporated under vacuum to give a yellow
oil which was dried under high vacuum to afford the title compound
as a yellow solid (50 mg, 0.095 mmol, 35% yield). .sup.1H NMR (400
MHz, MeOH-d.sub.4) .delta. 8.49 (s, 1H), 7.66 (d, J=9 Hz, 1H), 7.47
(d, J=9 Hz, 2H), 7.23-7.41 (m, 3H), 6.85 (s, 1H), 5.04 (br. s.,
1H), 4.57 (t, J=7 Hz, 1H), 3.77 (s, 3H), 3.51 (d, J=7 Hz, 2H), 3.28
(br. s., 2H), 3.17 (br. s., 2H), 2.59 (s, 3H), 2.07 (br. s., 2H),
1.93 (br. s., 2H). LC/MS (M(.sup.35Cl)+H).sup.+ 480; RT 0.78 min
(using LC/MS apparatus c)).
Example 49:
2-[(4S)-6-(4-chlorophenyl)-8-hydroxy-1-methyl-4H-[1,2,4]triazolo[4,3-a][1-
,4]benzodiazepin-4-yl]-N-ethylacetamide
##STR00095##
[0390] Intermediate 22 (383 mg, 1.00 mmol), ethylamine
hydrochloride (163 mg, 2.0 mmol), HATU (457 mg, 1.20 mmol) and
DIPEA (0.349 mL, 2.0 mmol) were dissolved in DMF (2 mL) and stirred
at RT overnight. More ethylamine hydrochloride (163 mg, 2.0 mmol)
and DIPEA (0.349 mL, 2.0 mmol) were added and the reaction stirred
for a further 2 h. The reaction mixture was diluted with EtOAc (20
mL) and washed with saturated aqueous sodium bicarbonate solution
(10 mL), water and brine. The organic phase was dried and
evaporated under vacuum. Purification of the residue by flash
chromatography on silica gel (DCM-methanol 10 to 20% gradient) gave
the title compound as an off-white solid (200 mg, 0.488 mmol, 49%
yield). LC/MS (M(.sup.35Cl)+H).sup.+ 410, (M(.sup.37Cl)+H).sup.+
412; RT 0.76 min (using LC/MS apparatus c)).
Example 50:
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[2-(dimethylamino)ethyl]acetamide
formic acid salt
##STR00096##
[0392] Intermediate 2 (105 mg, 0.265 mmol),
N,N-dimethyl-1,2-ethanediamine (0.071 mL, 0.278 mmol), COMU (136
mg, 0.318 mmol) and DIPEA (0.116 mL, 0.661 mmol) were dissolved in
DMF (3 mL) and stirred at 25.degree. C. On addition of amine turned
orange and allowed to stir at RT overnight. The solvent was
evaporated to give a yellow oil which was partitioned between DCM
and saturated aqueous sodium bicarbonate solution. The mixture was
separated on a hydrophobic frit and the chlorinated phase
evaporated under vacuum to give a yellow oil which was dissolved in
MeOH (1 ml) and purified by Mass Directed AutoPrep to give the
title compound as a white solid (48 mg, 0.094 mmol, 35.4% yield).
.sup.1H NMR (400 MHz, Methanol-d.sub.4) .delta. 8.40 (br. s., 1H),
7.70 (d, J=9 Hz, 1H), 7.52 (d, J=9 Hz, 2H), 7.26-7.45 (m, 3H), 6.87
(s, 1H), 4.62 (t, J=7 Hz, 1H), 3.65 (s, 3H), 3.65-3.92 (m, 3H),
3.32-3.55 (m, 3H), 3.24 (br. s., 1H), 2.90 (s, 6H), 2.62 (s, 3H).
LC/MS (M(.sup.35Cl)+H).sup.+ 467, (M(.sup.37Cl)+H).sup.+ 469; RT
0.76 min (using LC/MS apparatus c)).
Example 51:
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[3-(dimethylamino)propyl]acetamide
formic acid salt
##STR00097##
[0394] Intermediate 2 (100 mg, 0.252 mmol),
N,N-dimethyl-1,3-propanediamine (0.033 mL, 0.265 mmol), HATU (115
mg, 0.302 mmol) and DIPEA (0.106 mL, 0.605 mmol) were dissolved in
DMF (3 mL) and stirred at 25.degree. C. overnight. The solvent was
evaporated to give a yellow oil which was partitioned between DCM
and saturated aqueous sodium bicarbonate solution. The mixture was
separated on a hydrophobic frit and the chlorinated phase
evaporated under vacuum to give a yellow oil which was dissolved in
MeOH (1 ml) and purified by Mass Directed AutoPrep to give the
title compound as a white solid (95 mg, 0.180 mmol, 71.5% yield).
.sup.1H NMR (400 MHz, Methanol-d.sub.4) .delta. 8.41 (br. s., 1H),
7.74 (d, J=9 Hz, 1H), 7.56 (d, J=9 Hz, 2H), 7.37-7.46 (m, 3H), 6.91
(d, J=3 Hz, 1H), 4.66 (t, J=7 Hz, 1H), 3.83 (s, 3H), 3.35-3.57 (m,
3H), 3.28-3.18 (m, 4H), 2.87 (s, 6H), 2.66 (s, 3H), 1.89-2.09 (m,
2H). LC/MS (M(.sup.35Cl)+H).sup.+ 481, (M(.sup.37Cl)+H).sup.+ 483;
RT 0.79 min (using LC/MS apparatus c)).
Example 52:
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[3-(4-morpholinyl)propyl]acetamide
##STR00098##
[0396] Intermediate 2 (150 mg, 0.378 mmol),
[3-(4-morpholinyl)propyl]amine (57.2 mg, 0.397 mmol), HATU (172 mg,
0.454 mmol) and DIPEA (0.158 mL, 0.907 mmol) were dissolved in DMF
(3 mL) and stirred at 25.degree. C. overnight. The solvent was
evaporated to give a yellow oil which was partitioned between DCM
and saturated aqueous sodium bicarbonate solution. The mixture was
separated on a hydrophobic frit and the chlorinated phase
evaporated under vacuum to give a yellow oil. Purification by flash
chromatography on silica gel (DCM-methanol 5 to 20% gradient) gave
the title compound as a pale cream solid (77 mg, 0.147 mmol, 38.9%
yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.54 (t, J=5 Hz,
1H), 7.42 (d, J=9 Hz, 2H), 7.19-7.37 (m, 3H), 7.13 (dd, J=9, 3 Hz,
1H), 6.78 (d, J=3 Hz, 1H), 4.58 (t, J=7 Hz, 1H), 3.73 (s, 3H), 3.66
(t, J=5 Hz, 3H), 3.36-3.48 (m, 2H), 3.22-3.36 (m, 3H), 2.46-2.76
(m, 4H), 2.17-2.46 (m, 5H), 1.56-1.85 (m, 2H). LC/MS
(M(.sup.35Cl)+H).sup.+ 523, (M(.sup.37Cl)+H).sup.+ 525; RT 0.74 min
(using LC/MS apparatus c)).
Example 53:
2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3--
a][1,4]benzodiazepin-4-yl]-N-[2-(1,1-dioxido-4-thiomorpholinyl)ethyl]aceta-
mide
##STR00099##
[0398] Intermediate 2 (150 mg, 0.378 mmol),
[2-(1,1-dioxido-4-thiomorpholinyl)ethyl]amine (70.7 mg, 0.397
mmol), HATU (172 mg, 0.454 mmol) and DIPEA (0.158 mL, 0.907 mmol)
were dissolved in DMF (3 mL) and stirred at 25.degree. C.
overnight. The solvent was evaporated to give a yellow oil which
was partitioned between DCM and saturated aqueous sodium
bicarbonate solution. The mixture was separated on a hydrophobic
frit and the chlorinated phase evaporated under vacuum to give an
orange oil. Purification by flash chromatography on silica gel
(DCM-methanol 5 to 20% gradient) gave the title compound as a pale
cream solid (84 mg, 0.151 mmol, 39.9% yield). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.41-7.70 (m, 4H), 7.30-7.41 (m, 2H), 7.25 (dd,
J=9, 3 Hz, 1H), 6.87 (d, J=3 Hz, 1H), 4.64 (dd, J=8, 6 Hz, 1H),
3.82 (s, 3H), 3.65-3.78 (m, 1H), 3.56 (dd, J=15, 8 Hz, 1H),
3.26-3.50 (m, 2H), 3.19 (q, J=7 Hz, 1H), 2.86-3.11 (m, 5H), 2.63
(s, 3H), 1.23-1.53 (m, 4H). LC/MS (M(.sup.35Cl)+H).sup.+ 557,
(M(.sup.37Cl)+H).sup.+ 559; RT 0.84 min (using LC/MS apparatus
c)).
Examples 54 to 57
[0399] Examples 54 to 57 were prepared from Intermediate 2 and the
appropriate amine using the procedure described in Example 1.
TABLE-US-00006 ##STR00100## Example R.sup.3 Config. A Physical Data
54 Cl (S) ##STR00101## RT 2.85 min (M(.sup.35Cl) + H).sup.+ 474,
(M(.sup.37Cl) + H).sup.+ 476 55 Cl (S) ##STR00102## RT 3.02 min
(M(.sup.35Cl) + H).sup.+ 477, (M(.sup.37Cl) + H).sup.+ 479 56 Cl
(S) ##STR00103## RT 2.65 min (M(.sup.35Cl) + H).sup.+ 480,
(M(.sup.37Cl) + H).sup.+ 482 57 Cl (R) ##STR00104## RT 4.23 min
(M(.sup.35Cl) + H).sup.+ 501, (M(.sup.37Cl) + H).sup.+ 503
[0400] LC/MS was carried out using apparatus c).
[0401] Further Example compounds of the invention include:
TABLE-US-00007 Ex. Structure 58 ##STR00105## 59 ##STR00106## 60
##STR00107## 61 ##STR00108## 62 ##STR00109## 63 ##STR00110## 64
##STR00111## 65 ##STR00112## 66 ##STR00113## 67 ##STR00114## 68
##STR00115## 69 ##STR00116## 70 ##STR00117## 71 ##STR00118## 72
##STR00119## 73 ##STR00120## 74 ##STR00121## 75 ##STR00122## 77
##STR00123## 77 ##STR00124## 78 ##STR00125## 79 ##STR00126## 80
##STR00127## 81 ##STR00128## 82 ##STR00129##
[0402] It will be understood that in the context of the examples
used to illustrate the invention that information about how the
compounds were prepared cannot be drawn from the format used to
present the information, for example, the intermediates and final
products may have been prepared by different individuals, and/or at
different timepoints, employing various batches and appropriate
techniques.
[0403] Experimental details of LC/MS methods D and F as referred to
herein are as follows:
[0404] LC/MS (Method D) was conducted on a Supelcosil LCABZ+PLUS
column (3 .mu.m, 3.3 cm.times.4.6 mm ID) eluting with 0.1%
HCO.sub.2H and 0.01 M ammonium acetate in water (solvent A), and
95% acetonitrile and 0.05% HCO.sub.2H in water (solvent B), using
the following elution gradient 0-0.7 minutes 0% B, 0.7-4.2 minutes
0.fwdarw.100% B, 4.2-5.3 minutes 100% B, 5.3-5.5 minutes
100.fwdarw.0% B at a flow rate of 3 mL/minute. The mass spectra
(MS) were recorded on a Fisons VG Platform mass spectrometer using
electrospray positive ionisation [(ES+ve to give [M+H].sup.+ and
[M+NH.sub.4].sup.+ molecular ions] or electrospray negative
ionisation [(ES-ve to give [M-H]- molecular ion] modes. Analytical
data from this apparatus are given with the following format:
[M+H].sup.+ or [M-H].sup.-.
[0405] LC/MS (Method F) was conducted on an Sunfire C18 column (30
mm.times.4.6 mm i.d. 3.5 .mu.m packing diameter) at 30 degrees
centigrade, eluting with 0.1% v/v solution of Trifluoroacetic Acid
in Water (Solvent A) and 0.1% v/v solution of Trifluoroacetic Acid
in Acetonitrile (Solvent B) using the following elution gradient
0-0.1 min 3% B, 0.1-4.2 min 3-100% B, 4.2-4.8 min 100% B, 4.8-4.9
min 100-3% B, 4.9-5.0 min 3% B at a flow rate of 3 ml/min. The UV
detection was an averaged signal from wavelength of 210 nm to 350
nm and mass spectra were recorded on a mass spectrometer using
positive electrospray ionization. Ionisation data was rounded to
the nearest integer.
[0406] LC/HRMS: Analytical HPLC was conducted on a Uptisphere-hsc
column (3 .mu.m 33.times.3 mm id) eluting with 0.01M ammonium
acetate in water (solvent A) and 100% acetonitrile (solvent B),
using the following elution gradient 0-0.5 minutes 5% B, 0.5-3.75
minutes 5.fwdarw.100% B, 3.75-4.5 100% B, 4.5-5 100.fwdarw.5% B,
5-5.5 5% B at a flow rate of 1.3 mL/minute. The mass spectra (MS)
were recorded on a micromass LCT mass spectrometer using
electrospray positive ionisation [ES+ve to give MH.sup.+ molecular
ions] or electrospray negative ionisation [ES-ve to give (M-H)-
molecular ions] modes.
[0407] TLC (thin layer chromatography) refers to the use of TLC
plates sold by Merck coated with silica gel 60 F254.
[0408] Silica chromatography techniques include either automated
(Flashmaster or Biotage SP4) techniques or manual chromatography on
pre-packed cartridges (SPE) or manually-packed flash columns.
[0409] When the name of a commercial supplier is given after the
name of a compound or a reagent, for instance "compound X
(Aldrich)" or "compound X/Aldrich", this means that compound X is
obtainable from a commercial supplier, such as the commercial
supplier named.
Reference compound A:
2-methyl-6-(methyloxy)-4H-3,1-benzoxazin-4-one
##STR00130##
[0411] A solution of 5-methoxyanthranilic acid (Lancaster) (41.8 g,
0.25 mol) was refluxed in acetic anhydride (230 mL) for 3.5 h
before being concentrated under reduced pressure. The crude
compound was then concentrated twice in the presence of toluene
before being filtered and washed twice with ether to yield to the
title compound (33.7 g, 71% yield) as a brown solid; LC/MS (Method
D): m/z 192 [M+H].sup.+, Rt 1.69 min.
Reference compound B:
[2-amino-5-(methyloxy)phenyl](4-chlorophenyl)methanone
##STR00131##
[0413] To a solution of
2-methyl-6-(methyloxy)-4H-3,1-benzoxazin-4-one (for a preparation
see Reference compound A) (40.0 g, 0.21 mol) in a toluene/ether
(2/1) mixture (760 mL) at 0.degree. C. was added dropwise a
solution of 4-chlorophenylmagnesium bromide (170 mL, 1M in
Et.sub.2O, 0.17 mol). The reaction mixture was allowed to warm to
room temperature and stirred for 1 h before being quenched with 1N
HCl (200 mL). The aqueous layer was extracted with EtOAc
(3.times.150 mL) and the combined organics were washed with brine
(100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The crude compound was then dissolved in
EtOH (400 mL) and 6N HCl (160 mL) was added. The reaction mixture
was refluxed for 2 h before being concentrated to one-third in
volume. The resulting solid was filtered and washed twice with
ether before being suspended in EtOAc and neutralised with 1N NaOH.
The aqueous layer was extracted with EtOAc (3.times.150 mL) and the
combined organics were washed with brine (150 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The title compound was obtained as a yellow solid (39 g, 88%
yield); LC/MS (Method D): m/z 262 [M+H]+, Rt 2.57 min.
Reference Compound C: Methyl
N'-[2-[(4-chlorophenyl)carbonyl]-4-(methyloxy)phenyl]-N.sup.2-{[(9H-fluor-
en-9-ylmethyl)oxy]carbonyl}-L-.alpha.-asparaginate
##STR00132##
[0415] Methyl
N-{[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-.alpha.-aspartyl
chloride (Int. J. Peptide Protein Res. 1992, 40, 13-18) (93 g, 0.24
mol) was dissolved in CHCl.sub.3 (270 mL) and
[2-amino-5-(methyloxy)phenyl](4-chlorophenyl)methanone (for a
preparation see Reference compound B) (53 g, 0.2 mol) was added.
The resulting mixture was stirred at 60.degree. C. for 1 h before
being cooled and concentrated at 60% in volume. Ether was added at
0.degree. C. and the resulting precipitate was filtered and
discarded. The filtrate was concentrated under reduced pressure and
used without further purification.
Reference compound D: Methyl
[(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-oxo-2,3-dihydro-1H-1,4-benzodiaz-
epin-3-yl]acetate
##STR00133##
[0417] To a solution of Methyl
N1-[2-[(4-chlorophenyl)carbonyl]-4-(methyloxy)phenyl]-N2-{[(9H-fluoren-9--
ylmethyl)oxy]carbonyl}-L-.alpha.-asparaginate (for a preparation
see Reference compound C) (assumed 0.2 mol) in DCM (500 mL) was
added Et.sub.3N (500 mL, 3.65 mol) and the resulting mixture was
refluxed for 24 h before being concentrated. The resulting crude
amine was dissolved in 1,2-DCE (1.5 L) and AcOH (104 mL, 1.8 mol)
was added carefully. The reaction mixture was then stirred at
60.degree. C. for 2 h before being concentrated in vacuo and
dissolved in DCM. The organic layer was washed with 1N HCl and the
aqueous layer was extracted with DCM (.times.3). The combined
organic layers were washed twice with water, and brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The crude solid was recrystallised in MeCN leading to the title
compound (51 g) as a pale yellow solid. The filtrate could be
concentrated and recrystallised in MeCN to give to another 10 g of
the desired product R.sub.f=0.34 (DCM/MeOH:95/5). HRMS (M+H).sup.+
calculated for C.sub.19H.sub.18.sup.35ClN.sub.2O.sub.4 373.0955;
found 373.0957.
Reference compound E: Methyl
[(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-thioxo-2,3-dihydro-1H-1,4-benzod-
iazepin-3-yl]acetate
##STR00134##
[0419] A suspension of P.sub.4S.sub.10 (36.1 g, 81.1 mmol) and
Na.sub.2CO.sub.3 (8.6 g, 81.1 mmol) in 1,2-DCE (700 mL) at room
temperature was stirred for 2 h before Methyl
[(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-oxo-2,3-dihydro-1H-1,4-benzodiaz-
epin-3-yl]acetate (for a preparation see Reference compound D)
(16.8 g, 45.1 mmol) was added. The resulting mixture was stirred at
70.degree. C. for 2 h before being cooled and filtered. The solid
was washed twice with DCM and the filtrate washed with sat.
NaHCO.sub.3 and brine. The organic layer was dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The crude product was purified by flash-chromatography on silica
gel (DCM/MeOH:99/1) to afford the title compound (17.2 g, 98%
yield) as a yellowish solid. LC/MS (Method D): m/z 389
[M(.sup.35Cl)+H].sup.+, Rt 2.64 min
[0420] HRMS (M+H).sup.+ calculated for
C.sub.19H.sub.18.sup.35ClN.sub.2O.sub.3S 389.0727; found
389.0714.
Reference compound F: Methyl
[(3S)-2-[(1Z)-2-acetylhydrazino]-5-(4-chlorophenyl)-7-(methyloxy)-3H-1,4--
benzodiazepin-3-yl]acetate
##STR00135##
[0422] To a suspension of Methyl
[(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-thioxo-2,3-dihydro-1H-1,4-benzod-
iazepin-3-yl]acetate (for a preparation see Reference compound E
(9.0 g, 23.2 mmol) in THF (300 mL) at 0.degree. C. was added
hydrazine monohydrate (3.4 mL, 69.6 mmol) dropwise. The reaction
mixture was stirred for 5 h between 5.degree. C. and 15.degree. C.
before being cooled at 0.degree. C. Et.sub.3N (9.7 mL, 69.6 mmol)
was then added slowly and acetyl chloride (7.95 mL, 69.6 mmol) was
added dropwise. The mixture was then allowed to warm to room
temperature for 16 h before being concentrated under reduced
pressure. The crude product was dissolved in DCM and washed with
water. The organic layer was dried over Na.sub.2SO.sub.4, filtered
and concentrated in vacuo to give the crude title compound (9.7 g,
98% yield) which was used without further purification.
R.sub.f=0.49 (DCM/MeOH:90/10).
Reference compound G: Methyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate
##STR00136##
[0424] The crude Methyl [(3S)-2-[(1
Z)-2-acetylhydrazino]-5-(4-chlorophenyl)-7-(methyloxy)-3H-1,4-benzodiazep-
in-3-yl]acetate (for a preparation see Reference compound F)
(assumed 9.7 g) was suspended in THF (100 ml) and AcOH (60 mL) was
added at room temperature. The reaction mixture was stirred at this
temperature for 2 days before being concentrated under reduced
pressure. The crude solid was triturated in i-Pr.sub.2O and
filtered to give the title compound (8.7 g, 91% over 3 steps) as an
off-white solid.
[0425] HRMS (M+H).sup.+ calculated for
C.sub.21H.sub.20ClN.sub.4O.sub.3 411.1229; found 411.1245.
Reference compound H:
[(4S)-6-(4-Chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetic acid
##STR00137##
[0427] To a solution of Methyl
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetate (for a preparation see Reference
compound G)(7.4 g, 18.1 mmol) in THF (130 mL) at room temperature
was added 1N NaOH (36.2 mL, 36.2 mmol). The reaction mixture was
stirred at this temperature for 5 h before being quenched with 1N
HCl (36.2 mL) and concentrated in vacuo. Water is then added and
the aqueous layer was extracted with DCM (.times.3) and the
combined organic layers were dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give the title compound
(7 g, 98% yield) as a pale yellow solid.
Reference compound I: 1,1-dimethylethyl
[5-({[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4-
,3-a][1,4]benzodiazepin-4-yl]acetyl}amino)pentyl]carbamate
##STR00138##
[0429] A mixture of
[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a]-
[1,4]benzodiazepin-4-yl]acetic acid (for a preparation see
Reference compound H) (1.0 g, 2.5 mmol), HATU (1.9 g, 5 mmol) and
DIPEA (0.88 ml, 5 mmol) was stirred for 80 minutes at room
temperature, to this was added 1,1-dimethylethyl
(4-aminobutyl)carbamate (1.05 ml, 5.0 mmol, available from
Aldrich). The reaction mixture was stirred at room temperature for
2 h before it was concentrated. The residue was taken up in
dichloromethane and washed with 1N HCl. The aqueous layer was
extracted with dichloromethane twice. Organic layer was washed with
1N sodium hydroxide, followed by a saturated solution of sodium
chloride, dried over sodium sulphate and concentrated. The residue
was purified by flash-chromatography on silica using
dichloromethane/methanol 95/5 to give the title compound as a
yellow solid (1.2 g). LC/MS (Method D): rt=3.04 min.
Reference compound J:
N-(5-aminopentyl)-2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1-
,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamide
trifluoroacetate
##STR00139##
[0431] To a solution of 1,1-dimethylethyl
[5-({[(4S)-6-(4-chlorophenyl)-1-methyl-8-methyloxy)-4H-[1,2,4]triazolo[4,-
3-a][1,4]benzodiazepin-4-yl]acetyl}amino)pentyl]carbamate (for a
preparation see Reference compound H) (0.2 g, 0.34 mmol) in
dichloromethane (3 ml) was added trifluoroacetic acid (0.053 ml,
0.68 mmol) dropwise at 00.degree. C. The reaction mixture was
stirred for 3 h from 00.degree. C. to room temperature. The
reaction mixture was concentrated to dryness to afford the title
compound as a hygroscopic yellow oil (200 mg) LC/MS (Method D):
rt=2.33 min.
[0432] HRMS (M+H).sup.+ calculated for
C.sub.25H.sub.29ClN.sub.6O.sub.2 481.2119; found 481.2162.
Reference compound K: Mixture of 5- and 6-isomers of Alexa Fluor
488-N-(5-aminopentyl)-2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4-
H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamide
##STR00140##
[0434]
N-(5-aminopentyl)-2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-
-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamide
trifluoroacetate (for a preparation see Reference compound J)(7.65
mg, 0.013 mmol) was dissolved in N,N-Dimethylformamide (DMF) (300
.mu.l) and added to Alexa Fluor 488 carboxylic acid succinimidyl
ester (5 mg, 7.77 .mu.mol, mixture of 5 and 6 isomers, available
from Invitrogen, product number A-20100) in an Eppendorf centrifuge
tube. Hunig's base (7.0 .mu.l, 0.040 mmol) was added and the
mixture vortex mixed overnight. After 18 h the reaction mixture was
evaporated to dryness and the residue redissolved in DMSO/water
(50%, <1 ml total), applied to a preparative Phenomenex Jupiter
C18 column and eluted with a gradient of 95% A: 5% B to 100% B
(A=0.1% trifluoroacetic acid in water, B=0.1% TFA/90%
acetonitrile/10% water) at a flow rate of 10 ml/min over 150
minutes. Impure fractions were combined and re-purified using the
same system. Fractions were combined and evaporated to yield the
title product (2.8 mg) as a mixture of the 2 regioisomers shown.
LC/MS (Method F): MH+=999, rt=1.88 min.
Biological Test Methods
Fluorescence Anisotropy Binding Assay
[0435] The binding of the compounds of formula (I) to Bromodomain
2, 3 and 4 was assessed using a Fluorescence Anisotropy Binding
Assay.
[0436] The Bromodomain protein, fluorescent ligand (Reference
compound K see above) and a variable concentration of test compound
are incubated together to reach thermodynamic equilibrium under
conditions such that in the absence of test compound the
fluorescent ligand is significantly (>50%) bound and in the
presence of a sufficient concentration of a potent inhibitor the
anisotropy of the unbound fluorescent ligand is measurably
different from the bound value.
[0437] All data was normalized to the mean of 16 high and 16 low
control wells on each plate. A four parameter curve fit of the
following form was then applied:
y=a+((b-a)/(1+(10 x/10 c) d)
[0438] Where `a` is the minimum, `b` is the Hill slope, `c` is the
pIC50 and `d` is the maximum.
[0439] Recombinant Human Bromodomains (Bromodomain 2 (1-473),
Bromodomain 3 (1-435) and Bromodomain 4 (1-477)) were expressed in
E. coli cells (in pET15b vector) with a six-His tag at the
N-terminal. The His-tagged Bromodomain was extracted from E. coli
cells using 0.1 mg/ml lysozyme and sonication. The Bromodomain was
then purified by affinity chromatography on a HisTRAP HP column,
eluting with a linear 10-500 mM Imidazole gradient, over 20 Cv.
Further purification was completed by Superdex 200 prep grade size
exclusion column. Purified protein was stored at -80.degree. C. in
20 mM HEPES pH 7.5 and 100 mM NaCl.
[0440] Protocol for Bromodomain 2: All components were dissolved in
buffer composition of 50 mM HEPES pH7.4, 150 mm NaCl and 0.5 mM
CHAPS with final concentrations of Bromodomain 2, 75 nM,
fluorescent ligand 5 nM. 10 .mu.l of this reaction mixture was
added using a micro multidrop to wells containing 100 nl of various
concentrations of test compound or DMSO vehicle (1% final) in
Greiner 384 well Black low volume microtitre plate and equilibrated
in the dark for 60 mins at room temperature. Fluorescence
anisotropy was read in Envision (.lamda.ex=485 nm, .lamda.EM=530
nm; Dichroic -505 nM).
[0441] Protocol for Bromodomain 3: All components were dissolved in
buffer of composition 50 mM HEPES pH7.4, 150 mm NaCl and 0.5 mM
CHAPS with final concentrations of Bromodomain 3, 75 nM,
fluorescent ligand 5 nM. 10 .mu.l of this reaction mixture was
added using a micro multidrop to wells containing 100 nl of various
concentrations of test compound or DMSO vehicle (1% final) in
Greiner 384 well Black low volume microtitre plate and equilibrated
in the dark for 60 mins at room temperature. Fluorescence
anisotropy was read in Envision (.lamda.ex=485 nm, .lamda.EM=530
nm; Dichroic -505 nM).
[0442] Protocol for Bromodomain 4: All components were dissolved in
buffer of composition 50 mM HEPES pH7.4, 150 mm NaCl and 0.5 mM
CHAPS with final concentrations of Bromodomain 4, 75 nM,
fluorescent ligand 5 nM. 10 .mu.l of this reaction mixture was
added using a micro multidrop to wells containing 100 nl of various
concentrations of test compound or DMSO vehicle (1% final) in
Greiner 384 well Black low volume microtitre plate and equilibrated
in the dark for 60 mins at room temperature. Fluorescence
anisotropy was read in Envision (.lamda.ex=485 nm, .lamda.EM=530
nm; Dichroic -505 nM).
[0443] All examples (with the exception of examples 5-8, 18, 19,
30, 32, 33, 35, 49, 52, 53, 70-80 and 82) were tested in the assays
described above. All tested compounds with the exception of Example
81 had a pIC50.gtoreq.5.0 in one or more of the BRD2, BRD3 and
BRD4. Examples 1, 2, 4, 9-17, 22-24, 26, 34, 36-48, 50, 51, 54-56
and 58-65 had a pIC50.gtoreq.6.0 in one or more of the BRD2, BRD3
and BRD4 assays described above.
LPS Stimulated Whole Blood Measuring TNF.alpha. Levels Assay
[0444] Activation of monocytic cells by agonists of toll-like
receptors such as bacterial lipopolysaccharide (LPS) results in
production of key inflammatory mediators including TNF.alpha.. Such
pathways are widely considered to be central to the pathophysiology
of a range of auto-immune and inflammatory disorders.
[0445] Compounds to be tested are diluted to give a range of
appropriate concentrations and 1 ul of the dilution stocks is added
to wells of a 96 plate. Following addition of whole blood (130 ul)
the plates are incubated at 37 degrees (5% CO2) for 30 min before
the addition of 10 ul of 2.8 ug/ml LPS, diluted in complete RPMI
1640 (final concentration=200 ng/ml), to give a total volume of 140
ul per well. After further incubation for 24 hours at 37 degrees,
140 ul of PBS are added to each well. The plates are sealed, shaken
for 10 minutes and then centrifuged (2500 rpm.times.10 min). 100 ul
of the supernatant are removed and TNF.alpha. levels assayed by
immunoassay (typically by MesoScale Discovery technology) either
immediately or following storage at -20 degrees. Dose response
curves for each compound was generated from the data and an IC50
value was calculated.
[0446] Examples 1, 12, 14, 34, 36, 37 and 81 were tested in the
above assay and were found to have a pIC50.gtoreq.5.0. Examples 1,
12, 14, 34, 36 and 37 were tested in the above assay and were found
to have a pIC50.gtoreq.6.0.
Measurement of LPS Induced IL-6 Secretion from Whole Blood
[0447] Activation of monocytic cells by agonists of toll-like
receptors such as bacterial lipopolysaccharide (LPS) results in
production of key inflammatory mediators including IL-6. Such
pathways are widely considered to be central to the pathophysiology
of a range of auto-immune and inflammatory disorders.
[0448] Compounds to be tested are diluted to give a range of
appropriate concentrations of which 1 ul of the diluted stocks is
added to a 96 well plate. Following addition of whole blood (130
ul) the plates are incubated at 37 degrees (5% CO2) for 30 min
before the addition of 10 ul of 2.8 ug/ml LPS, diluted in complete
RPMI 1640 (final concentration=200 ng/ml), to give a total volume
of 140 ul per well. After further incubation for 24 hours at 37
degrees, 140 ul of PBS are added to each well. The plates are
sealed, shaken for 10 minutes and then centrifuged (2500
rpm.times.10 min). 100 ul of the supernatant are removed and IL-6
levels assayed by immunoassay (typically by MesoScale Discovery
technology) either immediately or following storage at -20 degrees.
Concentration response curves for each compound was generated from
the data and an IC50 value was calculated.
[0449] Examples 1-4, 9-11, 13, 15, 17, 26, 31, 34, 38-48, 50, 51,
54, 55, 56, 58-64, 67, 69 and 81 were tested in the above assay and
all, with the exception of Example 81, were found to have a
pIC50.gtoreq.5.0.
[0450] These data demonstrate that bromodomain inhibitors tested in
the above two whole blood assays inhibited the production of the
key inflammatory mediators TNF.alpha. and/or IL6. This suggests
that such compounds have an anti-inflammatory profile.
In Vivo Mouse Endotoxemia Model
[0451] High doses of Endotoxin (bacterial lipopolysaccharide)
administered to animals produce a profound shock syndrome including
a strong inflammatory response, dysregulation of cardiovascular
function, organ failure and ultimately mortality. This pattern of
response is very similar to human sepsis and septic shock, where
the body's response to a significant bacterial infection can be
similarly life threatening.
[0452] To test the compounds of formula (I) and pharmaceutically
acceptable salts thereof groups of eight Balb/c male mice were
given a lethal dose of 15 mg/kg LPS by intraperitoneal injection.
Ninety minutes later, animals were dosed intravenously with vehicle
(20% cyclodextrin 1% ethanol in apyrogen water) or compound (10
mg/kg). The survival of animals was monitored at 4 days.
[0453] Numbers of animals surviving at 4 days (summed across
multiple repeat experiments)
TABLE-US-00008 Vehicle 4/66 (6%) Compound of Example 1 24/56
(52%)
[0454] These data demonstrate that the bromodomain inhibitor tested
in the above model gave rise to a significant animal survival
effect following intravenous administration.
Cell Growth Assay
[0455] Human cell lines (n=33 comprising 15 heme cell lines, 14
breast cell lines and 4 other cell lines) were cultured in
RPMI-1640 containing 10% fetal bovine serum, 1000 viable cells per
well were plated in 384-well black flat bottom polystyrene plates
(Greiner #781086) in 48 .mu.l of culture media. All plates were
placed at 5% CO.sub.2, 37.degree. C. overnight. The following day
one plate was harvested with CellTiter-Glo (CTG, Promega #G7573)
for a time equal to 0 (T0) measurement and compound (20 point
titration from 14.7 uM to 7 pM) was added to the remaining plates.
The final concentration of DMSO in all wells was 0.15%. Cells were
incubated for 72 hours or the indicated time and each plate was
developed with CellTiter-Glo reagent using a volume equivalent to
the cell culture volume in the wells. Plates were shaken for
approximately 2 minutes and chemiluminescent signal was read on the
Analyst GT (Molecular Devices) or EnVision Plate Reader (Perkin
Elmer).
[0456] Results were expressed as a percent of the T0 and plotted
against the compound concentration. The T0 value was normalized to
100% and represents the number of cells at time of compound
addition and the concentration response data were fit with a 4
parameter curve fit using XLfit software (model 205). The
concentration that inhibited cell growth by 50% (gIC.sub.50) is the
midpoint of the `growth window` (between the T0 and DMSO control).
The Ymin-T0 value is determined by subtracting the T0 value (100%)
from the Ymin value (%) determined from the fit of the
concentration response curve. Values from the wells with no cells
were subtracted from all samples for background correction.
[0457] Example 1 was tested in accordance with the above assay and
found to have a gIC.sub.50 in the range 163-29300 nM across all
cell lines, more specifically in the range 163-2480 nM for heme
cell lines and 410-29300 nM for breast cell lines.
[0458] These data demonstrate that the bromodomain inhibitor tested
in the above assay inhibited cell growth in a panel of oncology
cell lines.
[0459] All publications, including but not limited to patents and
patent applications, cited in this specification are herein
incorporated by reference as if each individual publication were
specifically and individually indicated to be incorporated by
reference herein as though fully set forth.
* * * * *