U.S. patent application number 11/403763 was filed with the patent office on 2006-11-23 for dna-pk inhibitors.
This patent application is currently assigned to KuDOS Pharmaceuticals Limited. Invention is credited to Hilary Alan Calvert, Xiao-ling Cockcroft, Nicola Jane Curtin, Marine Desage-El Murr, Mark Frigerio, Bernard Thomas Golding, Roger John Griffin, Ian Robert Hardcastle, Marc Geoffrey Hummersone, Niall Morrison Barr Martin, Keith Allan Menear, David Richard Newell, Graeme Cameron Murray Smith.
Application Number | 20060264623 11/403763 |
Document ID | / |
Family ID | 36579227 |
Filed Date | 2006-11-23 |
United States Patent
Application |
20060264623 |
Kind Code |
A1 |
Smith; Graeme Cameron Murray ;
et al. |
November 23, 2006 |
DNA-PK inhibitors
Abstract
Compounds of formula l: ##STR1## wherein A, B and D are
respectively selected from the group consisting of: (i) CH, NH, C;
(ii) CH, N, N;and (iii) CH, O, C; are disclosed for use in
inhibiting DNA-PK.
Inventors: |
Smith; Graeme Cameron Murray;
(Cambridge, GB) ; Martin; Niall Morrison Barr;
(Cambridge, GB) ; Menear; Keith Allan; (Cambridge,
GB) ; Hummersone; Marc Geoffrey; (Cambridge, GB)
; Cockcroft; Xiao-ling; (Cambridge, GB) ;
Frigerio; Mark; (Cambridge, GB) ; Griffin; Roger
John; (Newcastle upon Tyne, GB) ; Golding; Bernard
Thomas; (Newcastle upon Tyne, GB) ; Hardcastle; Ian
Robert; (Newcastle upon Tyne, GB) ; Newell; David
Richard; (Newcastle upon Tyne, GB) ; Calvert; Hilary
Alan; (Newcastle upon Tyne, GB) ; Curtin; Nicola
Jane; (Newcastle upon Tyne, GB) ; Desage-El Murr;
Marine; (Paris, FR) |
Correspondence
Address: |
MICHAEL BEST & FRIEDRICH, LLP
ONE SOUTH PINCKNEY STREET
P O BOX 1806
MADISON
WI
53701
US
|
Assignee: |
KuDOS Pharmaceuticals
Limited
Cambridge
GB
Cancer Research Technology Limited
London
GB
|
Family ID: |
36579227 |
Appl. No.: |
11/403763 |
Filed: |
April 13, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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60671830 |
Apr 15, 2005 |
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60671886 |
Apr 15, 2005 |
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60696064 |
Jul 1, 2005 |
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60718904 |
Sep 20, 2005 |
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Current U.S.
Class: |
544/114 |
Current CPC
Class: |
A61P 31/14 20180101;
A61P 35/00 20180101; C07D 407/14 20130101; C07D 405/04 20130101;
C07D 409/04 20130101; A61P 31/18 20180101; C07D 413/00 20130101;
C07D 471/04 20130101; A61P 43/00 20180101; C07D 407/04 20130101;
C07D 401/14 20130101 |
Class at
Publication: |
544/114 |
International
Class: |
C07D 413/00 20060101
C07D413/00 |
Foreign Application Data
Date |
Code |
Application Number |
Apr 18, 2005 |
GB |
0507831.6 |
Claims
1. A compound of formula I: ##STR144## and isomers, salts,
solvates, chemically protected forms, and prodrugs thereof,
wherein: A, B and D are respectively selected from the group
consisting of: (i) CH, NH, C; (ii) CH, N, N;and (iii) CH, O, C; the
dotted lines represent two double bonds in the appropriate
locations; R.sup.N1 and R.sup.N2 are independently selected from
hydrogen, an optionally substituted C.sub.1-7 alkyl group,
C.sub.3-20 heterocyclyl group, or C.sub.5-20 aryl group, or may
together form, along with the nitrogen atom to which they are
attached, an optionally substituted heterocyclic ring having from 4
to 8 ring atoms; Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5 and Z.sup.6,
together with the carbon atom to which they are bound, form an
aromatic ring; Z.sup.2 is selected from the group consisting of
CR.sup.2, N, NH, S, and O; Z.sup.3 is CR.sup.3; Z.sup.4 is selected
from the group consisting of CR.sup.4, N, NH, S, and O; Z.sup.5 is
a direct bond, or is selected from the group consisting of O, N,
NH, S, and CH; Z.sup.6 is selected from the group consisting of O,
N, NH, S, and CH; R.sup.2 is H; R.sup.3 is selected from halo or
optionally substituted C.sub.5-20 aryl; R.sup.4 is selected from
the group consisting of H, OH, NO.sub.2, NH.sub.2 and Q-Y--X, where
Q is --NH--C(.dbd.O)-- or --O--; Y is an optionally substituted
C.sub.1-5 alkylene group; X is selected from SR.sup.S1 or
NR.sup.N3R.sup.N4, wherein, R.sup.S1, or R.sup.N3 and R.sup.N4 are
independently selected from hydrogen, optionally substituted
C.sub.1-7 alkyl, C.sub.5-20 aryl, or C.sub.3-20 heterocyclyl
groups, or R.sup.N3 and R.sup.N4 may together form, along with the
nitrogen atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms; if Q is --O--, X
may additionally be selected from --C(.dbd.O)--NR.sup.N5R.sup.N6,
wherein R.sup.N5 and R.sup.N6 are independently selected from
hydrogen, optionally substituted C.sub.1-7 alkyl, C.sub.5-20 aryl,
or C.sub.3-20 heterocyclyl groups, or R.sup.N5 and R.sup.N6 may
together form, along with the nitrogen atom to which they are
attached, an optionally substituted heterocyclic ring having from 4
to 8 ring atoms and if Q is --NH--C(.dbd.O)--, --Y--X may be
additionally selected from C.sub.1-7 alkyl.
2. A compound according to claim 1, wherein Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5 and Z.sup.6, together with the carbon atom to
which they are bound, form a substituted aryl group selected from
substituted phenyl, thiophenyl, furanyl, thiazolyl, imidazolyl,
pyridyl, pyrimidinyl, isoxazolyl, oxazolyl, isothiazolyl.
3. A compound according to claim 2, wherein Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5 and Z.sup.6, together with the carbon atom to
which they are bound form a group selected from substituted phenyl,
thiazolyl, thiophenyl, or pyridyl.
4. A compound according to claim 1, wherein R.sup.3 is selected
from C.sub.5 heteroaryl, pyridyl and phenyl.
5. A compound according to claim 1, wherein Z.sup.2 is CR.sup.2,
Z.sup.3 is CR.sup.3, Z.sup.4 is CR.sup.4 and Z.sup.5 and Z.sup.6
are both CH.
6. A compound according to claim 5, wherein R.sup.3 is
unsubstituted phenyl, R.sup.N1 and R.sup.N2 form a morpholino group
and R.sup.4 is not H.
7. A compound according to claim 1, wherein R.sup.4 is Q-Y--X.
8. A compound according to claim 7, wherein Q is --NH--C(.dbd.O)--
and X is NR.sup.N3R.sup.N4.
9. A compound according to claim 7, wherein Q is --O--, X is
NR.sup.N3R.sup.N4, and Y is an optionally substituted C.sub.1-3
alkylene group.
10. A compound according to claim 1, wherein R.sup.N1 and R.sup.N2
form, along with the nitrogen atom to which they are attached, a
heterocyclic ring having from 4 to 8 atoms.
11. A compound according to claim 1, wherein R.sup.N1 and R.sup.N2
form, along with the nitrogen atom to which they are attached, a
group selected from morpholino and thiomorpholino.
12. A composition comprising a compound according to claim 1, and a
pharmaceutically acceptable carrier or diluent.
13. A method of treatment of a subject suffering from a disease
ameliorated by the inhibition of DNA-PK, comprising administering
to said subject a compound according to claim 1.
14. A method of inhibiting DNA-PK in vitro or in vivo, comprising
contacting a cell with an effective amount of a compound according
to claim 1.
Description
[0001] The present invention relates to compounds which act as
DNA-PK inhibitors, their use and synthesis.
[0002] The DNA-dependent protein kinase (DNA-PK) is a nuclear
serine/threonine protein kinase that is activated upon association
with DNA. Biochemical and genetic data have revealed this kinase to
be composed of a large catalytic subunit, termed DNA-PKcs, and a
regulatory component termed Ku. DNA-PK has been shown to be a
crucial component of both the DNA double-strand break (DSB) repair
machinery and the V(D)J recombination apparatus. In addition,
recent work has implicated DNA-PK components in a variety of other
processes, including the modulation of chromatin structure and
telomere maintenance (Smith, G. C. M. and Jackson, S. P.,Genes and
Dev., 13, 916-934 (1999)).
[0003] DNA DSBs are regarded as the most lethal lesion a cell can
encounter. To combat the serious threats posed by DNA DSBs,
eukaryotic cells have evolved several mechanisms to mediate their
repair. In higher eukaryotes, the predominant of these mechanisms
is DNA non-homologous end-joining (NHEJ), also known as
illegitimate recombination. DNA-PK plays a key role in this
pathway. Increased DNA-PK activity has been demonstrated both in
vitro and in vivo and correlates with the resistance of tumour
cells to IR and bifunctional alkylating agents (Muller C., et al.,
Blood, 92, 2213-2219 (1998), Sirzen F., et al., Eur. J. Cancer, 35,
111-116 (1999)). Therefore, increased DNA-PK activity has been
proposed as a cellular and tumour resistance mechanism. Hence,
inhibition of DNA-PK with a small molecule inhibitor may prove
efficacious in tumours where over-expression is regarded as a
resistance mechanism.
[0004] It also has been previously found that the PI 3-kinase
inhibitor LY294002: ##STR2##
[0005] is able to inhibit DNA-PK function in vitro (Izzard, R. A.,
et al., Cancer Res., 59, 2581-2586 (1999)). The IC.sub.50
(concentration at which 50% of enzyme activity is lost) for
LY294002 towards DNA-PK is, at .about.1 .mu.M, the same as that for
PI 3-kinase. Furthermore it has been shown that LY294002 is also
able to weakly sensitise cells to the effects of IR (Rosenzweig, K.
E., et al., Clin. Cancer Res., 3, 1149-1156 (1999)).
[0006] WO 03/024949 describes a number of classes of compounds
useful as DNA-PK inhibitors, including 2-amino-chromen-4-ones of
the general structure: ##STR3##
[0007] of which: ##STR4##
[0008] was one example. This compound exhibited an IC.sub.50 of
0.76 .mu.M and an SER of 1.5 (500 nM) (see below for methods).
[0009] Other examples of DNA-PK inhibitors include
1(2-hydroxy-4-morpholin-4-yl-phenyl)-ethanone (Kashishian, A., et
al., Mol. Cancer Ther, 2, 1257-1264 (2003)): ##STR5##
[0010] and SU11752 (Ismail, I. H., et al., Oncogene, 23, 873-882
(2004)) ##STR6##
[0011] Given the involvement of DNA-PK in DNA repair processes, and
that small molecule inhibitors have been shown to radio- and
chemo-sensitise mammalian cells in culture, an application of
specific DNA-PK inhibitory drugs would be to act as agents that
will enhance the efficacy of both cancer chemotherapy and
radiotherapy. DNA-PK inhibitors may also prove useful in the
treatment of retroviral mediated diseases. For example it has been
demonstrated that loss of DNA-PK activity severely represses the
process of retroviral integration (Daniel R, et al., Science, 284,
644-7 (1999)).
[0012] The present inventors have now discovered further compounds
which exhibit similar or improved levels of DNA-PK inhibition,
whilst possessing other useful properties for use as active
pharmaceuticals, in particular improved solubility and cellular
efficacy.
[0013] Accordingly, the first aspect of the invention provides a
compound of formula l: ##STR7##
[0014] and isomers, salts, solvates, chemically protected forms,
and prodrugs thereof, wherein: A, B and D are respectively selected
from the group consisting of:
[0015] (i) CH, NH, C;
[0016] (ii) CH, N, N;and
[0017] (iii) CH, 0, C;
[0018] the dotted lines represent two double bonds in the
appropriate locations; R.sup.N1 and R.sup.N2 are independently
selected from hydrogen, an optionally substituted C.sub.1-7 alkyl
group, C.sub.3-20 heterocyclyl group, or C.sub.5-20 aryl group, or
may together form, along with the nitrogen atom to which they are
attached, an optionally substituted heterocyclic ring having from 4
to 8 ring atoms;
[0019] Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5 and Z.sup.6, together
with the carbon atom to which they are bound, form an aromatic
ring;
[0020] Z.sup.2 is selected from the group consisting of CR.sup.2,
N, NH, S, and O; Z.sup.3 is CR.sup.3; Z.sup.4 is selected from the
group consisting of CR.sup.4, N, NH, S, and O; Z.sup.5 is a direct
bond, or is selected from the group consisting of O, N, NH, S, and
CH; Z.sup.6 is selected from the group consisting of O, N, NH, S,
and CH;
[0021] R.sup.2 is H;
[0022] R.sup.3 is selected from halo or optionally substituted
C.sub.5-20 aryl;
[0023] R.sup.4 is selected from the group consisting of H, OH,
NO.sub.2, NH.sub.2 and Q-Y--X, where Q is --NH--C(.dbd.O)-- or
--O--;
[0024] Y is an optionally substituted C.sub.1-5 alkylene group;
[0025] X is selected from SR.sup.S1 or NR.sup.N3R.sup.N4,
wherein,
[0026] R.sup.S1, or R.sup.N3 and R.sup.N4 are independently
selected from hydrogen, optionally substituted C.sub.1-7 alkyl,
C.sub.5-20 aryl, or C.sub.3-20 heterocyclyl groups, or R.sup.N3 and
R.sup.N4 may together form, along with the nitrogen atom to which
they are attached, an optionally substituted heterocyclic ring
having from 4 to 8 ring atoms;
[0027] if Q is --O--, X may additionally be selected from
--C(.dbd.O)--NR.sup.N5R.sup.N6, wherein R.sup.N5 and R.sup.N6 are
independently selected from hydrogen, optionally substituted
C.sub.1-7 alkyl, C.sub.5-20 aryl, or C.sub.3-20 heterocyclyl
groups, or R.sup.N5 and R.sup.N6 may together form, along with the
nitrogen atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms and
[0028] if Q is --NH--C(.dbd.O)--, --Y--X may be additionally
selected from C.sub.1-7 alkyl.
[0029] Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5 and Z.sup.6 are selected
such that the group they form including the carbon atom to which
Z.sup.2 and Z.sup.6 are bound is aromatic.
[0030] In some embodiments of the present invention, the compounds
of the first aspect may be compounds of formula II or more
particularly formula IIa, in which Z.sup.2 is CR.sup.2, Z.sup.3 is
CR.sup.3, Z.sup.4 is CR.sup.4 and Z.sup.5 and Z.sup.6 are both CH:
##STR8##
[0031] wherein A, B, D, R.sup.N1, R.sup.N2, R.sup.2, R.sup.3 and
R.sup.4 are as described above. In these particular embodiments, if
R.sup.3 is unsubstituted phenyl, and R.sup.N1 and R.sup.N2 form a
morpholino group, R.sup.4 is not H.
[0032] The options for A, B and D result in compounds of the
following formulae, where Ar represents the aromatic ring formed by
Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5 and Z.sup.6: TABLE-US-00001
Formula A B D Structure Ia CH NH C ##STR9## Ib CH N N ##STR10## Ic
CH O C ##STR11##
[0033] A second aspect of the invention provides a composition
comprising a compound of the first aspect and a pharmaceutically
acceptable carrier or diluent.
[0034] A third aspect of the invention provides a compound of the
first aspect for use in a method of therapy.
[0035] A fourth aspect of the invention provides for the use of a
compound of the first aspect in the preparation of a medicament for
treating a disease ameliorated by the inhibition of DNA-PK.
[0036] It is preferred that the medicament of the fourth aspect
selectivity inhibits the activity of DNA-PK compared to PI 3-kinase
and/or ATM. Selectivity is an important issue as inhibition of
other PI 3-kinase family members may lead to unwanted side-effects
associated with the loss of function of those enzymes.
[0037] In particular, the compounds may be used in the preparation
of a medicament for:
[0038] (a) use as an adjunct in cancer therapy or for potentiating
tumour cells for treatment with ionising radiation or
chemotherapeutic agents; or
[0039] (b) the treatment of retroviral mediated diseases.
[0040] A further aspect of the invention provides an active
compound as described herein for use in a method of treatment of
the human or animal body, preferably in the form of a
pharmaceutical composition.
[0041] Another aspect of the invention provides a method of
inhibiting DNA-PK in vitro or in vivo, comprising contacting a cell
with an effective amount of an active compound as described
herein.
DEFINITIONS
[0042] C.sub.1-7 alkyl: The term "C.sub.1-7alkyl", as used herein,
pertains to a monovalent moiety obtained by removing a hydrogen
atom from a C.sub.1-7 hydrocarbon compound having from 1 to 7
carbon atoms, which may be aliphatic or alicyclic, or a combination
thereof, and which may be saturated, partially unsaturated, or
fully unsaturated.
[0043] Examples of saturated linear C.sub.1-7 alkyl groups include,
but are not limited to, methyl, ethyl, n-propyl, n-butyl, and
n-pentyl (amyl).
[0044] Examples of saturated branched C.sub.1-7 alkyl groups
include, but are not limited to, iso-propyl, iso-butyl, sec-butyl,
tert-butyl, and neo-pentyl.
[0045] Examples of saturated alicyclic C.sub.1-7 alkyl groups (also
referred to as "C.sub.3-7 cycloalkyl" groups) include, but are not
limited to, groups such as cyclopropyl, cyclobutyl, cyclopentyl,
and cyclohexyl, as well as substituted groups (e.g., groups which
comprise such groups), such as methylcyclopropyl,
dimethylcyclopropyl, methylcyclobutyl, dimethylcyclobutyl,
methylcyclopentyl, dimethylcyclopentyl, methylcyclohexyl,
dimethylcyclohexyl, cyclopropylmethyl and cyclohexylmethyl.
[0046] Examples of unsaturated C.sub.1-7 alkyl groups which have
one or more carbon-carbon double bonds (also referred to as
"C.sub.2-7alkenyl" groups) include, but are not limited to, ethenyl
(vinyl, --CH.dbd.CH.sub.2), 2-propenyl (allyl,
--CH--CH.dbd.CH.sub.2), isopropenyl (--C(CH.sub.3).dbd.CH.sub.2),
butenyl, pentenyl, and hexenyl.
[0047] Examples of unsaturated C.sub.1-7 alkyl groups which have
one or more carbon-carbon triple bonds (also referred to as
"C.sub.2-7 alkynyl" groups) include, but are not limited to,
ethynyl (ethinyl) and 2-propynyl (propargyl).
[0048] Examples of unsaturated alicyclic (carbocyclic) C.sub.1-7
alkyl groups which have one or more carbon-carbon double bonds
(also referred to as "C.sub.3-7cycloalkenyl" groups) include, but
are not limited to, unsubstituted groups such as cyclopropenyl,
cyclobutenyl, cyclopentenyl, and cyclohexenyl, as well as
substituted groups (e.g., groups which comprise such groups) such
as cyclopropenylmethyl and cyclohexenylmethyl.
[0049] C.sub.3-20 heterocyclyl: The term "C.sub.3-20 heterocyclyl",
as used herein, pertains to a monovalent moiety obtained by
removing a hydrogen atom from a ring atom of a C.sub.3-20
heterocyclic compound, said compound having one ring, or two or
more rings (e.g., spiro, fused, bridged), and having from 3 to 20
ring atoms, atoms, of which from 1 to 10 are ring heteroatoms, and
wherein at least one of said ring(s) is a heterocyclic ring.
Preferably, each ring has from 3 to 7 ring atoms, of which from 1
to 4 are ring heteroatoms. Ring heteroatoms may preferably be
selected from the group consisting of O, N, S and P. "C.sub.3-20"
denotes ring atoms, whether carbon atoms or heteroatoms.
[0050] Examples of C.sub.3-20 heterocyclyl groups having one
nitrogen ring atom include, but are not limited to, those derived
from aziridine, azetidine, pyrrolidines (tetrahydropyrrole),
pyrroline (e.g., 3-pyrroline, 2,5-dihydropyrrole), 2H-pyrrole or
3H-pyrrole (isopyrrole, isoazole), piperidine, dihydropyridine,
tetrahydropyridine, and azepine.
[0051] Examples of C.sub.3-20 heterocyclyl groups having one oxygen
ring atom include, but are not limited to, those derived from
oxirane, oxetane, oxolane (tetrahydrofuran), oxole (dihydrofuran),
oxane (tetrahydropyran), dihydropyran, pyran (C.sub.6), and oxepin.
Examples of substituted C.sub.3-20 heterocyclyl groups include
sugars, in cyclic form, for example, furanoses and pyranoses,
including, for example, ribose, lyxose, xylose, galactose, sucrose,
fructose, and arabinose.
[0052] Examples of C.sub.3-20 heterocyclyl groups having one
sulphur ring atom include, but are not limited to, those derived
from thiirane, thietane, thiolane (tetrahydrothiophene), thiane
(tetrahydrothiopyran), and thiepane.
[0053] Examples of C.sub.3-20 heterocyclyl groups having two oxygen
ring atoms include, but are not limited to, those derived from
dioxolane, dioxane, and dioxepane.
[0054] Examples of C.sub.3-20 heterocyclyl groups having two
nitrogen ring atoms include, but are not limited to, those derived
from imidazolidine, pyrazolidine (diazolidine), imidazoline,
pyrazoline (dihydropyrazole), and piperazine.
[0055] Examples of C.sub.3-20 heterocyclyl groups having one
nitrogen ring atom and one oxygen ring atom include, but are not
limited to, those derived from tetrahydrooxazole, dihydrooxazole,
tetrahydroisoxazole, dihydroisoxazole, morpholine,
tetrahydrooxazine, dihydrooxazine, and oxazine.
[0056] Examples of C.sub.3-20 heterocyclyl groups having one oxygen
ring atom and one sulphur ring atom include, but are not limited
to, those derived from oxathiolane and oxathiane (thioxane).
[0057] Examples of C.sub.3-20 heterocyclyl groups having one
nitrogen ring atom and one sulphur ring atom include, but are not
limited to, those derived from thiazoline, thiazolidine, and
thiomorpholine.
[0058] Other examples of C.sub.3-20heterocyclyl groups include, but
are not limited to, oxadiazine and oxathiazine.
[0059] Examples of heterocyclyl groups which additionally bear one
or more oxo (.dbd.O) groups, include, but are not limited to, those
derived from:
[0060] C.sub.5 heterocyclics, such as furanone, pyrone, pyrrolidone
(pyrrolidinone), pyrazolone (pyrazolinone), imidazolidone,
thiazolone, and isothiazolone;
[0061] C.sub.6 heterocyclics, such as piperidinone (piperidone),
piperidinedione, piperazinone, piperazinedione, pyridazinone, and
pyrimidinone (e.g., cytosine, thymine, uracil), and barbituric
acid;
[0062] fused heterocyclics, such as oxindole, purinone (e.g.,
guanine), benzoxazolinone, benzopyrone (e.g., coumarin);
[0063] cyclic anhydrides (--C(.dbd.O)--O--C(.dbd.O)-- in a ring),
including but not limited to maleic anhydride, succinic anhydride,
and glutaric anhydride; cyclic carbonates (--O--C(.dbd.O)--O-- in a
ring), such as ethylene carbonate and 1,2-propylene carbonate;
[0064] imides (--C(.dbd.O)--NR--C(.dbd.O)-- in a ring), including
but not limited to, succinimide, maleimide, phthalimide, and
glutarimide;
[0065] lactones (cyclic esters, --O--C(.dbd.O)-- in a ring),
including, but not limited to, .beta.-propiolactone,
.gamma.-butyrolactone, .delta.-valerolactone (2-piperidone), and
.epsilon.-caprolactone;
[0066] lactams (cyclic amides, --NR--C(.dbd.O)-- in a ring),
including, but not limited to, .beta.-propiolactam,
.gamma.-butyrolactam (2-pyrrolidone), .delta.-valerolactam, and
.epsilon.-caprolactam;
[0067] cyclic carbamates (--O--C(.dbd.O)--NR-- in a ring), such as
2-oxazolidone;
[0068] cyclic ureas (--NR--C(.dbd.O)--NR-- in a ring), such as
2-imidazolidone and pyrimidine-2,4-dione (e.g., thymine,
uracil).
[0069] C.sub.5-20 aryl: The term "C.sub.5-20 aryl", as used herein,
pertains to a monovalent moiety obtained by removing a hydrogen
atom from an aromatic ring atom of a C.sub.5-20 aromatic compound,
said compound having one ring, or two or more rings (e.g., fused),
and having from 5 to 20 ring atoms, and wherein at least one of
said ring(s) is an aromatic ring. Preferably, each ring has from 5
to 7 ring atoms.
[0070] The ring atoms may be all carbon atoms, as in "carboaryl
groups", in which case the group may conveniently be referred to as
a "C.sub.5-20 carboaryl" group.
[0071] Examples of C.sub.5-20 aryl groups which do not have ring
heteroatoms (i.e. C.sub.5-20 carboaryl groups) include, but are not
limited to, those derived from benzene (i.e. phenyl) (C.sub.6),
naphthalene (C.sub.10), anthracene (C.sub.14), phenanthrene
(C.sub.14), naphthacene (C.sub.18), and pyrene (C.sub.16).
[0072] Examples of aryl groups which comprise fused rings, one of
which is not an aromatic ring, include, but are not limited to,
groups derived from indene and fluorene.
[0073] Alternatively, the ring atoms may include one or more
heteroatoms, including but not limited to oxygen, nitrogen, and
sulphur, as in "heteroaryl groups". In this case, the group may
conveniently be referred to as a "C.sub.5-20 heteroaryl" group,
wherein "C.sub.5-20" denotes ring atoms, whether carbon atoms or
heteroatoms. Preferably, each ring has from 5 to 7 ring atoms, of
which from 0 to 4 are ring heteroatoms.
[0074] Examples of C.sub.5-20 heteroaryl groups include, but are
not limited to, C.sub.5 heteroaryl groups derived from furan
(oxole), thiophene (thiole), pyrrole (azole), imidazole
(1,3-diazole), pyrazole (1,2-diazole), triazole, oxazole,
isoxazole, thiazole, isothiazole, oxadiazole, and oxatriazole; and
C.sub.6 heteroaryl groups derived from isoxazine, pyridine (azine),
pyridazine (1,2-diazine), pyrimidine (1,3-diazine; e.g., cytosine,
thymine, uracil), pyrazine (1,4-diazine), triazine, tetrazole, and
oxadiazole (furazan).
[0075] Examples of C.sub.5-20 heterocyclic groups (some of which
are C.sub.5-20 heteroaryl groups) which comprise fused rings,
include, but are not limited to, C.sub.9 heterocyclic groups
derived from benzofuran, isobenzofuran, indole, isoindole, purine
(e.g., adenine, guanine), benzothiophene, benzimidazole; C.sub.10
heterocyclic groups derived from quinoline, isoquinoline,
benzodiazine, pyridopyridine, quinoxaline; C.sub.13 heterocyclic
groups derived from carbazole, dibenzothiophene, dibenzofuran;
C.sub.14 heterocyclic groups derived from acridine, xanthene,
phenoxathiin, phenazine, phenoxazine, phenothiazine.
[0076] The above C.sub.1-7 alkyl, C.sub.3-20 heterocyclyl and
C.sub.5-20 aryl groups whether alone or part of another
substituent, may themselves optionally be substituted with one or
more groups selected from themselves and the additional
substituents listed below.
[0077] Halo: --F, --Cl, --Br, and --I.
[0078] Hydroxy: --OH.
[0079] Ether: --OR, wherein R is an ether substituent, for example,
a C.sub.1-7 alkyl group (also referred to as a C.sub.1-7 alkoxy
group, discussed below), a C.sub.3-20 heterocyclyl group (also
referred to as a C.sub.3-20 heterocyclyloxy group), or a C.sub.5-20
aryl group (also referred to as a C.sub.5-20 aryloxy group),
preferably a C.sub.1-7 alkyl group.
[0080] C.sub.1-7 alkoxy: --OR, wherein R is a C.sub.1-7 alkyl
group. Examples of C17 alkoxy groups include, but are not limited
to, --OCH.sub.3 (methoxy), --OCH.sub.2CH.sub.3 (ethoxy) and
--OC(CH.sub.3).sub.3 (tert-butoxy).
[0081] Oxo (keto, -one): .dbd.O. Examples of cyclic compounds
and/or groups having, as a substituent, an oxo group (.dbd.O)
include, but are not limited to, carbocyclics such as
cyclopentanone and cyclohexanone; heterocyclics, such as pyrone,
pyrrolidone, pyrazolone, pyrazolinone, piperidone, piperidinedione,
piperazinedione, and imidazolidone; cyclic anhydrides, including
but not limited to maleic anhydride and succinic anhydride; cyclic
carbonates, such as propylene carbonate; imides, including but not
limited to, succinimide and maleimide; lactones (cyclic esters,
--O--C(.dbd.O)-- in a ring), including, but not limited to,
.beta.-propiolactone, .gamma.-butyrolactone, .delta.-valerolactone,
and .epsilon.-caprolactone; and lactams (cyclic amides,
--NH--C(.dbd.O)-- in a ring), including, but not limited to,
.beta.-propiolactam, .gamma.-butyrolactam (2-pyrrolidone),
.delta.-valerolactam, and .epsilon.-caprolactam.
[0082] Imino (imine): .dbd.NR, wherein R is an imino substituent,
for example, hydrogen, C.sub.1-7 alkyl group, a
C.sub.3-20heterocyclyl group, or a C.sub.5-20 aryl group,
preferably hydrogen or a C.sub.1-7 alkyl group. Examples of ester
groups include, but are not limited to, .dbd.NH, .dbd.NMe,
.dbd.NEt, and .dbd.NPh.
[0083] Formyl (carbaldehyde, carboxaldehyde): --C(.dbd.O)H.
[0084] Acyl (keto): --C(.dbd.O)R, wherein R is an acyl substituent,
for example, a C.sub.1-7alkyl group (also referred to as C.sub.1-7
alkylacyl or C.sub.1-7 alkanoyl), a C.sub.3-20 heterocyclyl group
(also referred to as C.sub.3-20 heterocyclylacyl), or a C.sub.5-20
aryl group (also referred to as C.sub.5-20 arylacyl), preferably a
C.sub.1-7 alkyl group. Examples of acyl groups include, but are not
limited to, --C(.dbd.O)CH.sub.3 (acetyl),
--C(.dbd.O)CH.sub.2CH.sub.3 (propionyl),
--C(.dbd.O)C(CH.sub.3).sub.3 (butyryl), and --C(.dbd.O)Ph (benzoyl,
phenone).
[0085] Carboxy (carboxylic acid): --COOH.
[0086] Ester (carboxylate, carboxylic acid ester, oxycarbonyl):
--C(.dbd.O)OR, wherein R is an ester substituent, for example, a
C.sub.1-7 alkyl group, a C.sub.3-20 heterocyclyl group, or a
C.sub.5-20 aryl group, preferably a C.sub.1-7alkyl group. Examples
of ester groups include, but are not limited to,
--C(.dbd.O)OCH.sub.3, --C(.dbd.O)OCH.sub.2CH.sub.3,
--C(.dbd.O)OC(CH.sub.3).sub.3, and --C(.dbd.O)OPh.
[0087] Acyloxy (reverse ester): --OC(.dbd.O)R, wherein R is an
acyloxy substituent, for example, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably a C.sub.1-7alkyl group. Examples of acyloxy groups
include, but are not limited to, --OC(.dbd.O)CH.sub.3 (acetoxy),
--OC(.dbd.O)CH.sub.2CH.sub.3, --OC(.dbd.O)C(CH.sub.3).sub.3,
--OC(.dbd.O)Ph, and --OC(.dbd.O)CH.sub.2Ph.
[0088] Amido (carbamoyl, carbamyl, aminocarbonyl, carboxamide):
--C(.dbd.O)NR.sup.1R.sup.2, wherein R.sup.1 and R.sup.2 are
independently amino substituents, as defined for amino groups.
Examples of amido groups include, but are not limited to,
--C(.dbd.O)NH.sub.2, --C(.dbd.O)NHCH.sub.3,
--C(.dbd.O)N(CH.sub.3).sub.2, --C(.dbd.O)NHCH.sub.2CH.sub.3, and
--C(.dbd.O)N(CH.sub.2CH.sub.3).sub.2, as well as amido groups in
which R.sup.1 and R.sup.2, together with the nitrogen atom to which
they are attached, form a heterocyclic structure as in, for
example, piperidinocarbonyl, morpholinocarbonyl,
thiomorpholinocarbonyl, and piperazinocarbonyl.
[0089] Acylamido (acylamino): --NR.sup.1C(.dbd.O)R.sup.2, wherein
R.sup.1 is an amide substituent, for example, hydrogen, a C.sub.1-7
alkyl group, a C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl
group, preferably hydrogen or a C.sub.1-7 alkyl group, and R.sup.2
is an acyl substituent, for example, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably hydrogen or a C.sub.1-7 alkyl group. Examples of
acylamide groups include, but are not limited to,
--NHC(.dbd.O)CH.sub.3, --NHC(.dbd.O)CH.sub.2CH.sub.3, and
--NHC(.dbd.O)Ph. R.sup.1 and R.sup.2 may together form a cyclic
structure, as in, for example, succinimidyl, maleimidyl and
phthalimidyl: ##STR12##
[0090] Acylureido: --N(R.sup.1)C(O)NR.sup.2C(O)R.sup.3 wherein
R.sup.1 and R.sup.2 are independently ureido substituents, for
example, hydrogen, a C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably hydrogen
or a C.sub.1-7 alkyl group. R.sup.3 is an acyl group as defined for
acyl groups. Examples of acylureido groups include, but are not
limited to, --NHCONHC(O)H, --NHCONMeC(O)H, --NHCONEtC(O)H,
--NHCONMeC(O)Me, --NHCONEtC(O)Et, --NMeCONHC(O)Et, --NMeCONHC(O)Me,
--NMeCONHC(O)Et, --NMeCONMeC(O)Me, --NMeCONEtC(O)Et, and
--NMeCONHC(O)Ph.
[0091] Carbamate: --NR.sup.1--C(O)--OR.sup.2 wherein R.sup.1 is an
amino substituent as defined for amino groups and R.sup.2 is an
ester group as defined for ester groups. Examples of carbamate
groups include, but are not limited to, --NH--C(O)--O--Me,
--NMe--C(O)--O--Me, --NH--C(O)--O-Et, --NMe-C(O)--O-t-butyl, and
--NH--C(O)--O--Ph.
[0092] Thioamido (thiocarbamyl): --C(.dbd.S)NR.sup.1R.sup.2,
wherein R.sup.1 and R.sup.2 are independently amino substituents,
as defined for amino groups. Examples of amido groups include, but
are not limited to, --C(.dbd.S)NH.sub.2, --C(.dbd.S)NHCH.sub.3,
--C(.dbd.S)N(CH.sub.3).sub.2, and
--C(.dbd.S)NHCH.sub.2CH.sub.3.
[0093] Tetrazolyl: a five membered aromatic ring having four
nitrogen atoms and one carbon atom, ##STR13##
[0094] Amino: --NR.sup.1R.sup.2, wherein R.sup.1 and R.sup.2 are
independently amino substituents, for example, hydrogen, a
C.sub.1-7 alkyl group (also referred to as C.sub.1-7 alkylamino or
di-C.sub.1-7 alkylamino), a C.sub.3-20 heterocyclyl group, or a
C.sub.5-20 aryl group, preferably H or a C.sub.1-7alkyl group, or,
in the case of a "cyclic" amino group, R.sup.1 and R.sup.2, taken
together with the nitrogen atom to which they are attached, form a
heterocyclic ring having from 4 to 8 ring atoms. Examples of amino
groups include, but are not limited to, --NH.sub.2, --NHCH.sub.3,
--NHC(CH.sub.3).sub.2, --N(CH.sub.3).sub.2,
--N(CH.sub.2CH.sub.3).sub.2, and --NHPh. Examples of cyclic amino
groups include, but are not limited to, aziridino, azetidino,
pyrrolidino, piperidino, piperazino, morpholino, and
thiomorpholino.
[0095] Imino: .dbd.NR, wherein R is an imino substituent, for
example, for example, hydrogen, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably H or a C.sub.1-7 alkyl group.
[0096] Amidine: --C(.dbd.NR)NR.sub.2, wherein each R is an amidine
substituent, for example, hydrogen, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably H or a C.sub.1-7 alkyl group. An example of an amidine
group is --C(.dbd.NH)NH.sub.2.
[0097] Carbazoyl (hydrazinocarbonyl): --C(O)--NN--R.sup.1 wherein
R.sup.1 is an amino substituent as defined for amino groups.
Examples of azino groups include, but are not limited to,
--C(O)--NN--H, --C(O)--NN-Me, --C(O)--NN-Et, --C(O)--NN--Ph, and
--C(O)--NN--CH.sub.2--Ph.
[0098] Nitro: --NO.sub.2.
[0099] Nitroso: --NO.
[0100] Azido: --N.sub.3.
[0101] Cyano (nitrile, carbonitrile): --CN.
[0102] Isocyano: --NC.
[0103] Cyanato: --OCN.
[0104] Isocyanato: --NCO.
[0105] Thiocyano (thiocyanato): --SCN.
[0106] Isothiocyano (isothiocyanato): --NCS.
[0107] Sulfhydryl (thiol, mercapto): --SH.
[0108] Thioether (sulfide): --SR, wherein R is a thioether
substituent, for example, a C.sub.1-7 alkyl group (also referred to
as a C.sub.1-7 alkylthio group), a C.sub.3-20 heterocyclyl group,
or a C.sub.5-20 aryl group, preferably a C.sub.1-7 alkyl group.
Examples of C.sub.1-7 alkylthio groups include, but are not limited
to, --SCH.sub.3 and --SCH.sub.2CH.sub.3.
[0109] Disulfide: --SS--R, wherein R is a disulfide substituent,
for example, a C.sub.1-7 alkyl group, a C.sub.3-20 heterocyclyl
group, or a C.sub.5-20 aryl group, preferably a C.sub.1-7 alkyl
group (also referred to herein as C.sub.1-7 alkyl disulfide).
Examples of C.sub.1-7 alkyl disulfide groups include, but are not
limited to, --SSCH.sub.3 and --SSCH.sub.2CH.sub.3.
[0110] Sulfone (sulfonyl): --S(.dbd.Q).sub.2R, wherein R is a
sulfone substituent, for example, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably a C.sub.1-7 alkyl group. Examples of sulfone groups
include, but are not limited to, --S(.dbd.O).sub.2CH.sub.3
(methanesulfonyl, mesyl), --S(.dbd.O).sub.2CF.sub.3 (triflyl),
--S(.dbd.O).sub.2CH.sub.2CH.sub.3, --S(.dbd.O).sub.2C.sub.4F.sub.9
(nonaflyl), --S(.dbd.O).sub.2CH.sub.2CF.sub.3 (tresyl),
--S(.dbd.O).sub.2Ph(phenylsulfonyl), 4-methylphenylsulfonyl(tosyl),
4-bromophenylsulfonyl(brosyl), and 4-nitrophenyl (nosyl).
[0111] Sulfine (sulfinyl, sulfoxide): --S(.dbd.O)R, wherein R is a
sulfine substituent, for example, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably a C.sub.1-7 alkyl group. Examples of sulfine groups
include, but are not limited to, --S(.dbd.O)CH.sub.3 and
--S(.dbd.O)CH.sub.2CH.sub.3.
[0112] Sulfonyloxy: --OS(.dbd.O).sub.2R, wherein R is a sulfonyloxy
substituent, for example, a C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably a
C.sub.1-7 alkyl group. Examples of sulfonyloxy groups include, but
are not limited to, --OS(.dbd.O).sub.2CH.sub.3 and
--OS(.dbd.O).sub.2CH.sub.2CH.sub.3.
[0113] Sulfinyloxy: --OS(.dbd.O)R, wherein R is a sulfinyloxy
substituent, for example, a C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably a
C.sub.1-7 alkyl group. Examples of sulfinyloxy groups include, but
are not limited to, --OS(.dbd.O)CH.sub.3 and
--OS(.dbd.O)CH.sub.2CH.sub.3.
[0114] Sulfamino: --NR.sup.1S(.dbd.O).sub.2OH, wherein R.sup.1 is
an amino substituent, as defined for amino groups. Examples of
sulfamino groups include, but are not limited to,
--NHS(.dbd.O).sub.2OH and --N(CH.sub.3)S(.dbd.O).sub.2OH.
[0115] Sulfinamino: --NR.sup.1S(.dbd.O)R, wherein R.sup.1 is an
amino substituent, as defined for amino groups, and R is a
sulfinamino substituent, for example, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably a C.sub.1-7 alkyl group. Examples of sulfinamino groups
include, but are not limited to, --NHS(.dbd.O)CH.sub.3 and
--N(CH.sub.3)S(.dbd.O)C.sub.6H.sub.5.
[0116] Sulfamyl: --S(.dbd.O)NR.sup.1R.sup.2, wherein R.sup.1 and
R.sup.2 are independently amino substituents, as defined for amino
groups. Examples of sulfamyl groups include, but are not limited
to, --S(.dbd.O)NH.sub.2, --S(.dbd.O)NH(CH.sub.3),
--S(.dbd.O)N(CH.sub.3).sub.2, --S(.dbd.O)NH(CH.sub.2CH.sub.3),
--S(.dbd.O)N(CH.sub.2CH.sub.3).sub.2, and --S(.dbd.O)NHPh.
[0117] Sulfonamino: --NR.sup.1S(.dbd.O).sub.2R, wherein R.sup.1 is
an amino substituent, as defined for amino groups, and R is a
sulfonamino substituent, for example, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably a C.sub.1-7 alkyl group. Examples of sulfonamino groups
include, but are not limited to, --NHS(.dbd.O).sub.2CH.sub.3 and
--N(CH.sub.3)S(.dbd.O).sub.2C.sub.6H.sub.5. A special class of
sulfonamino groups are those derived from sultams--in these groups
one of R.sup.1 and R is a C.sub.5-20 aryl group, preferably phenyl,
whilst the other of R.sup.1 and R is a bidentate group which links
to the C.sub.5-20 aryl group, such as a bidentate group derived
from a C.sub.1-7 alkyl group. Examples of such groups include, but
are not limited to: ##STR14##
[0118] Phosphoramidite: --OP(OR.sup.1)--NR.sup.2.sub.2, where
R.sup.1 and R.sup.2 are phosphoramidite substituents, for example,
--H, a (optionally substituted) C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably --H, a
C.sub.1-7 alkyl group, or a C.sub.5-20 aryl group. Examples of
phosphoramidite groups include, but are not limited to,
--OP(OCH.sub.2CH.sub.3)--N(CH.sub.3).sub.2,
--OP(OCH.sub.2CH.sub.3)--N(i-Pr).sub.2, and
--OP(OCH.sub.2CH.sub.2CN)--N(i-Pr).sub.2.
[0119] Phosphoramidate: --OP(.dbd.O)(OR.sup.1)--NR.sup.2.sub.2,
where R.sup.1 and R.sup.2 are phosphoramidate substituents, for
example, --H, a (optionally substituted) C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably --H, a C.sub.1-7 alkyl group, or a C.sub.5-20 aryl
group. Examples of phosphoramidate groups include, but are not
limited to, --OP(.dbd.O)(OCH.sub.2CH.sub.3)--N(CH.sub.3).sub.2,
--OP(.dbd.O)(OCH.sub.2CH.sub.3)--N(i-Pr).sub.2, and
--OP(.dbd.O)(OCH.sub.2CH.sub.2CN)--N(i-Pr).sub.2.
[0120] In many cases, substituents may themselves be substituted.
For example, a C.sub.1-7 alkoxy group may be substituted with, for
example, a C.sub.1-7 alkyl (also referred to as a C.sub.1-7
alkyl-C.sub.1-7alkoxy group), for example, cyclohexylmethoxy, a
C.sub.3-20 heterocyclyl group (also referred to as a C.sub.5-20
aryl-C.sub.1-7 alkoxy group), for example phthalimidoethoxy, or a
C.sub.5-20 aryl group (also referred to as a
C.sub.5-20aryl-C.sub.1-7alkoxy group), for example, benzyloxy.
[0121] C.sub.1-5 Alkylene: The term "C.sub.1-5 alkylene", as used
herein, pertains to a bidentate moiety obtained by removing two
hydrogen atoms, either both from the same carbon atom, or one from
each of two different carbon atoms, of an aliphatic linear
hydrocarbon compound having from 1 to 5 carbon atoms (unless
otherwise specified), which may be saturated, partially
unsaturated, or fully unsaturated. Thus, the term "alkylene"
includes the sub-classes alkenylene, alkynylene, etc., discussed
below.
[0122] Examples of saturated C.sub.1-5 alkylene groups include, but
are not limited to, --(CH.sub.2).sub.n-- where n is an integer from
1 to 5, for example, --CH.sub.2-(methylene),
--CH.sub.2CH.sub.2-(ethylene),
--CH.sub.2CH.sub.2CH.sub.2-(propylene), and
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2-(butylene).
[0123] Examples of partially unsaturated C.sub.1-5 alkylene groups
include, but is not limited to, --CH.dbd.CH-(vinylene),
--CH.dbd.CH--CH.sub.2--, --CH.sub.2--CH.dbd.CH.sub.2--,
--CH.dbd.CH--CH.sub.2--CH.sub.2--,
--CH.dbd.CH--CH.sub.2--CH.sub.2--CH.sub.2--,
--CH.dbd.CH--CH.dbd.CH-- and
--CH.dbd.CH--CH.dbd.CH--CH.sub.2--.
[0124] The substituent groups listed above may be substituents on
an alkylene group.
Includes Other Forms
[0125] Included in the above are the well known ionic, salt,
solvate, and protected forms of these substituents. For example, a
reference to carboxylic acid (--COOH) also includes the anionic
(carboxylate) form (--COO.sup.-), a salt or solvate thereof, as
well as conventional protected forms. Similarly, a reference to an
amino group includes the protonated form
(--N.sup.+HR.sup.1R.sup.2), a salt or solvate of the amino group,
for example, a hydrochloride salt, as well as conventional
protected forms of an amino group. Similarly, a reference to a
hydroxyl group also includes the anionic form (--O.sup.-), a salt
or solvate thereof, as well as conventional protected forms of a
hydroxyl group.
Isomers, Salts, Solvates, Protected Forms, and Prodrugs
[0126] Certain compounds may exist in one or more particular
geometric, optical, enantiomeric, diasteriomeric, epimeric,
stereoisomeric, tautomeric, conformational, or anomeric forms,
including but not limited to, cis- and trans-forms; E- and Z-forms;
c-, t-, and r-forms; endo- and exo-forms; R--, S--, and meso-forms;
D- and L-forms; d- and l-forms; (+) and (-) forms; keto-, enol-,
and enolate-forms; syn- and anti-forms; synclinal- and
anticlinal-forms; .alpha.- and .beta.-forms; axial and equatorial
forms; boat-, chair-, twist-, envelope-, and halfchair-forms; and
combinations thereof, hereinafter collectively referred to as
"isomers" (or "isomeric forms").
[0127] Note that, except as discussed below for tautomeric forms,
specifically excluded from the term "isomers", as used herein, are
structural (or constitutional) isomers (i.e. isomers which differ
in the connections between atoms rather than merely by the position
of atoms in space). For example, a reference to a methoxy group,
--OCH.sub.3, is not to be construed as a reference to its
structural isomer, a hydroxymethyl group, --CH.sub.2OH. Similarly,
a reference to ortho-chlorophenyl is not to be construed as a
reference to its structural isomer, meta-chlorophenyl. However, a
reference to a class of structures may well include structurally
isomeric forms falling within that class (e.g., C.sub.1-7 alkyl
includes n-propyl and iso-propyl; butyl includes n-, iso-, sec-,
and tert-butyl; methoxyphenyl includes ortho-, meta-, and
para-methoxyphenyl).
[0128] The above exclusion does not pertain to tautomeric forms,
for example, keto-, enol-, and enolate-forms, as in, for example,
the following tautomeric pairs: keto/enol (illustrated below),
imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime,
thioketone/enethiol, N-nitroso/hyroxyazo, and nitro/aci-nitro.
##STR15##
[0129] Note that specifically included in the term "isomer" are
compounds with one or more isotopic substitutions. For example, H
may be in any isotopic form, including .sup.1H, .sup.2H (D), and
.sup.3H (T); C may be in any isotopic form, including .sup.12C,
.sup.13C, and .sup.14C; 0 may be in any isotopic form, including
.sup.16O and .sup.18O; and the like.
[0130] Unless otherwise specified, a reference to a particular
compound includes all such isomeric forms, including (wholly or
partially) racemic and other mixtures thereof. Methods for the
preparation (e.g. asymmetric synthesis) and separation (e.g.,
fractional crystallisation and chromatographic means) of such
isomeric forms are either known in the art or are readily obtained
by adapting the methods taught herein, or known methods, in a known
manner.
[0131] Unless otherwise specified, a reference to a particular
compound also includes ionic, salt, solvate, and protected forms of
thereof, for example, as discussed below.
[0132] It may be convenient or desirable to prepare, purify, and/or
handle a corresponding salt of the active compound, for example, a
pharmaceutically-acceptable salt. Examples of pharmaceutically
acceptable salts are discussed in Berge, et al., J. Pharm. Sci.,
66, 1-19 (1977).
[0133] For example, if the compound is anionic, or has a functional
group which may be anionic (e.g., --COOH may be --COO.sup.-), then
a salt may be formed with a suitable cation. Examples of suitable
inorganic cations include, but are not limited to, alkali metal
ions such as Na.sup.+ and K.sup.+, alkaline earth cations such as
Ca.sup.2+ and Mg.sup.2+, and other cations such as Al.sup.3+.
Examples of suitable organic cations include, but are not limited
to, ammonium ion (i.e., NH.sub.4.sup.+) and substituted ammonium
ions (e.g., NH.sub.3R.sup.+, NH.sub.2R.sub.2.sup.+, NHR.sub.3+,
NR.sub.4.sup.+). Examples of some suitable substituted ammonium
ions are those derived from: ethylamine, diethylamine,
dicyclohexylamine, triethylamine, butylamine, ethylenediamine,
ethanolamine, diethanolamine, piperazine, benzylamine,
phenylbenzylamine, choline, meglumine, and tromethamine, as well as
amino acids, such as lysine and arginine. An example of a common
quaternary ammonium ion is N(CH.sub.3).sub.4.sup.+.
[0134] If the compound is cationic, or has a functional group which
may be cationic (e.g., --NH.sub.2 may be --NH.sub.3.sup.+), then a
salt may be formed with a suitable anion. Examples of suitable
inorganic anions include, but are not limited to, those derived
from the following inorganic acids: hydrochloric, hydrobromic,
hydroiodic, sulphuric, sulphurous, nitric, nitrous, phosphoric, and
phosphorous. Examples of suitable organic anions include, but are
not limited to, those derived from the following organic acids:
acetic, propionic, succinic, glycolic, stearic, palmitic, lactic,
malic, pamoic, tartaric, citric, gluconic, ascorbic, maleic,
hydroxymaleic, phenylacetic, glutamic, aspartic, benzoic, cinnamic,
pyruvic, salicyclic, sulfanilic, 2-acetyoxybenzoic, fumaric,
phenylsulfonic, toluenesulfonic, methanesulfonic, ethanesulfonic,
ethane disulfonic, oxalic, pantothenic, isethionic, valeric,
lactobionic, and gluconic. Examples of suitable polymeric anions
include, but are not limited to, those derived from the following
polymeric acids: tannic acid, carboxymethyl cellulose.
[0135] It may be convenient or desirable to prepare, purify, and/or
handle a corresponding solvate of the active compound. The term
"solvate" is used herein in the conventional sense to refer to a
complex of solute (e.g. active compound, salt of active compound)
and solvent. If the solvent is water, the solvate may be
conveniently referred to as a hydrate, for example, a mono-hydrate,
a di-hydrate, a tri-hydrate, etc.
[0136] It may be convenient or desirable to prepare, purify, and/or
handle the active compound in a chemically protected form. The term
"chemically protected form", as used herein, pertains to a compound
in which one or more reactive functional groups are protected from
undesirable chemical reactions, that is, are in the form of a
protected or protecting group (also known as a masked or masking
group or a blocked or blocking group). By protecting a reactive
functional group, reactions involving other unprotected reactive
functional groups can be performed, without affecting the protected
group; the protecting group may be removed, usually in a subsequent
step, without substantially affecting the remainder of the
molecule. See, for example, Protective Groups in Organic Synthesis
(T. Green and P. Wuts, Wiley, 1 999).
[0137] For example, a hydroxy group may be protected as an ether
(--OR) or an ester (--OC(.dbd.O)R), for example, as: a t-butyl
ether; a benzyl, benzhydryl (diphenylmethyl), or trityl
(triphenylmethyl) ether; a trimethylsilyl or t-butyldimethylsilyl
ether; or an acetyl ester (--OC(.dbd.O)CH.sub.3, --OAc).
[0138] For example, an aldehyde or ketone group may be protected as
an acetal or ketal, respectively, in which the carbonyl group
(>C.dbd.O) is converted to a diether (>C(OR).sub.2), by
reaction with, for example, a primary alcohol. The aldehyde or
ketone group is readily regenerated by hydrolysis using a large
excess of water in the presence of acid.
[0139] For example, an amine group may be protected, for example,
as an amide or a urethane, for example, as: a methyl amide
(--NHCO--CH.sub.3); a benzyloxy amide
(--NHCO--OCH.sub.2C.sub.6H.sub.5, --NH--Cbz); as a t-butoxy amide
(--NHCO--OC(CH.sub.3).sub.3, --NH--Boc); a 2-biphenyl-2-propoxy
amide (--NHCO--OC(CH.sub.3).sub.2C.sub.6H.sub.4C.sub.6H.sub.5,
--NH--Bpoc), as a 9-fluorenylmethoxy amide (--NH--Fmoc), as a
6-nitroveratryloxy amide (--NH--Nvoc), as a
2-trimethylsilylethyloxy amide (--NH--Teoc), as a
2,2,2-trichloroethyloxy amide (--NH-Troc), as an allyloxy amide
(--NH--Alloc), as a 2(-phenylsulphonyl)ethyloxy amide (--NH--Psec);
or, in suitable cases, as an N-oxide (>NO$).
[0140] For example, a carboxylic acid group may be protected as an
ester for example, as: an C.sub.1-7 alkyl ester (e.g. a methyl
ester; a t-butyl ester); a C.sub.1-7 haloalkyl ester (e.g., a
C.sub.1-7 trihaloalkyl ester); a triC.sub.1-7 alkylsilyl-C.sub.1-7
alkyl ester; or a C.sub.5-20 aryl-C.sub.1-7 alkyl ester (e.g. a
benzyl ester; a nitrobenzyl ester); or as an amide, for example, as
a methyl amide.
[0141] For example, a thiol group may be protected as a thioether
(--SR), for example, as: a benzyl thioether; an acetamidomethyl
ether (--S--CH.sub.2NHC(.dbd.O)CH.sub.3).
[0142] It may be convenient or desirable to prepare, purify, and/or
handle the active compound in the form of a prodrug. The term
"prodrug", as used herein, pertains to a compound which, when
metabolised (e.g. in vivo), yields the desired active compound.
Typically, the prodrug is inactive, or less active than the active
compound, but may provide advantageous handling, administration, or
metabolic properties.
[0143] For example, some prodrugs are esters of the active compound
(e.g. a physiologically acceptable metabolically labile ester).
During metabolism, the ester group (--C(.dbd.O)OR) is cleaved to
yield the active drug. Such esters may be formed by esterification,
for example, of any of the carboxylic acid groups (--C(.dbd.O)OH)
in the parent compound, with, where appropriate, prior protection
of any other reactive groups present in the parent compound,
followed by deprotection if required. Examples of such
metabolically labile esters include those wherein R is C.sub.1-7
alkyl (e.g. -Me, -Et); C.sub.1-7 aminoalkyl (e.g. aminoethyl;
2-(N,N-diethylamino)ethyl; 2-(4-morpholino)ethyl); and
acyloxy-C.sub.1-7 alkyl (e.g. acyloxymethyl; acyloxyethyl; e.g.
pivaloyloxymethyl; acetoxymethyl; 1-acetoxyethyl;
1-(1-methoxy-1-methyl)ethyl-carbonxyloxyethyl; 1-(benzoyloxy)ethyl;
isopropoxy-carbonyloxymethyl; 1-isopropoxy-carbonyloxyethyl;
cyclohexyl-carbonyloxymethyl; 1-cyclohexyl-carbonyloxyethyl;
cyclohexyloxy-carbonyloxymethyl; 1-cyclohexyloxy-carbonyloxyethyl;
(4-tetrahydropyranyloxy)carbonyloxymethyl;
1-(4-tetrahydropyranyloxy)carbonyloxyethyl;
(4-tetrahydropyranyl)carbonyloxymethyl; and
1-(4-tetrahydropyranyl)carbonyloxyethyl).
[0144] Also, some prodrugs are activated enzymatically to yield the
active compound, or a compound which, upon further chemical
reaction, yields the active compound. For example, the prodrug may
be a sugar derivative or other glycoside conjugate, or may be an
amino acid ester derivative.
Selective Inhibition
[0145] `Selective inhibition` means the inhibition of one enzyme to
a greater extent than the inhibition of one or more other enzymes.
This selectivity is measurable by comparing the concentration of a
compound required to inhibit 50% of the activity (IC.sub.50) of one
enzyme against the concentration of the same compound required to
inhibit 50% of the activity (IC.sub.50) of the other enzyme (see
below). The result is expressed as a ratio. If the ratio is greater
than 1, then the compound tested exhibits some selectivity in its
inhibitory action.
[0146] The compounds of the present invention preferably exhibit a
selectivity of greater than 3, 10, 20 or 50 against DNA-PK over PI
3-kinase.
[0147] The compounds of the present invention preferably exhibit a
selectivity of greater than 5, 10, 50 or 100 against DNA-PK over
ATM.
[0148] It is preferred that the IC.sub.50 values used to assess
selectivity are determined using the methods described in WO
03/024949, which is herein incorporated by reference.
Further Preferences
[0149] Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5 and Z6
[0150] When Z.sup.5 is not a single bond, Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5 and Z.sup.6 and the carbon atom to which Z.sup.2
and Z.sup.6 are bound, form a six-memebered aromatic ring, and it
is preferred that one or two of Z.sup.2, Z.sup.4, Z.sup.5 and
Z.sup.6 are N and the rest are CH. When Z.sup.5 is a single bond,
Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5 and Z.sup.6 and the carbon atom
to which Z.sup.2 and Z.sup.6 are bound, form a five-memebered
aromatic ring, and it is preferred that one or two of Z.sup.2,
Z.sup.4 and Z.sup.6 are selected from S, O and N and that the rest
are CH. It may be preferred that one of Z.sup.2, Z.sup.4 and
Z.sup.6 is selected from O and S, and that the others are both CH
or one is N and the other CH.
[0151] Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5 and Z.sup.6, together
with the carbon atom to which they are bound, preferably form a
substituted aryl group selected from substituted phenyl,
thiophenyl, furanyl, thiazolyl, imidazolyl, pyridyl, pyrimidinyl,
isoxazolyl, oxazolyl, isothiazolyl. More preferably they form a
group selected from substituted phenyl, thiazolyl, thiophenyl, or
pyridyl.
[0152] Z.sup.2 is preferably S or CR.sup.2, where R.sup.2 is
preferably H.
[0153] Z.sup.3 is preferably CR.sup.3. R.sup.3is preferably
optionally substituted C.sub.5-20 aryl, more preferably C.sub.5-6
aryl.
[0154] Some preferred embodiments have R.sup.3 as C.sub.5
heteroaryl, pyridyl and phenyl, of which phenyl is most preferred.
R.sup.3 is preferably unsubstituted.
[0155] In embodiments where R.sup.3 is C.sub.5-20 aryl, it may
include one or more fused rings. In these embodiments, R.sup.3 may
preferably be selected from naphthyl, indolyl, quinolinyl,
isoquinolinyl
[0156] In embodiments where R.sup.3 is C.sub.5 heteroaryl, it is
preferably selected from groups derived from furan, thiophen,
2-methyl-thiophene, 2-nitrothiophene, thiophen-2-ylamine, thiazole,
imidazole, and 1-methyl-1H-imidazole.
[0157] In embodiments where R.sup.3 is substituted aryl, the
optional substituents are preferably selected from halo (most
preferably fluoro), C.sub.5-20 aryl, R, OR, SO.sub.2R and COR,
where R is C.sub.1-7 alkyl.
[0158] Z.sup.4 is preferably N or CR.sup.4, where R.sup.4is H or
Q-Y--X
[0159] Z.sup.5 is preferably a direct bond or CH.
[0160] Z.sup.6 is preferably N, S or CH.
[0161] R.sup.4
[0162] When Z.sup.2, Z.sup.3, Z.sup.5 and Z.sup.6 all represent CH,
and Z.sup.4 represents CR.sup.4, it is preferred that R.sup.4 is
Q-Y--X. If at least one of Z.sup.2, Z.sup.3, Z.sup.5 and Z.sup.6 is
O, N or S, it is preferred that R.sup.4 is H.
[0163] When Q is --NH--C(.dbd.O)--, X is preferably
NR.sup.N3R.sup.N4. It is further preferred that Y is an optionally
substituted C.sub.1-3 alkylene group, more preferably an optionally
substituted C.sub.1-2 alkylene group and most preferably a
C.sub.1-2 alkylene group.
[0164] When Q is --O-- and X is NR.sup.N3R.sup.N4, then Y is
preferably an optionally substituted C.sub.1-3 alkylene group, more
preferably an optionally substituted C.sub.1-2 alkylene group and
most preferably a C.sub.1-2 alkylene group.
[0165] In some embodiments, R.sup.N3 and R.sup.N4 are preferably
independently selected from H and optionally substituted C.sub.1-7
alkyl, more preferably H and optionally substituted C.sub.1-4 alkyl
and most preferably H and optionally substituted C.sub.1-3 alkyl
(e.g. methyl, ethyl, iso-propyl, n-propyl). Preferred optional
substitutents include, but are not limited to, hydroxy, methoxy,
--NH.sub.2, optionally substituted C.sub.6-aryl and optionally
substituted C.sub.5-6 heterocyclyl.
[0166] In other embodiments, R.sup.N3 and R.sup.N4 form, together
with the nitrogen atom to which they are attached, an optionally
substituted nitrogen containing heterocylic ring having from 4 to 8
ring atoms. Preferably, the heterocyclic ring has 5 to 7 ring
atoms. Examples of preferred groups include, morpholino,
piperidinyl, piperazinyl, homopiperazinyl and tetrahydropyrrolo,
with piperazinyl being particularly preferred. These groups may be
substituted, and a particularly preferred group is optionally
substituted piperazinyl, where the substituent is preferably on the
4-nitrogen atom. Preferred N-substituents include optionally
substituted C.sub.1-4 alkyl, optionally substituted C.sub.6 aryl
and acyl (with a C.sub.1-4 alkyl group as the acyl
substituent).
[0167] R.sup.N5 and R.sup.N6
[0168] The preferences for R.sup.N5 and R.sup.N6 may be the same as
for R.sup.N3 and R.sup.N4 expressed above.
R.sup.N1 nad R.sup.N2
[0169] In compounds of formula I, when R.sup.N1 and R.sup.N2 form,
along with the nitrogen atom to which they are attached, a
heterocyclic ring having from 4 to 8 atoms, this may form part of a
C.sub.4-20 heterocyclyl group defined above (except with a minimum
of 4 ring atoms), which must contain at least one nitrogen ring
atom. It is preferred that R.sup.N1 and R.sup.N2 form, along with
the nitrogen atom to which they are attached, a heterocyclic ring
having 5, 6 or 7 atoms, more preferably 6 ring atoms.
[0170] Single rings having one nitrogen atom include azetidine,
azetidine, pyrrolidine (tetrahydropyrrole), pyrroline (e.g.,
3-pyrroline, 2,5-dihydropyrrole), 2H-pyrrole or 3H-pyrrole
(isopyrrole, isoazole), piperidine, dihydropyridine,
tetrahydropyridine, and azepine; two nitrogen atoms include
imidazolidine, pyrazolidine (diazolidine), imidazoline, pyrazoline
(dihydropyrazole), and piperazine; one nitrogen and one oxygen
include tetrahydrooxazole, dihydrooxazole, tetrahydroisoxazole,
dihydroisoxazole, morpholine, tetrahydrooxazine, dihydrooxazine,
and oxazine; one nitrogen and one sulphur include thiazoline,
thiazolidine, and thiomorpholine.
[0171] Preferred rings are those containing one heteroatom in
addition to the nitrogen, and in particular, the preferred
heteroatoms are oxygen and sulphur. Thus preferred groups include
morpholino, thiomorpholino, thiazolinyl. Preferred groups without a
further heteroatom include pyrrolidino.
[0172] The most preferred groups are morpholino and
thiomorpholino.
[0173] As mentioned above, these heterocyclic groups may themselves
be substituted; a preferred class of substituent is a C.sub.1-7
alkyl group. When the heterocyclic group is morpholino, the
substituent group or groups are preferably methyl or ethyl, and
more preferably methyl. A sole methyl substituent is most
preferably in the 2 position.
[0174] As well as the single ring groups listed above, rings with
bridges or cross-links are also envisaged. Examples of these types
of ring where the group contains a nitrogen and an oxygen atom are:
##STR16##
[0175] These are named 8-oxa-3-aza-bicyclo[3.2.1]oct-3-yl,
6-oxa-3-aza-bicyclo[3.1.0]hex-3-yl, 2-oxa-5-aza-bicyclo[2.2.1
]hept-5-yl, and 7-oxa-3-aza-bicyclo[4.1.0]hept-3-yl,
respectively.
General Synthesis Methods
[0176] Compounds of formula II, where R.sup.4 is Q-Y--X and Q is
--NH--C(.dbd.O)-- can be represented as Formula 1: ##STR17##
[0177] These compounds, where --Y--X is not C.sub.1-7 alkyl, can be
made from compounds of Formula 2: ##STR18##
[0178] wherein L is a leaving group, for example chloro or bromo,
by reacting with the appropriate amine or thiol. This reaction can
be carried at room temperature, or may be heated, if necessary.
[0179] Compounds of Formula 2 can be synthesised by the reaction of
a compound of Formula 3: ##STR19##
[0180] with a compound of Formula 4: ##STR20##
[0181] in the presence of an organic base, for example,
triethylamine.
[0182] Compounds of Formula 1 where --Y--X is C.sub.1-7 alkyl can
be synthesised by the reaction of a compound of Formula 3 with a
compound of Formula 4a: ##STR21##
[0183] in the presence of an organic base, for example,
triethylamine.
[0184] The compounds of Formula 3 may be synthesised by reducing a
compound of Formula 5: ##STR22##
[0185] using an appropriate reducing agent, for example, zinc in
acetic acid.
[0186] Compounds of Formula 5 can be synthesised by the
Suzuki-Miyaura coupling of compounds of Formulae 6 and 7:
##STR23##
[0187] where X' is a group such as bromo or OTf. The coupling
moieities may be reversed.
[0188] The above methods for the synthesis of compounds of formula
11 may be adapted to the synthesis of compounds of general formula
I.
[0189] Compounds of Formula 7 may be synthesised as follows.
[0190] Compounds of Formula 7a: ##STR24##
[0191] may be synthesised from compounds of Formula 8a:
##STR25##
[0192] by cyclocondensation via pyrolysis.
[0193] The compounds of Formula 8a may be synthesised from a
compound of Formula 9a: ##STR26##
[0194] by reaction with the appropriate amine of formula
HNR.sup.N1R.sup.N2, in an appropriate solvent.
[0195] The compound of Formula 9a can be synthesised from a
compound of Formula 10a: ##STR27##
[0196] by reaction with a Meldrum's acid derivative of Formula 11a:
##STR28##
[0197] in an appropriate solvent.
[0198] Compounds of Formula 7b: ##STR29##
[0199] may be synthesised by reacting a compound of Formula 8b:
##STR30##
[0200] with triflic anhydride, in a solvent such as DCM, in the
presence of a base, such as triethylamine.
[0201] Compounds of Formula 8b may be synthesised from a compound
of Formula 9b: ##STR31##
[0202] by nuceleophilic substitution of the chloride by an amine of
formula HNR.sup.1R.sup.2.
[0203] The compound of Formula 9b can be synthesised from a
compound of Formula 10b: ##STR32##
[0204] by chlorination using a chlorinating agent, e.g. POCl.sub.3.
The compound of Formula 10b may be synthesised from a compound of
Formula 11b: ##STR33##
[0205] by reaction with diethyl malonate, or an equivalent
thereof.
[0206] Compounds of Formula 7c: ##STR34##
[0207] Routes to compounds of Formula 7c are described in WO
03/024949 (Synthesis Route 6). Compounds of formula 11, where
R.sup.4 is Q-Y--X, Q is --O-- and X is selected from SR.sup.S1 or
NR.sup.N3R.sup.N4 can be represented as Formula 13: ##STR35##
[0208] wherein X'' represents SR.sup.S1 or NR.sup.N3R.sup.N4. These
compounds may be synthesised from compounds of Formula 14:
##STR36##
[0209] wherein L is a leaving group, for example chloro or bromo,
by reacting with the appropriate 10 amine or thiol. This reaction
can be carried at room temperature, or may be heated, if
necessary.
[0210] Compounds of Formula 14 may be synthesised by the reaction
of a compound of Formula 15: ##STR37##
[0211] with a compound of Formula 16: ##STR38##
[0212] wherein, if Y is non-symmetrical, it is preferred that L is
not Br, in the presence of, for example, potassium carbonate.
[0213] Compounds of Formula 15 can be synthesised from compounds of
Formula 3 using a diazotisation-hydrolysis procedure. This first
converts the amino group into the diazonium fluoroborate salt, for
example, using HBF.sub.4 and butyl nitrite, which is then
hydrolysed using, for example, aqueous copper (I) oxide-copper (II)
nitrate.
[0214] Compounds of formula II, where Q is --O-- and X is
--C(.dbd.O)--NR.sup.N5R.sup.N6 can be represented as Formula 17:
##STR39##
[0215] wherein X''' represents NR.sup.N5R.sup.N6. These compounds
may be synthesised from compounds of Formula 18: ##STR40##
[0216] by reaction with the appropriate amine in the presence of
HBTU and HOBT.
[0217] Compounds of Formula 18 may be made from compounds of
Formula 19: ##STR41##
[0218] by reaction with sodium hydroxide in methanol. The compounds
of Formula 19 may be synthesised from compounds of Formula 15 by
reaction with a compound of Formula 20: ##STR42##
[0219] in the presence of, for example, potassium carbonate.
[0220] Compounds of the present invention where R.sup.4 is H may be
made by the coupling of an appropriate boronic acid to a compound
of Formula 7, in an analagous way to that described above.
[0221] Compounds of the present invention where Z.sup.3 is CR.sup.3
and R.sup.3 is a C.sub.5-6 aryl group may be synthesised by a
double Suzuki coupling of compounds of Formulae 7, 21 and 22:
##STR43##
[0222] where Ar' represents the C.sub.5-6 aryl group and X.sup.2 is
an appropriate halide.
[0223] In particular, compounds of the present invention where
Z.sup.3 is CR.sup.3; Z.sup.2, Z.sup.4, Z.sup.5 and Z.sup.6 are all
CH; A, B, D represent CH, O, C respectively; and R.sup.3 is a
C.sub.5-6 aryl group may be synthesised by a double Suzuki coupling
of compounds 7c, 21a and 22a: ##STR44##
[0224] where Ar represents the C.sub.5-6 aryl group and X is an
appropriate halide.
USE OF COMPOUNDS OF THE INVENTION
[0225] The present invention provides active compounds,
specifically, active 8-aryl-2-amin-4-yl-quinolin-4-ones,
pyridopyrimidine-4-ones, and chromen-4-ones.
[0226] The term "active", as used herein, pertains to compounds
which are capable of inhibiting DNA-PK activity, and specifically
includes both compounds with intrinsic activity (drugs) as well as
prodrugs of such compounds, which prodrugs may themselves exhibit
little or no intrinsic activity.
[0227] One assay which may be used in order to assess the DNA-PK
inhibition offered by a particular compound is described in the
examples below.
[0228] The present invention further provides a method of
inhibiting DNA-PK inhibition in a cell, comprising contacting said
cell with an effective amount of an active compound, preferably in
the form of a pharmaceutically acceptable composition. Such a
method may be practised in vitro or in vivo.
[0229] For example, a sample of cells (e.g. from a tumour) may be
grown in vitro and an active compound brought into contact with
said cells in conjunction with agents that have a known curative
effect, and the enhancement of the curative effect of the compound
on those cells observed.
[0230] The present invention further provides active compounds
which inhibit DNA-PK activity as well as methods of methods of
inhibiting DNA-PK activity comprising contacting a cell with an
effective amount of an active compound, whether in vitro or in
vivo.
[0231] Active compounds may also be used as cell culture additives
to inhibit DNA-PK, for example, in order to sensitize cells to
known chemotherapeutic agents or ionising radiation treatments in
vitro.
[0232] Active compounds may also be used as part of an in vitro
assay, for example, in order to determine whether a candidate host
is likely to benefit from treatment with the compound in
question.
[0233] The invention further provides active compounds for use in a
method of treatment of the human or animal body. Such a method may
comprise administering to such a subject a
therapeutically-effective amount of an active compound, preferably
in the form of a pharmaceutical composition.
[0234] The term "treatment", as used herein in the context of
treating a condition, pertains generally to treatment and therapy,
whether of a human or an animal (e.g. in veterinary applications),
in which some desired therapeutic effect is achieved, for example,
the inhibition of the progress of the condition, and includes a
reduction in the rate of progress, a halt in the rate of progress,
amelioration of the condition, and cure of the condition. Treatment
as a prophylactic measure (i.e. prophylaxis) is also included.
[0235] The term "therapeutically-effective amount" as used herein,
pertains to that amount of an active compound, or a material,
composition or dosage from comprising an active compound, which is
effective for producing some desired therapeutic effect,
commensurate with a reasonable benefit/risk ratio.
[0236] The term "adjunct" as used herein relates to the use of
active compounds in conjunction with known therapeutic means. Such
means include cytotoxic regimes of drugs and/or ionising radiation
as used in the treatment of different cancer types. Examples of
adjunct anti-cancer agents that could be combined with compounds
from the invention include, but are not limited to, the following:
alkylating agents: nitrogen mustards, mechlorethamine,
cyclophosphamide, ifosfamide, melphalan, chlorambucil:
Nitrosoureas: carmustine (BCNU), lomustine (CCNU), semustine
(methyl-CCNU), ethylenimine/methylmelamine, thriethylenemelamine
(TEM), triethylene thiophosphoramide (thiotepa), hexamethylmelamine
(HMM, altretamine): Alkyl sufonates; busulfan; Triazines,
dacarbazine (DTIC): Antimetabolites; folic acid analogs,
methotrexate, trimetrexate, pyrimidine analogs, 5-fluorouracil,
fluorodeoxyuridine, gemcitabine, cytosine arabinoside (AraC,
cytarabine), 5-azacytidine, 2,2'-difluorodeoxycytidine: Purine
analogs; 6-mercaptopurine, 6-thioguanine, azathioprine,
2'-deoxycoformycin (pentostatin, erythrohydroxynonyladenine (EHNA),
fludarabine phosphate, 2-Chlorodeoxyadenosine (cladribine, 2-CdA):
Topoisomerase I inhibitors; camptothecin, topotecan, irinotecan,
rubitecan: Natural products; antimitotic drugs, paclitaxel, vinca
alkaloids, vinblastine (VLB), vincristine, vinorelbine,
Taxotere.TM. (docetaxel), estramustine, estramustine phosphate;
epipodophylotoxins, etoposide, teniposide: Antibiotics; actimomycin
D, daunomycin (rubidomycin), doxorubicin (adriamycin),
mitoxantrone, idarubicin, bleomycins, plicamycin (mithramycin),
mitomycin C, dactinomycin: Enzymes; L-asparaginase, RNAse A:
Biological response modifiers; interferon-alpha, IL-2, G-CSF,
GM-CSF: Differentiation Agents; retinoic acid derivatives:
Radiosensitizers;, metronidazole, misonidazole,
desmethylmisonidazole, pimonidazole, etanidazole, nimorazole, RSU
1069, E09, RB 6145, SR4233, nicotinamide, 5-bromodeozyuridine,
5-iododeoxyuridine, bromodeoxycytidine: Platinium coordination
complexes; cisplatin, carboplatin: Anthracenedione; mitoxantrone,
AQ4N Substituted urea, hydroxyurea; Methylhydrazine derivatives,
N-methylhydrazine (MIH), procarbazine; Adrenocortical suppressant,
mitotane (o.p'-DDD), aminoglutethimide: Cytokines; interferon
(.alpha., .beta., .gamma.), interleukin; Hormones and antagonists;
adrenocorticosteroids/antagonists, prednisone and equivalents,
dexamethasone, aminoglutethimide; Progestins, hydroxyprogesterone
caproate, medroxyprogesterone acetate, megestrol acetate;
Estrogens, diethylstilbestrol, ethynyl estradiol/equivalents;
Antiestrogen, tamoxifen; Androgens, testosterone propionate,
fluoxymesterone/equivalents; Antiandrogens, flutamide,
gonadotropin-releasing hormone analogs, leuprolide; Nonsteroidal
antiandrogens, flutamide; EGFR inhibitors, VEGF inhibitors;
Proteasome inhibitors.
Cancer
[0237] The present invention provides active compounds which are
anticancer agents or adjuncts for treating cancer. One of ordinary
skill in the art is readily able to determine whether or not a
candidate compound treats a cancerous condition for any particular
cell type, either alone or in combination.
[0238] Examples of cancers include, but are not limited to, lung
cancer, small cell lung cancer, gastrointestinal cancer, bowel
cancer, colon cancer, breast carinoma, ovarian carcinoma, prostate
cancer, testicular cancer, liver cancer, kidney cancer, bladder
cancer, pancreas cancer, brain cancer, sarcoma, osteosarcoma,
Kaposi's sarcoma, melanoma and leukemias.
[0239] Any type of cell may be treated, including but not limited
to, lung, gastrointestinal (including, e.g., bowel, colon), breast
(mammary), ovarian, prostate, liver (hepatic), kidney (renal),
bladder, pancreas, brain, and skin.
[0240] The anti cancer treatment defined hereinbefore may be
applied as a sole therapy or may involve, in addition to the
compound of the invention, conventional surgery or radiotherapy or
chemotherapy. Such chemotherapy may include one or more of the
following categories of anti-tumour agents:--
[0241] (i) other antiproliferative/antineoplastic drugs and
combinations thereof, as used in medical oncology, such as
alkylating agents (for example cis platin, oxaliplatin,
carboplatin, cyclophosphamide, nitrogen mustard, melphalan,
chlorambucil, busulphan, temozolamide and nitrosoureas);
antimetabolites (for example gemcitabine and antifolates such as
fluoropyrimidines like 5 fluorouracil and tegafur, raltitrexed,
methotrexate, cytosine arabinoside, and hydroxyurea); antitumour
antibiotics (for example anthracyclines like adriamycin, bleomycin,
doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C,
dactinomycin and mithramycin); antimitotic agents (for example
vinca alkaloids like vincristine, vinblastine, vindesine and
vinorelbine and taxoids like taxol and taxotere and polokinase
inhibitors); and topoisomerase inhibitors (for example
epipodophyllotoxins like etoposide and teniposide, amsacrine,
topotecan and camptothecin);
[0242] (ii) cytostatic agents such as antioestrogens (for example
tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene and
iodoxyfene), antiandrogens (for example bicalutamide, flutamide,
nilutamide and cyproterone acetate), LHRH antagonists or LHRH
agonists (for example goserelin, leuprorelin and buserelin),
progestogens (for example megestrol acetate), aromatase inhibitors
(for example as anastrozole, letrozole, vorazole and exemestane)
and inhibitors of 5*-reductase such as finasteride;
[0243] (iii) anti-invasion agents (for example c-Src kinase family
inhibitors like
4-(6-chloro-2,3-methylenedioxyanilino)-7-[2-(4-methylpiperazin-1-yl)ethox-
y]-5-tetrahydropyran-4-yloxyquinazoline (AZD0530; International
Patent Application WO 01/94341) and
N-(2-chloro-6-methylphenyl)-2-{6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-met-
hylpyrimidin4-ylamino}thiazole-5-carboxamide (dasatinib,
BMS-354825; J. Med. Chem., 2004, 47, 6658-6661), and
metalloproteinase inhibitors like marimastat, inhibitors of
urokinase plasminogen activator receptor function or antibodies to
Heparanase);
[0244] (iv) inhibitors of growth factor function: for example such
inhibitors include growth factor antibodies and growth factor
receptor antibodies (for example the anti erbB2 antibody
trastuzumab [HerceptinT], the anti-EGFR antibody panitumumab, the
anti erbB1 antibody cetuximab [Erbitux, C225] and any growth factor
or growth factor receptor antibodies disclosed by Stern et al.
Critical reviews in oncology/haematology, 2005, Vol. 54, pp11-29);
such inhibitors also include tyrosine kinase inhibitors, for
example inhibitors of the epidermal growth factor family (for
example EGFR family tyrosine kinase inhibitors such as
N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-
-amine (gefitinib, ZD 1839),
N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine
(erlotinib, OSI 774) and
6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)-quinazol-
in-4-amine (Cl 1033), erbB2 tyrosine kinase inhibitors such as
lapatinib, inhibitors of the hepatocyte growth factor family,
inhibitors of the platelet-derived growth factor family such as
imatinib, inhibitors of serine/threonine kinases (for example
Ras/Raf signalling inhibitors such as farnesyl transferase
inhibitors, for example sorafenib (BAY 43-9006)), inhibitors of
cell signalling through MEK and/or AKT kinases, inhibitors of the
hepatocyte growth factor family, c-kit inhibitors, abl kinase
inhibitors, IGF receptor (insulin-like growth factor) kinase
inhibitors; aurora kinase inhibitors (for example AZD1 152,
PH739358, VX-680, MLN8054, R763, MP235, MP529, VX-528 AND AX39459)
and cyclin dependent kinase inhibitors such as CDK2 and/or CDK4
inhibitors;
[0245] (v) antiangiogenic agents such as those which inhibit the
effects of vascular endothelial growth factor, [for example the
anti vascular endothelial cell growth factor antibody bevacizumab
(AvastinT) and VEGF receptor tyrosine kinase inhibitors such as
4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)qu-
inazoline (ZD6474; Example 2 within WO 01/32651),
4-(4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-1-ylpropoxy)-
quinazoline (AZD2171; Example 240 within WO 00/47212), vatalanib
(PTK787; WO 98/35985) and SU11248 (sunitinib; WO 01/60814),
compounds such as those disclosed in International Patent
Applications WO97/22596, WO 97/30035, WO 97/32856 and WO 98/13354
and compounds that work by other mechanisms (for example linomide,
inhibitors of integrin avb3 function and angiostatin)];
[0246] (vi) vascular damaging agents such as Combretastatin A4 and
compounds disclosed in International Patent Applications WO
99/02166, WO 00/40529, WO 00/41669, WO 01/92224, WO 02/04434 and WO
02/08213;
[0247] (vii) antisense therapies, for example those which are
directed to the targets listed above, such as ISIS 2503, an
anti-ras antisense;
[0248] (viii) gene therapy approaches, including for example
approaches to replace aberrant genes such as aberrant p53 or
aberrant BRCA1 or BRCA2, GDEPT (gene directed enzyme pro drug
therapy) approaches such as those using cytosine deaminase,
thymidine kinase or a bacterial nitroreductase enzyme and
approaches to increase patient tolerance to chemotherapy or
radiotherapy such as multi drug resistance gene therapy; and
[0249] (ix) immunotherapy approaches, including for example ex vivo
and in vivo approaches to increase the immunogenicity of patient
tumour cells, such as transfection with cytokines such as
interleukin 2, interleukin 4 or granulocyte macrophage colony
stimulating factor, approaches to decrease T cell anergy,
approaches using transfected immune cells such as cytokine
transfected dendritic cells, approaches using cytokine transfected
tumour cell lines and approaches using anti idiotypic
antibodies
Administration
[0250] The active compound or pharmaceutical composition comprising
the active compound may be administered to a subject by any
convenient route of administration, whether
systemically/peripherally or at the site of desired action,
including but not limited to, oral (e.g. by ingestion); topical
(including e.g. transdermal, intranasal, ocular, buccal, and
sublingual); pulmonary (e.g. by inhalation or insufflation therapy
using, e.g. an aerosol, e.g. through mouth or nose); rectal;
vaginal; parenteral, for example, by injection, including
subcutaneous, intradermal, intramuscular, intravenous,
intraarterial, intracardiac, intrathecal, intraspinal,
intracapsular, subcapsular, intraorbital, intraperitoneal,
intratracheal, subcuticular, intraarticular, subarachnoid, and
intrasternal; by implant of a depot, for example, subcutaneously or
intramuscularly.
[0251] The subject may be a eukaryote, an animal, a vertebrate
animal, a mammal, a rodent (e.g. a guinea pig, a hamster, a rat, a
mouse), murine (e.g. a mouse), canine (e.g. a dog), feline (e.g. a
cat), equine (e.g. a horse), a primate, simian (e.g. a monkey or
ape), a monkey (e.g. marmoset, baboon), an ape (e.g. gorilla,
chimpanzee, orang-utan, gibbon), or a human.
Formulations
[0252] While it is possible for the active compound to be
administered alone, it is preferable to present it as a
pharmaceutical composition (e.g. formulation) comprising at least
one active compound, as defined above, together with one or more
pharmaceutically acceptable carriers, adjuvants, excipients,
diluents, fillers, buffers, stabilisers, preservatives, lubricants,
or other materials well known to those skilled in the art and
optionally other therapeutic or prophylactic agents.
[0253] Thus, the present invention further provides pharmaceutical
compositions, as defined above, and methods of making a
pharmaceutical composition comprising admixing at least one active
compound, as defined above, together with one or more
pharmaceutically acceptable carriers, excipients, buffers,
adjuvants, stabilisers, or other materials, as described
herein.
[0254] The term "pharmaceutically acceptable" as used herein
pertains to compounds, materials, compositions, and/or dosage forms
which are, within the scope of sound medical judgement, suitable
for use in contact with the tissues of a subject (e.g. human)
without excessive toxicity, irritation, allergic response, or other
problem or complication, commensurate with a reasonable
benefit/risk ratio. Each carrier, excipient, etc. must also be
"acceptable" in the sense of being compatible with the other
ingredients of the formulation.
[0255] Suitable carriers, excipients, etc. can be found in standard
pharmaceutical texts, for example, Remington's Pharmaceutical
Sciences, 18th edition, Mack Publishing Company, Easton, Pa.,
1990.
[0256] The formulations may conveniently be presented in unit
dosage form and may be prepared by any methods well known in the
art of pharmacy. Such methods include the step of bringing into
association the active compound with the carrier which constitutes
one or more accessory ingredients. In general, the formulations are
prepared by uniformly and intimately bringing into association the
active compound with liquid carriers or finely divided solid
carriers or both, and then if necessary shaping the product.
[0257] Formulations may be in the form of liquids, solutions,
suspensions, emulsions, elixirs, syrups, tablets, losenges,
granules, powders, capsules, cachets, pills, ampoules,
suppositories, pessaries, ointments, gels, pastes, creams, sprays,
mists, foams, lotions, oils, boluses, electuaries, or aerosols.
[0258] Formulations suitable for oral administration (e.g. by
ingestion) may be presented as discrete units such as capsules,
cachets or tablets, each containing a predetermined amount of the
active compound; as a powder or granules; as a solution or
suspension in an aqueous or non-aqueous liquid; or as an
oil-in-water liquid emulsion or a water-in-oil liquid emulsion; as
a bolus; as an electuary; or as a paste.
[0259] A tablet may be made by conventional means, e.g.,
compression or moulding, optionally with one or more accessory
ingredients. Compressed tablets may be prepared by compressing in a
suitable machine the active compound in a free-flowing form such as
a powder or granules, optionally mixed with one or more binders
(e.g. povidone, gelatin, acacia, sorbitol, tragacanth,
hydroxypropylmethyl cellulose); fillers or diluents (e.g. lactose,
microcrystalline cellulose, calcium hydrogen phosphate); lubricants
(e.g. magnesium stearate, talc, silica); disintegrants (e.g. sodium
starch glycolate, cross-linked povidone, cross-linked sodium
carboxymethyl cellulose); surface-active or dispersing or wetting
agents (e.g. sodium lauryl sulfate); and preservatives (e.g. methyl
p-hydroxybenzoate, propyl p-hydroxybenzoate, sorbic acid). Moulded
tablets may be made by moulding in a suitable machine a mixture of
the powdered compound moistened with an inert liquid diluent. The
tablets may optionally be coated or scored and may be formulated so
as to provide slow or controlled release of the active compound
therein using, for example, hydroxypropylmethyl cellulose in
varying proportions to provide the desired release profile. Tablets
may optionally be provided with an enteric coating, to provide
release in parts of the gut other than the stomach.
[0260] Formulations suitable for topical administration (e.g.
transdermal, intranasal, ocular, buccal, and sublingual) may be
formulated as an ointment, cream, suspension, lotion, powder,
solution, past, gel, spray, aerosol, or oil. Alternatively, a
formulation may comprise a patch or a dressing such as a bandage or
adhesive plaster impregnated with active compounds and optionally
one or more excipients or diluents.
[0261] Formulations suitable for topical administration in the
mouth include losenges comprising the active compound in a
flavoured basis, usually sucrose and acacia or tragacanth;
pastilles comprising the active compound in an inert basis such as
gelatin and glycerin, or sucrose and acacia; and mouthwashes
comprising the active compound in a suitable liquid carrier.
[0262] Formulations suitable for topical administration to the eye
also include eye drops wherein the active compound is dissolved or
suspended in a suitable carrier, especially an aqueous solvent for
the active compound.
[0263] Formulations suitable for nasal administration, wherein the
carrier is a solid, include a coarse powder having a particle size,
for example, in the range of about 20 to about 500 microns which is
administered in the manner in which snuff is taken, i.e. by rapid
inhalation through the nasal passage from a container of the powder
held close up to the nose. Suitable formulations wherein the
carrier is a liquid for administration as, for example, nasal
spray, nasal drops, or by aerosol administration by nebuliser,
include aqueous or oily solutions of the active compound.
[0264] Formulations suitable for administration by inhalation
include those presented as an aerosol spray from a pressurised
pack, with the use of a suitable propellant, such as
dichlorodifluoromethane, trichlorofluoromethane,
dichoro-tetrafluoroethane, carbon dioxide, or other suitable
gases.
[0265] Formulations suitable for topical administration via the
skin include ointments, creams, and emulsions. When formulated in
an ointment, the active compound may optionally be employed with
either a paraffinic or a water-miscible ointment base.
Alternatively, the active compounds may be formulated in a cream
with an oil-in-water cream base. If desired, the aqueous phase of
the cream base may include, for example, at least about 30% w/w of
a polyhydric alcohol, i.e., an alcohol having two or more hydroxyl
groups such as propylene glycol, butane-1,3-diol, mannitol,
sorbitol, glycerol and polyethylene glycol and mixtures thereof.
The topical formulations may desirably include a compound which
enhances absorption or penetration of the active compound through
the skin or other affected areas. Examples of such dermal
penetration enhancers include dimethylsulfoxide and related
analogues.
[0266] When formulated as a topical emulsion, the oily phase may
optionally comprise merely an emulsifier (otherwise known as an
emulgent), or it may comprises a mixture of at least one emulsifier
with a fat or an oil or with both a fat and an oil. Preferably, a
hydrophilic emulsifier is included together with a lipophilic
emulsifier which acts as a stabiliser. It is also preferred to
include both an oil and a fat. Together, the emulsifier(s) with or
without stabiliser(s) make up the so-called emulsifying wax, and
the wax together with the oil and/or fat make up the so-called
emulsifying ointment base which forms the oily dispersed phase of
the cream formulations.
[0267] Suitable emulgents and emulsion stabilisers include Tween
60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl
monostearate and sodium lauryl sulphate. The choice of suitable
oils or fats for the formulation is based on achieving the desired
cosmetic properties, since the solubility of the active compound in
most oils likely to be used in pharmaceutical emulsion formulations
may be very low. Thus the cream should preferably be a non-greasy,
non-staining and washable product with suitable consistency to
avoid leakage from tubes or other containers. Straight or branched
chain, mono- or dibasic alkyl esters such as di-isoadipate,
isocetyl stearate, propylene glycol diester of coconut fatty acids,
isopropyl myristate, decyl oleate, isopropyl palmitate, butyl
stearate, 2-ethylhexyl palmitate or a blend of branched chain
esters known as Crodamol CAP may be used, the last three being
preferred esters. These may be used alone or in combination
depending on the properties required.
[0268] Alternatively, high melting point lipids such as white soft
paraffin and/or liquid paraffin or other mineral oils can be
used.
[0269] Formulations suitable for rectal administration may be
presented as a suppository with a suitable base comprising, for
example, cocoa butter or a salicylate.
[0270] Formulations suitable for vaginal administration may be
presented as pessaries, tampons, creams, gels, pastes, foams or
spray formulations containing in addition to the active compound,
such carriers as are known in the art to be appropriate.
[0271] Formulations suitable for parenteral administration (e.g. by
injection, including cutaneous, subcutaneous, intramuscular,
intravenous and intradermal), include aqueous and non-aqueous
isotonic, pyrogen-free, sterile injection solutions which may
contain anti-oxidants, buffers, preservatives, stabilisers,
bacteriostats, and solutes which render the formulation isotonic
with the blood of the intended recipient; and aqueous and
non-aqueous sterile suspensions which may include suspending agents
and thickening agents, and liposomes or other microparticulate
systems which are designed to target the compound to blood
components or one or more organs. Examples of suitable isotonic
vehicles for use in such formulations include Sodium Chloride
Injection, Ringer's Solution, or Lactated Ringer's Injection.
Typically, the concentration of the active compound in the solution
is from about 1 ng/ml to about 10 .mu.g/ml, for example from about
10 ng/ml to about 1 .mu.g/ml. The formulations may be presented in
unit-dose or multi-dose sealed containers, for example, ampoules
and vials, and may be stored in a freeze-dried (lyophilised)
condition requiring only the addition of the sterile liquid
carrier, for example water for injections, immediately prior to
use. Extemporaneous injection solutions and suspensions may be
prepared from sterile powders, granules, and tablets. Formulations
may be in the form of liposomes or other microparticulate systems
which are designed to target the active compound to blood
components or one or more organs.
Dosage
[0272] It will be appreciated that appropriate dosages of the
active compounds, and compositions comprising the active compounds,
can vary from patient to patient. Determining the optimal dosage
will generally involve the balancing of the level of therapeutic
benefit against any risk or deleterious side effects of the
treatments of the present invention. The selected dosage level will
depend on a variety of factors including, but not limited to, the
activity of the particular compound, the route of administration,
the time of administration, the rate of excretion of the compound,
the duration of the treatment, other drugs, compounds, and/or
materials used in combination, and the age, sex, weight, condition,
general health, and prior medical history of the patient. The
amount of compound and route of administration will ultimately be
at the discretion of the physician, although generally the dosage
will be to achieve local concentrations at the site of action which
achieve the desired effect without causing substantial harmful or
deleterious side-effects.
[0273] Administration in vivo can be effected in one dose,
continuously or intermittently (e.g. in divided doses at
appropriate intervals) throughout the course of treatment. Methods
of determining the most effective means and dosage of
administration are well known to those of skill in the art and will
vary with the formulation used for therapy, the purpose of the
therapy, the target cell being treated, and the subject being
treated. Single or multiple administrations can be carried out with
the dose level and pattern being selected by the treating
physician.
[0274] In general, a suitable dose of the active compound is in the
range of about 100 .mu.g to about 250 mg per kilogram body weight
of the subject per day. Where the active compound is a salt, an
ester, prodrug, or the like, the amount administered is calculated
on the basis of the parent compound and so the actual weight to be
used is increased proportionately.
EXAMPLES
[0275] The following are examples are provided solely to illustrate
the present invention and are not intended to limit the scope of
the invention, as described herein.
Acronyms
[0276] For convenience, many chemical moieties are represented
using well known abbreviations, including but not limited to,
methyl (Me), ethyl (Et), n-propyl (nPr), iso-propyl (iPr), n-butyl
(nBu), tert-butyl (tBu), n-hexyl (nHex), cyclohexyl (cHex), phenyl
(Ph), biphenyl (biPh), benzyl (Bn), naphthyl (naph), methoxy (MeO),
ethoxy (EtO), benzoyl (Bz), and acetyl (Ac).
[0277] For convenience, many chemical compounds are represented
using well known abbreviations, including but not limited to,
methanol (MeOH), ethanol (EtOH), iso-propanol (i-PrOH), methyl
ethyl ketone (MEK), ether or diethyl ether (Et.sub.2O), acetic acid
(AcOH), dichloromethane (methylene chloride, DCM), trifluoroacetic
acid (TFA), dimethylfonmamide (DMF), tetrahydrofuran (THF), and
dimethylsulfoxide (DMSO).
General Experimental Details
[0278] Chemicals were purchased from the Aldrich Chemical Company,
Lancaster Synthesis Ltd and Acros Organics (Fisher Scientific UK
Ltd). THF was freshly distilled from sodium/benzophenone. Methanol
and ethanol were distilled from magnesium/iodine. DCM was dried by
distillation over phosphorus pentoxide. Acetone was dried by
distillation over calcium hydride. All solvents not used
immediately were stored over molecular sieves (4 .ANG., 3-5 mm
beads), under nitrogen. Anhydrous DMF was obtained from Aldrich in
SureSeal.TM. bottles. Triethylamine was dried by distillation over
calcium hydride and stored over potassium hydroxide, under
nitrogen.
[0279] Thin layer chromatography (TLC), was performed using Merck
silica gel 60F.sub.254 pre-coated on aluminium sheets which were
subsequently dried and visualised using either short wave (254 nm)
ultraviolet light or by treatment with either ninhydrin or
sulphuric acid then vanillin. `Flash` column chromatography was
carried out at medium pressure using Davisil silica gel (40-63
.mu.m).
[0280] Melting points were determined using a Stuart Scientific
SMP3 apparatus and are uncorrected. .sup.1H and .sup.13C nuclear
magnetic resonance (NMR) spectra were obtained using a Bruker
Spectrospin AC 300E spectrometer (.sup.1H 300 MHz or .sup.13C 75
MHz) or a Bruker
[0281] Spectrospin AC 500E spectrometer (.sup.1H 500 MHz or
.sup.13C 125 MHz). Chemical shifts are reported in parts per
million (.delta.) downfield of teramethylsulfone using residual
solvent peaks as internal standards. Multiplicities are indicated
by s (singlet), d (doublet), t (triplet), q (quartet), m
(multiplet), br (broad) or combinations thereof. LC/MS spectra were
obtained using a Micromass Platform instrument running in positive
or negative ion electrospray mode. Separation was achieved using a
C18 column (50.times.4.6 mm; Supelco Discovery or Waters Symmetry)
and a 15 minute gradient elution of 0.05% formic acid and methanol
(10-90%). IR spectra were recorded on a Bio-Rad FTS 3000MX diamond
ATR as a neat sample.
Synthesis of Key Intermediates
(ii) Synthesis of
9-triflyl-2-morpholin-4-yl-pyrido[2,1-a]pyrimidin4-one (5)
[0282] ##STR45##
(a) 2,9-dihydroxy-pyrido[2,1-a]pyrimidin-4-one (2)
[0283] A mixture of malonic acid bis-(2,4,6-trichloro-phenyl) ester
(17.33 g; 37.5 mmol) and 3-hydroxy-2-amino-pyridine (1)(4.12 g;
37.5 mmol) dissolved in bromobenzene (37 mL) was heated at reflux
for 3 hours. Upon cooling, the reaction mixture was filtered and
the solid was washed with ethanol. The solid was solubilised in 1 M
NaOH and drops of AcOH were added to precipitate the product as a
pale yellow solid (6.53 g). Yield=98%. m.p.: 320.degree. C.
(degradation); R.sub.1=0.11, MeOH:DCM (3:17); UV:
.lamda..sub.max=252 nm; IR: (cm.sup.-1) 2862, 1688, 1564, 1374,
1295, 1102, 783; .sup.1H NMR, (DMSO, 300 MHz), .delta. (ppm): 5.22
(1H, s, CH-3), 7.12 (1H, t, J.sub.H6-H7=7 Hz, H.sub.arom-7), 7.27
(1H, d, J.sub.H7-H8=8 Hz, H.sub.arom-8), 8.43 (1H, d, J.sub.H6-H7=7
Hz, H.sub.arom-6); .sup.13C NMR, (CDCl.sub.3, 75 MHz), .delta.
(ppm): 103.25, 116.46, 117.05, 119.03, 143.97, 148.82, 157.26,
157.50.
(b) 2-Chloro-9-hydroxy-pyrido[2,1-a]pyrimidin-4-one (3)
[0284] In a round bottom flask,
2,9-dihydroxy-pyrido[2,1-a]pyrimidin-4-one (2)(1.07 g; 6.0 mmol)
was dissolved in phosphorous oxychloride (7.5 mL). This solution
was heated to reflux for 48 hours. Upon cooling, the reaction
mixture was poured carefully into ice cold water (100 mL) and
adjusted to pH 7 by addition of a saturated solution of sodium
carbonate. The aqueous layer was extracted with dichloromethane.
The organic layer was dried over magnesium sulphate and evaporated
to yield a brown solid. This solid was purified by flash
chromatography using dichloromethane as eluant to furnish the title
compound as a white solid (712 mg). Yield.dbd.60%. m.p.:
162.degree. C.; R.sub.f=0.34, MeOH:DCM (1:19); Mass Spec.: (m/z)
196.93 [M+1].sup.+ (Rt=4.67 min, 12 min gradient); UV:
.lamda..sub.max=210 nm; IR: (cm.sup.-1) 3103, 1684, 1630, 1511,
1458, 1297, 1105; .sup.1H NMR, (CDCl.sub.3, 300 MHz), .delta.
(ppm): 6.4 (1H, s, CH-3), 7.11 (1H, t, J.sub.H6-H7=7 Hz,
H.sub.arom-7), 7.25 (1H, d, J.sub.H7-H8=8 Hz, H.sub.arom-8), 8.51
(1H, d, J.sub.H6-H7=7 Hz, H.sub.arom-6); .sup.13C NMR, (CDCl.sub.3,
75 MHz), .delta. (ppm): 103.25, 116.46, 117.05, 119.03, 143.97,
148.82, 157.26, 157.50.
(c) 9-Hydroxy-2-morpholin-4-yl-pyrido[2,1-a]pyrimidin-4-one (4)
[0285] In a round bottom flask,
2-chloro-9-hydroxy-pyrido[2,1-a]pyrimidin-4-one (3)(141.7 mg; 0.721
mmol) and morpholine (314 .mu.L; 3.605 mmol) were dissolved in
ethanol (5 mL). This solution was heated to reflux for 18 hours
with vigorous stirring. Upon cooling, the solvent was evaporated.
The yellow raw solid was recrystallised in ethanol, giving 173.8 mg
of white crystals. Yield=97%. m.p.: 245.degree. C.; R.sub.f=0.27,
MeOH:DCM (1:19); Mass Spec.: (m/z) 248.08 [M+1].sup.+ (Rt=4.92 min,
12 min gradient); UV: .lamda..sub.max=267 nm ; IR: (cm.sup.-1)
3302, 1690, 1644, 1551, 1427, 1224, 1110.; .sup.1H NMR,
(CDCl.sub.3, 500 MHz), .delta. (ppm): 3.56 (4H, m,
N--CH.sub.2-morpholine), 3.75 (4H, m, O--CH.sub.2-morpholine), 5.55
(1H, s, CH-3), 6.80 (1H, t, J.sub.H6-H7=7 Hz, H.sub.arm-7), 7.02
(1H, dd, J.sub.H7-H8=8 Hz, J.sub.H6-H8=1.3 Hz, H.sub.arom-8), 7.33
(1H, s, OH), 8.37 (1H, dd, J.sub.H6-H7=7 Hz, J.sub.H6-H8=1.3 Hz,
H.sub.arom-6); .sup.13C NMR, (CDCl.sub.3, 125 MHz), .delta. (ppm):
45.27, 67.02, 82.16, 113.46, 114.18, 119.50, 143.00, 147.51,
159.00, 161.00.
(d) 9-Triflate-2-morpholin-4-yl-pyrido[2,1-a]pyrimidin-4-one
(5)
[0286] In a three-neck round bottom flask with a thermometer,
9-hydroxy-2-morpholin-4-yl-pyrido[2,1-a]pyrimidin-4-one (4) (2.11
g; 8.54 mmol) was solubilised in DCM (70 mL), cooled to -30.degree.
C. and triethylamine (3.572 mL; 25.63 mmol) added. After 5 minutes
triflic anhydride (2.101 mL; 12.81 mmol), solubilised in 10 ml of
DCM, was added dropwise to the reaction mixture over 30 minutes,
via an addition funnel. The temperature of the reaction mixture was
kept under -20.degree. C. during the addition. After 3 hours, the
reaction mixture was washed with a saturated solution of
Na.sub.2CO.sub.3 (50 mL) and extracted with DCM (3.times.30 mL).
The organic layer was dried over magnesium sulphate and evaporated
to yield a brown solid. This solid was purified by flash
chromatography using dichloromethane as eluant to furnish the title
compound as an orange solid (2.91 g). Yield=90%. m.p.:
146-147.degree. C.; R.sub.f=0.42; MeOH:DCM (1:19); Mass Spec.:
(m/z) 380.16 [M+1].sup.+ (Rt=3.34 min, 12 min gradient); UV:
.lamda..sub.max=271 nm; IR: (cm.sup.-1) 1705, 1644, 1551, 1189,
1112, 939, 769; .sup.1H NMR, (CDCl.sub.3, 300 MHz), .delta. (ppm):
3.56 (4H, m, N--CH.sub.2-morpholine), 3.71 (4H, m,
O--CH.sub.2-morpholine), 5.53 (1H, s, CH-3), 6.80 (1H, t,
J.sub.H6-H7=7 Hz, H.sub.arom-7), 7.46 (1H, dd, J.sub.H7-H8=8 Hz,
J.sub.H6-H8=1.3 Hz, H.sub.arom-8), 8.79 (1H, dd, J.sub.H6-H7=7 Hz,
J.sub.H6-H8=1.3 Hz, H.sub.arom-6); .sup.13C NMR, (CDCl.sub.3, 75
MHz), .delta. (ppm): 45.19, 66.87, 81.76, 110.16, 112.61, 116.85,
121.10, 125.34, 127.86, 128.13, 141.46, 145.79, 158.07, 160.42.
(iv) Synthesis of 8-bromo-2-morpholin-4-yl-1H-quinolin-4-one
(12)
[0287] ##STR46##
(a)
5-(Bis-methylsulfanyl-methylene)-2,2-dimethyl-[1,3]dioxane-4,6-dione
(8)
[0288] In a 250 mL two neck flask, a well stirred solution of
2,2-dimethyl-1,3-dioxane-4,6-dione (6) (Meldrum's acid) (4.09 g;
28.4 mmol) in DMSO (14 mL) was formed. Triethylamine (7.92 mL; 56.8
mmol) and carbon disulfide (1.71 mL; 28.4 mmol) were added to this
mixture in quick succession. The mixture was then stirred
vigorously for 1 hour at room temperature before being cooled in an
ice-bath. Iodomethane (3.54 mL; 56.8 mL) was slowly added to the
reaction mixture with cooling (ice-bath). When the addition was
completed the reaction mixture was allowed to warm to room
temperature and was stirred for a further 4 hours before being
diluted with ice cold water (25 mL). Scratching of the mixture
precipitated the product which was filtered off and washed with
petrol. The product was obtained as a yellow solid (2.76 g) and was
pure enough for use in subsequent reactions. Yield=45%. m.p.:
118.degree. C. (literature.sup.28: 116-118.degree. C.); IR:
(cm.sup.-1) 3728, 1668, 1373, 1302, 1264, 1199; .sup.1H NMR,
(CDCl.sub.3, 300 MHz), .delta. (ppm): 1.54 (6H, s, 2CH.sub.3), 2.58
(6H, s, 2CH.sub.3--S); .sup.12C NMR, (CDCl.sub.3, 75 MHz), .delta.
(ppm): 21.86, 27.22, 103.32, 160.33, 194.
(b)
5-[2-Bromoanilino-(methylthio)-methylene]-2,2-dimethyl-4,6-dione
(10)
[0289] In a 10 mL round bottom flask with cooler and nitrogen
bubbler, isopropylidene bismethylthiolidene malonate (8)(900 mg;
3.63 mmol) and 2-bromoaniline (15)(624 mg; 3.63 mmol) were
dissolved in 2,2,2-trifluoroethanol (3.6 mL). The mixture was
stirred and heated to reflux for 22 hours. Upon cooling, the
solvent was evaporated. The residue was recrystallised from
methanol to yield the title compound as white crystals (1.192 g).
Yield=88%. m.p.: 159.degree. C.; R.sub.f=0.31, DCM; IR: (cm.sup.-1)
2990, 1706, 1653, 1535, 1370, 1199; UV: .lamda..sub.max=313 nm;
.sup.1H NMR, (CDCl.sub.3, 300 MHz), .delta. (ppm): 1.69 (6H, s,
2CH.sub.3), 2.15 (3H, s, CH.sub.3--S), 7.8 (1H, dt, J.sub.H4-H5=8
Hz, J.sub.H4-H6=2 Hz, H.sub.arom-4), 7.35 (2H, m, H.sub.arom-5 and
H.sub.arom-6), 7,61 (1H, dd, J.sub.H3-H4=8 Hz, J.sub.H3-H5=1.2 Hz,
H.sub.arom-3), 12.51 (1H, s, N--H); .sup.13C NMR, (CDCl.sub.3, 75
MHz), .delta. (ppm): 18.75, 26.48, 87.54, 103.32, 120.45, 127.78,
128.46, 129.48, 133.57, 136.91, 163.87, 178.70.
(c)
5-[(2-Bromo-anilino)-morpholin-4-yl-methylene]-2,2-dimethyl-[1,3]dioxa-
ne-4,6-dione (11)
[0290] In a 10 mL round bottom flask with cooler and nitrogen
bubbler,
5-[2-bromoanilino-(methylthio)-methylene]-2,2-dimethyl-4,6-dione
(10)(234 mg; 0.629 mmol) and morpholine (110 .mu.L; 1.257 mmol)
were dissolved in 2,2,2-trifluoroethanol (1 mL). The mixture was
stirred and heated to reflux for 18 hours. Upon cooling, the
solvents were evaporated. The residue was recrystallised from
methanol to yield the title compound as white crystals (0.124 g).
Yield=50%. m.p.: 212-213.degree. C.; R.sub.f=0.05; DCM; IR:
(cm.sup.-1) 1627, 1342, 1305, 1100, 1022, 934; UV:
.lamda..sub.max=241 nm; .sup.1H NMR, (CDCl.sub.3, 300 MHz), .delta.
(ppm): 1.77 (6H, s, 2CH.sub.3), 3.24 (4H, t, J.sub.ab=5 Hz,
2CH.sub.2--N morpholine), 3.66 (4H, t, J.sub.ab=5 Hz, 2CH.sub.2--O
morpholine), 7.18 (2H, m, H.sub.arom-4 and H.sub.arom-6), 7.40 (1H,
t, J.sub.H5-H6=8 Hz, H.sub.arom-5), 7.69 (1H, dd, J.sub.H3-H4=8 Hz,
J.sub.H3-H5=1.4 Hz, H.sub.arom-3), 9.62 (1H, s, N--H); .sup.13C
NMR, (CDCl.sub.3, 75 MHz), .delta. (ppm): 26.83, 51.14, 65.62,
87.54, 102.84, 120.45, 127.15, 128.92, 129.03, 134.48, 138.46,
164.92, 178.70.
(d) 8-Bromo-2-morpholin-4-yl-1H-quinolin-4-one (12)
[0291] In a Schlenk tube,
5-[(2-bromo-anilino)-morpholin-4-yl-methylene]-2,2-dimethyl-[1,3]dioxane--
4,6-dione (11)(103.3 mg; 0.2513 mmol) was dissolved in diphenyl
ether (0.7 mL). The mixture was stirred and heated to reflux for 4
hours. Upon cooling, petroleum ether was added. The product was
collected by suction. The residue was purified by flash
chromatography using dichloromethane/methanol (95:5) as eluant. The
product was obtained as a brown oil (65.1 mg). Yield=84%.
R.sub.f=0.25, MeOH:DCM (1:19); Mass Spec.: (m/z) 310.98 [M+1].sup.+
(Rt=5.24 min, 12 min gradient); IR: (cm.sup.-1) 3395, 2959, 2849,
1617, 1577, 1487, 1421, 1384, 1327, 1263, 1229, 1188, 1152, 1111,
1066, 999, 902, 785; UV: .lamda..sub.max=254 nm; .sup.1H NMR,
(CDCl.sub.3, 300 MHz), .delta. (ppm): 3.72 (4H, t, J.sub.ab=5 Hz,
2CH.sub.2--N morpholine), 3.75 (4H, t, J.sub.ab=5 Hz, 2CH.sub.2--O
morpholine), 5.95 (1H, s, H-3), 7.04 (1H, t, J.sub.H6-H7=8 Hz,
H.sub.arom-6), 7.69 (1H, dd, J.sub.H6-H7=8 Hz, J.sub.H5-H7=1.3 Hz,
H.sub.arom-7), 8.09 (1H, d, J.sub.H5-H6=8 Hz, H.sub.arom-5);
.sup.13C NMR, (CDCl.sub.3, 75 MHz), .delta. (ppm): 46.35, 66.59,
92.50, 114.53, 123, 123.50, 124.73, 134.45, 138, 156.06, 172.6.
Example 1
Parallel Synthesis from 9-Triflate-2-morpholin-4-yl-pyrido[2,1-a]
pyrimidin-4-one (5)
[0292] ##STR47##
[0293] An appropriate boronic acid (0.395 mmol) and potassium
carbonate (109.3 mg; 0.7914 mmol) were introduced in to a carousel
tube. The flask was evacuated and purged with argon. This operation
was performed 3 times. In a Schlenk tube,
9-triflate-2-morpholin-4-yl-pyrido[2,1-a]pyrimidin-4-one (5)(100
mg; 0.2638 mmol; per carousel tube) was solubilized in dioxane (2
mL; per carousel tube). Argon was bubbled in to the solution, which
was sonicated for 15 minutes. In another Schlenk tube,
terakis-(triphenylphosphine)-palladium (15.2 mg; 0.013 mmol; per
carousel tube) was solubilized in dioxane (2 mL; per carousel
tube). Argon was bubbled in to the solution, which was sonicated
for 15 minutes. 2 mL of each solution was mixed together in the
carousel tube, stirred and heated at 95.degree. C. for 48 hours.
Upon cooling, the solution was filtered through a Radleys Discovery
Technologie solid phase extraction column of 500 mg of silica,
which was placed on a stalker parallel purification system. The
column was washed with ethyl acetate (20 mL), and collected as
Phase 1. The column was then washed with dichloromethane/methanol
(85:15) (20 mL) and collected as Phase 2. Both phases were checked
for product via LC/MS. In some cases, phase 2 contained only
impurities, in other cases both phases were combined and
evaporated. Depending on the product, the purification was
performed by HPLC or by flash chromatography. TABLE-US-00002
Compound Ar 13a ##STR48##
NMR Results
[0294] 13a: .sup.1H NMR, (CDCl.sub.3, 300 MHz), .delta. (ppm): 3.42
(4H, m, N--CH.sub.2-morpholine); 3.56 (4H, m,
O--CH.sub.2-morpholine); 5.40 (1H, s, CH-3); 6.87 (1H, t,
J.sub.H6-H7=7 Hz, H.sub.arom-7); 7.26-7.83 (10H, m,
H.sub.arom-biphenyl and H.sub.arom-8); 8.87 (1H, dd,
J.sub.H6-H7=7.1 Hz, J.sub.H6-H8=1.6 Hz, H.sub.arom-6).
[0295] .sup.13C--NMR, (CDCl.sub.3, 300 MHz), .delta. (ppm): 44.92
(CH.sub.2--N-morpholine); 66.87 (CH--O--morpholine);81.24 (CH-3);
112.90 (CH-7); 127.22; 127.35; 127.56; 127.91; 128.81; 129.01;
129.24; 129.39; 135.74; 136.74; 137.59; 141.11; 149.46; 159.41(Cq);
160.72 (Cq).
Example 2
Parallel Synthesis from 8-Bromo-2-morpholin-4-yl-1H-quinolin-4-one
(12)
[0296] ##STR49##
[0297] An appropriate boronic acid (0.486 mmol) and potassium
carbonate (269.2 mg; 1.946 mmol) were introduced in to a carousel
tube. The flask was evacuated and purged with argon. This operation
was performed 3 times. In a Schlenk tube,
8-bromo-2-morpholin-4-yl-1H-quinolin-4-one (12)(100 mg; 0.324 mmol;
per carousel tube) was solubilized in dioxane (2 mL; per carousel
tube). Argon was bubbled in to the solution, which was sonicated
for 15 minutes. In another Schlenk tube,
terakis-(triphenylphosphine)-palladium (18.7 mg; 0.016 mmol; per
carousel tube) was solubilized in dioxane (2 mL; per carousel
tube). Argon was bubbled in to the solution, which was sonicated
for 15 minutes. 2 mL of each solution was mixed together in the
carousel tube, stirred and heated at 95.degree. C. for 48 hours.
Upon cooling, the solution was filtered through a Radleys Discovery
Technologie solid phase extraction column of 500 mg of silica,
which was placed on a stalker parallel purification system. The
column was washed with ethyl acetate (20 mL), and collected as
Phase 1. The column was then washed with dichloromethane/methanol
(85:15) (20 mL), and collected as Phase 2.
[0298] Both phases were checked for product via LC/MS. In some
cases, phase 2 contained only impurities, in other cases both
phases were combined and evaporated. Depending on the product, the
purification was performed by HPLC or by flash chromatography.
TABLE-US-00003 14a ##STR50##
NMR Results
[0299] 14a: .sup.1H NMR, (CDCl.sub.3, 300 MHz), .delta. (ppm): 3.13
(4H, s, N--CH.sub.2-morpholine); 3.70 (4H, s,
O--CH.sub.2-morpholine); 5.78 (1H, s, CH-3); 7.28-7.47 (5H, m,
Ham-biphenyl and H.sub.arom-7); 7.47-7.72 (6H, m,
H.sub.arom-biphenyl); 8.14 (1H, s, H.sub.arom-8); 8.32 (1H, s,
H.sub.arom-6).
[0300] .sup.13C-NMR, (CDCl.sub.3, 300 MHz), .delta. (ppm): 46.73
(CH.sub.2--N-morpholine); 66.27 (CH--O-morpholine); 92.86 (CH-3);
122.72; 123.58; 124.17; 125.93; 127.39; 127.54; 127.81; 128.01;
128.45; 128.98; 129.16; 129.43; 129.81; 130.51; 132.65; 135.56;
137.42; 140.29; 143.16; 154.31; 178.88.
Example 3
(a) 8-(6-Hydroxy-biphenyl-3-yl)-2-morpholin-4-yl-chromen-4-one
(17)
[0301] ##STR51##
(i) 5-Iodo-biphenyl-2-ol (15)
[0302] A solution of 6.02 g (35.37 mmol, 1 eq.) of 2-phenylphenol,
5.30 g of sodium iodide (35.37 mmol, 1 eq.), 1.56 g of sodium
hydroxide (35.37 mmol, 1 eq.) in 150 mL methanol was cooled to
0.degree. C. 26.32 g of NaOCl were added dropwise via syringe over
30 minutes. Upon addition of each drop, the solution develops an
orange colouration that fades instantly. The reaction mixture was
left to stir at 0.degree. C. during 1 hour before heating with 10%
aqueous sodium thiosulfate (60 mL). The pH of the reaction mixture
was adjusted to pH 7 using HCl and diethyl ether (150 mL) was
added. The layers were separated and the organic layer was washed
with brine, dried over MgSO.sub.4, filtered and evaporated.
Chromatography over silica gel (elution: DCM) afforded the title
compound as a white solid (9.27g, 89%). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 5.51 (broad s, 1H); 6.79 (d, J=8.5 Hz, 1H);
7.37-7.53 (m, 7H); .sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 83.9;
118.8; 129.2; 129.5; 129.8; 130.9; 135.8; 138.7; 139.5; 152.3.
(ii)
2-Morpholin-4-yl-8-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-chr-
omen-4-one (16)
[0303] A round bottom flask containing 500 mg (1.32 mmol, 1 eq.) of
chromenone triflate (see WO 03/024949), 402 mg (1.58 mmol, 1.2 eq.)
of bis(pinacolato)diboron, 388 mg (3.95 mmol, 3 eq.) of potassium
acetate and 15 mL 1,4-dioxane was sonicated for 15 minutes. To the
reaction mixture were added PdCl.sub.2dppf (54 mg, 0.07 mmol, 5 mol
%) and dppf (39 mg, 0.07 mmol, 5 mol %). The reaction was heated at
90.degree. C. under argon overnight. Upon completion, the reaction
mixture was diluted with diethyl ether and the organic layer was
washed with a saturated NaCl solution, dried over MgSO.sub.4,
filtered and evaporated. Chromatography over silica gel
(elution:ethyl acetate/methanol, gradient) afforded the title
compound (378 mg, 1.06 mmol, 80%). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 1.19 (s, 12H); 3.61-3.64 (m, 4H); 3.74-3.78 (m,
4H); 5.51 (s, 3H); 7.27 (t, J=7.5 Hz, 1H); 7.94 (d, J=7.2 Hz); 8.20
(d, J=6.3 Hz, 1H); .sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 24.9;
25.4; 45.1; 66.5; 83.5; 84.3; 86.8; 124.7; 129.6; 140.6; 158.1;
162.0; 178.0.
(iii) 8-(6-Hydroxy-biphenyl-3-yl)-2-morpholin-4-yl-chromen-4-one
(17)
[0304] In a Schlenk tube were placed 107 mg (0.30 mmol) of boronic
ester (16), 80 mg (0.27 mmol) of iodide, 4 mg (0.023 mmol) of
PdCl.sub.2(PPh.sub.3).sub.2 and 3 mL of aqueous 2M
Na.sub.2CO.sub.3. THF (12 mL) was degassed prior to addition and
the reaction mixture was heated to reflux for 2 hours. Water (10
mL) was added and the reaction mixture was extracted with DCM (60
mL). The combined organic layers were dried over magnesium sulfate,
filtered and concentrated under vacuum. Purification by
reprecicpitation afforded 89 mg (83% yield) of the title compound.
M.p. 243-247.degree. C.; UV .lamda..sub.max=304 nm; IR (cm.sup.-1)
2854, 1618, 1556, 1406, 1243, 1114, 989, 777, 557; .sup.H NMR (300
MHz, CDCl.sub.3) .delta. 3.32 (m, 4H); 3.57 (m, 4H); 5.55 (s, 1H);
7.20-7.65 (m, 10H); 8.15 (d; J=7.3 Hz; 1H); .sup.13C NMR (75 MHz,
CDCl.sub.3) .delta. 44.7; 60.4; 65.09; 86.4; 116.5; 123.3; 124.4;
124.9; 127.7; 128.4; 128.8; 129.0; 129.7; 130.0; 132.1; 133.1;
137.6; 150.6; 153.8; 162.6; 171.3; 177.6.
(b) 2-Morpholin-4-yl-8-(3-thiophen-3-yl-phenyl)-chromen-4-one (20)
and 2-Morpholin-4-yl-8-(3-thiophen-2-yl-phenyl)-chromen-4-one
(21)
[0305] ##STR52##
(i) 8-(3-Hydroxy-phenyl)-2-morpholin-4-yl-chromen-4-one (18)
[0306] In a Schlenk tube were placed equimolar amounts of boronic
ester and triflate, along with PdCl.sub.2(PPh.sub.3).sub.2 and
aqueous 2M Na.sub.2CO.sub.3. THF was degassed prior to addition and
the reaction mixture was heated to reflux for 2 hours. Water was
added and the reaction mixture was extracted with DCM. The combined
organic layers were dried over magnesium sulfate, filtered and
concentrated under vacuum. Purification by reprecipitation afforded
693 mg (90% yield). M.p. 203-205.degree. C.; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 3.29 (s (broad), 4H); 3.64 (s (broad), 4H);
5.51 (s, 1H); 6.95 (d, J=7.3 Hz, 1H); 7.00 (d, J=8.6 Hz, 1H);
7.27-7.42 (m, 3H); 7.49 (d, J=7.5 Hz; 1H); 8.11 (d; J=7.5 Hz; 1H);
.sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 44.9; 66.2; 116.0; 117.1;
120.7; 125.0; 125.4; 130.3; 130.6; 131.0; 134.1; 137.3; 140.3;
151.2; 157.5; 162.7; UV .lamda..sub.max=302 nm; IR (cm.sup.-1)
2862, 1614, 1553, 1414, 1244, 1112, 993, 779.
(ii) Trifluoro-methanesulfonic acid
3-(2-morpholin-4-yl-4-oxo-4H-chromen-8-yl)-phenyl ester (19)
[0307] In a round bottom flask were placed
8-(3-Hydroxy-phenyl)-2-morpholin-4-yl-chromen-4-one (18)(277 mg,
0.86 mmol, 1 eq.), N-phenyl triflimide (1.224 g, 3.43 mmol, 4 eq.),
Et.sub.3N (480 .mu.L, 3.43 mmol, 4 eq.) and 10 mL THF. The reaction
mixture was stirred at room temperature overnight and water was
added (10 mL). The mixture was extracted with DCM and the organic
layers were combined, dried over MgSO.sub.4, filtered and
concentrated. Chromatography over silica gel (elution: ethyl
acetate/methanol gradient) afforded the title compound as a white
solid (300 mg, 0.66 mmol, 77%). .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 3.28-3.31 (m, 4H); 3.69-3.72 (m, 4H); 5.49 (s, 1H);
7.28-7.60 (m, 6H); 8.18 (d; J=7.8 Hz, 1H); .sup.13C NMR (75 MHz,
CDCl.sub.3) .delta. 44.6; 65.4; 87.6; 120.8; 121.6; 124.6; 124.9;
126.0; 129.1; 130.9; 133.1; 139.5; 149.1; 150.4; 162.5; 171.1;
177.1.
(iii) 2-Morpholin-4-yl-8-(3-thiophen-3-yl-phenyl)-chromen-4-one 46
(20)
[0308] Suzuki-Miyaura coupling was carried out as above to yield
the desired compound. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
3.31 (m; 4H); 3.64 (m; 4H); 5.54 (s; 1H); 7.43-7.60 (m; 6H);
7.64-7.68 (m; 2H); 7.80 (s; 1H); 8.22 (d; J=6.0 Hz, 1H); .sup.13C
NMR (75 MHz, CDCl.sub.3) .delta. 44.7; 65.8; 87.0; 120.6; 123.4;
124.9; 125.2; 125.9; 126.2; 126.3; 126.8; 127.7; 130.1; 133.3;
135.9; 136.8; 141.6; 150.6; 162.5; 177.2; UV .lamda..sub.max=301
nm; IR (cm.sup.-1) 3094, 2847, 1615, 1556, 1404, 1244, 1066, 985,
769.
(iv) 2-Morpholin-4-yl-8-(3-thiophen-2-yl-phenyl)-chromen-4-one 47
(21)
[0309] Suzuki-Miyaura coupling was carried out as above to yield
the desired compound. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
3.34 (m; 4H); 3.65 (m; 4H); 5.56 (s; 1H); 7.09-7.12 (m; 1H);
7.26-7.52 (m; 5H); 7.62-7.68 (m; 2H); 7.92 (s; 1H); 8.20 (d; J=6.0
Hz, 1H); .sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 44.7; 65.8;
87.0; 123.3; 123.4; 124.9; 125.3; 125.9; 127.1; 127.5; 128.3;
128.4; 129.2; 129.9; 133.4; 134.5; 136.9; 143.6; 150.5; 162.5;
177.1; UV .lamda..sub.max=296 nm; IR (cm.sup.-1) 2921, 2855, 1639,
1570, 1396, 1240, 1115, 1074, 765. ##STR53##
[0310] Hydroxy derivative (17) was dissolved in dry DMF. Potassium
carbonate and methyl bromoacetate were added and the reaction
mixture was stirred at 60.degree. C. overnight. The mixture was
poured in water and extracted with ethyl acetate. The organic layer
was washed with water, brine, dried over magnesium sulphate and
concentrated to give the methyl ester. This ester was dissolved in
methanol and aqueous NaOH was added. The reaction mixture was
stirred at 60.degree. C. for one hour until completed before being
evaporated to dryness to give the sodium salt. The sodium salt was
dissolved in DMF and divided in equal portions in tubes containing
appropriate amines, HOBT and TBTU. The reaction mixtures were
stirred at room temperature overnight and diluted in methanol
before being submitted for purification. TABLE-US-00004 ##STR54##
Rt Compound R M/z (min) Purity (%) 24a ##STR55## 568 4.64 89 24b
##STR56## 554 4.12 68 24c ##STR57## 545 4.39 86 24d ##STR58## 570
4.03 78 24e ##STR59## 525 5.49 90 24f ##STR60## 547.9 4.16 70 24g
##STR61## 541 5.13 86 24h ##STR62## 555 3.69 90 24i ##STR63## 527
3.67 87 (d) ##STR64## ##STR65## ##STR66##
[0311] In a round-bottom flask were introduced compound 17,
1,2-dibromoethane, potassium carbonate and dry DMF. The reaction
mixture was stirred at room temperature. Upon completion, the
reaction mixture was divided in aliquots that were added, together
with the appropriate amines in reactions tubes. The resulting
mixtures were stirred at room temperature overnight and diluted
with methanol before being submitted for purification.
TABLE-US-00005 ##STR67## Compound R M/z Rt (min) Purity (%) 25a
##STR68## 531.5 3.9 90
Example 4
(a) 8-(6-Amino-biphenyl-3-yl)-2-morpholin-4-yl-chromen-4-one
(27)
[0312] ##STR69##
(i) 5-Bromo-biphenyl-2-ylamine (26)
[0313] In a two-neck round-bottom flask were placed the starting
material 2-phenylaniline (2.035 g, 12.03 mmol, 1 eq.), 1.295 g
NH.sub.4Br (13.23 mmol, 1.1 eq.) and 24 mL glacial acetic acid.
Hydrogen peroxide (1.64 g of 27.5% w/w solution, 13.23 mmol, 1.1
eq.) was added dropwise via a syringe and the reaction mixture was
left to stir for 24 hours. Upon disappearance of the starting
material, a saturated solution of Na.sub.2CO.sub.3 was added and
the mixture was extracted with DCM. The combined organic layers
were dried over MgSO.sub.4, filtered and concentrated under vacuum.
The crude mixture was purified by flash chromatography over silica
gel (elution petrol/DCM 4:1, then gradient) to afford the expected
product as a beige solid (2.076 g, 70%).
[0314] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 3.76 (broad s,
2H); 6.67 (d, J=7.3 Hz, 1H); 7.27 (d, J=7.9 Hz, 1H); 7.37-7.52 (m,
6H).
[0315] .sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 110.6; 117.5;
128.1; 129.3; 129.4; 129.8; 131.5; 133.2; 138.5; 143.0.
(ii) 8-(6-Amino-biphenyl-3-yl)-2-morpholin-4-yl-chromen-4-one
(27)
[0316] In a schlenk tube were placed 145 mg (0.41 mmol) of
chromenone boronate (16), 101 mg (0.41 mmol) of biphenylamine
bromide, 48 mg (0.041 mmol) of Pd(PPh.sub.3).sub.4 and 338 mg (2.44
mmol) of K.sub.2CO.sub.3 dioxane (10 mL) was degassed prior to
addition and the reaction mixture was heated to reflux for 12
hours. Water (10 mL) was added and the reaction mixture was
extracted with DCM (60 mL). The combined organic layers were dried
over magnesium sulfate, filtered and concentrated under vacuum.
Silica gel chromatography (elution Ethyl acetate/methanol 95:5 then
gradient) afforded the title compound as a white powder in 52%
yield.
[0317] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 3.26-3.33 (m; 4H);
3.67-3.71 (m; 4H); 5.49 (s; 1H); 7.28-7.58 (m, 10H), 8.14 (d; J=7.4
Hz; 1H).
[0318] .sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 45.0; 60.7; 66.3;
87.2; 116.7; 123.7; 124.4; 125.0; 127.9; 128.6; 128.7; 129.1;
129.8; 130.5; 132.0; 133.3; 136.6; 150.9; 154.0; 162.8; 171.5;
177.7.
[0319] UV: .lamda..sub.max=305 nm
[0320] IR: (cm.sup.-1) 1616, 1560, 1400, 1242, 983, 905, 700.
[0321] Mass Spectrum: (m/z) 399 [M+1].sup.+.
[0322] HPLC assay system: LCMS R.sub.7=3.82 min, 5 min
gradient.
[0323] TLC: R.sub.f=0.54 (EA/MeOH, 9:1) ##STR70##
[0324] Amino derivative (27) was dissolved in dry DMA.
Triethylamine and chloroacetyl chloride were added and the reaction
mixture was stirred at room temperature for 4 hours. Aliquots of
the reaction mixture were added to tubes containing the appropriate
amines. The reaction mixtures were stirred at room temperature
overnight and diluted in methanol before being submitted for
purification. TABLE-US-00006 ##STR71## Compound R M/z Rt (min)
Purity (%) 29a ##STR72## 526.4 4.3 96 29b ##STR73## 554.4 4.62 95
29c ##STR74## 567.4 4.33 95 29d ##STR75## 553.5 4.12 99 29e
##STR76## 544.5 3.94 99 29f ##STR77## 540.6 4.04 100 (c) ##STR78##
##STR79## ##STR80##
[0325] In a round-bottom flask were introduced compound 27,
triethylamine, 3-bromopropionyl chloride and dry DMA. The reaction
mixture was stirred at room temperature overnight. Upon completion,
the reaction mixture was divided in aliquots that were added,
together with the corresponding amines in reactions tubes. The
resulting mixtures were stirred at room temperature overnight and
diluted with methanol before being submitted for purification.
TABLE-US-00007 ##STR81## Compound R M/z Rt (min) Purity (%) 31a
##STR82## 583.5 3.94 99
Example 5
[0326] ##STR83##
[0327] In order to synthesise compounds having a heteroaryl-phenyl
substituent, a double Suzuki-coupling was used. The reaction was
carried out in a microwave at 150.degree. C. for 5 minutes, with
the following reagents in addition to the reactive compounds:
PdCl.sub.2(PPh.sub.3).sub.2, Na.sub.2CO.sub.3 aq.,
DME/H.sub.2O/EtOh (7:3:2). The compounds were then submitted for
purification. TABLE-US-00008 ##STR84## Compound R M/z Rt (min)
Purity (%) 32a ##STR85## 385 3.78 90 32b ##STR86## 386 4.34 99 32c
##STR87## 388 3.5 98 32d ##STR88## 403.9 6.11 96 32e ##STR89##
390.9 5.05 75 32f ##STR90## 404 3.96 100 32g ##STR91## 404 3.63 100
32h ##STR92## 385 3.38 100
Example 6
[0328] ##STR93##
a) 8-(2-Chloro-pyridin-4-yl)-2-morpholin-4-yl-chromen-4-one
(33)
[0329] To a solution of trifluoro-methanesulfonic acid
2-morpholin-4-yl-4-oxo-4H-chromen-8-yl ester (1.00 g, 2.64 mmol) in
anhydrous dioxane (40 mL) under a N2 atmosphere was added
2-chloropyridine-4-boronic acid (0.436 g, 2.77 mmol) in one
portion, followed by the addition of
tetrakis(triphenylphosphine)palladium (0) (0.147 g, 0.132 mmol) and
crushed potassium carbonate (1.093 g, 7.91 mmol) in one portion
each successively. A condensor was fitted and the reaction was
heated to 110.degree. C. After 48 hours, the reaction was cooled to
RT and filtered through celite with dichloromethane washings. The
combined organics were washed with a saturated brine solution
(1.times.10 mL), dried over MgSO.sub.4, filtered and concentrated
in vacuo. On addition of hexanes, the precipitation of a solid
occurred over 12 hours which was collected by filtration to yield
the title compound (assumed 100%) and required no further
purification.
b) 8-(2--Substituted-pyridin-4-yl)-2-morpholin-4-yl-chromen-4-one
(34)
[0330] 8-(2-Chloro-pyridin-4-yl)-2-morpholin-4-yl-chromen-4-one
(0.30 g, 0.875 mmol) was dissolved in N,N-dimethylformamide (10
mL). Aliquots (1 mL) were then added to vials containing crushed
potassium carbonate (0.029 g, 0.210 mmol), the required boronic
acid (0.096 mmol) and bis(tri-.sup.tbutylphosphine)palladium (0) (6
mg, 0.003 mmol). The mixtures were then subjected to microwave
radiation at 150.degree. C. for 300s. The resulting solutions were
filtered through an SEP silica cartridge with methanol washing, and
then subjected to purification via HPLC. TABLE-US-00009 ##STR94##
Compound R M/z (M + H) Rt (min) Purity (%) 34a ##STR95## 385.4 8.98
97 34b ##STR96## 436.3 6.56 98 34c ##STR97## 469.3 12.51 99 34d
##STR98## 427.3 9.52 92 34e ##STR99## 504.3 10.57 89 34f ##STR100##
403.3 10.01 99 34g ##STR101## 480.3 11.07 99 34h ##STR102## 399.3
9.55 93 34i ##STR103## 476.3 10.59 95 34j ##STR104## 512.4 11.08 93
34k ##STR105## 424.4 7.31 96 34l ##STR106## 415.3 8.52 99 34m
##STR107## 492.4 9.61 98 34n ##STR108## 404.3 4.52 98 34o
##STR109## 386.4 3.70 94
Example 7
[0331] ##STR110##
a) 8-(4-Bromo-pyridin-2-yl)-2-morpholin-4-yl-chromen-4-one (35)
[0332] To a solution of
2-Morpholin-4-yl-8-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-chromen-
-4-one (0.321 g, 0.90 mmol) in a 1:1 mixture of toluene:ethanol (15
mL) was added 2,4-dibromopyridine (0.315 g, 0.99 mmol) in one
portion, followed by the addition of
trans-dichlorobis(triphenylphosphine)palladium (II) (0.011 g, 0.015
mmol) and crushed potassium carbonate (0.435 g, 3.15 mmol) in one
portion each successively. The reaction mixture was then subjected
to microwave radiation at 150.degree. C. for 300s. The resulting
solution was filtered with ethanol washings and the combined
organics were concentrated in vacuo. The crude product was purified
by flash chromatography (SiO2) (EtOAc:EtOH 100:1, then DCM:MeOH
20:1) to yield the title compound as a brown foam (0.275 g,
79%).
b) 8-(4-Substituted-pyridin-2-yl)-2-morpholin-4-yl-chromen-4-one
(36)
[0333] 8-(4-Bromo-pyridin-2-yl)-2-morpholin-4-yl-chromen-4-one
(0.275 g, 0.710 mmol) was dissolved in anhydrous
N,N-dimethylformamide (10 mL). Aliquots (1 mL) were then added to
vials containing crushed potassium carbonate (0.029 g, 0.210 mmol),
the required boronic acid (0.105 mmol) and
bis(tri-.sup.tbutylphosphine)palladium (0) (2 mg, 0.004 mmol). The
mixtures were then subjected to microwave radiation at 150.degree.
C. for 300s. The resulting solutions were filtered through a SEP
silica cartridge with methanol washing, and then subjected to
purification via HPLC. TABLE-US-00010 ##STR111## Compound R M/z (M
+ H) Rt (min) Purity (%) 36a ##STR112## 309.3 3.41 99 36b
##STR113## 385.5 9.29 99 36c ##STR114## 391.3 4.47 91 36d
##STR115## 391.3 4.59 71 36e ##STR116## 386.4 3.85 98 36f
##STR117## 375.4 4.26 85 36g ##STR118## 387.3 3.91 99 36h
##STR119## 404.3 4.40 96
Example 8
[0334] ##STR120##
a) 8-(5-Chloro-thiophen-2-yl)-2-morpholin-4-yl-chromen-4-one
(37)
[0335] To a solution of trifluoro-methanesulfonic acid
2-morpholin-4-yl-4-oxo-4H-chromen-8-yl ester (0.341 g, 0.90 mmol)
in a 10:1 mixture of 1,4-dioxane:N,N-dimethylacetamide (15 mL) was
added a catalytic amount of triethylamine, followed by the addition
of 5-chloro-2-thiopheneboronic acid (0.292 g, 1.80 mmol). The
mixture was then subjected to microwave radiation at 150.degree. C.
for 120s. The addition of a further amount of
5-chloro-2-thiopheneboronic acid (0.292 g, 1.80 mmol), followed by
microwave irradiation at 150.degree. C. for 120s was repeated
twice. After filtration, the filtrate was concentrated in vacuo and
the resulting solid triturated with tetrahydrofuran. A second
filtration revealed the title compound (0.222 g, 71%) and required
no further purification.
b) 8-(5-Substituted-thiophen-2-yl)-2-morpholin-4-yl-chromen-4-one
(38)
[0336] 8-(5-Chloro-thiophen-2-yl)-2-morpholin-4-yl-chromen-4-one
(0.222 g, 0.64 mmol) was dissolved in N,N-dimethylformamide (10
mL). Aliquots (1 mL) were then added to vials containing crushed
potassium carbonate (0.026 g, 0.19 mmol), the required boronic acid
(0.066 mmol) and bis(tri-.sup.tbutylphosphine)palladium (0) (1-2
mg, 0.003 mmol). The mixtures were then subjected to microwave
radiation at 150.degree. C. for 120s. The resulting solutions were
filtered through an SEP silica cartridge with methanol washing, and
then subjected to purification via HPLC. TABLE-US-00011 ##STR121##
Rt Compound R M/z (M + H) (min) Purity (%) 38a ##STR122## 390.3
13.47 98 38b ##STR123## 441.3 8.82 100 38c ##STR124## 468.3 10.65
100 38d ##STR125## 474.3 15.50 99 38e ##STR126## 432.3 12.18 100
38f ##STR127## 408.3 13.66 100 38g ##STR128## 404.3 14.74 96 38h
##STR129## 440.3 15.29 94 38i ##STR130## 429.3 12.60 92 38j
##STR131## 420.3 13.49 100
Example 9
[0337] ##STR132##
a) 8-(5-Bromo-thiazol-2-yl)-2-morpholin-4-yl-chromen-4-one (39)
[0338] To a solution of
2-Morpholin-4-yl-8-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-chromen-
-4-one (0.535 g, 1.50 mmol) in a 1:1 mixture of toluene:ethanol (25
mL) was added 2,5-dibromothiazole (0.400 g, 1.65 mmol) in one
portion, followed by the addition of tetrakis(triphenyl
phosphine)palladium (0) (0.087 g, 0.075 mmol) and crushed potassium
carbonate (0.621 g, 4.50 mmol) in one portion each successively.
The reaction mixture was then subjected to microwave radiation at
130.degree. C. for 3600s. The resulting solution was filtered with
ethanol washings and the combined organics were concentrated in
vacuo. The crude product was purified by flash chromatography
(SiO2) (EtOAc:EtOH 20:1, then 20:3) to yield the title compound as
a dark solid (0.255 g, 43%).
b) 8-(5--Substituted-thiazol-2-yl)-2-morpholin-4-yl-chromen-4-one
(40)
[0339] 8-(5-Bromo-thiazol-2-yl)-2-morpholin-4-yl-chromen-4-one
(0.255 g, 0.650 mmol) was dissolved in anhydrous
N,N-dimethylformamide (10 mL). Aliquots (1 mL) were then added to
vials containing crushed potassium carbonate (0.029 g, 0.210 mmol),
the required boronic acid (0.098 mmol) and
bis(tri-.sup.tbutylphosphine)palladium (0) (2 mg, 0.004 mmol). The
mixtures were then subjected to microwave radiation at 150.degree.
C. for 300s. The resulting solutions were filtered through a SEP
silica cartridge with methanol washing, and then subjected to
purification via HPLC. TABLE-US-00012 ##STR133## Compound R M/z (M
+ H) Rt (min) Purity (%) 40a ##STR134## 390.3 6.10 93 40b
##STR135## 396.2 5.93 96
Example 10
[0340] ##STR136##
a) 8-(5-Bromo-thiazol-2-yl)-2-morpholin-4-yl-chromen-4-one (41)
[0341] To a solution of
2-Morpholin-4-yl-8-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-chromen-
-4-one (0.535 g, 1.50 mmol) in a 1:1 mixture of toluene:ethanol (25
mL) was added 2,5-dibromothiazole (0.400 g, 1.65 mmol) in one
portion, followed by the addition of tetrakis(triphenyl
phosphine)palladium (0) (0.087 9, 0.075 mmol) and crushed potassium
carbonate (0.621 g, 4.50 mmol) in one portion each successively.
The reaction mixture was then subjected to microwave radiation at
130.degree. C. for 3600s. The resulting solution was filtered with
ethanol washings and the combined organics were concentrated in
vacuo. The crude product was purified by flash chromatography
(SiO2) (EtOAc:EtOH 20:1, then 20:3) to yield the title compound as
a dark solid (0.255 g, 43%).
b) 8-(5--Substituted-thiazol-2-yl)-2-morpholin-4-yl-chromen-4-one
(42)
[0342] 8-(5-Bromo-thiazol-2-yl)-2-morpholin-4-yl-chromen-4-one
(0.255 9, 0.650 mmol) was dissolved in anhydrous
N,N-dimethylformamide (10 mL). Aliquots (1 mL) were then added to
vials containing crushed potassium carbonate (0.029 g, 0.210 mmol),
the required boronic acid (0.098 mmol) and
bis(tri-.sup.tbutylphosphine)palladium (0) (2 mg, 0.004 mmol). The
mixtures were then subjected to microwave radiation at 150.degree.
C. for 300s. The resulting solutions were filtered through a SEP
silica cartridge with methanol washing, and then subjected to
purification via HPLC. TABLE-US-00013 ##STR137## Compound R M/z (M
+ H) Rt (min) Purity (%) 42a ##STR138## 397.3 5.55 99 42b
##STR139## 397.3 5.72 92 42c ##STR140## 392.3 4.34 82 42d
##STR141## 381.3 5.26 97 42e ##STR142## 392.3 3.92 99 42f
##STR143## 391.3 5.76 95
Biological Examples DNA-PK Inhibition
[0343] In order to assess the inhibitory action of the compounds
against DNA-PK in vitro, the following assay was used to determine
IC.sub.50 values.
[0344] Mammalian DNA-PK (500ng/ml) was isolated from HeLa cell
nuclear extract (Gell, D. and Jackson S. P., Nucleic Acids Res.
27:3494-3502 (1999)) following chromatography utilising
Q-sepharose, S-sepharose and Heparin agarose. DNA-PK (250 ng)
activity was measured at 30.degree. C., in a final volume of 40
.mu.l, in buffer containing 25 mM Hepes, pH7.4, 12.5 mM MgCl.sub.2,
50 mM KCl, 1 mM DTT, 10% Glycerol, 0.1% NP-40 and 1 mg of the
substrate GST-p53N66 (the amino terminal 66 amino acid resiudes of
human wild type p53 fused to glutathione S-transferase) in
polypropylene 96 well plates. To the assay mix, varying
concentrations of inhibitor (in DMSO at a final concentration of
1%) were added. After 10 minutes of incubation, ATP was added to
give a final concentration of 50 .mu.M along with a 30mer double
stranded DNA oligonucleotide (final concentraion of 0.5 ng/ml) to
initiate the reaction. After 1 hour with shaking, 150 .mu.l of
phosphate buffered saline (PBS) was added to the reaction and 5
.mu.l then transferred to a 96 well opaque white plate containing
45 .mu.l of PBS per well where the GSTp53N66 substrate was allowed
to bind to the wells for 1 hour. To detect the phosphorylation
event on the serine 15 residue of p53 elicited by DNA-PK a p53
phosphoserine-15 antibody (Cell Signaling Technology) was used in a
basic ELISA procedure. An anti-rabbit HRP conjugated secondary
antibody (Pierce) was then employed in the ELISA before the
addition of chemiluminescence reagent (NEN Renaissance) to detect
the signal as measured by chemiluminescent counting via a TopCount
NXT (Packard).
[0345] The enzyme activity for each compound is then calculated
using the following equation: % .times. .times. Inhibition = 100 -
( ( cpm .times. .times. .times. of .times. .times. .times. unknown
- mean .times. .times. .times. negative .times. .times. cpm ) 100 (
mean .times. .times. positive .times. .times. cpm - mean .times.
.times. .times. negative .times. .times. cpm ) ) ##EQU1##
[0346] The results are discussed below as IC.sub.50 values (the
concentration at which 50% of the enzyme activity is inhibited).
These are determined over a range of different concentrations,
normally from 10 .mu.M down to 0.001 .mu.M. Such IC.sub.50 values
are used as comparative values to identify increased compound
potencies.
Survival Enhancement Ratio
[0347] The Survival Enhancement Ratio (SER) is a ratio of the
enhancement of cell kill elicited by the DNA-PK inhibitor after 2
Grays of irradiation compared to unirradiated control cells. DNA-PK
inhibitors were used at a fixed concentration of 500 nM. Radiation
was delivered by a Faxitron 43855D machine at a dose rate of 1 Gy
pre minute The SER at 2 Gray irradiation was calculated from the
formula: SER = Cell .times. .times. survival .times. .times. in
.times. .times. presence .times. .times. .times. of .times. .times.
DNA - PK .times. .times. inhibitor Cell .times. .times. .times.
survival .times. .times. .times. of .times. .times. .times. control
.times. .times. .times. cells Cell .times. .times. survival .times.
.times. after .times. .times. IR Cell .times. .times. survival
.times. .times. .times. after .times. .times. .times. IR .times.
.times. in .times. .times. presence .times. .times. .times. of
.times. .times. DNA - PK .times. .times. inhibitor ##EQU2##
[0348] The degree of cell killing was monitored by a standard
clonogenic survival assay. Briefly, tissue culture treated 6-well
plates were seeded with HeLa cells at an appropriate concentration
to give 100-200 colonies per well and returned to the incubator in
order to allow the cells to attach. Four hours later, compound or
vehicle control was added to the cells. The cells were then
incubated for 1 hour in the presence of inhibitor prior to
irradiation at 2 Gray using a Faxitron 43855D cabinet X-ray
machine. The cells were then incubated for a further 16 hours
before the media was replaced with fresh media in the absence of
DNA-PK inhibitor. After 8 days, colonies formed were fixed and
stained with Giemsa (Sigma, Poole, UK) and scored using an
automated colony counter (Oxford Optronics Ltd, Oxford, UK). The
data was calculated as described above.
Results
[0349] All the compounds showed activity in DNA-PK inhibition, some
exhibiting an IC.sub.50 of less than about 500 nM.
[0350] All the compounds showed an SER of 1 or more.
* * * * *