U.S. patent application number 12/233736 was filed with the patent office on 2009-02-12 for atm inhibitors.
This patent application is currently assigned to KUDOS PHARMACEUTICALS LIMITED. Invention is credited to Xiao-Ling Fan Cockcroft, Roger John Griffin, Marc Geoffrey Hummersone, Niall Morrison Barr Martin, Ian Timothy Williams Matthews, Keith Allan Menear, Laurent Jean Martin Rigoreau, Graeme Cameron Murray Smith.
Application Number | 20090043091 12/233736 |
Document ID | / |
Family ID | 34193236 |
Filed Date | 2009-02-12 |
United States Patent
Application |
20090043091 |
Kind Code |
A1 |
Smith; Graeme Cameron Murray ;
et al. |
February 12, 2009 |
ATM INHIBITORS
Abstract
A compound of formula I: ##STR00001## and isomers, salts,
solvates, chemically protected forms, and prodrugs thereof,
wherein: R.sup.1 and R.sup.2 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 R.sup.N1 is
selected from hydrogen, an optionally substituted C.sub.1-7 alkyl
group, an optionally substituted C.sub.3-20 heterocyclyl group, an
optionally substituted C.sub.5-20 aryl group, an acyl group, an
ester group and an amido group, and its use as a
pharmaceutical.
Inventors: |
Smith; Graeme Cameron Murray;
(Cambridge, GB) ; Martin; Niall Morrison Barr;
(Cambridge, GB) ; Cockcroft; Xiao-Ling Fan;
(Horsham, GB) ; Matthews; Ian Timothy Williams;
(Horsham, GB) ; Menear; Keith Allan; (Horsham,
GB) ; Rigoreau; Laurent Jean Martin; (Horsham,
GB) ; Hummersone; Marc Geoffrey; (Horsham, GB)
; Griffin; Roger John; (Morpeth, GB) |
Correspondence
Address: |
MICHAEL BEST & FRIEDRICH LLP
ONE SOUTH PINCKNEY STREET, P O BOX 1806
MADISON
WI
53701
US
|
Assignee: |
KUDOS PHARMACEUTICALS
LIMITED
Cambridge
GB
|
Family ID: |
34193236 |
Appl. No.: |
12/233736 |
Filed: |
September 19, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10918180 |
Aug 13, 2004 |
7429660 |
|
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12233736 |
|
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60494776 |
Aug 13, 2003 |
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Current U.S.
Class: |
540/575 ;
540/598; 544/121; 544/124; 544/129; 544/134; 544/137; 544/139;
544/140; 544/145; 544/35; 544/82 |
Current CPC
Class: |
A61P 43/00 20180101;
A61P 31/14 20180101; A61P 35/02 20180101; A61P 21/02 20180101; C07D
409/14 20130101; A61P 25/00 20180101; C07D 409/04 20130101; C07D
417/04 20130101; C07D 491/04 20130101; C07D 495/04 20130101; A61P
31/12 20180101; C07D 417/14 20130101; C07D 413/04 20130101; A61P
35/00 20180101; A61P 31/18 20180101; A61P 25/08 20180101 |
Class at
Publication: |
540/575 ;
544/145; 544/121; 544/82; 544/124; 544/137; 544/140; 544/129;
544/134; 544/139; 540/598; 544/35 |
International
Class: |
C07D 243/08 20060101
C07D243/08; C07D 413/14 20060101 C07D413/14; C07D 223/02 20060101
C07D223/02; C07D 279/18 20060101 C07D279/18 |
Claims
1. A compound of formula III: ##STR00586## and isomers, salts,
solvates, chemically protected forms, and prodrugs thereof,
wherein: R.sup.1 and R.sup.2 together form, along with the nitrogen
atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms; R.sup.C3 is of
formula: ##STR00587## wherein r is 0 or 1, and q can be 1 or 2 when
r is 0 and q is 1 when r is 1, and wherein R.sup.N4 and R.sup.N5
are independently selected from hydrogen, optionally substituted
C.sub.1-7 alkyl groups, optionally substituted C.sub.3-20
heterocyclyl groups and optionally substituted C.sub.5-20 aryl
groups, or together form, along with the nitrogen atom to which
they are attached, an optionally substituted heterocyclic ring
having from 4 to 8 ring atoms.
2. A compound according to claim 1, wherein r=0.
3. A compound according to claim 1, wherein R.sup.N4 and R.sup.N5
are independently selected from hydrogen, optionally substituted
C.sub.1-7 alkyl groups, or together form, along with the nitrogen
atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms.
4. A compound according to claim 3, wherein one of R.sup.N4 and
R.sup.N5 is an optionally substituted C.sub.1-7 alkyl group, the
other is preferably H, and the optional substituents are selected
from hydroxy, amino, cyano, C.sub.3-20 heterocylyl and C.sub.5-20
aryl (more preferably C.sub.5-7 aryl).
5. A compound according to claim 1, wherein one of R.sup.N4 and
R.sup.N5 is of formula IIb: ##STR00588## wherein r is from 1 to 3;
and R.sup.N2 and R.sup.N3 are independently selected from hydrogen,
optionally substituted C.sub.1-7 alkyl groups, optionally
substituted C.sub.3-20 heterocyclyl groups and optionally
substituted C.sub.5-20 aryl groups, or together form, along with
the nitrogen atom to which they are attached, an optionally
substituted heterocyclic ring having from 4 to 8 ring atoms.
6. A compound according to claim 5, wherein R.sup.N2 and R.sup.N3
are independently selected from hydrogen, optionally substituted
C.sub.1-7 alkyl groups or together form, along with the nitrogen
atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms.
7. A compound according to claim 1, wherein R.sup.N4 and R.sup.N5
together form, along with the nitrogen atom to which they are
attached, a ring selected from piperidine, homopiperidine,
piperazine, homopiperazine and morpholino.
8. A compound according to claim 7, wherein the ring is piperazine
or homopiperazine, and the other nitrogen ring atom is substituted
by a group selected from an optionally substituted C.sub.1-7 alkyl
group, an optionally substituted C.sub.3-20 heterocyclyl group, an
optionally substituted C.sub.5-20 aryl group, an acyl group, an
ester group or an amido group.
9. A compound of formula VI: ##STR00589## and isomers, salts,
solvates, chemically protected forms, and prodrugs thereof,
wherein: R.sup.1 and R.sup.2 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 R.sup.C4 is of
formula: ##STR00590## wherein r is 0 or 1, and q can be 1 or 2 when
r is 0 and q is 1 when r is 1, and wherein R.sup.N4 and R.sup.N5
are independently selected from hydrogen, optionally substituted
C.sub.1-7 alkyl groups, optionally substituted C.sub.3-20
heterocyclyl groups and optionally substituted C.sub.5-20 aryl
groups, or together form, along with the nitrogen atom to which
they are attached, an optionally substituted heterocyclic ring
having from 4 to 8 ring atoms.
10. A compound according to claim 9, wherein r=0.
11. A compound according to claim 9, wherein R.sup.N4 and R.sup.N5
are independently selected from hydrogen, optionally substituted
C.sub.1-7 alkyl groups, or together form, along with the nitrogen
atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms.
12. A compound according to claim 11, wherein one of R.sup.N4 and
R.sup.N5 is an optionally substituted C.sub.1-7 alkyl group, the
other is preferably H and the optional substituents are selected
from hydroxy, halo, C.sub.1-7 alkoxy, thiol, C.sub.1-7 thioether,
amino, ester, cyano, C.sub.3-20 heterocylyl and C.sub.5-20
aryl.
13. A compound according to claim 9, wherein, one of R.sup.N4 and
R.sup.N5 is of formula IIb: ##STR00591## wherein r is from 1 to 3;
and R.sup.N2 and R.sup.N3 are independently selected from hydrogen,
optionally substituted C.sub.1-7 alkyl groups, optionally
substituted C.sub.3-20 heterocyclyl groups and optionally
substituted C.sub.5-20 aryl groups, or together form, along with
the nitrogen atom to which they are attached, an optionally
substituted heterocyclic ring having from 4 to 8 ring atoms.
14. A compound according to claim 13, wherein R.sup.N2 and R.sup.N3
are independently selected from hydrogen, optionally substituted
C.sub.1-7 alkyl groups or together form, along with the nitrogen
atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms.
15. A compound according to claim 14, wherein R.sup.N4 and R.sup.N5
together form, along with the nitrogen atom to which they are
attached, a ring selected from piperidine, homopiperidine,
piperazine, homopiperazine and morpholino.
16. A compound according to claim 15, wherein the ring is
piperazine or homopiperazine, and the other nitrogen ring atom is
substituted by a group selected from an optionally substituted
C.sub.1-7 alkyl group, an optionally substituted C.sub.3-20
heterocyclyl group, an optionally substituted C.sub.5-20 aryl
group, an acyl group, an ester group and an amido group.
17. A compound according to claim 1, wherein R.sup.1 and R.sup.2
form, along with the nitrogen atom to which they are attached, a
ring selected from morpholino and thiomorpholino.
18. A compound according to claim 9, wherein R.sup.1 and R.sup.2
form, along with the nitrogen atom to which they are attached, a
ring selected from morpholino and thiomorpholino.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of U.S. application Ser.
No. 10/918,180 filed on Aug. 13, 2004, which claims priority
benefits to U.S. Provisional Application No. 60/494,776 filed on
Aug. 13, 2003, which are incorporated herein by reference in their
entireties.
[0002] The present invention relates to compounds which act as ATM
inhibitors, their use and synthesis.
[0003] Human DNA is constantly under attack from reactive oxygen
intermediates principally from by-products of oxidative metabolism.
Reactive oxygen species are capable of producing DNA single-strand
breaks and, where two of these are generated in close proximity,
DNA double strand breaks (DSBs). In addition, single- and
double-strand breaks can be induced when a DNA replication fork
encounters a damaged template, and are generated by exogenous
agents such as ionising radiation (IR) and certain anti-cancer
drugs (e.g. bleomycin, etoposide, camptothecin). DSBs also occur as
intermediates in site-specific V(D)J recombination, a process that
is critical for the generation of a functional vertebrate immune
system. If DNA DSBs are left unrepaired or are repaired
inaccurately, mutations and/or chromosomal aberrations are induced,
which in turn may lead to cell death. To combat the serious threats
posed by DNA DSBs, eukaryotic cells have evolved several mechanisms
to mediate their repair. Critical to the process of DNA repair is
the slowing down of cellular proliferation to allow time for the
cell to repair the damage. A key protein in the detection of DNA
DSBs and in the signalling of this information to the cell cycle
machinery is the kinase ATM (ataxia telangiectasia mutated)
(Durocher and Jackson (2001) DNA-PK, ATM and ATR as sensors of DNA
damage: variations on a theme? Curr Opin Cell Biol., 13:225-31,
Abraham (2001) Cell cycle checkpoint signaling through the ATM and
ATR kinases. Genes Dev., 15; 2177-96).
[0004] The ATM protein is an .about.350 kDa polypeptide that is a
member of the phosphatidylinositol (PI) 3-kinase family of proteins
by virtue of a putative kinase domain in its carboxyl-terminal
region (Savitsky et al (1995) A single ataxia telangiectasia gene
with a product similar to PI-3 kinase. Science, 268:1749-53).
Classical PI 3-kinases, such as PI 3-kinase itself, are involved in
signal transduction and phosphorylate inositol lipids that act as
intracellular second messengers (reviewed in Toker and Cantley
(1997) Signalling through the lipid products of
phosphoinositide-3-OH kinase, Nature, 387: 673-6). However, ATM
bears most sequence similarity with a subset of the PI 3-kinase
family that comprises proteins which, like ATM, are involved in
cell cycle control and/or in the detection and signalling of DNA
damage (Keith and Schreiber (1995) PIK-related kinases: DNA repair,
recombination, and cell cycle checkpoints, Science, 270; 50-1,
Zakian (1995) ATM-related genes: what do they tell us about
functions of the human gene? Cell, 82; 685-7). Notably there is no
evidence to date that any members of this subset of the PI 3-kinase
family are able to phosphorylate lipids. However, all members of
this family have been shown to possess serine/threonine kinase
activity. ATM phosphorylates key proteins involved in a variety of
cell-cycle checkpoint signalling pathways that are initiated in
response to DNA DSBs production (see below). These downstream
effector proteins include p53, Chk2, NBS1/nibrin, BRCA1 and Rad 17
(Abraham, 2001)
[0005] ATM is the product of the gene mutated in
ataxia-telangiectasia (A-T) (Savitsky et al (1995)). A-T is a human
autosomal recessive disorder present at an incidence of around 1 in
100,000 in the population. A-T is characterised by a number of
debilitating symptoms, including progressive cerebellar
degeneration, occulocutaneous telangiectasia, growth retardation,
immune deficiencies, cancer predisposition and certain
characteristics of premature ageing (Lavin and Shiloh (1997), The
genetic defect in ataxia-telangiectasia.
[0006] Annu. Rev. Immunol., 15:177-202; Shiloh (2001), ATM and ATR:
networking cellular responses to DNA damage, Curr. Opin. Genet.
Dev., 11:71-7). At the cellular level, A-T is characterised by a
high degree of chromosomal instability, radio-resistant DNA
synthesis, and hypersensitivity to ionizing radiation (IR) and
radiomimetic drugs. In addition, A-T cells are defective in the
radiation induced G.sub.1-S, S, and
G.sub.2-M cell cycle checkpoints that are thought to arrest the
cell cycle in response to DNA damage in order to allow repair of
the genome prior to DNA replication or mitosis (Lavin and Shiloh,
1997). This may in part reflect the fact that A-T cells exhibit
deficient or severely delayed induction of p53 in response to IR.
Indeed, p53-mediated downstream events are also defective in A-T
cells following IR exposure. ATM therefore acts upstream of p53 in
an IR-induced DNA damage signalling pathway. A-T cells have also
been shown to accumulate DNA double-strand breaks (dsbs) after
ionizing radiation, suggesting a defect in dsb repair.
[0007] It is clear that ATM is a key regulator of the cellular
response to DNA DSBs. Therefore the inhibition of this kinase
through small molecules will sensitise cells to both ionising
radiation and to chemotherapeutics that induce DNA DSBs either
directly or indirectly. ATM inhibitors may thus be used as adjuncts
in cancer radiotherapy and chemotherapy. To date the only reported
inhibitors of ATM (caffeine and wortmannin; Sarkaria, et al.,
(1999) Inhibition of ATM and ATR kinase activities by the
radiosensitizing agent, caffeine. Cancer Res., 59:4375-82; Banin,
et al., (1998) Enhanced phosphorylation of p53 by ATM in response
to DNA damage. Science, 281:1674-1677) do cause radiosensitisation
but it is unclear whether this mechanism of action is mediated
through ATM inhibition as these small molecules are very
non-specific in action as kinase inhibitors.
[0008] ATM function in response to ionising radiation induced DNA
damage has been shown to be tissue specific. For example, while
fibroblasts derived from Atm null mice are radiosensitive Atm null
neurons are radioresistant through a lack of IR induced apoptosis
(Herzog et al., (1998) Requirement for Atm in ionizing
radiation-induced cell death in the developing central nervous
system. Science, 280: 1089-91). Therefore, inhibitors of ATM have
the potential to be radio-protective in specific cellular
contexts.
[0009] ATM inhibitors may also prove useful in the treatment of
retroviral mediated diseases. It has been demonstrated that ATM
function is required to allow stable retroviral DNA transduction
under certain conditions (Daniel et al. (2001) Wortmannin
potentiates integrase-mediated killing of lymphocytes and reduces
the efficiency of stable transduction by retroviruses. Mol. Cell.
Biol., 21: 1164-72). Therefore ATM inhibitors have the potential to
block retroviral DNA integration.
[0010] ATM is known to play a crucial role in controlling the
length of telomeric chromosomal ends (Metcalfe et al. (1996)
Accelerated telomere shortening in ataxia telangiectasia. Nat.
Genet., 13:350-3). Telomeric ends in most normal cell types shorten
at each cell division. Cells with excessively shortened telomeres
are unable to divide. Inhibitors of ATM may therefore, have utility
in preventing cancer progression by limiting the growth potential
of cancerous or pre-cancerous cells. Furthermore, ATM does not
appear to be part of the telomerase enzyme itself (Metcalfe et al.
(1996)) Therefore it is likely that ATM inhibitors will work
synergistically with anti-telomerase drugs.
[0011] Cells derived from A-T patients or from mice null for ATM
grow slower in culture than genetically matched ATM positive cells.
Therefore an ATM inhibitor may have growth
inhibitory/anti-proliferative properties in its own right.
Therefore an ATM inhibitor may be used as a cytostatic agent in the
treatment of cancer.
[0012] A-T patients display immuno-deficiencies, demonstrating that
ATM is required for generation of a fully functional immune system.
Inhibitors of ATM may, therefore, be used in modulating the immune
system.
[0013] In summary ATM inhibitors have the potential to sensitise
tumour cells to ionising radiation or DNA DSB inducing
chemotherapeutics, to modulate telomere length control mechanisms,
to block retroviral integration, modulate the immune system and to
protect certain cell types from DNA damage induced apoptosis.
[0014] Some of the present inventors have previously described a
broad class of compounds which exhibit inhibition of ATM. These are
described in an International Patent application (PCT/GB03/001817,
filed 29 Apr. 2003) and a US patent application Ser. No.
(10/426,147, filed 29 Apr. 2003).
[0015] The present inventors have now discovered within that broad
class of compounds, further specific classes compounds which
exhibit inhibition of ATM. Accordingly, the first aspect of the
invention provides a compound of formula I:
##STR00002##
and isomers, salts, solvates, chemically protected forms, and
prodrugs thereof, wherein: R.sup.1 and R.sup.2 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
R.sup.N1 is selected from hydrogen, an optionally substituted
C.sub.1-7 alkyl group, an optionally substituted C.sub.3-20
heterocyclyl group, an optionally substituted C.sub.5-20 aryl
group, an acyl group, an ester group and an amido group.
[0016] The second aspect of the invention provides a compound of
formula II:
##STR00003##
and isomers, salts, solvates, chemically protected forms, and
prodrugs thereof, wherein: R.sup.1 and R.sup.2 together form, along
with the nitrogen atom to which they are attached, an optionally
substituted heterocyclic ring having from 4 to 8 ring atoms;
R.sup.c1 is --NR.sup.3R.sup.4, where R.sup.3 and R.sup.4 are
independently selected from hydrogen, optionally substituted
C.sub.1-7 alkyl groups, optionally substituted C.sub.3-20
heterocyclyl groups and optionally substituted C.sub.5-20 aryl
groups, or together form, along with the nitrogen atom to which
they are attached, an optionally substituted heterocyclic ring
having from 4 to 8 ring atoms, or R.sup.c1 is of formula IIa:
##STR00004##
wherein R.sup.C2 is selected from an optionally substituted
C.sub.1-7 alkyl group, an optionally substituted C.sub.3-20
heterocyclyl group, an optionally substituted C.sub.5-20 aryl
group, an ester group, an ether group and an amino group.
[0017] A third aspect of the invention provides a compound of
formula III:
##STR00005##
and isomers, salts, solvates, chemically protected forms, and
prodrugs thereof, wherein: R.sup.1 and R.sup.2 together form, along
with the nitrogen atom to which they are attached, an optionally
substituted heterocyclic ring having from 4 to 8 ring atoms;
R.sup.C3 is of formula:
##STR00006##
wherein r is 0 or 1, and q can be 1 or 2 when r is 0 and q is 1
when r is 1, and wherein R.sup.N4 and R.sup.N5 are independently
selected from hydrogen, optionally substituted C.sub.1-7 alkyl
groups, optionally substituted C.sub.3-20 heterocyclyl groups and
optionally substituted C.sub.5-20 aryl groups, or together form,
along with the nitrogen atom to which they are attached, an
optionally substituted heterocyclic ring having from 4 to 8 ring
atoms.
[0018] A fourth aspect of the invention provides a compound of
formula IV:
##STR00007##
and isomers, salts, solvates, chemically protected forms, and
prodrugs thereof, wherein: R.sup.1 and R.sup.2 together form, along
with the nitrogen atom to which they are attached, an optionally
substituted heterocyclic ring having from 4 to 8 ring atoms;
R.sup.N6 is of formula:
##STR00008##
wherein R.sup.N7 and R.sup.N8 are independently selected from
hydrogen, optionally substituted C.sub.1-7 alkyl groups, optionally
substituted C.sub.3-20 heterocyclyl groups and optionally
substituted C.sub.5-20 aryl groups, or 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] A fifth aspect of the invention provides a compound of
formula V:
##STR00009##
and isomers, salts, solvates, chemically protected forms, and
prodrugs thereof, wherein: R.sup.1 and R.sup.2 together form, along
with the nitrogen atom to which they are attached, an optionally
substituted heterocyclic ring having from 4 to 8 ring atoms; n is 1
or 2; and R.sup.N9 is selected from hydrogen, an optionally
substituted C.sub.1-7 alkyl group, an optionally substituted
C.sub.3-20 heterocyclyl group, an optionally substituted C.sub.5-20
aryl group, an acyl group, an ester group or an amido group; with
the proviso that R.sup.N9 is not an unsubstituted methyl group.
[0020] A sixth aspect of the present invention provides a compound
of formula VI:
##STR00010##
and isomers, salts, solvates, chemically protected forms, and
prodrugs thereof, wherein: R.sup.1 and R.sup.2 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
R.sup.C4 is of formula:
##STR00011##
wherein r is 0 or 1, and q can be 1 or 2 when r is 0 and q is 1
when r is 1, and wherein R.sup.N4 and R.sup.N5 are independently
selected from hydrogen, optionally substituted C.sub.1-7 alkyl
groups, optionally substituted C.sub.3-20 heterocyclyl groups and
optionally substituted C.sub.5-20 aryl groups, or together form,
along with the nitrogen atom to which they are attached, an
optionally substituted heterocyclic ring having from 4 to 8 ring
atoms.
[0021] A seventh aspect of the present invention provides a
compound of formula VII:
##STR00012##
and isomers, salts, solvates, chemically protected forms, and
prodrugs thereof, wherein: R.sup.1 and R.sup.2 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
R.sup.N10 and R.sup.N11 are independently selected from hydrogen,
optionally substituted C.sub.1-7 alkyl groups, optionally
substituted C.sub.3-20 heterocyclyl groups and optionally
substituted C.sub.5-20 aryl groups, or together form, along with
the nitrogen atom to which they are attached, an optionally
substituted heterocyclic ring having from 4 to 8 ring atoms.
[0022] An eight aspect of the present invention provides a compound
of formula VIII:
##STR00013##
and isomers, salts, solvates, chemically protected forms, and
prodrugs thereof, wherein: R.sup.1 and R.sup.2 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
R.sup.N12 and R.sup.N13 are independently selected from hydrogen,
optionally substituted C.sub.1-7 alkyl groups, optionally
substituted C.sub.3-20 heterocyclyl groups and optionally
substituted C.sub.5-20 aryl groups, or together form, along with
the nitrogen atom to which they are attached, an optionally
substituted heterocyclic ring having from 4 to 8 ring atoms.
[0023] A ninth aspect of the present invention provides a compound
of formula IX:
##STR00014##
and isomers, salts, solvates, chemically protected forms, and
prodrugs thereof, wherein: R.sup.1 and R.sup.2 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
R.sup.C5 is of formula:
##STR00015##
wherein q is 1 or 2, and wherein R.sup.N4 and R.sup.N5 are
independently selected from hydrogen, optionally substituted
C.sub.1-7 alkyl groups, optionally substituted C.sub.3-20
heterocyclyl groups and optionally substituted C.sub.5-20 aryl
groups, or together form, along with the nitrogen atom to which
they are attached, an optionally substituted heterocyclic ring
having from 4 to 8 ring atoms.
[0024] A tenth aspect of the invention provides a composition
comprising a compound of any one of the first to ninth aspects and
a pharmaceutically acceptable carrier or diluent.
[0025] An eleventh aspect of the invention provides the use of a
compound of any one of the first to ninth aspects in a method of
therapy.
[0026] A twelfth aspect of the invention provides the use of a
compound of any one of the first to ninth aspects in the
preparation of a medicament for inhibiting the activity of ATM.
[0027] A thirteenth aspect of the invention provides for the use of
a compound as defined in one of the first to ninth aspects of the
invention in the preparation of a medicament for use as an adjunct
in cancer therapy or for potentiating tumour cells for treatment
with ionising radiation or chemotherapeutic agents.
[0028] A fourteenth aspect of the invention provides for the use of
a compound as defined in any one of the first to ninth aspects of
the invention in the preparation of a medicament for the treatment
of retroviral mediated diseases or disease ameliorated by the
inhibition of ATM, which include acquired immunodeficiency
syndrome.
[0029] 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.
[0030] Another aspect of the invention provides a method of
inhibiting ATM in vitro or in vivo, comprising contacting a cell
with an effective amount of an active compound as described
herein.
DEFINITIONS
[0031] C.sub.1-7 alkyl: The term "C.sub.1-7 alkyl", 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.
[0032] 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).
[0033] 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.
[0034] 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.
[0035] 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.
[0036] 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).
[0037] 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.3-7 cycloalkenyl" 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.
[0038] 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. "C.sub.3-20" denotes ring atoms, whether
carbon atoms or heteroatoms.
[0039] 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.
[0040] 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.
[0041] 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.
[0042] 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.
[0043] 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.
[0044] 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.
[0045] 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).
[0046] 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.
[0047] Other examples of C.sub.3-20 heterocyclyl groups include,
but are not limited to, oxadiazine and oxathiazine.
[0048] Examples of heterocyclyl groups which additionally bear one
or more oxo (.dbd.O) groups, include, but are not limited to, those
derived from: [0049] C.sub.5 heterocyclics, such as furanone,
pyrone, pyrrolidone (pyrrolidinone), pyrazolone (pyrazolinone),
imidazolidone, thiazolone, and isothiazolone; [0050] C.sub.6
heterocyclics, such as piperidinone (piperidone), piperidinedione,
piperazinone, piperazinedione, pyridazinone, and pyrimidinone
(e.g., cytosine, thymine, uracil), and barbituric acid; [0051]
fused heterocyclics, such as oxindole, purinone (e.g., guanine),
benzoxazolinone, benzopyrone (e.g., coumarin); [0052] cyclic
anhydrides (--C(.dbd.O)--O--C(.dbd.O)-- in a ring), including but
not limited to maleic anhydride, succinic anhydride, and glutaric
anhydride; [0053] cyclic carbonates (--O--C(.dbd.O)--O-- in a
ring), such as ethylene carbonate and 1,2-propylene carbonate;
[0054] imides (--C(.dbd.O)--NR--C(.dbd.O)-- in a ring), including
but not limited to, succinimide, maleimide, phthalimide, and
glutarimide; [0055] 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; [0056] 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; [0057] cyclic
carbamates (--O--C(.dbd.O)--NR-- in a ring), such as 2-oxazolidone;
[0058] cyclic ureas (--NR--C(.dbd.O)--NR-- in a ring), such as
2-imidazolidone and pyrimidine-2,4-dione (e.g., thymine,
uracil).
[0059] 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.
[0060] 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.
[0061] 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).
[0062] 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.
[0063] 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.
[0064] 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).
[0065] Examples of 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.
[0066] 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.
Halo: --F, --Cl, --Br, and --I.
Hydroxy: --OH.
[0067] 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. C.sub.1-7 alkoxy: --OR, wherein
R is a C.sub.1-7 alkyl group. Examples of C.sub.1-7 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). C.sub.1-2 alkdioxylene: The term "C.sub.1-2
alkdioxylene," as used herein, pertains to a bidentate moiety
obtained by removing two hydrogen atoms from each of two different
alcohol groups of a C.sub.1-2 hydrocarbon diol compound having from
1 or 2 carbon atoms, i.e. CH.sub.2(OH).sub.2 and
HO--CH.sub.2--CH.sub.2--OH, to form --O--CH.sub.2--O-- and
--O--CH.sub.2--CH.sub.2--O--. This bidentate moiety may be the
substituent group of a single atom or of two adjacent atoms. 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. 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. Formyl (carbaldehyde,
carboxaldehyde): --C(.dbd.O)H. 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) Carboxy (carboxylic acid): --COOH. 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. 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. 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. 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:
##STR00016##
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.
Tetrazolyl: a five membered aromatic ring having four nitrogen
atoms and one carbon atom,
##STR00017##
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. Imino: .dbd.NR, wherein R is an imino 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. 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.
Nitro: --NO.sub.2.
Nitroso: --NO.
Azido: --N.sub.3.
[0068] Cyano (nitrile, carbonitrile): --CN.
Isocyano: --NC.
Cyanato: --OCN.
Isocyanato: --NCO.
[0069] Thiocyano (thiocyanato): --SCN. Isothiocyano
(isothiocyanato): --NCS. Sulfhydryl (thiol, mercapto): --SH.
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.
[0070] 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.
Sulfone (sulfonyl): --S(.dbd.O).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).
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. 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. 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. 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. 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. 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.
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.
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:
##STR00018##
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. 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.
[0071] 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.
Includes Other Forms
[0072] 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
[0073] 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").
[0074] 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).
[0075] 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.
##STR00019##
Note that specifically included in the term "isomer" are compounds
with one or more isotopic substitutions. For example, H may be in
any isotopic form, including .sup.1H, .sup.2H (D), and .sup.3H (T);
C may be in any isotopic form, including .sup.12C, .sup.13C, and
.sup.14C; O may be in any isotopic form, including .sup.16O and
.sup.18O; and the like.
[0076] 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.
[0077] Unless otherwise specified, a reference to a particular
compound also includes ionic, salt, solvate, and protected forms of
thereof, for example, as discussed below.
[0078] 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., 1977,
"Pharmaceutically Acceptable Salts", J. Pharm. Sci., Vol. 66, pp.
1-19.
[0079] 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.sup.+, NR.sub.4.sup.+). Examples of some suitable
substituted ammonium ions are those derived from: ethylamine,
diethylamine, dicyclohexylamine, triethylamine, butylamine,
ethylenediamine, ethanolamine, diethanolamine, piperazine,
benzylamine, phenylbenzylamine, choline, meglumine, and
tromethamine, as well as amino acids, such as lysine and arginine.
An example of a common quaternary ammonium ion is
N(CH.sub.3).sub.4.sup.+.
[0080] 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.
[0081] 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.
[0082] 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, 1999).
[0083] 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).
[0084] 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.
[0085] 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.cndot.).
[0086] 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.
[0087] 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).
[0088] 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.
[0089] 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).
[0090] 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.
Further Preferences
[0091] The following preferences may be different for different
aspects of the present invention, and may be combined together.
[0092] In formulae I, II, III, IV, V, VI, VII, VIII and IX, R.sup.1
and R.sup.2 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.1 and R.sup.2 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.
[0093] Single rings having one nitrogen atom include azetidine,
azetidine, pyrrolidine (tetrahydropyrrole), pyrroline (e.g.,
3-pyrroline, 2,5-dihydropyrroe), 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.
[0094] 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.
[0095] The most preferred groups are morpholino and
thiomorpholino.
[0096] 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.
[0097] 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:
##STR00020##
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.
First Aspect
[0098] Preferably R.sup.N1 is selected from hydrogen, an optionally
substituted C.sub.1-7 alkyl group, an optionally substituted
C.sub.5-20 aryl group, an acyl group and an ester group.
[0099] If R.sup.N1 is an optionally substituted C.sub.1-7 alkyl
group, it is preferably an optionally substituted C.sub.1-4 alkyl
group, and more preferably an optionally substituted C.sub.1-2
alkyl group. The optional substituents are preferably selected from
hydroxyl, C.sub.1-7 alkoxy (e.g. methoxy, ethoxy), acyl and
amido.
[0100] If R.sup.N1 is an optionally substituted C.sub.5-7 aryl
group, it is preferably an optionally substituted carboaryl or
heteroaryl (containing 1 or 2 nitrogen ring atoms) group and more
preferably an optionally substituted phenyl, pyridyl or pyrimidyl
group. It is further preferred that the C.sub.5-7 aryl group is
unsubstituted.
[0101] If R.sup.N1 is an acyl group, then the acyl substituent is
preferably a C.sub.1-7 alkyl group (more preferably C.sub.1-4
alkyl, e.g. methyl) or a C.sub.5-7 aryl group (more preferably
C.sub.5-6 aryl, e.g. furanyl, thiophenyl, phenyl, pyridyl).
[0102] If R.sup.N1 is an ester group, the ester substituents is
preferably a C.sub.1-7 alkyl group (more preferably C.sub.1-4
alkyl, e.g. methyl, t-butyl).
[0103] Particularly preferred R.sup.N1 groups include, but are not
limited to, 2-fluoro phenyl and furan-2-yl.
Second Aspect
[0104] R.sup.c1 is preferably of formula IIa.
[0105] If R.sup.c1 is --NR.sup.3R.sup.4, then R.sup.3 and R.sup.4
are preferably selected from optionally substituted C.sub.1-7 alkyl
groups and more preferably optionally substituted C.sub.1-4 alkyl
groups. It is most preferred that one, or both, of R.sup.3 and
R.sup.4 is hydrogen.
[0106] R.sup.C2 is preferably selected from an optionally
substituted C.sub.1-7 alkyl group, an optionally substituted
C.sub.3-20 heterocyclyl group, an optionally substituted C.sub.5-20
aryl group and an ester group, and is more preferably selected from
an optionally substituted C.sub.1-7 alkyl group and an optionally
substituted C.sub.3-20 heterocyclyl group.
[0107] If R.sup.C2 is an ester group, the ester substituent is
preferably a C.sub.1-7 alkyl group (more preferably a C.sub.1-4
alkyl group, e.g. methyl).
[0108] If R.sup.C2 is an optionally substituted C.sub.3-20
heterocyclyl group, it is preferably an optionally substituted
C.sub.5-7 heterocyclyl group, and more preferably contains at least
one nitrogen ring atom (e.g. 4-piperidyl).
[0109] If R.sup.C2 is an optionally substituted C.sub.5-20 aryl
group, then it is preferably an optionally substituted C.sub.5-6
aryl group. More preferably, it is selected from optionally
substituted phenyl or a C.sub.5-6 heteroaryl group containing one
or two heteroatoms selected from nitrogen, oxygen and sulphur (e.g.
pyridyl, thiazolyl, furanyl, thiophenyl, isoxazolyl,
1,2-diazolyl).
[0110] If R.sup.C2 is an optionally substituted C.sub.1-7 alkyl
group, it is preferably an optionally substituted C.sub.1-4 alkyl
group (e.g. methyl, ethyl, propyl, cyclopropyl, cyclobutyl).
Preferable optional substituents include, but are not limited to,
amino, thioether, ester, C.sub.5-20 aryl, C.sub.3-20 heterocyclyl,
acyloxy, ether and alkoxy.
[0111] In one embodiment, R.sup.C2 is of formula IIb:
##STR00021##
wherein r is from 1 to 3 (more preferably 1 or 2); and R.sup.N2 and
R.sup.N3 are independently selected from hydrogen, optionally
substituted C.sub.1-7 alkyl groups, optionally substituted
C.sub.3-20 heterocyclyl groups and optionally substituted
C.sub.5-20 aryl groups, or together form, along with the nitrogen
atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms.
[0112] More preferably R.sup.N2 and R.sup.N3 are independently
selected from hydrogen, optionally substituted C.sub.1-7 alkyl
groups or together form, along with the nitrogen atom to which they
are attached, an optionally substituted heterocyclic ring having
from 4 to 8 ring atoms.
[0113] If one of R.sup.N2 and R.sup.N3 is an optionally substituted
C.sub.1-7 alkyl group, the other is preferably H. The optionally
substituted C.sub.1-7 alkyl group, is preferably an optionally
substituted C.sub.1-4 alkyl group (e.g. methyl, ethyl, n-propyl,
n-butyl). The optional substituents are preferably selected from
hydroxy, NH.sub.2, C.sub.3-20 heterocylyl (more preferably
C.sub.5-7 heterocylyl) and C.sub.5-20 aryl (more preferably
C.sub.5-7 aryl).
[0114] If R.sup.N2 and R.sup.N3 together form, along with the
nitrogen atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms, this ring
preferably has 6 or 7 ring atoms, and is more preferably selected
from piperidine, piperazine, homopiperazine and morpholino. Where
the heterocyclic ring is piperazine or homopiperazine, the other
nitrogen ring atom may be substituted by, for example, an
optionally substituted C.sub.1-7 alkyl group, an optionally
substituted C.sub.3-20 heterocyclyl group, an optionally
substituted C.sub.5-20 aryl group, an acyl group, an ester group or
an amido group. Of these possible substituents, the optionally
substituted C.sub.1-7 alkyl group (e.g. methyl), the optionally
substituted C.sub.5-20 aryl group (e.g. phenyl) and the acyl group
are more preferred.
Third Aspect
[0115] R.sup.C3 may be of formula:
##STR00022##
It is preferred that r=0.
[0116] More preferably R.sup.N4 and R.sup.N5 are independently
selected from hydrogen, optionally substituted C.sub.1-7 alkyl
groups, or together form, along with the nitrogen atom to which
they are attached, an optionally substituted heterocyclic ring
having from 4 to 8 ring atoms.
[0117] If one of R.sup.N4 and R.sup.N5 is an optionally substituted
C.sub.1-7 alkyl group, the other is preferably H. The optionally
substituted C.sub.1-7 alkyl group, is preferably an optionally
substituted C.sub.1-4 alkyl group (e.g. methyl, ethyl, n-propyl,
n-butyl). The optional substituents are preferably selected from
hydroxy, amino, cyano, C.sub.3-20 heterocylyl (more preferably
C.sub.5-7 heterocylyl) and C.sub.5-20 aryl (more preferably
C.sub.5-7 aryl).
[0118] In one embodiment, one of R.sup.N4 and R.sup.N5 is of
formula IIb:
##STR00023##
as defined above.
[0119] If R.sup.N4 and R.sup.N5 together form, along with the
nitrogen atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms, this ring
preferably has 6 or 7 ring atoms, and is more preferably selected
from piperidine, homopiperidine, piperazine, homopiperazine and
morpholino. Where the heterocyclic ring is piperazine or
homopiperazine, the other nitrogen ring atom may be substituted by,
for example, an optionally substituted C.sub.1-7 alkyl group, an
optionally substituted C.sub.3-20 heterocyclyl group, an optionally
substituted C.sub.5-20 aryl group, an acyl group, an ester group or
an amido group. Of these possible substituents, the optionally
substituted C.sub.1-7 alkyl group (e.g. methyl), the optionally
substituted C.sub.5-20 aryl group (e.g. phenyl) and the acyl group
are more preferred.
Fourth Aspect
[0120] R.sup.N7 and R.sup.N8 are preferably independently selected
from hydrogen, optionally substituted C.sub.1-7 alkyl groups and
optionally substituted C.sub.3-20 heterocyclyl groups, or together
form, along with the nitrogen atom to which they are attached, an
optionally substituted heterocyclic ring having from 4 to 8 ring
atoms.
[0121] More preferably R.sup.N7 and R.sup.N8 are independently
selected from hydrogen, optionally substituted C.sub.1-7 alkyl
groups or together form, along with the nitrogen atom to which they
are attached, an optionally substituted heterocyclic ring having
from 4 to 8 ring atoms.
[0122] If one of R.sup.N7 and R.sup.N8 is an optionally substituted
C.sub.1-7 alkyl group, the other is preferably H. The optionally
substituted C.sub.1-7 alkyl group, is preferably an optionally
substituted C.sub.1-4 alkyl group (e.g. methyl, ethyl, n-propyl,
n-butyl). The optional substituents are preferably selected from
hydroxy, halo, amino, C.sub.4-7 cycloalkyl, C.sub.3-20 heterocylyl
(more preferably C.sub.5-7 heterocylyl) and C.sub.5-20 aryl (more
preferably C.sub.5-7 aryl).
[0123] In one embodiment, one of R.sup.N7 and R.sup.N8 is of
formula IIb:
##STR00024##
as defined above.
[0124] If R.sup.N7 and R.sup.N8 together form, along with the
nitrogen atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms, this ring
preferably has 5 to 7 ring atoms, and is more preferably selected
from pyrrole, piperidine, piperazine, homopiperazine and
morpholino. Where the heterocyclic ring is piperazine or
homopiperazine, the other nitrogen ring atom may be substituted by,
for example, an optionally substituted C.sub.1-7 alkyl group, an
optionally substituted C.sub.3-20 heterocyclyl group, an optionally
substituted C.sub.5-20 aryl group, an acyl group, an ester group or
an amido group. Of these possible substituents, the optionally
substituted C.sub.1-7 alkyl group (e.g. methyl), the optionally
substituted C.sub.5-20 aryl group (e.g. phenyl) and the acyl group
are more preferred. Where the heterocyclic ring does not include a
further nitrogen ring atom, the ring may be further substituted by
one or more substituent groups, for example, but not limited to,
C.sub.1-7 alkyl, amido, hydroxy and ester.
Fifth Aspect
[0125] R.sup.N9 is preferably selected from hydrogen, an optionally
substituted C.sub.1-7 alkyl group, an optionally substituted
C.sub.3-20 heterocyclyl group, an optionally substituted C.sub.5-20
aryl group, or an acyl group; with the proviso that R.sup.N9 is not
an unsubstituted methyl group.
[0126] If R.sup.N9 is an optionally substituted C.sub.1-7 alkyl
group, it is preferably an optionally substituted C.sub.1-4 alkyl
group, and more preferably an optionally substituted C.sub.1-2
alkyl group. The optional substituents are preferably selected from
hydroxyl, C.sub.1-7 alkoxy (e.g. methoxy, ethoxy), acyl and
amido.
[0127] If R.sup.N9 is an optionally substituted C.sub.5-7 aryl
group, it is preferably an optionally substituted carboaryl or
heteroaryl (containing 1 or 2 nitrogen ring atoms) group and more
preferably an optionally substituted phenyl, pyridyl or pyrimidyl
group. It is further preferred that the C.sub.5-7 aryl group is
unsubstituted.
[0128] If R.sup.N9 is an acyl group, then the acyl substituent is
preferably a C.sub.1-7 alkyl group (more preferably C.sub.1-4
alkyl, e.g. methyl) or a C.sub.5-7 aryl group (more preferably
C.sub.5-6 aryl, e.g. furanyl, thiophenyl, phenyl, pyridyl).
[0129] Particularly preferred R.sup.N9 groups include, but are not
limited to, 4-fluoro phenyl, ethyl and
2-(2'-hydroxy-ethoxy)-ethyl.
Sixth Aspect
[0130] R.sup.C4 may be of formula:
##STR00025##
It is preferred that r=0.
[0131] More preferably R.sup.N4 and R.sup.N5 are independently
selected from hydrogen, optionally substituted C.sub.1-7 alkyl
groups, or together form, along with the nitrogen atom to which
they are attached, an optionally substituted heterocyclic ring
having from 4 to 8 ring atoms.
[0132] If one of R.sup.N4 and R.sup.N5 is an optionally substituted
C.sub.1-7 alkyl group, the other is preferably H. The optionally
substituted C.sub.1-7 alkyl group, is preferably an optionally
substituted C.sub.1-4 alkyl group (e.g. methyl, ethyl, n-propyl,
n-butyl). The optional substituents are preferably selected from
hydroxy, halo, C.sub.1-7 alkoxy (e.g. methoxy), thiol, C.sub.1-7
thioether (e.g. --SMe), amino, ester (preferably C.sub.1-7 alkyl
ester, e.g. --C(.dbd.O)OMe), cyano, C.sub.3-20 heterocylyl (more
preferably C.sub.5-7 heterocylyl) and C.sub.5-20 aryl (more
preferably C.sub.5-7 aryl).
[0133] In one embodiment, one of R.sup.N4 and R.sup.N5 is of
formula IIb:
##STR00026##
as defined above.
[0134] If R.sup.N4 and R.sup.N5 together form, along with the
nitrogen atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms, this ring
preferably has 6 or 7 ring atoms, and is more preferably selected
from piperidine, homopiperidine, piperazine, homopiperazine and
morpholino. Where the heterocyclic ring is piperazine or
homopiperazine, the other nitrogen ring atom may be substituted by,
for example, an optionally substituted C.sub.1-7 alkyl group, an
optionally substituted C.sub.3-20 heterocyclyl group, an optionally
substituted C.sub.5-20 aryl group, an acyl group, an ester group or
an amido group. Of these possible substituents, the optionally
substituted C.sub.1-7 alkyl group (e.g. methyl), the optionally
substituted C.sub.5-20 aryl group (e.g. phenyl) and the acyl group
are more preferred.
Seventh Aspect
[0135] Preferably R.sup.N10 and R.sup.N11 are independently
selected from hydrogen, optionally substituted C.sub.1-7 alkyl
groups or together form, along with the nitrogen atom to which they
are attached, an optionally substituted heterocyclic ring having
from 4 to 8 ring atoms.
[0136] If one of R.sup.N10 and R.sup.N11 is an optionally
substituted C.sub.1-7 alkyl group, the other is preferably H. The
optionally substituted C.sub.1-7 alkyl group, is preferably an
optionally substituted C.sub.1-4 alkyl group (e.g. methyl, ethyl,
n-propyl, n-butyl). The optional substituents are preferably
selected from hydroxy, halo, cyano, NH.sub.2, C.sub.1-7 alkoxy
(e.g. methoxy), C.sub.1-7 thioether (e.g. --SMe), C.sub.3-20
heterocylyl (more preferably C.sub.5-7 heterocylyl) and C.sub.5-20
aryl (more preferably C.sub.5-7 aryl).
[0137] If R.sup.N10 and R.sup.N11 together form, along with the
nitrogen atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms, this ring
preferably has 6 or 7 ring atoms, and is more preferably selected
from piperidine, homopiperadine, piperazine, homopiperazine,
morpholino and thiomorpholino. These groups may be optionally
substituted, for example by an optionally substituted C.sub.1-7
alkyl or amido groups. Where the heterocyclic ring is piperazine or
homopiperazine, the other nitrogen ring atom may be substituted by,
for example, an optionally substituted C.sub.1-7 alkyl group, an
optionally substituted C.sub.3-20 heterocyclyl group, an optionally
substituted C.sub.5-20 aryl group, an acyl group, an ester group or
an amido group. Of these possible substituents, the optionally
substituted C.sub.1-7 alkyl group (e.g. methyl), the optionally
substituted C.sub.5-20 aryl group (e.g. phenyl) and the acyl group
are more preferred.
Eighth Aspect
[0138] Preferably R.sup.N12 and R.sup.N13 are independently
selected from hydrogen, optionally substituted C.sub.1-7 alkyl
groups or together form, along with the nitrogen atom to which they
are attached, an optionally substituted heterocyclic ring having
from 4 to 8 ring atoms.
[0139] If one of R.sup.N12 and R.sup.N13 is an optionally
substituted C.sub.1-7 alkyl group, the other is preferably H. The
optionally substituted C.sub.1-7 alkyl group, is preferably an
optionally substituted C.sub.1-4 alkyl group (e.g. methyl, ethyl,
n-propyl, n-butyl). The optional substituents are preferably
selected from hydroxy, halo, cyano, NH.sub.2, C.sub.1-7 alkoxy
(e.g. methoxy), C.sub.1-7 thioether (e.g. --SMe), C.sub.3-20
heterocylyl (more preferably C.sub.5-7 heterocylyl) and C.sub.5-20
aryl (more preferably C.sub.5-7 aryl).
[0140] If R.sup.N12 and R.sup.N13 together form, along with the
nitrogen atom to which they are attached, an optionally substituted
heterocyclic ring having from 4 to 8 ring atoms, this ring
preferably has 6 or 7 ring atoms, and is more preferably selected
from piperidine, homopiperadine, piperazine, homopiperazine,
morpholino and thiomorpholino. These groups may be optionally
substituted, for example by an optionally substituted C.sub.1-7
alkyl or amido groups. Where the heterocyclic ring is piperazine or
homopiperazine, the other nitrogen ring atom may be substituted by,
for example, an optionally substituted C.sub.1-7 alkyl group, an
optionally substituted C.sub.3-20 heterocyclyl group, an optionally
substituted C.sub.5-20 aryl group, an acyl group, an ester group or
an amido group. Of these possible substituents, the optionally
substituted C.sub.1-7 alkyl group (e.g. methyl), the optionally
substituted C.sub.5-20 aryl group (e.g. phenyl) and the acyl group
are more preferred.
Ninth Aspect
[0141] R.sup.C5 may be of formula:
##STR00027##
It is preferred that q=2.
[0142] More preferably R.sup.N4 and R.sup.N5 are independently
selected from hydrogen, optionally substituted C.sub.1-7 alkyl
groups.
[0143] If one of R.sup.N4 and R.sup.N5 is an optionally substituted
C.sub.1-7 alkyl group, the other is preferably H. The optionally
substituted C.sub.1-7 alkyl group, is preferably an optionally
substituted C.sub.1-4 alkyl group (e.g. methyl, ethyl, n-propyl,
n-butyl). The optional substituents are preferably selected from
hydroxy, amino, cyano, C.sub.3-20 heterocylyl (more preferably
C.sub.5-7 heterocylyl) and C.sub.5-20 aryl (more preferably
C.sub.5-7 aryl).
Acronyms
[0144] 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), acetyl (Ac), 1,3-bis(diphenylphosphino)
propane (dppf).
[0145] 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), dimethylformamide (DMF), tetrahydrofuran (THF), and
dimethylsulfoxide (DMSO).
Synthesis Routes
[0146] Compounds of the present invention may be synthesised by the
coupling of a 2-chloro-6-amino-pyran-4-one to an appropriate
arylboronic acid or arylboronate ester using a palladium catalysed
coupling reaction, e.g. Suzuki coupling.
Synthesis of 2-chloro-6-amino-pyran-4-ones
[0147] These may be synthesised by the following route:
##STR00028##
In step (a) CCl.sub.4 is added across the carbon-carbon double bond
of diketene by free-radical addition to yield
4-chloro-4(2,2,2,-trichloro-ethyl)-oxetan-2-one (1). Suitable
initiators include peroxide, such as BCHPO
((bis-4-t-butylcyclohexyl)peroxydicarbonate).
[0148] In step (b), the amine R.sup.1R.sup.2NH opens the lactone
ring by nucleophilic attack at the carbonyl centre. The oxy anion
generated then displaces the chlorine atom on the .alpha.-carbon to
give rise to a .beta.-keto-amide intermediate. Further elimination
of HCl finally give the 5,5-dichloro-1-amino-pent-4-ene-1,3-dione.
Suitable conditions for this step include inorganic base such as
sodium hydrogen carbonate and solvent such as dry
dichloromethane.
[0149] In step (c), ring closure takes place by displacement of one
of the 5-chloro groups by the oxygen of the amide moiety to form
the pyran-4-one ring, which reaction is catalysed by a Lewis acid,
such as perchloric acid.
Arylboronic Acids and Aryl Boronate Esters
[0150] The appropriate arylboronic acids and arylboronate esters
may be synthesised by using one of the routes described in the
examples below. General synthesis steps are shown below.
Synthesis of Aryl Boronate Esters
##STR00029##
[0151] Aryl boronate esters may be formed by Pd(0)-catalysed cross
coupling reaction of the appropriate aryl triflate or aryl halide
with tetra(alkoxy)diboron, e.g. pinacol diboron. Suitable
conditions include the use of a catalyst, such as PdCl.sub.2dppf,
extra ligands, such as dppf, potassium acetate as a base, in a
solvent such as dioxane, DMF or DMSO.
[0152] Examples of this method are to be found in T Ishiyama, et
al., Tet. Lett., vol. 38, no. 19, 3447-3450, 1997 and A Giroux, et
al., Tet. Lett., vol. 38, no. 22, 3841-3844, 1997.
Synthesis of Aryl Boronic Acids
##STR00030##
[0153] Boronic acids may be generated via lithiation of the
aromatic ring by tert-butyl lithium followed by the reaction of the
anion formed with alkyl borate such as triethyl borate to give the
desired aryl boronic acid.
Palladium Catalysed Coupling
[0154] The coupling of the arylboronic acid or arylboronate ester
to the 2-chloro-6-amino-pyran-4-one can be carried out using the
normal conditions, e.g. a palladium catalyst (Pd(PPh.sub.3).sub.4,
Pd(dppf)Cl.sub.2) and base (Na.sub.2CO.sub.3, NaOCH.sub.2CH.sub.3,
TlOH, N(CH.sub.2CH.sub.3).sub.3, K.sub.3PO.sub.4).
Use of Compounds of the Invention
[0155] The present invention provides active compounds,
specifically, active 2-aryl-6-amino-pyran-4-ones.
[0156] The term "active", as used herein, pertains to compounds
which are capable of inhibiting ATM 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.
[0157] One assay which may be used in order to assess the ATM
inhibition offered by a particular compound is described in the
examples below.
[0158] The present invention further provides a method of
inhibiting ATM 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.
[0159] 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.
[0160] The present invention further provides active compounds
which inhibit ATM activity as well as methods of inhibiting ATM
activity comprising contacting a cell with an effective amount of
an active compound, whether in vitro or in vivo.
[0161] 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.
[0162] 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.
[0163] The term "therapeutically-effective amount" as used herein,
pertains to that amount of an active compound, or a material,
composition or dosage form comprising an active compound, which is
effective for producing some desired therapeutic effect,
commensurate with a reasonable benefit/risk ratio.
[0164] The term "adjunct" as used herein relates to the use of
active compounds in conjunction with known therapeutic means. Such
means include cytotoxic regimens of drugs and/or ionising radiation
as used in the treatment of different cancer types. In particular,
the active compounds are known to potentiate the actions of a
number of cancer chemotherapy treatments, which include, but are
not limited to, the topoisomerase class of poisons and any
chemotherapeutic that will induce a DNA double strand break used in
treating cancer. Topoisomerase I inhibitors that may be used in
combination with compounds of the invention include the
camptothecin compounds, e.g. topotecan (Hycamtin), irinotecan
(CPT11-Camptosar), rubitecan and exatecan. Dual Topoisomerase I and
II inhibitors that may be used in combination with compounds of the
invention include benzophenainse, XR 11576/MLN 576 and
benzopyridoindoles. Topoisomerase II inhibitors that may be used in
combination with compounds of the invention include the
intercalators and DNA binders Doxorubicin, Danorubicin, and other
rubicins, the acridines (Amsacrine, m-AMSA), plus Mitoxantrone and
AQ4. Non-intercalators which are topoisomerase II inhibitors
include Etopside and Teniposide (epipodophyllotoxins).
[0165] The present inventors have previously found that ATM
inhibitory compounds of similar structure to those of the present
invention can efficiently repress retroviral vector transduction in
one-step, cell based integration assays (termed LUCIA) and inhibit
HIV-1 infection in 4-day replication assays at sub-micromolar
concentrations. Further, in contrast to the observations of Daniel
et al., where it was concluded that the effect of ATM on retroviral
integration would only be seen in a DNA-PK-deficient background,
this effect works in the presence of functional DNA-PK
activity.
[0166] Initial linkage of linear retroviral DNA with host cell
chromosomal DNA is catalysed by viral integrase (IN) and results in
short staggered DNA strand breaks in the host cell DNA at the site
of attachment (Brown, P. O. (1990) Integration of retroviral DNA.
Curr Top Microbiol Immunol, 157, 19-48). These gapped DNA
intermediates are shown to be sensed as sites of DNA damage by the
host cell and repaired by the ATM pathway to complete the process
of integration and allow productive infection to occur. Compounds
of the invention would be able to prevent the repair of gapped DNA
intermediates by the ATM pathway and thus prevent complete
integration of retroviral DNA into the host genome.
[0167] As described above, the invention provides a compound as
defined in the first to ninth aspects of the invention for use in
the treatment of retroviral infection and the use of such a
compound in the manufacture of a medicament for use in the
treatment of retroviral infection.
[0168] Also provided by the invention is a method of treatment of a
retroviral infection comprising administering a compound as defined
in the first to ninth aspects of the invention to an individual in
need thereof.
[0169] Retroviral mediated diseases which may be treated as
described above include HIV infection and acquired immunodeficiency
syndrome (AIDS) and Human T-cell Leukaemia virus (HTLV) infection
and its associated diseases adult T-cell leukaemia/lymphoma (ATLL)
and tropical spastic paraparesis/HTLV-1 associated myelopathy
(TSP/HAM).
[0170] Compounds of the invention may be used in combination with
other retroviral therapies to suppress virus replication, for
example in a `highly active anti-retroviral therapy` or HAART
treatment.
[0171] The invention provides a pharmaceutical composition
comprising a compound as described herein and one or more other
anti-retroviral agents.
[0172] The invention also provides a composition comprising a
compound as defined in the first to ninth aspects of the invention
and one or more other anti-retroviral agents for treatment of a
retroviral infection and the use of such a composition in the
manufacture of a medicament for use in the treatment of a
retroviral infection.
[0173] Suitable anti-retroviral agents which inhibit retroviral
replication, for example retroviral protease inhibitors (PI) such
as Sequinavir, Indinavir, Ritonavir and Nelfinavir, nucleoside
retroviral reverse transcriptase inhibitors such as
3'-azido-3'deoxythymidine (AZT; Zidovudine), 2',3'-Dideoxycytosine
(ddC; Zalcitabine), 2',3'-Dideoxyinosine (ddI; Didanosine) and 3TC;
(Lamivudine), and non-nucleoside retroviral reverse transcriptase
inhibitors such as Nevirapine, Delavirdine and Efavirenz.
Administration
[0174] 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.
[0175] 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, orangutan, gibbon), or a human.
Formulations
[0176] 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.
[0177] 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.
[0178] 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.
[0179] 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.
[0180] 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.
[0181] 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.
[0182] 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.
[0183] 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.
[0184] 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.
[0185] 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.
[0186] 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.
[0187] 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.
[0188] 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.
[0189] 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.
[0190] 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.
[0191] 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.
[0192] Alternatively, high melting point lipids such as white soft
paraffin and/or liquid paraffin or other mineral oils can be
used.
[0193] Formulations suitable for rectal administration may be
presented as a suppository with a suitable base comprising, for
example, cocoa butter or a salicylate.
[0194] 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.
[0195] 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
[0196] 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.
[0197] 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.
[0198] 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
[0199] The following examples are provided solely to illustrate the
present invention and are not intended to limit the scope of the
invention, as described herein.
A) Chemical Examples
General Experimental Methods
[0200] Thin layer chromatography was carried out using Merck
Kieselgel 60 F.sub.254 glass backed plates. The plates were
visualized by the use of a UV lamp (254 nm). Silica gel 60
(particle sizes 40-63.mu.) supplied by E.M. Merck was employed for
flash chromatography. .sup.1H NMR spectra were recorded at 300 MHz
on a Bruker DPX-300 instrument. Chemical shifts were referenced to
tetramethylsilane.
Purification and Identification of Libraries Samples
[0201] The samples were purified on Gilson LC units.
[0202] Mobile phase A--0.1% aqueous TFA, Mobile phase
B--Acetonitrile, Flow rate 6 ml/min., Gradient--typically starting
at 90% A/10% B for one minute, rising to 97% B after 15 minutes,
holding there for 2 minutes, then back to the starting conditions.
Column: Jones Chromatography Genesis 4.mu. C18 column, 10
mm.times.250 mm. Peak acquisition based on UV detection at 254
nm.
[0203] Mass Specs were recorded on a Finnegan LCQ instrument in
positive ion mode.
[0204] Mobile phase A--0.1% aqueous formic acid, Mobile phase
B--Acetonitrile, Flow rate 2 ml/min., Gradient--starting at 95%
A/5% B for one minute, rising to 98% B after 5 minutes, holding
there for 3 minutes, then back to the starting conditions.
Column--Phenomenex 5.mu. Luna C18 column, 4.6 mm.times.50 mm
[0205] UV detection at 254 nm, PDA detection scanning from 210 to
600 nm.
Mass Spectra of Other Compounds
[0206] Mass spectra of non-library compounds and building blocks
were recorded on a Micromass ZQ instrument (single quadrupole,
operating in electrospray ionisation mode), using a Waters 600 HPLC
Pump and 2700 Autosampler.
[0207] Mobile Phase A: 0.1% Formic acid in water, Mobile phase B:
0.1% Formic acid in acetonitrile, Flow rate: 2.0 ml/min., Gradient:
5% B to 95% B over 3 mins, hold 3 mins. Column: Varies, but always
C18 50 mm.times.4.6 mm (Currently Genesis C18 4.mu.. Jones
Chromatography). PDA detection: Waters 996, scan range 210-400
nm.
Synthesis of 2-Chloro-6-morpholin-4-yl-pyran-4-one (3)
##STR00031##
[0208] 4-Chloro-4-(2,2,2-trichloro-ethyl)-oxetan-2-one (1)
[0209] A solution of BCHPO
(bis-4-t-butylcyclohexyl)peroxydicarbonate (11.8 g) and diketene
(83.5 ml) in CCl.sub.4 (300 ml) was added dropwise over 120 minutes
to a refluxing solution of CCl.sub.4, and was stirred for a further
1 hour. The resulting pale yellow solution was cooled and
azeotroped with DCM. The resulting residue was stirred with hexane
(3.times.150 ml) for 10 minutes and the liquor was decanted off
through a celite pad. The filtered liquors were combined and
concentrated in vacuo to give 1 as a pale yellow oil (125.0 g,
52.9%).
5,5-Dichloro-1-morpholin-4-yl-pent-4-ene-1,3-dione (2)
[0210] Two separate solutions of 1 (62.5 g, 0.26 mmol) and
morpholine (24.0 g, 0.28 mol) in DCM (120 ml) were added
simultaneously to a mixture of NaHCO.sub.3 (44.0 g, 0.52 mol) in
dry DCM (300 ml). The reaction was maintained at 15.degree. C. over
140 minutes with stirring. The reaction was filtered, washed with
DCM (3.times.100 ml) and the combined organic layers were
concentrated in vacuo to a slurry which was then passed through a
short silica pad, and further washed with DCM (4.times.100 ml). The
combined organic layers were concentrated in vacuo, suspended in
hexane (400 ml) and stirred for 1 hour, filtered and dried to give
a cream solid. The solid was suspended in TBME (100 ml), stirred
for 15 minutes, filtered, washed with TBME and dried to give 2 as a
white powder (47.8 g, 72%). m/z (LC-MS, ESP): 252 (M.sup.++1).
2-Chloro-6-morpholin-4-yl-pyran-4-one (3)
[0211] To a suspension of 2 (11.3 g, 44.9 mmol) in dioxane was
added perchloric acid (11.4 ml, 0.14 mol) and the reaction was
heated at 90.degree. C. under N.sub.2 for 1 hour. The reaction was
cooled, neutralised with 2M NaOH (75 ml) and filtered. The aqueous
layer was extracted with DCM (4.times.30 ml) and the organic layers
were combined and dried over MgSO.sub.4. The organic layer was
further treated with charcoal and filtered through celite. The dark
yellow filtrate was evaporated in vacuo, and the resulting solid
was triturated with hexane (50 ml) and dried to give 3 (7.3 g, 75%)
as a light yellow powder. m/z (LC-MS, ESP): 216 (M.sup.++1).
.sup.1H-NMR (300 MHz, DMSO-d.sub.6): 3.3 (t, 4H), 3.65 (t, 4H), 5.4
(d, 1H), 6.25 (d, 1H).
Example 1
6-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthrene-2-carboxylic
acid amide Derivatives
##STR00032## ##STR00033##
[0212] 3-Chlorosulfonyl-4-fluoro-benzoic acid
[0213] Chlorosulphonic acid (100 ml, 1.5 mol) was gradually added
to 4-fluorobenzoic acid (43 g, 0.307 mol) with stirring. The clear
dark yellow mixture was heated to 150.degree. C. for 24 hours. The
yellow solution was cooled back to room temperature and poured onto
ice with vigorous stirring. The white precipitate was filtered and
pressed dry. The solid was dried overnight in a desiccator under
vacuum and over activated silica (54.65 g, 75%). Mp:
116-117.degree. C.; m/z (LC-MS, ESP), RT=4.03 min,
(M.sup.--1)=237-239 (ratio 1:3).
4-Fluoro-3-sulfino-benzoic acid
[0214] Sodium sulphite (130 g, 1.034 mol) was added slowly to a
solution of 3-chlorosulfonyl-4-fluoro-benzoic acid (49.39 g, 0.207
mol) in water (150 ml) at 0.degree. C. with a vigorous stirring.
After the addition was completed the reaction was warmed back to
room temperature for 1 hour and the pH of the solution was kept
around pH 6-7 with 2N sodium hydroxide solution. The white milky
suspension was filtered and the solid washed with 2N sodium
hydroxide solution (150 ml) and then water (100 ml). The filtrate
was then cooled in an ice bath and concentrated HCl was added until
no more precipitate was formed (pH<1). The white precipitate was
then filtered, pressed dry and left in a dessicator overnight under
vacuum and over activated silica (27.92 g, 66%). m/z (LC-MS, ESP),
RT=0.98 min, (M.sup.--1)=203
4-(2-Bromo-phenylsulfanyl)-3-sulfino-benzoic acid
[0215] 2-Bromobenzenethiol (25 g, 132 mmol) was added to a solution
of 4-fluoro-3-sulfino-benzoic acid (13.5 g, 66 mmol) and NaOH
pellets (11 g, 264 mmol) in water (30 ml). The yellow mixture was
then degassed for 10 minutes and then heated to 140.degree. C. for
48 hours. The reaction was then cooled to 0.degree. C. and
acidified to pH 4-5 (pH paper) with concentrated HCl. The
precipitate formed was filtered, washed with hexane and was dried
in a vacuum dessicator over activated silica overnight (20.69 g,
84%). m/z (LC-MS, ESP), RT=3.67 min, (M.sup.--1)=373.
6-Bromo-thianthrene-2-carboxylic acid
[0216] 4-(2-bromo-phenylsulfanyl)-3-sulfino-benzoic acid (14 g, 38
mmol) was added slowly to a stirred solution of methanesulphonic
acid (160 ml). The purple solution was heated to 60.degree. C. for
3 hours. The reaction was cooled down to room temperature and was
poured into ice (300 ml) where an off-white precipitate appeared.
The solid was filtered and washed with water (100 ml) and then
dried in a vacuum dessicator over activated silica (9.48 g, 73%).
.sup.1HNMR (300 MHz, CDCl.sub.3): .delta..sub.H=7.29 (1H, t), 7.59
(1H, dd), 7.70 (1H, dd) 7.74 (1H, d), 7.87 (1H, dd), 8.03 (1H, d).
m/z (LC-MS, ESP), RT=4.99 min, (M.sup.--1)=339
6-Bromo-thianthrene-2-carboxylic acid methyl ester
[0217] To 6-bromo-thianthrene-2-carboxylic acid (9 g, 28 mmol) in
methanol (180 ml) was slowly added conc. H.sub.2SO.sub.4 (5 ml).
The milky white suspension was heated to 80.degree. C. until all
the solid had gone into solution (2 hrs). The suspension was
concentrated in vacuo. Water (100 ml) was added and the organics
were then extracted with dichloromethane (3.times.70 ml), dried
over MgSO.sub.4 and evaporated in vacuo, yielding to a yellow
solid. (4.48 g, 45%). .sup.1HNMR (300 MHz, CDCl.sub.3):
.delta..sub.H=3.94 (3H, s); 7.13 (1H, t), 7.44 (1H, dd), 7.54 (1H,
dd) 7.61 (1H, d), 7.93 (1H, dd), 8.13 (1H, d).
6-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-thianthrene-2-carboxylic
acid methyl ester
[0218] 6-Bromo-thianthrene-2-carboxylic acid methyl ester (1 g, 2.8
mmol), bis(pinacolato)diboron (0.86 g, 3.4 mmol) and potassium
acetate (0.12 g, 0.14 mmol) in 1,4-dioxane (15 ml) was degassed for
15 minutes. To the yellow suspension was then added
PdCl.sub.2(dppf) (78 mg, 0.14 mmol) and dppf (0.83 g, 8.5 mmol).
The dark red mixture was heated to 90.degree. C. under a N.sub.2
atmosphere for 48 hours. The crude mixture was purified by flash
chromatography (dichloromethane) to give viscous brown oil (1.13
g), which was used without any further purification.
6-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthrene-2-carboxylic
acid methyl ester (4)
[0219]
6-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-thianthrene-2-carb-
oxylic acid methyl ester (1.1 g, 2.83 mmol),
2-chloro-6-morpholin-4-yl-pyran-4-one (3) (0.73 g, 3.4 mmol) and
K.sub.2CO.sub.3 (0.8 g, 5.66 mmol) were dissolved in dry
1,4-dioxane (7 ml). The mixture was degassed for 15 mins and
Pd(PPh.sub.3).sub.4 (0.16 g, 5 mol %) was then added The dark brown
mixture was heated to 90.degree. C. under an atmosphere of N.sub.2
for 24 hour. The reaction mixture was concentrated in vacuo and
water (100 ml) was added. The brown solid was filtered and washed
with water (1.23 g, 96%). m/z (LC-MS, ESP), RT=4.49 min,
(M.sup.++1)=454.
6-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthrene-2-carboxylate
sodium salt (5)
[0220]
6-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthrene-2-carboxylic
acid methyl ester (1.1 g, 2.43 mmol) and NaOH Pellets (97 mg, 2.43
mmol) were dissolved in methanol (40 ml). The brown suspension was
heated to 80.degree. C. under N.sub.2 for 24 hours. The solvent was
removed in vacuo and the residue was triturated with diethyl ether.
The product was collected by filtration as a fine dark brown powder
(1.11 g, 99%). m/z (LC-MS, ESP), RT=3.90 min, (M.sup.++1)=438
6-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthrene-2-carboxylic
acid amide Derivatives
[0221]
6-(6-morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthrene-2-carboxylate
sodium salt (138) (20 mg, 0.04 mmol), HBTU (18 mg, 0.05 mmol),
di-isopropylethylamine (9 .mu.l, 0.05 mmol), the appropriate amine
(0.04 mmol) and dry dimethylacetamide (0.5 ml). The dark brown
mixture was stirred at room temp for 2 hours and then purified by
preparative HPLC to give the desired products, which are shown
below:
TABLE-US-00001 ##STR00034## Purity Retention Time Compound R (%)
(Mins) M.sup.+ + 1 6 ##STR00035## 95 3.14 522 7 ##STR00036## 90
4.92 598 8 ##STR00037## 90 3.37 585 9 ##STR00038## 90 3.28 585 10
##STR00039## 95 3.12 596 11 ##STR00040## 95 3.52 550 12
##STR00041## 85 5.16 612 13 ##STR00042## 95 4.08 586 14
##STR00043## 95 3.28 607 15 ##STR00044## 95 4.66 602 16
##STR00045## 95 4.36 608 17 ##STR00046## 90 3.18 566 18
##STR00047## 95 3.81 602 19 ##STR00048## 90 3.12 508 20
##STR00049## 90 3.16 635
Example 2
2-(7-Amino-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one amide
derivatives
##STR00050##
[0222] 2-Chloro-5-nitro-benzenesulfinic acid
[0223] A solution of 1-chloro-4-nitrobenzene (476 g, 3.02 mol) in
chlorosulfonic acid (1 litre) was stirred and heated at 120.degree.
C. for 22 hours. The reaction was cooled to room temperature and
was poured slowly with stirring onto ice (10 litres). The resulting
dark brown precipitate was filtered, pressed dry, dried in a vacuum
desiccators over night and was used without any further
purification (770 g). The resulting
2-chloro-5-nitro-benzenesulfonyl chloride was suspended in water
and cooled to 0.degree. C. (ice bath). Sodium sulphite (1.9 Kg, 15
mol) was added to the stirred solution by portion (100 g portion
and in a manner that the temperature remains under 25.degree. C.).
After the addition the mixture was allowed to warm back to room
temperature and left stirring for one hour. During that time the pH
of the solution was kept at pH 7-8 (Whattman pH paper) by addition
of 8M solution of sodium hydroxide (400 ml). The solution was then
diluted with water (6 L) and sodium hydroxide was added to it (8M,
0.5 L). The resulting mixture was filtered through a sintered
funnel (number 3). The filtrate was cooled to 0.degree. C. (ice
bath) and acidify with concentrated HCl to pH<1. The brown
solution then turn green and a silvery green plates precipitate.
The solid was filtered, pressed dry and dried in a vacuum
desiccator to give the title compound (213 g, 32%).
2-(2-Bromo-phenylsulfanyl)-5-nitro-benzenesulfinic acid
[0224] To a solution of NaOH (80.18 g, 2.004 mol) in water (600 ml
sonicated and degassed) was added 2-bromothiophenol (108.23 ml,
1.002 mol) and 2-chloro-5-nitro-benzenesulfinic acid (222.11 g,
1.002 mol). The mixture was stirred and heated at 125.degree. C.
for 19 hours. The reaction was cooled down at 0.degree. C. and
concentrated HCl added dropwise from a dropping funnel while
stirring until pH<1. The precipitate formed was filtered, dried
in a desiccators overnight and was used without any further
purification (417.77 g). m/z (LC-MS, ESP), RT=3.51 min,
(M.sup.--1)=372-374, (1:1, bromine isotope ratio present).
1-Bromo-7-nitro-thianthrene
[0225] 2-(2-Bromo-phenylsulfanyl)-5-nitro-benzenesulfinic acid
(208.5 g, 557 mmol) thinly ground was dissolved slowly in
methanesulfonic acid (2 litre) with stirring. The mixture was
stirred and heated at 50.degree. C. for 17 hours. The solution was
poured onto ice (5 L) with stirring, then filtered and the solid
collected. The residue was suspended in water (500 ml) and then
basified to pH 8 with concentrated ammonia. The solid was then
filtered, suspended into methanol (1 L) and filtered again. The
solid was then dried in a desiccator under vacuum to give the title
compound (115 g, 61%). m/z (LC-MS, ESP), RT=5.53 min (no
ionisation). .sup.1HNMR (300 MHz, CDCl.sub.3): .delta..sub.H=7.15
(1H, t), 7.45 (1H, dd), 7.58 (1H, dd), 7.67 (1H, d), 8.11 (1H, dd),
8.31 (1H, d).
6-Bromo-thianthren-2-ylamine
[0226] Zn dust (144.1 g, 2.2 mol) was added to a stirred solution
of 1-bromo-7-nitro-thianthrene (125 g, 367.4 mmol) in glacial
acetic acid (500 ml at 0.degree. C. (ice bath). After one hour the
ice bath was removed and the solution was left to react overnight
at room temperature. The mixture was filtered through a pad of
Celite and washed with copious amount of dichloromethane and the
filtrate was evaporated in vacuo. Water was added (500 ml) to the
residue and the pH was adjusted to pH 8 by addition of concentrated
ammonia. The solid formed was filtered and dried in a vacuum
desiccator to give the title compound (118.5 g) and was used
without any further purification. m/z (LC-MS, ESP), RT=4.92,
(M.sup.++1)=310-312 (1:1, bromine isotope ratio present)
(6-Bromo-thianthren-2-yl)-carbamic acid tert-butyl ester
[0227] Di-tert-butyl dicarbonate (100 g, 0.458 mol) was added to a
solution of 6-bromo-thianthren-2-ylamine (118.51 g, 0.382 mol) in
dry THF (500 ml). The mixture was stirred and heated at 50.degree.
C. overnight. The solvent was removed in vacuo and the residue was
triturated in methanol to give the title compound as a pale brown
solid (40.87 g, 26%). .sup.1HNMR (300 MHz, CDCl.sub.3):
.delta..sub.H=1.51 (9H, s); 6.50 (1H, bs); 7.08 (1H, t); 7.16 (1H,
dd); 7.43 (2H, m); 7.51 (1H, dd); 7.71 (1H, d).
[6-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-2-yl]-carbamic
acid tert-butyl ester
[0228] (6-Bromo-thianthren-2-yl)-carbamic acid tert-butyl ester
(7.48 g, 18.24 mmol), bis(pinacolato)diboron (6.48 g, 25.54 mmol)
and potassium acetate (6.27 g, 63.84 mmol) in 1,4-dioxane (50 ml).
To the yellow suspension was then added PdCl.sub.2(dppf) (745 mg,
0.91 mmol) and dppf (506 mg, 0.91 mmol). The dark red mixture was
heated to 110.degree. C. under a N.sub.2 atmosphere for 24 hours.
The crude mixture was purified by flash chromatography (silica,
dichloromethane) to give
[6-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-thianthren-2-yl]-carbam-
ic acid tert-butyl ester as a viscous brown oil which was used
without any further purification.
[0229]
[6-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-thianthren-2-yl]--
carbamic acid tert-butyl ester (8.34 g, 18.24 mmol),
2-chloro-6-morpholin-4-yl-pyran-4-one (3) (4.72 g, 21.89 mmol) and
grinded K.sub.2CO.sub.3 (5.04 g, 36.48 mmol) were dissolved in dry
1,4-dioxane (100 ml). The mixture was degassed for 15 minutes and
Pd(PPh.sub.3).sub.4 (1.02 g, 0.91 mmol) was then added The dark
brown mixture was heated to 100.degree. C. under an atmosphere of
N.sub.2 for 24 hour. The reaction mixture was concentrated in vacuo
and water (100 ml) was added. The brown solid was filtered, washed
with water, dried overnight in a vacuum desiccator and was used
without any further purification to give the title compound (11.87
g). m/z (LC-MS, ESP), RT=4.61 min, (M.sup.++1)=511.2
2-(7-Amino-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one (21)
[0230] To a solution of
[6-(6-morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-2-yl]-carbamic
acid tert-butyl ester (11.95 g, 23.4 mmol) in dichloromethane (150
ml) was added trifluoroacetic acid (30 ml) and left under stirring
at room temperature overnight. The solvent was dried in vacuo
revealing a viscous dark brown liquid. Saturated sodium bicarbonate
solution (400 ml) was added to the residue, which was left to stir
for 20 minutes. The brown precipitate was filtered, washing with
water and left to dry in a vacuum desiccator overnight. The solid
was then purified by column chromatography (silica,
MeOH/Dichloromethane, 3:97, Rf=0.28). .sup.1HNMR (300 MHz,
CDCl.sub.3): .delta..sub.H=3.24 (2H, bs); 3.46 (4H, bs); 3.81 (4H,
bs); 6.33 (1H, s); 6.58 (1H, m); 6.86 (1H, d); 7.21 (1H, d); 7.30
(1H, t); 7.38 (1H, d); 7.63 (1H, d); m/z (LC-MS, ESP), RT=3.8 min,
(M.sup.++1)=411
2-(7-Amino-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one amide
derivatives
[0231] To a small test tube was added
2-(7-amino-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one (21) (20
mg, 0.05 mmol), dry dimethylacetamide (0.5 ml), triethylamine (0.01
ml, 0.08 mmol) and the desired acid chloride (0.08 mmol) with
stirring overnight. The reaction was purified by preparative HPLC
to give the desired products, which are shown below:
TABLE-US-00002 ##STR00051## Retention Com- Purity Time pound R (%)
(Mins) M.sup.+ + 1 22 ##STR00052## 95 3.73 453 23 ##STR00053## 95
3.91 483 24 ##STR00054## 95 4.08 479 25 ##STR00055## 95 4.45 559 26
##STR00056## 95 4.03 539 27 ##STR00057## 95 4.44 540 28
##STR00058## 95 3.82 516 29 ##STR00059## 95 4.18 505 30
##STR00060## 95 3.77 516 31 ##STR00061## 95 4.73 545 32
##STR00062## 95 4.42 521 33 ##STR00063## 95 4.33 535 34
##STR00064## 95 4.31 493 35 ##STR00065## 95 4.19 534 36
##STR00066## 95 3.93 497 37 ##STR00067## 95 4.18 535 38
##STR00068## 90 3.68 564 39 ##STR00069## 95 4.28 573 40
##STR00070## 95 4.43 605 41 ##STR00071## 95 3.96 525 42
##STR00072## 95 3.88 511 43 ##STR00073## 95 4.42 587 44
##STR00074## 95 4.67 533 45 ##STR00075## 910 4.62 573 46
##STR00076## 95 5.16 604 47 ##STR00077## 95 4.03 506 48
##STR00078## 95 4.00 527
2-(7-Amino-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one Biotin
Derivatives
[0232] To a small test tube was added
2-(7-amino-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one (21) (49
mg, 0.12 mmol), HBTU (51 mg, 0.13 mmol), di-isopropylethylamine
(26.mu., 0.15 mmol), the appropriate biotin derivative (0.122 mmol)
and dry dimethylacetamide (0.5 ml) with stirring overnight. The
reaction was purified by preparative HPLC to give the desired
product, which is shown below:
TABLE-US-00003 ##STR00079## Purity Retention Time Compound R (%)
(Mins) M.sup.+ + 1 49 ##STR00080## 85 3.73 637 50 ##STR00081## 90
3.68 750
2-Amino-N-[6-(6-morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-2-yl]-aceta-
mide derivatives
[0233] To a small test tube was added
2-(7-amino-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one (21) (20
mg, 0.05 mmol), dry dimethylacetamide (0.5 ml), triethylamine (8
.mu.l, 0.06 mmol) and chloroacetyl chloride (4 .mu.l, 0.06 mmol)
with stirring overnight. The appropriate amine or thiol (20 mg or
20 .mu.l) was then added and left to stir at room temperature
overnight. The reaction was purified by preparative HPLC to give
the desired products, which are shown below:
TABLE-US-00004 ##STR00082## Purity Retention Time Compound R (%)
(Mins) M.sup.+ + 1 51 ##STR00083## 95 3.23 538 52 ##STR00084## 95
3.30 536 53 ##STR00085## 95 3.18 537 54 ##STR00086## 95 3.24 551 55
##STR00087## 95 3.15 556 56 ##STR00088## 95 3.46 584 57
##STR00089## 95 3.11 512 58 ##STR00090## 95 2.91 511 59
##STR00091## 95 3.23 522 60 ##STR00092## 90 3.17 565 61
##STR00093## 95 3.11 551 62 ##STR00094## 85 3.43 550 63
##STR00095## 95 2.94 595 64 ##STR00096## 90 3.49 588 65
##STR00097## 90 3.64 552 66 ##STR00098## 95 3.30 524 67
##STR00099## 95 3.08 581 68 ##STR00100## 95 3.32 552 69
##STR00101## 95 3.19 496 70 ##STR00102## 95 3.40 566 71
##STR00103## 95 3.12 559 72 ##STR00104## 95 3.40 587 73
##STR00105## 90 3.23 553 74 ##STR00106## 85 3.13 580 75
##STR00107## 85 2.93 579 76 ##STR00108## 95 3.25 526 77
##STR00109## 95 3.24 579 78 ##STR00110## 95 3.28 595 79
##STR00111## 95 3.21 625 80 ##STR00112## 95 3.28 579 81
##STR00113## 95 3.25 565 82 ##STR00114## 95 3.35 614 83
##STR00115## 95 3.72 631 84 ##STR00116## 95 3.46 615 85
##STR00117## 95 3.33 636 86 ##STR00118## 95 2.96 608 87
##STR00119## 95 3.13 579 88 ##STR00120## 90 3.04 579 89
##STR00121## 95 3.18 581 90 ##STR00122## 90 3.12 595
3-Amino-N-[6-(6-morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-2-yl]-propi-
onamide derivatives
[0234] To a small test tube was added
2-(7-amino-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one (21) (20
mg, 0.05 mmol), dry dimethylacetamide (0.5 ml), triethylamine (8
.mu.l, 0.06 mmol) and 3-bromopropionyl chloride (5 .mu.l, 0.05
mmol) with stirring overnight. The appropriate amine or thiol (20
mg or 20 .mu.l, hydrochloride salts were freed by addition of
triethylamine) was then added and left to stir at room temperature
overnight. The reaction was purified by preparative HPLC to give
the desired products, which are shown below:
TABLE-US-00005 ##STR00123## Purity Retention Time Compound R (%)
(Mins) M.sup.+ + 1 91 ##STR00124## 95 3.21 552 92 ##STR00125## 95
3.37 550 93 ##STR00126## 90 3.11 551 94 ##STR00127## 95 3.16 565 95
##STR00128## 95 3.17 570 96 ##STR00129## 95 3.18 526 97
##STR00130## 95 2.93 525 98 ##STR00131## 95 3.28 536 99
##STR00132## 95 2.98 579 100 ##STR00133## 90 2.99 565 101
##STR00134## 95 3.43 564 102 ##STR00135## 95 3.01 609 103
##STR00136## 95 3.54 602 104 ##STR00137## 95 3.68 566 105
##STR00138## 95 3.33 538 106 ##STR00139## 95 3.10 595 107
##STR00140## 95 3.37 566 108 ##STR00141## 90 3.20 510 109
##STR00142## 95 3.38 580 110 ##STR00143## 95 3.13 573 111
##STR00144## 95 3.23 601 112 ##STR00145## 95 2.99 567 113
##STR00146## 95 3.20 594 114 ##STR00147## 95 2.98 593 115
##STR00148## 90 3.25 540 116 ##STR00149## 95 3.20 593 117
##STR00150## 95 3.18 639 118 ##STR00151## 85 3.20 593 119
##STR00152## 85 3.18 579 120 ##STR00153## 95 3.28 628 121
##STR00154## 95 3.69 645 122 ##STR00155## 95 3.43 629 123
##STR00156## 95 3.33 650 124 ##STR00157## 95 2.98 622 125
##STR00158## 95 3.14 539 126 ##STR00159## 95 2.98 593 127
##STR00160## 90 2.95 593 128 ##STR00161## 95 3.13 595 129
##STR00162## 90 2.97 609
Example 3
2-(4-Hydroxy-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one
derivatives
##STR00163##
[0235] Thianthren-1-ol
[0236] To a cooled (-78.degree. C.) solution of thianthrene (10 g,
46.23 mmol) in anhydrous THF (200 ml) was added t-BuLi (1.7M in
pentane, 40.81 ml, 69.34 mmol), under a nitrogen atmosphere, over a
period of 10 minutes and the resulting yellow solution was then
allowed to warm to room temperature and stirred vigorously for a
further 16 hours. EtMgBr (3M in THF, 23 ml, 69.34 mmol) was then
added in a dropwise fashion to the cooled (0.degree. C.) reaction
mixture which was then stirred for a further 45 minutes before
oxygen was bubbled through the solution. After two hours the
reaction was quenched by dropwise addition of 1M NaOH (100 ml) and
washed with EtOAc (1.times.300 ml) before being acidified to pH4
with aqueous 1M HCl. The mixture was then extracted with EtOAc
(3.times.100 mL), dried (MgSO.sub.4), filtered and concentrated in
vacuo to give a mild amber slurry that was purified by flash
chromatography (SiO.sub.2) (10:1, Ethyl acetate/Hexanes), to give
the title compound (3.82 g, 36%) as a yellow solid. m/z (LC-MS,
ESP): RT=4.62 min, (M.sup.++1)=231
4-Bromo-thianthren-1-ol
[0237] Thianthren-1-ol (6.5 g, 27.98 mmol) was dissolved in glacial
acetic acid (100 ml) and to this solution was added bromine (17.1
.mu.l, 13.99 mmol) in a dropwise fashion over 30 minutes. Water
(200 ml) was added to the amber solution and the mixture extracted
with EtOAc (3.times.100 ml). The combined organic extracts were
then washed with saturated potassium bicarbonate solution
(3.times.100 ml), dried using MgSO.sub.4, filtered and concentrated
in vacuo to give the title compound. (7.56 g, 87%). m/z (LC-MS,
ESP): RT=4.92 min, (M.sup.++1)=309.
1-Bromo-4-methoxymethoxy-thianthrene
[0238] To a cooled (0.degree. C.) solution of
4-Bromo-thianthren-1-ol (1.0 g, 3.21 mmol) in DMF (12 ml) was added
NaH (60% dispersed in mineral oil, 0.23 g, 9.63 mmol) portionwise
over 30 minutes and stirred for a further 30 minutes at this
temperature. Chloromethylmethylether (0.26 g, 3.241 mmol) was then
added and the reaction allowed to stir at room temperature for 2
hours. Water was added to the mixture which was then extracted
using EtOAc (3.times.50 ml). The combined organic extracts were
then dried (MgSO.sub.4), filtered and concentrated in vacuo to give
a mild amber liquid. The crude residue was purified by flash
chromatography (SiO.sub.2) (10:1, Hexanes/EtOAc) to give the title
compound as a mild amber oil that crystallised upon standing (1.11
g, 97%). m/z (LC-MS, ESP): RT=5.55 min, (M.sup.++1)=355.
2-(4-Methoxymethoxy-thianthren-1-yl)-4,4,5,5-tetramethyl-[1,3,2]dioxaborol-
ane
[0239] 1-Bromo-4-methoxymethoxy-thianthrene (2.06 g, 5.80 mmol) was
dissolved in anhydrous dioxane (10 ml) and to this solution was
added bis(pinacolato)diboron (1.75 g, 6.96 mmol),
1,1-bis(diphenylphosphino)ferrocene (0.18 g, 0.05 mmol) and
potassium acetate (1.7 g, 17.40 mmol) under a nitrogen atmosphere.
The mixture was degassed for 10 minutes and
(1,1'-bis(diphenylphosphino)ferrocene-dichloropalladium(II)
dichloromethane adduct (0.24 g, 5% eq) was added. The reaction was
heated at 90.degree. C. under a nitrogen atmosphere for 24 hours.
The dark brown reaction mixture was then allowed to cool to room
temperature before it was applied to a thick silica pad prepared in
hexanes and eluted with hexanes:CH.sub.2Cl.sub.2-(2:1). The eluent
was concentrated in vacuo to give a dark brown oil (2.33 g, 100%)
that was used for the next transformation with no further
purification. RT=5.63 min.
2-(4-Methoxymethoxy-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one
[0240]
2-(4-Methoxymethoxy-thianthren-1-yl)-4,4,5,5-tetramethyl-[1,3,2]dio-
xaborolane (6.23 g, 17.14 mmol),
2-chloro-6-morpholin-4-yl-pyran-4-one (3) (6.99 g, 17.40 mmol) and
powdered potassium carbonate (7.20 g, 52.20 mmol) were suspended in
anhydrous dioxane (60 ml) under a nitrogen atmosphere. The mixture
was degassed for 15 minutes before the addition of
tetrakis(triphenylphosphino)palladium (1.2 g, 5% eq) and then
degassed for a further 15 minutes after addition. The reaction was
then heated at 90.degree. C. for 24 hours. Water (60 ml) was added
and the mixture extracted with EtOAc (3.times.30 ml). The organic
extracts were then dried using MgSO.sub.4, filtered and
concentrated in vacuo to yield a dark yellow oil. The crude residue
was then purified by flash chromatography (SiO.sub.2)
(9:1-EtOAc/MeOH) to give the title compound (7.93 g, 79%) as a
green powder. m/z (LC-MS, ESP): RT=4.39 min, (M.sup.++1)=456
2-(4-Hydroxy-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one
(130)
[0241] To a solution of
2-(4-Methoxymethoxy-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one
(1.1 g, 2.42 mmol) in acetone (50 ml) was added sodium iodide (0.40
g, 2.66 mmol) and concentrated HCl (3 ml). The dark suspension was
stirred at room temperature for 72 hours before the careful
addition of 1M sodium hydroxide solution until pH 12 was attained.
The mixture was washed with EtOAc (2.times.50 ml) and then
acidified with concentrated HCl until pH 1 whereupon a brown
precipitate formed which was filtered off (0.90 g, 90%) and
corresponded to the title compound. m/z (LC-MS, ESP): RT=4.39 min,
(M.sup.++1)=456
2-(4-Hydroxy-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one
derivatives
[0242] To a solution of
2-(4-Hydroxy-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one (130)
(25 mg, 0.06 mmol) in anhydrous Dimethylformamide (0.66 ml) was
added powdered potassium carbonate (33 mg, 0.24 mmol) and
1,2-dibromoethane (26 .mu.l, 0.3 mmol). After stirring the reaction
mixture for 24 hours the appropriate amine or thiol was added to
the solution and stirred at room temperature for a further 24
hours. The crude mixture was then purified by preparative HPLC to
give the desired compounds, which are shown below:
TABLE-US-00006 ##STR00164## Purity Retention Time Compound R (%)
(Mins) M.sup.+ + 1 131 ##STR00165## 95 3.13 470 132 ##STR00166## 95
3.20 484 133 ##STR00167## 95 3.21 484 134 ##STR00168## 95 3.43 512
135 ##STR00169## 95 3.33 498 136 ##STR00170## 90 3.41 512 137
##STR00171## 95 3.24 494 138 ##STR00172## 95 3.31 540 139
##STR00173## 95 3.38 536 140 ##STR00174## 95 3.45 538 141
##STR00175## 95 3.35 554 142 ##STR00176## 85 3.33 510 143
##STR00177## 95 3.61 552 144 ##STR00178## 95 3.38 524 145
##STR00179## 95 3.23 510 146 ##STR00180## 95 3.28 524 147
##STR00181## 95 3.23 526 148 ##STR00182## 90 3.41 538 149
##STR00183## 60 3.52 553 150 ##STR00184## 90 3.52 567 151
##STR00185## 95 3.66 567 152 ##STR00186## 85 3.02 554 153
##STR00187## 85 3.31 541 154 ##STR00188## 90 2.99 567 155
##STR00189## 95 3.00 553 156 ##STR00190## 95 2.97 567 157
##STR00191## 95 3.66 589 158 ##STR00192## 95 3.40 547 159
##STR00193## 90 3.16 547 160 ##STR00194## 95 3.43 561 161
##STR00195## 95 3.47 575 162 ##STR00196## 95 2.95 550 163
##STR00197## 95 3.17 569 164 ##STR00198## 95 3.12 658 165
##STR00199## 90 2.97 555 166 ##STR00200## 85 2.97 569 167
##STR00201## 95 3.06 583 168 ##STR00202## 85 3.15 541 169
##STR00203## 95 3.07 527 170 ##STR00204## 85 3.03 653
[0243] To a solution of
2-(4-Hydroxy-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one (130)
(25 mg, 0.06 mmol) in anhydrous dimethylformamide (0.66 ml) was
added powdered potassium carbonate (33 mg, 0.24 mmol) and
1,3-dibromopropane (23 .mu.l, 0.3 mmol). After stirring the
reaction mixture for 24 hours the appropriate amine or thiol was
added to the solution and stirred at room temperature for a further
24 hours. The crude mixture was then purified by preparative HPLC
to give the desired compounds, which are shown below:
TABLE-US-00007 ##STR00205## Purity Retention Time Compound R (%)
(Mins) M.sup.+ + 1 171 ##STR00206## 95 3.33 526 172 ##STR00207## 95
3.24 498 173 ##STR00208## 95 3.48 554 174 ##STR00209## 85 3.48 526
175 ##STR00210## 95 3.46 526 176 ##STR00211## 95 3.33 512 177
##STR00212## 95 3.64 552 178 ##STR00213## 95 3.36 554 179
##STR00214## 85 3.39 550 180 ##STR00215## 85 3.53 552 181
##STR00216## 95 3.44 568 182 ##STR00217## 85 3.40 524 183
##STR00218## 85 3.70 566 184 ##STR00219## 95 3.47 538 185
##STR00220## 95 3.31 524 186 ##STR00221## 95 3.42 538 187
##STR00222## 95 3.24 540 188 ##STR00223## 95 3.49 552 189
##STR00224## 95 4.08 589 190 ##STR00225## 95 3.42 567 191
##STR00226## 90 2.95 568 192 ##STR00227## 95 3.24 555 193
##STR00228## 95 2.96 581 194 ##STR00229## 85 2.94 581 195
##STR00230## 90 3.33 561 196 ##STR00231## 90 3.13 561 197
##STR00232## 95 3.36 589 198 ##STR00233## 85 2.89 564 199
##STR00234## 90 2.93 583 200 ##STR00235## 90 2.99 597 201
##STR00236## 85 2.94 555 202 ##STR00237## 90 3.10 631 203
##STR00238## 90 2.91 541
##STR00239##
[4-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-1-yloxy]-acetic
acid methyl ester
[0244] To a solution of
2-(4-Hydroxy-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one (130)
(1.20 g, 2.91 mmol) in dimethylformamide (10 ml), under and inert
atmosphere, was added powdered potassium carbonate (1.21 g, 8.75
mmol) and methylbromoacetate (0.55 ml, 5.82 mmol). The temperature
of the reaction was raised to 60.degree. C. for 48 hours and then
cooled to room temperature before the addition of water (20 mL).
The mixture was then extracted using ethyl acetate (3.times.30 ml).
The combined organics were dried (MgSO.sub.4), filtered and
concentrated in vacuo to give the title compound as an off white
solid (1.40 g, 99%) that was suitably clan and required no further
purification. m/z (LC-MS, ESP): RT=4.48 min, (M.sup.++1)=484
[4-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-1-yloxy]-acetic
acid sodium salt
[0245] To a solution of
[4-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-1-yloxy]-acetic
acid methyl ester (1.41 g, 2.91 mmol) in anhydrous methanol (10 ml)
was added sodium hydroxide (120 mg, 2.91 mmol) in a single portion.
The solution was then stirred at room temperature for 24 hours
whereupon the solvent was removed in vacuo to give the title
compound as an off white paste (1.43 g, 98%) that required no
further purification. m/z (LC-MS, ESP): RT=3.79 min,
(M.sup.--23)=468
[4-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-1-yloxy]-acetic
acid derivatives
[0246] To a solution of
[4-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-1-yloxy]-acetic
acid sodium salt (20 mg, 0.04 mmol) in dimethylacetamide (1 ml) was
added O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (25 mg, 0.07 mmol) followed by
N,N-diisopropylethylamine (16 .mu.l, 0.09 mmol). Finally, the
appropriate amine or thiol was added to the solution which was
stirred at room temperature for 24 hours. The crude mixture was
then purified by preparative HPLC to give the desired compounds,
which are shown below:
TABLE-US-00008 ##STR00240## Purity Retention Time Compound R (%)
(Mins) M.sup.+ + 1 204 ##STR00241## 95 4.23 526 205 ##STR00242## 90
3.98 498 206 ##STR00243## 90 4.33 554 207 ##STR00244## 90 4.15 526
208 ##STR00245## 90 4.32 526 209 ##STR00246## 95 4.11 512 210
##STR00247## 85 3.98 552 211 ##STR00248## 95 4.20 554 212
##STR00249## 95 4.17 550 213 ##STR00250## 95 4.16 552 214
##STR00251## 95 4.77 568 215 ##STR00252## 90 4.43 524 216
##STR00253## 95 4.02 566 217 ##STR00254## 95 4.29 538 218
##STR00255## 85 4.38 524 219 ##STR00256## 85 4.23 538 220
##STR00257## 85 3.83 540 221 ##STR00258## 90 3.35 552 222
##STR00259## 95 3.30 569 223 ##STR00260## 90 3.31 585 224
##STR00261## 95 3.64 589 225 ##STR00262## 95 3.33 567 226
##STR00263## 95 3.41 568 227 ##STR00264## 90 3.49 555 228
##STR00265## 90 3.46 581 229 ##STR00266## 95 3.27 581 230
##STR00267## 95 3.28 561 231 ##STR00268## 95 3.51 561 232
##STR00269## 95 3.33 589 233 ##STR00270## 95 3.43 564 234
##STR00271## 95 3.34 583 235 ##STR00272## 95 3.21 597 236
##STR00273## 990 3.63 555 237 ##STR00274## 95 3.25 631 238
##STR00275## 95 3.57 541
Example 4
2-Morpholin-4-yl-6-(10H-phenothiazin-4-yl)-pyran-4-one alkyl
derivatives
##STR00276##
[0247] 10H-Phenothiazin-4-ol
[0248] To a solution of 3-phenylamino-phenol (5 g, 26.99 mmol) in
1,2-dichlorobenzene (50 ml) was added S.sub.8 sulfur (1.82 g, 56.76
mmol) in a single portion and iodine (0.1 g, 0.39 mmol) which was
added in three portions over 10 minutes. A reflux condenser was
attached to the reaction vessel which was heated to 185.degree. C.
under a nitrogen atmosphere. The mixture was stirred at this
temperature for 4 hours and then allowed to cool to room
temperature. The reaction mixture was filtered to remove a black
precipitate and the filtrate diluted with Et.sub.2O (100 ml) and
washed with water (2.times.100 ml). The organic layer was separated
and the volatile solvents removed to give a deep green oil that was
purified by flash column chromatography (SiO.sub.2) (Hexanes then
8:1-Hexanes:EtOAc) to give a pale yellow solid (2.38 g, 40.96%) m/z
(LC-MS, ESP) 216 [M+H].sup.+, RT=4.12 mins.
4-Hydroxy-phenothiazine-10-carboxylic acid tert-butyl ester
[0249] To a solution of 10H-phenothiazin-4-ol (0.77 g, 3.58 mmol)
in anhydrous pyridine (10 ml) was added di-tertiary butyl
dicarbonate (3.12 g, 14.31 mmol) in a single portion. The solution
was heated to 80.degree. C. and stirred under a nitrogen atmosphere
for 60 minutes before being allowed to cool to room temperature and
treated with water (20 ml) and extracted with EtOAc (2.times.30
ml). The organic layers were then washed with water (20 ml), dried
using MgSO.sub.4, filtered and concentrated in vacuo to give an
amber oil. The crude residue was treated with MeOH (15 ml) and
solid NaOH (0.65 g, 16.25 mmol). The mixture was heated to
80.degree. C. for 60 minutes then cooled to room temperature and
neutralised to pH7 with 1M HCl solution. The resulting suspension
was then filtered and dried to give the title compound as a beige
solid (1.13 g, 100%) that was used without further purification.
m/z (LC-MS, ESP): 315 [M-H].sup.-, RT=4.72 mins.
4-Trifluoromethanesulfonyloxy-phenothiazine-10-carboxylic acid
tert-butyl ester
[0250] Trifluoromethanesulfonic anhydride (2.95 ml, 17.09 mmol) was
added in a dropwise fashion over 10 minutes to a cooled (0.degree.
C.) stirred solution of 4-hydroxy-phenothiazine-10-carboxylic acid
tert-butyl ester (3.60 g, 11.41 mmol) in pyridine (40 ml). The
reaction mixture was stirred at 0.degree. C. for 1 hour before the
addition of water (80 ml). The mixture was extracted using EtOAc
(2.times.60 ml). The organic extracts were then dried using
MgSO.sub.4, filtered and concentrated in vacuo to give a dark brown
oil. The crude residue was then purified by flash chromatography
(SiO.sub.2) (4:1-Hexanes:EtOAc) to yield a yellow oil (5.02 g,
98.24%) m/z (LC-MS, ESP): 348 [M+H-BOC].sup.+, RT=5.61 mins.
4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenothiazine-10-carboxyl-
ic acid tert-butyl ester
[0251] To a stirred solution of
4-trifluoromethanesulfonyloxy-phenothiazine-10-carboxylic acid
tert-butyl ester (3.0 g, 6.7 mmol) in anhydrous dioxane (10 ml) was
added bis(pinacolato)diboron (2.05 g, 8.06 mmol) and potassium
acetate (1.96 g, 20.01 mmol). The reaction was then degassed
(sonication for 20 minutes then saturated with N.sub.2) before the
addition of
dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium(II)
dichloromethane adduct (0.27 g, 0.33 mmol). The reaction mixture
was degassed for a further 20 minutes before a reflux condenser was
attached to the reaction vessel which was then heated to 90.degree.
C. and stirred vigorously for 72 hours. The dark brown reaction
mixture was then allowed to cool to room temperature before it was
applied to a thick silica pad prepared in hexanes and eluted with
hexanes:CH.sub.2Cl.sub.2-(2:1). The eluent was concentrated in
vacuo to give a dark brown oil (2.85 g, 100%) that was used for the
next transformation with no further purification. m/z (LC-MS, ESP):
326 [M+H-BOC].sup.+, RT=5.86 mins.
4-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-phenothiazine-10-carboxylic
acid tert-butyl ester
[0252] Powdered potassium carbonate (2.03 g, 14.68 mmol) and
2-chloro-6-morpholin-4-yl-pyran-4-one (3) (1.44 g, 6.70 mmol) were
added to a stirred solution of
4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenothiazine-10-carboxy-
lic acid tert-butyl ester (2.85 g, 6.70 mmol) in anhydrous dioxane
(20 ml) and the mixture degassed (sonication for 20 minutes then
saturated with N.sub.2) thoroughly. Tetrakis (triphenylphosphine)
palladium was then added in a single portion and the mixture
degassed (sonication for 20 minutes then saturated with N.sub.2)
once again before a reflux condenser was attached and the mixture
heated to 100.degree. C. under a nitrogen atmosphere for 20 hours.
Water (30 ml) was added and the mixture extracted with EtOAc
(3.times.30 ml). The organic extracts were then dried using
MgSO.sub.4, filtered and concentrated in vacuo to yield a dark
brown, crystalline solid (3.21 g, 100%) that was taken forward with
no further purification. m/z (LC-MS, ESP): 479 [M+H].sup.+, RT=4.55
mins.
2-Morpholin-4-yl-6-(10H-phenothiazin-4-yl)-pyran-4-one (239)
[0253] To a stirred solution of
4-(6-morpholin-4-yl-4-oxo-4H-pyran-2-yl)-phenothiazine-10-carboxylic
acid tert-butyl ester (3.65 g, 7.63 mmol), in CH.sub.2Cl.sub.2 (30
ml) was added trifluoroacetic acid in a single portion. The mixture
was stirred at room temperature for 20 hours whereupon the reaction
was concentrated in vacuo to give a thick syrup that was basified
in a dropwise fashion with saturated NaHCO.sub.3 (40 ml). The dark
green mixture was then stirred at room temperature for 18 hours.
The mixture was filtered and the filtrant retained, washed with
water and dried to give the title compound as a dark green solid
(2.89 g, 83.74% over 3 steps) m/z (LC-MS, ESP): 479 [M+H].sup.+,
RT=4.05 mins.
2-Morpholin-4-yl-6-(10H-phenothiazin-4-yl)-pyran-4-one alkyl
derivatives
[0254] To a cooled (0.degree. C.) solution of
2-morpholin-4-yl-6-(10H-phenothiazin-4-yl)-pyran-4-one (239) (20
mg, 0.05 mmol) in dimethylformamide (0.5 ml), was added sodium
hydride (60% in min oil, 6 mg, 0.15 mmol). The resulting brown
solution was stirred for 1 hour whereupon 1-bromo-3-chloropropane
(6.6 .mu.l, 0.06 mmol) was added in a single portion and stirred at
room temperature for 1 hour. The appropriate amine was then added
and the reaction mixture heated to 60.degree. C. for 48 hours and
then purified by preparative HPLC to give the desired products,
which are shown below:
TABLE-US-00009 ##STR00277## Purity Retention Time Compound R (%)
(Mins) M.sup.+ + 1 240 ##STR00278## 85 3.29 521 241 ##STR00279## 95
3.18 465 242 ##STR00280## 90 3.32 493 243 ##STR00281## 85 3.27 479
244 ##STR00282## 95 3.26 493 245 ##STR00283## 85 3.38 493 246
##STR00284## 95 3.23 479 247 ##STR00285## 85 3.53 519 248
##STR00286## 85 3.14 451 249 ##STR00287## 95 3.18 465 250
##STR00288## 90 3.31 517 251 ##STR00289## 95 3.45 519 252
##STR00290## 90 3.29 491 253 ##STR00291## 95 3.56 533 254
##STR00292## 95 3.37 505 255 ##STR00293## 95 3.23 491 256
##STR00294## 95 3.28 505 257 ##STR00295## 95 3.3 535 258
##STR00296## 95 3.33 535 259 ##STR00297## 95 3.2 507 260
##STR00298## 95 3.38 519 261 ##STR00299## 85 3.34 534 262
##STR00300## 95 3.17 552 263 ##STR00301## 85 2.95 548 264
##STR00302## 95 2.98 534 265 ##STR00303## 95 2.95 548 266
##STR00304## 85 3.33 528 267 ##STR00305## 90 3.13 528 268
##STR00306## 90 3.18 556 269 ##STR00307## 85 3.33 556 270
##STR00308## 95 2.93 577 271 ##STR00309## 95 2.93 531 272
##STR00310## 95 3.07 550 273 ##STR00311## 95 2.93 569 274
##STR00312## 90 3.08 639 275 ##STR00313## 85 3.47 649 276
##STR00314## 90 2.93 536 277 ##STR00315## 95 2.94 550 278
##STR00316## 95 2.98 564 279 ##STR00317## 95 2.95 522 280
##STR00318## 95 2.91 480 281 ##STR00319## 85 3.76 598 282
##STR00320## 95 2.93 508 283 ##STR00321## 95 2.93 508 284
##STR00322## 90 2.92 522 285 ##STR00323## 95 3.28 509 286
##STR00324## 95 3.38 523 287 ##STR00325## 90 3.16 549 288
##STR00326## 95 3.1 520 289 ##STR00327## 50 3.71 596 290
##STR00328## 95 3.24 583 291 ##STR00329## 85 2.95 583 292
##STR00330## 85 3.07 594 293 ##STR00331## 85 3.15 548 294
##STR00332## 85 3.8 610 295 ##STR00333## 95 3.36 584 296
##STR00334## 95 3.31 605 297 ##STR00335## 95 3.61 600 298
##STR00336## 95 3.53 606 299 ##STR00337## 95 3.66 606 300
##STR00338## 95 3.14 550 301 ##STR00339## 95 3.22 564 302
##STR00340## 95 3.32 600 303 ##STR00341## 95 3.06 506 304
##STR00342## 95 2.98 577 305 ##STR00343## 95 3.19 633 306
##STR00344## 95 3.18 535 307 ##STR00345## 95 2.93 534 308
##STR00346## 90 3.41 549 309 ##STR00347## 90 3.23 510 310
##STR00348## 95 3.55 591 311 ##STR00349## 95 3.63 620 312
##STR00350## 95 3.46 577 313 ##STR00351## 95 3.21 535 314
##STR00352## 95 3.22 535 315 ##STR00353## 85 3.43 577 316
##STR00354## 85 3.14 521 317 ##STR00355## 85 3.14 548 318
##STR00356## 85 3.63 622 319 ##STR00357## 85 3.26 565 320
##STR00358## 90 3.36 553 321 ##STR00359## 85 3.11 551 322
##STR00360## 95 3.1 541 323 ##STR00361## 95 3.12 511 324
##STR00362## 95 3.2 539 325 ##STR00363## 95 3.46 571 326
##STR00364## 95 3.45 563 327 ##STR00365## 95 3.13 525 328
##STR00366## 95 3.38 549 329 ##STR00367## 95 3.18 495 330
##STR00368## 90 4.74 597 331 ##STR00369## 95 2.93 534 332
##STR00370## 95 2.93 520 333 ##STR00371## 95 3.56 620 334
##STR00372## 95 2.91 564 335 ##STR00373## 95 3.23(recorded
onWatersMicromass) 604 336 ##STR00374## 99 -- 819
Example 5
2-(7-Amino-9H-thioxanthen-4-yl)-6-morpholin-4-yl-pyran-4-one
N-amide derivatives
##STR00375##
[0255] 2-(2-Bromo-phenylsulfanyl)-5-nitro-benzoic acid
[0256] 2-Bromobenzenethiol (7.2 ml, 85.9 mmol) was added to a
solution of KOH (9.6 g, 172 mmol) in water (50 ml) degassed for 15
minutes. 2-Fluoro-5-nitrobenzoic acid (15.9 g, 85.9 mmol) was added
to the reaction mixture, which was refluxed under a nitrogen
atmosphere overnight. The reaction was cooled to room temperature
and was acidified (pH 1) with conc. HCl. The precipitate formed was
filtered and dried overnight in a vacuum oven (50.degree. C.) to
give the crude title compound as a pale yellow solid (30 g, 99%).
The product was used without further purification. m/z (LC-MS,
ESP), RT=4.51 min, (M.sup.--1)=352-354, (1:1, bromine isotope ratio
present).
5-Bromo-2-nitro-thioxanthen-9-one
[0257] 2-(2-Bromo-phenylsulfanyl)-5-nitro-benzoic acid (34 g, 96
mmol) was suspended in methanesulphonic acid (400 ml) and heated at
150.degree. C. The crude mixture was slowly poured onto ice with
vigorous stirring and the precipitate formed was filtered. The
solid was suspended into water (50 ml) quenched to pH 7-8 with
conc. ammonia solution and filtered. The yellow/lime colored solid
was dried under vacuum at 50.degree. C. to give the crude title
compound, which was used without any further purification (23.09 g,
72%).
5-Bromo-2-nitro-9H-thioxanthene
[0258] To a cooled (0.degree. C.) suspension of
5-bromo-2-nitro-thioxanthen-9-one (23.03 g, 68.5 mmol) in anhydrous
tetrahydrofuran (100 ml) under nitrogen atmosphere, was added drop
wise borane-THF complex (151 ml, 1.0M in THF). The mixture was
allowed to warm to room temperature with stirring overnight. The
reaction mixture was cooled (0.degree. C.) and the excess borane
was quenched with acetone. The solvent was evaporated in vacuo and
the residue was triturated in saturated sodium bicarbonate (100
ml). The solid was filtered, washed with water and dried overnight
in a vacuum dissicator to give the title compound (22.07 g, 100%).
.sup.1HNMR (300 MHz, CDCl.sub.3): .delta..sub.H=4.02 (2H, s), 7.17
(1H, m), 7.27 (1H, m), 7.51 (1H, m), 7.62 (1H, m), 8.09 (1H, m),
8.20 (1H, d).
5-Bromo-9H-thioxanthen-2-ylamine
[0259] Zn dust (34.87 g, 533 mmol) was added to a stirred solution
of 5-bromo-2-nitro-9H-thioxanthene (28.64 g, 88.9 mmol) in glacial
acetic acid (300 ml) at 0.degree. C. (ice bath). After one hour the
ice bath was removed and the solution was left to react overnight
at room temperature. The mixture was filtered through a pad of
Celite and washed with copious amount of dichloromethane and the
filtrate was evaporated in vacuo. Water was added (800 ml) to the
residue and the pH was adjusted to pH 8 by addition of concentrated
ammonia (100 ml) The solid formed was filtered and dried in a
vacuum desiccator to give the title compound (26.64 g) and was used
without any further purification. m/z (LC-MS, ESP), RT=4.53 min,
(M.sup.++1)=294-292, (1:1, bromine isotope ratio present)
(5-Bromo-9H-thioxanthen-2-yl)-carbamic acid tert-butyl ester
[0260] Di-tert-butyl dicarbonate (22 g, 99 mmol) was added to a
solution of 5-bromo-9H-thioxanthen-2-ylamine (19.28 g, 66 mmol) in
dry THF (150 ml). The mixture was stirred and heated at 50.degree.
C. overnight. The solvent was removed in vacuo and the residue was
triturated in water to give a brown solid, which was purified by
column chromatography (silica, dichloromethane) to give the title
compound as a fluffy white solid (18.28 g, 71%). %). .sup.1HNMR
(300 MHz, CDCl.sub.3): .delta..sub.H=1.55 (9H, s), 3.88 (2H, s),
6.45 (1H, bs), 7.06 (2H, m), 7.23 (1H, d), 7.39 (1H, d), 7.45 (1H,
d), 7.58 (1H, bs).
[5-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthen-2-yl]-carbamic
acid tert-butyl ester
[0261] (5-Bromo-9H-thioxanthen-2-yl)-carbamic acid tert-butyl ester
(1 g, 2.55 mmol), bis(pinacolato)diboron (0.78 g, 3.06 mmol) and
potassium acetate (0.75 g, 7.65 mmol) in dry 1,4-dioxane (6 ml). To
the yellow suspension was then added PdCl.sub.2(dppf) (0.10 g, 0.13
mmol) and dppf (7 mg, 0.13 mmol). The dark red mixture was heated
to 110.degree. C. under a N.sub.2 atmosphere for 24 hours. The
crude mixture was purified by flash chromatography (silica,
dichloromethane) to give
[5-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-9H-thioxanthen-2-yl]-ca-
rbamic acid tert-butyl ester as a viscous brown oil which was used
without any further purification (1.12 g).
[0262]
[5-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-9H-thioxanthen-2--
yl]-carbamic acid tert-butyl ester (1.12 g),
2-chloro-6-morpholin-4-yl-pyran-4-one (0.66 g, 3.06 mmol) and
grinded K.sub.2CO.sub.3 (0.71 g, 5.10 mmol) were dissolved in dry
1,4-dioxane (5 ml). The mixture was degassed for 15 mins and
Pd(PPh.sub.3).sub.4 (0.15 g, 0.13 mmol) was then added The dark
brown mixture was heated to 100.degree. C. under an atmosphere of
N.sub.2 for 24 hour. The reaction mixture was concentrated in vacuo
and water (50 ml) was added. The brown solid was filtered, washed
with water, dried overnight in a vacuum desiccator and was used
without any further purification (1.51 g). m/z (LC-MS, ESP),
RT=4.47 min, (M.sup.++1)=493
2-(7-Amino-9H-thioxanthen-4-yl)-6-morpholin-4-yl-pyran-4-one
(337)
[0263] To a solution of
[5-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthen-2-yl]-carbamic
acid tert-butyl ester (19) (1.08 g, 2.19 mmol) in dichloromethane
(10 ml) was added trifluoroacetic acid (2 ml) and left under
stirring at room temperature overnight. The solvent was dried in
vacuo revealing a viscous dark brown liquid. Saturated sodium
bicarbonate solution (20 ml) was added to the residue, which was
left to stir for 20 mins. The brown precipitate was filtered,
washing with water and left to dry in the vacuum oven overnight
(0.77 g, 90%). .sup.1HNMR (300 MHz, DMSO-d.sub.6):
.delta..sub.H=3.40 (4H, t), 3.70 (4H, t), 3.77 (2H, s), 5.23 (2H,
bs), 5.50 (1H, d), 6.17 (1H, d), 6.44 (1H, dd), 6.65 (1H, d), 7.09
(1H, d), 7.35 (1H, t), 7.47-7.59 (2H, m); m/z (LC-MS, ESP), RT=3.51
minutes, (M.sup.++1)=392.
2-(7-Amino-9H-thioxanthen-4-yl)-6-morpholin-4-yl-pyran-4-one
N-amide derivatives
[0264] To a small test tube was added
2-(7-amino-9H-thioxanthen-4-yl)-6-morpholin-4-yl-pyran-4-one (337)
(20 mg, 0.05 mmol), dry dimethylacetamide (0.5 ml), triethylamine
(8 .mu.l, 0.06 mmol) and chloroacetyl chloride (4 .mu.l, 0.06 mmol)
with stirring overnight. The appropriate amine or thiol (20 mg or
20 .mu.l) was then added and left to stir at room temperature
overnight. The reaction was purified by preparative HPLC to give
the desired products, which are shown below:
TABLE-US-00010 ##STR00376## Purity Retention Time Compound R (%)
(Mins) M.sup.+ + 1 338 ##STR00377## 95 3.15 561 339 ##STR00378## 95
3.20 577 340 ##STR00379## 90 3.40 607 341 ##STR00380## 90 3.23 561
342 ##STR00381## 95 3.18 547 343 ##STR00382## 95 3.23 596 344
##STR00383## 95 3.69 613 345 ##STR00384## 90 3.37 597 346
##STR00385## 85 3.24 618 347 ##STR00386## 90 2.91 590 348
##STR00387## 90 3.13 563 349 ##STR00388## 85 3.12 575 350
##STR00389## 85 3.03 577 351 ##STR00390## 90 3.17 575 352
##STR00391## 90 3.58 561 353 ##STR00392## 90 3.29 610 354
##STR00393## 85 3.23 627 355 ##STR00394## 90 2.94 611 356
##STR00395## 90 2.94 604
Example 6
Synthesis of 2(-(7-hydroxy-thianthren-1-yl)-6-morpholin-4-yl
pyran-4-one ether and acetamide derivatives)
##STR00396##
[0265] 2-(7-Hydroxy-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one
(357)
[0266] To a solution of
2-(7-Amino-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one (21) (575
mg, 1.4 mmol) was suspended in ethanol (5 mL) was added
tetrafluoroboric acid (54 wt % in ether, 3 ml, 1.68 mmol). The
mixture was stirred at room temperature for 10 minutes before being
cooled to 0.degree. C. Butyl nitrite (220 .mu.l, 2.8 mmol) was then
added dropwise and the mixture stirred for 30 minutes before the
addition of diethyl ether (40 ml) which caused a precipitate to
form. The solid was collected by filtration and washed with cold
diethylether (30 ml) then added to a solution of cupric nitrate
trihidrate (210 g, 870 mmol) and cuprous oxide (190 mg, 1.31 mmol)
in water (300 ml). The reaction was stirred for 12 hours and then
filtered. The filtrant was washed with water, dried in a desiccator
and used without further purification give the title compound (0.58
g, 100%). NMR (300 MHz, CDCl.sub.3): .delta..sub.H=10.08 (1H, bs);
7.76 (1H, dd); 7.60 (1H, dd); 7.46 (1H, dd); 7.35 (1H, dd); 7.02
(1H, d); 6.74 (1H, dd); 6.24 (1H, d); 5.55 (1H, d); 3.69 (H, m),
3.36 (4H, m); m/z (LC-MS, ESP), RT=3.83 min, (M.sup.++1)=412
2-[7-(2-Bromo-ethoxy)-thianthren-1-yl]-6-morpholin-4-yl-pyran-4-one
ether derivatives
[0267] To a solution of
2-(7-Hydroxy-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one (357)
(20 mg, 0.049 mmol) in anhydrous DMF (1 ml) was added powdered
potassium carbonate (20 mg, 0.147 mmol) and dibromoethane (0.019
ml, 0.23 mmol). The mixture was stirred for 16 hours at 60.degree.
C. before diisopropylethylamine (0.29 mmol) the appropriate amine
(0.29 mmol) were added. The reaction was maintained, with stirring,
at 60.degree. C. for 24 hrs after which time the crude mixture was
purified by preparative HPLC to give the desired compounds, which
are shown below:
TABLE-US-00011 ##STR00397## Retention Purity Time Compound R (%)
(Mins) M.sup.+ + 1 358 ##STR00398## 85 3.19 566 359 ##STR00399## 90
3.61 553 360 ##STR00400## 90 3.55 582 361 ##STR00401## 90 3.36 538
362 ##STR00402## 85 3.61 602 363 ##STR00403## 90 3.36 568 364
##STR00404## 90 3.45 566 365 ##STR00405## 90 3.42 552 366
##STR00406## 90 3.31 546 367 ##STR00407## 95 3.31 499 368
##STR00408## 95 3.55 523 369 ##STR00409## 95 3.38 567 370
##STR00410## 85 3.31 524 371 ##STR00411## 85 3.16 552 372
##STR00412## 90 3.08 538 373 ##STR00413## 95 3.36 543 374
##STR00414## 95 3.62 571 375 ##STR00415## 90 3.64 537 376
##STR00416## 95 3.56 511 377 ##STR00417## 95 3.14 628 378
##STR00418## 95 3.29 566 379 ##STR00419## 85 4.36 537 380
##STR00420## 85 3.47 539 381 ##STR00421## 95 3.48 560 382
##STR00422## 90 3.60 511 383 ##STR00423## 90 3.46 497 384
##STR00424## 85 3.51 529 385 ##STR00425## 85 3.53 535 386
##STR00426## 95 3.42 513 387 ##STR00427## 95 3.48 495 388
##STR00428## 90 3.21 568 389 ##STR00429## 90 3.04 582 390
##STR00430## 95 3.28 559 391 ##STR00431## 90 3.03 549 392
##STR00432## 95 3.96 624 393 ##STR00433## 90 3.44 509 394
##STR00434## 95 3.66 595 395 ##STR00435## 95 3.51 618 396
##STR00436## 95 3.65 595 397 ##STR00437## 85 3.36 559 398
##STR00438## 90 3.56 574 399 ##STR00439## 95 3.46 601 400
##STR00440## 85 3.38 557 402 ##STR00441## 90 3.34 612 403
##STR00442## 95 525
[6-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-2-yloxy]-acetic
acid methyl ester
[0268] To a suspension of
2-(7-Hydroxy-thianthren-1-yl)-6-morpholin-4-yl-pyran-4-one (357)
(20 mg, 0.049 mmol) in anhydrous DMF was added powdered potassium
carbonate (20 mg, 0.145) followed by methyl bromoacetate (0.01 ml,
0.106 mmol). The mixture was warmed to 60.degree. C. and maintained
at this temperature with stirring for 2 hours. The reaction was
cooled, poured into water (5 ml) and extracted using ethyl acetate
(3*5 ml), then washed with water (2*5 ml). The combined organics
were then dried (MgSO4), filtered and concentrated in vacuo to give
a yellow solid (22 mg, 94%) which corresponded to the title
compound and was used without any further purification. m/z (LC-MS,
ESP), RT=4.13 min, (M.sup.++1)=484
[6-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-2-yloxy]-acetic
acid sodium salt
[0269] To a solution of
[6-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-2-yloxy]-acetic
acid methyl ester (22 mg, 0.045 mmol in methanol (2 ml) was added
solid sodium hydroxide (2 mg, 0.045 mmol). The mixture was stirred
vigorously and heated to 60.degree. C. for 1 hour. After this time,
the mixture was concentrated in vacuo to dryness to give the title
compound (22.4 mg, 100%) and used without any further
purification.
2-[6-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-2-yloxy]-acetamide
derivatives
[0270] To a solution of
[6-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-thianthren-2-yloxy]-acetic
acid sodium salt (22.4 mg, 0.045 mmol) in dry DMF (1 ml) was added
HBTU (25.9 mg, 0.068 mmol), diisopropylethyl amine (0.1 ml) and the
appropriate amine (0.137 mmol). The mixture was stirred at room
temperature for 72 hours after which time the crude mixture was
purified by preparative HPLC to give the desired compounds, which
are shown below:
TABLE-US-00012 ##STR00443## Retention Purity Time Compound R (%)
(Mins) M.sup.+ + 1 404 ##STR00444## 85 3.75 602 405 ##STR00445## 95
4.29 523 406 ##STR00446## 95 4.48 583 407 ##STR00447## 95 4.41 525
408 ##STR00448## 90 3.33 596 409 ##STR00449## 95 3.88 508 410
##STR00450## 95 3.50 573 411 ##STR00451## 85 3.46 543 412
##STR00452## 95 3.76 638 413 ##STR00453## 95 4.39 609 414
##STR00454## 95 3.97 632 415 ##STR00455## 95 4.39 609 416
##STR00456## 95 3.57 580 417 ##STR00457## 95 4.24 555 418
##STR00458## 95 3.81 567 419 ##STR00459## 85 3.63 546 420
##STR00460## 95 4.25 595 421 ##STR00461## 90 4.76 631 422
##STR00462## 95 3.67 597 423 ##STR00463## 95 4.54 670 424
##STR00464## 95 4.24 647 425 ##STR00465## 95 3.45 588 426
##STR00466## 90 3.52 615 427 ##STR00467## 95 3.70 571 428
##STR00468## 95 3.28 626 429 ##STR00469## 95 3.64 580 430
##STR00470## 90 4.38 655 431 ##STR00471## 95 5.05 628 432
##STR00472## 90 4.71 632 433 ##STR00473## 95 4.24 567 434
##STR00474## 90 3.36 554 435 ##STR00475## 90 3.41 596 436
##STR00476## 95 3.32 582 437 ##STR00477## 95 3.41 580 438
##STR00478## 95 3.35 566 439 ##STR00479## 85 3.44 560 440
##STR00480## 95 4.46 537 441 ##STR00481## 90 3.61 581 442
##STR00482## 95 3.35 566 443 ##STR00483## 90 3.34 552 444
##STR00484## 90 3.52 557 445 ##STR00485## 90 4.17 585 446
##STR00486## 95 4.56 551 447 ##STR00487## 95 4.34 525 448
##STR00488## 95 3.91 497 449 ##STR00489## 90 4.27 616 450
##STR00490## 90 3.34 552
Example 7
5-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthene-2-sulfonic
acid amide derivatives,
2-[5-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthen-2-yloxy]-aceta-
mide derivatives and
2-[7-(2-Amino-ethoxy)-9H-thioxanthen-4-yl]-6-morpholin-4-yl-pyran-4-one
derivatives
##STR00491##
[0271]
5-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthene-2-sulfonyl
chloride
[0272] Tetrafluorboric acid (supplied as 54 wt % in diethylether,
5.5 ml, 39.92 mmol) was added to a mixture of
2-(7-Amino-9H-thioxanthen-4-yl)-6-morpholin-4-yl-pyran-4-one (337)
(1.03 g, 2.61 mmol) suspended in ethanol (10 ml). The reaction was
stirred for 10 minutes at room temperature before being cooled to
0.degree. C. and butyl nitrite (600 .mu.l, 5.1 mmol) added. The
reaction was then stirred at room temperature for 1 hour before it
was poured into diethyl ether (80 ml) and the precipitate collected
by filtration. The collected solid was washed with cold ether (60
ml) and then added to a saturated solution of sulphur dioxide in
acetic acid*. The mixture was stirred at room temperature for 2
hours whereupon it was extracted into CH.sub.2Cl.sub.2 (3.times.30
ml). The combined dichloromethane extracts were washed with water
(1.times.20 m), dried using MgSO4, filtered and concentrated in
vacuo to give the title compound which was used without further
purification (1.19 g, 100%), m/z (LC-MS, ESP), RT=4.46 min,
(M.sup.++1)=478 *Acidic SO.sub.2 solution prepared by bubbling
SO.sub.2 gas through a vigorously stirred solution of acetic acid
(100 ml) until 10 g of gas had been dissolved. The solution was
then treated with a suspension of copper (II) chloride (4 g) in
water (10 ml). The mixture was then stirred and filtered to give an
emerald green solution which was used without any further
purification.
5-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthene-2-sulfonic
acid amide derivatives
[0273]
5-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthene-2-sulfonyl
chloride (20 mg, 0.02 mmol) was diluted in DMF (1 ml) and to this
solution was added the appropriate amine (0.04 mmol) and
diisopropylethyl amine (80 .mu.l, 0.46 mmol). After stirring the
reaction mixture for 24 hours at room temperature the crude mixture
was purified by preparative HPLC to give the desired compounds,
which are shown below:
TABLE-US-00013 ##STR00492## Retention Purity Time Compound R (%)
(Mins) M.sup.+ + 1 452 ##STR00493## 95 3.13 527 453 ##STR00494## 95
3.32 568 454 ##STR00495## 90 4.17 539 455 ##STR00496## 95 3.95 541
456 ##STR00497## 95 3.35 561 457 ##STR00498## 100 4.14 513 458
##STR00499## 95 4.20 499 459 ##STR00500## 90 4.13 531 460
##STR00501## 100 4.14 537 461 ##STR00502## 100 3.83 515 462
##STR00503## 95 4.03 497 463 ##STR00504## 95 3.29 584 464
##STR00505## 95 3.86 496 465 ##STR00506## 95 3.27 551 466
##STR00507## 90 3.38 531 467 ##STR00508## 90 3.78 542 468
##STR00509## 95 4.30 511 469 ##STR00510## 95 3.69 568 470
##STR00511## 90 4.38 543 471 ##STR00512## 100 3.98 555 472
##STR00513## 95 3.68 534 473 ##STR00514## 90 3.73 561 474
##STR00515## 85 3.65 559 475 ##STR00516## 85 3.81 541 476
##STR00517## 95 3.78 568 477 ##STR00518## 90 3.35 568 478
##STR00519## 90 4.44 555 479 ##STR00520## 90 3.33 542 480
##STR00521## 90 3.32 540 481 ##STR00522## 95 3.32 570 482
##STR00523## 90 3.38 554 483 ##STR00524## 90 3.33 548 484
##STR00525## 95 3.55 501 485 ##STR00526## 85 4.62 525 486
##STR00527## 95 3.50 569 487 ##STR00528## 85 3.33 526 488
##STR00529## 85 4.49 527 489 ##STR00530## 90 3.34 540 490
##STR00531## 85 3.56 545 491 ##STR00532## 90 4.53 513 492
##STR00533## 90 3.34 568 493 ##STR00534## 95 4.29 553
[5-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthen-2-yloxy]-acetic
acid methyl ester
[0274] To a solution of
2-(7-Amino-9H-thioxanthen-4-yl)-6-morpholin-4-yl-pyran-4-one (1.01
g, 2.56 mmol) in anhydrous DMF (25 ml) was added powdered potassium
carbonate (716 mg, 5.18 mmol) followed by methylbromoacetate (400
.mu.l, 4.1 mmol). The reaction was stirred at room temperature for
100 hours whereupon it was quenched by dropwise addition of water
(75 ml). The mixture was extracted using EtOAc (3.times.30 ml). The
organic extracts were then combined, dried using MgSO.sub.4,
filtered and concentrated in vacuo to give the title compound as a
brown oil that was used without further purification (1.02 g,
85.7%), m/z (LC-MS, ESP), RT=4.05 min, (M.sup.++1)=466
[5-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthen-2-yloxy]-acetic
acid sodium salt
[0275] To a solution of
[5-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthen-2-yloxy]-acetic
acid methyl ester (1.02 g, 2.18 mmol) in THF (25 ml) was added
aqueous NaOH (2.4 ml, 2.4 mmol, 1M solution). The mixture was
stirred at 40.degree. C. for 2 hours whereupon it was cooled to
room temperature and concentrated in vacuo to give a brown oil. The
residue was azeotroped with toluene (3.times.10 ml) to give the
title compound as a fine, dark brown powder that was used without
further purification (1.03 g, 100%), m/z (LC-MS, ESP), RT=3.72 min,
(M.sup.++1)-Na.sup.+=452
2-[5-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthen-2-yloxy]-acetam-
ide derivatives
[0276]
[5-(6-Morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthen-2-yloxy]-a-
cetic acid sodium salt (20 mg, 0.04 mmol) was diluted in anhydrous
DMF (0.5 ml). To this solution was then added HBTU (33 mg, 0.09
mmol), diisopropylethylamine (74 .mu.l, 0.2 mmol) and the
appropriate amine (0.08 mmol). After stirring the reaction mixture
for 24 hours at room temperature the crude mixture was purified by
preparative HPLC to give the desired compounds, which are shown
below:
TABLE-US-00014 ##STR00535## Retention Purity Time Compound R (%)
(Mins) M.sup.+ + 1 494 ##STR00536## 95 3.72 521 495 ##STR00537## 85
3.28 562 496 ##STR00538## 85 4.17 533 497 ##STR00539## 85 3.83 535
498 ##STR00540## 90 3.33 556 499 ##STR00541## 90 4.17 505 500
##STR00542## 95 4.05 525 501 ##STR00543## 95 4.06 531 502
##STR00544## 95 3.75 509 503 ##STR00545## 90 3.28 578 504
##STR00546## 95 3.81 490 505 ##STR00547## 85 3.42 555 506
##STR00548## 85 3.27 545 507 ##STR00549## 85 3.38 525 508
##STR00550## 85 3.58 536 509 ##STR00551## 90 3.50 562 510
##STR00552## 90 4.13 537 511 ##STR00553## 85 3.68 549 512
##STR00554## 85 3.55 528 513 ##STR00555## 90 4.13 577 514
##STR00556## 95 3.38 570 515 ##STR00557## 95 3.64 553 516
##STR00558## 95 3.55 562 517 ##STR00559## 90 3.35 562 518
##STR00560## 95 4.08 549 519 ##STR00561## 90 3.24 536 520
##STR00562## 90 3.29 578 521 ##STR00563## 95 3.23 534 522
##STR00564## 90 3.21 564 523 ##STR00565## 85 3.34 562 524
##STR00566## 90 3.23 548 525 ##STR00567## 85 3.30 542 526
##STR00568## 90 4.22 519 527 ##STR00569## 85 3.46 563 528
##STR00570## 95 3.23 520 529 ##STR00571## 95 3.26 548 530
##STR00572## 85 3.39 539 531 ##STR00573## 85 3.99 567 532
##STR00574## 95 4.37 533 533 ##STR00575## 95 4.13 507
2-[7-(2-Amino-ethoxy)-9H-thioxanthen-4-yl]-6-morpholin-4-yl-pyran-4-one
derivatives
[0277] To a solution of
2-(7-Hydroxy-9H-thioxanthen-4-yl)-6-morpholin-4-yl-pyran-4-one (20
mg, 0.05 mmol) in DMA (1 ml) was added NaH (60% dispersion in
mineral oil, 6 mg 0.15 mmol) at 0.degree. C. The mixture was
stirred at 0.degree. C. for 30 minutes before the addition of
1,2-dibromoethane (19 mg, 0.1 mmol) The mixture was then allowed to
warm to room temperature and stirred like this for 12 hours before
the addition of the appropriate amine. The reaction was stirred for
a further 24 hours the crude mixture was purified by preparative
HPLC to give the desire compounds, which are shown below:
TABLE-US-00015 ##STR00576## Retention Purity Time Compound R (%)
(Mins) M.sup.+ + 1 534 ##STR00577## 90 3.38 521 535 ##STR00578## 90
3.44 542 536 ##STR00579## 95 3.45 491 537 ##STR00580## 95 3.45 479
538 ##STR00581## 90 3.04 564 539 ##STR00582## 95 3.24 511 540
##STR00583## 95 3.49 523 541 ##STR00584## 85 3.72 584 542
##STR00585## 90 3.33 507
B) Biological Examples
Materials and Methods
In Vitro ATM Inhibition Assays
[0278] In order to assess the inhibitory action of the compounds
against ATM in vitro, the following assay was used to determine
IC.sub.50 values. ATM protein was immunoprecipitated from HeLa cell
nuclear extract using rabbit polyclonal anti-sera raised to the
C-terminal .about.500 amino-acid residues of the human ATM protein.
The immunoprecipitation was performed according to the methodology
described by Banin, S. et al. (1998). 10 .mu.l of
immunoprecipitated ATM in Buffer C (50 mM Hepes, pH 7.4, 6 mM
MgCl.sub.2, 150 mM NaCl, 0.1 mM sodium orthovanadate, 4 mM MnCl2,
0.1 mM dithiothreitol, 10% glycerol) was added to 32.5 .mu.l of
buffer C containing 1 .mu.g of the ATM substrate GSTp53N66 in a
V-bottomed 96 well polypropylene plate. The GSTp53N66 substrate is
the amino terminal 66 amino acid residues of human wild type p53
fused to glutathione S-transferase. ATM phosphorylates p53 on the
residue serine 15 (Banin, S. et al. (1998)). Varying concentrations
of inhibitor were then added. All compounds were diluted in DMSO to
give a final assay concentration of between 100 .mu.M and 0.1 nM,
with DMSO being at a final concentration of 1%. After 10 minutes of
incubation at 37.degree. C., the reactions were initiated by the
addition of 5 .mu.l of 500 .mu.M Na-ATP. After 1 hour with shaking
at 37.degree. C., 150 .mu.l of phosphate buffered saline (PBS) was
added to the reaction and the plate centrifuged at 1500 rpm for 10
minutes. 5 .mu.l of the reaction was then transferred to a 96 well
opaque white plate containing 45 .mu.l of PBS to allow the
GSTp53N66 substrate to bind to the plate wells. The plate was
covered and incubated at room temperature for 1 hour with shaking
before discarding the contents. The plate wells were washed twice
by the addition of PBS prior to the addition of 3% (w/v) bovine
serum albumin (BSA) in PBS. The plate was incubated at room
temperature for 1 hour with shaking before discarding the contents
and washing twice with PBS. To the wells, 50 .mu.l of a 1:10,000
dilution of primary phosphoserine-15 antibody (Cell Signaling
Technology, #9284L) in 3% BSA/PBS was added to detect the
phosphorylation event on the serine 15 residue of p53 elicited by
the ATM kinase. After 1 hour of incubation at room temperature with
shaking, the wells were washed four times with PBS prior to the
addition of an anti-rabbit HRP conjugated secondary antibody
(Pierce, 31462) with shaking for 1 hour at room temperature. The
wells were then washed four times with PBS before the addition of
chemiluminescence reagent (NEN Renaissance, NEL105). The plate was
then shaken briefly, covered with a transparent plate seal and
transferred to a TopCount NXT for chemiluminescent counting. Counts
per second, following a one second counting time, were recorded for
each reaction.
[0279] The enzyme activity for each compound is then calculated
using the following equation:
% Inhibition = 100 - ( ( cpm of unknown - mean negative cpm )
.times. 100 ( mean positive cpm - mean negative cpm ) )
##EQU00001##
Results
In Vitro ATM Assays
[0280] Compounds were assayed for ATM inhibition activity using the
method described above. The results are detailed 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 100 .mu.M down to 0.1 nM. Such
IC.sub.50 values are used as comparative values to identify
increased compound potencies.
[0281] The following compounds had IC.sub.50 values of less than
200 nM: 6, 8, 9, 11, 13-19, 21-129, 148, 153, 164, 170, 172, 181,
182, 200, 240-257, 259-262, 264, 266, 267, 270-272, 274, 277,
280-292, 294-307, 311, 312, 315, 321, 322, 324, 326-331, 333, 336,
338-356, 359, 361, 366, 367, 378, 379, 381-383, 395, 398, 399, 403,
405, 407, 409-411, 414, 416-419, 424, 426, 427, 430, 433-442,
444-446, 450, 454, 455, 457-459, 462, 465-466, 468-469, 471-476,
478, 480, 482, 484, 485, 488-490, 493, 494, 496-497, 499-501,
504-507, 509-513, 515, 516, 518, 520-526, 528, 532-533, 537,
540-542.
[0282] The following compounds had IC.sub.50 values of less than 10
.mu.M, in addition to those listed above: 7, 10, 12, 20, 131-147,
149-152, 154-163, 165-169, 171, 173-180, 183-199, 201-238, 256,
258, 263, 265, 268, 269, 273, 275, 276, 278, 279, 293, 308-310,
313, 314, 316-320, 322, 323, 325, 332, 334, 335, 358, 362, 363,
365, 368, 370-375, 377, 380, 384-392, 400-402, 406, 408, 412, 413,
415, 421-423, 425, 428, 429, 431-432, 443, 447-449, 452, 453, 456,
460, 461, 463, 464, 470, 477, 481, 483, 486, 487, 491, 492, 495,
498, 502, 503, 514, 517, 519, 527, 529-531, 534-536, 538.
* * * * *