U.S. patent application number 12/403288 was filed with the patent office on 2009-09-10 for novel boronic chalcone derivatives and uses thereof.
This patent application is currently assigned to JOHNS HOPKINS UNIVERSITY, JOHNS HOPKINS TECHNOLOGY TRANSFER. Invention is credited to Saeed R. Khan.
Application Number | 20090227542 12/403288 |
Document ID | / |
Family ID | 29740004 |
Filed Date | 2009-09-10 |
United States Patent
Application |
20090227542 |
Kind Code |
A1 |
Khan; Saeed R. |
September 10, 2009 |
Novel Boronic Chalcone Derivatives and Uses Thereof
Abstract
The present invention relates to novel boronic chalcone
derivatives which are useful as antitumor/anticancer agents. The
present compounds, which are inexpensive to synthesize, exhibit
unexpectedly good inhibitors of the growth of human breast cancer
cells. The present invention also relates to the use of the novel
boronic chalcone derivatives to treat cancer. The invention also
provides pharmaceutical compositions comprising the inhibitors of
the invention and methods of utilizing the inhibitors and
pharmaceutical compositions in the treatment and prevention of
cancer.
Inventors: |
Khan; Saeed R.; (Owings
Mills, MD) |
Correspondence
Address: |
Karen B. Dow;DLA PIPER LLP (US)
Suite 1100, 4365 Executive Drive
San Diego
CA
92121-2133
US
|
Assignee: |
JOHNS HOPKINS UNIVERSITY, JOHNS
HOPKINS TECHNOLOGY TRANSFER
|
Family ID: |
29740004 |
Appl. No.: |
12/403288 |
Filed: |
March 12, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10517781 |
Apr 20, 2005 |
7514579 |
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PCT/US03/18962 |
Jun 12, 2003 |
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12403288 |
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60388255 |
Jun 13, 2002 |
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60444429 |
Feb 3, 2003 |
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Current U.S.
Class: |
514/64 ; 548/405;
562/7 |
Current CPC
Class: |
A61P 35/04 20180101;
C07F 5/025 20130101; A61P 35/00 20180101 |
Class at
Publication: |
514/64 ; 562/7;
548/405 |
International
Class: |
C07F 5/02 20060101
C07F005/02; A61K 31/69 20060101 A61K031/69; A61P 35/00 20060101
A61P035/00; A61P 35/04 20060101 A61P035/04 |
Claims
1. A compound including resolved enantiomers, diastereomers,
solvates and pharmaceutically acceptable salts thereof, said
compound having the Formula (I): ##STR00010## where: Ar is aryl or
heteroaryl, each of which may be unsubstituted or substituted with
a substituent selected from the group consisting of F, CI, Br, I,
alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-OR.sup.1,
Z.sub.n-NO.sub.2, Z.sub.n-CN, Z.sub.n-CO.sub.2R.sup.1,
Z.sub.n-(C.dbd.O)R.sup.1, Z.sub.n-O(C.dbd.O)R.sup.1,
Z.sub.n--O-alkyl, Z.sub.n-OAr, Z.sub.n-SH, Z.sub.n-SR.sup.1,
Z.sub.n-SOR.sup.1, Z.sub.n-SO.sub.2R.sup.1, Z.sub.n-S--Ar,
Z.sub.n-SOAr, Z.sub.n-SO.sub.2Ar, Z.sub.n--Ar, Z.sub.n-heteroaryl,
Z.sub.n-(C.dbd.O)NR.sup.1R.sup.2, Z.sub.n-NR.sup.1(C.dbd.O)R.sup.1,
Z.sub.n-SO.sub.2NR.sup.1R.sup.2, PO.sub.3H.sub.2, and
SO.sub.3H.sub.2; W is H, Z.sub.n-F, Z.sub.n-Cl, Z.sub.n-Br,
Z.sub.n-I, Z.sub.n-CF.sub.3, Z.sub.n-NO.sub.2, Z.sub.n-OR.sup.1,
Z.sub.n-NR.sup.1R.sup.2, Z.sub.n-COOR.sup.1, Z.sub.n-SR.sup.1,
Z.sub.n-(C.dbd.O)R.sup.1, Z.sub.n-O(C.dbd.O)R.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O)R.sup.1, Z.sub.n-(C.dbd.O)NR.sup.1, alkyl,
allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein
said alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or
Z.sub.n-heteroaryl may be substituted or unsubstituted; X is
Z.sub.n, Z.sub.n-O, Z.sub.n-S, Z.sub.n-NR.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O), Z.sub.n-C.dbd.O, Z.sub.n-OC(.dbd.O), or
Z.sub.n-C(.dbd.O)O; R.sup.1 and R.sup.2 are independently H, an
amine protecting group, an alcohol protecting group, an acid
protecting group, a sulfur protecting group, alkyl, ally), alkenyl,
alkynyl, heteroalkyl, heteroallyl, heteroalkenyl, heteroalkynyl,
alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl,
Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein said alkyl, allyl,
alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl may be
substituted or unsubstituted, or R.sup.1 together with R.sup.2 and
N forms a saturated or partially unsaturated heterocycle ring
having 1 or more heteroatoms in said ring, wherein said heterocycle
may be substituted or unsubstituted and wherein said heterocycle
may be fused to an aromatic ring; Z is an alkylene having at least
1 carbon, or an alkenylene or alkynylene each having at least 2
carbons, wherein said alkylene, alkenylene, or alkynylene may be
substituted or unsubstituted; and n is zero or any integer.
2. The compound of claim 1 having the structure ##STR00011##
3. The compound of claim 1 having the structure ##STR00012##
4. The compound of claim 1 having the structure ##STR00013##
5. The compound of claim 1 having the structure ##STR00014##
6. The compound of claim 1 having the structure ##STR00015##
7. The compound of claim 1 having the structure ##STR00016##
8. The compound of claim 1 having the structure ##STR00017##
9. The compound of claim 1 having the structure ##STR00018##
10. A compound including resolved enantiomers, diastereomers,
solvates and pharmaceutically acceptable salts thereof, said
compound having the Formula (III): ##STR00019## where Ar is aryl or
heteroaryl, each of which may be substituted or unsubstituted;
R.sup.4 is H, an amine protecting group, Z.sub.n-OR.sup.1,
Z.sub.n-SR.sup.1, Z.sub.n-NR.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O)R.sup.1, Z.sub.n-C.dbd.OR.sup.1,
Z.sub.n-OC(.dbd.O)R.sup.1, Z.sub.n-C(.dbd.O)OR.sup.1, alkyl, allyl,
alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein
said alkyl, allyl, alkenyl, alkynyl, heteroalkyl,-heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or
Z.sub.n-heteroaryl may be substituted or unsubstituted; R.sup.1 is
H, alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or
Z.sub.n-heteroaryl, wherein said alkyl, allyl, alkenyl, alkynyl,
heteroalkyl, heteroallyl, heteroalkenyl, heteroalkynyl, alkoxy,
heteroalkoxy, Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, or
Z.sub.n-Ar or Z.sub.n-heteroaryl may be substituted or
unsubstituted, Z is an alkylene having at least 1 carbon, or an
alkenylene or alkynylene each having at least 2 carbons, wherein
said alkylene, alkenylene, or alkynylene may be substituted or
unsubstituted; and n is zero or any integer.
11. A method of treating a tumor or cancer in a patient in need
thereof comprising administering to said patient an effective
amount of a compound having the Formula (III): ##STR00020## where
Ar is aryl or heteroaryl, each of which may be substituted or
unsubstituted; R.sup.4 is H, an amine protecting group,
Z.sub.n-OR.sup.1, Z.sub.n-SR.sup.1, Z.sub.n-NR.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O)R.sup.1, Z.sub.n-C.dbd.OR.sup.1,
Z.sub.n-OC(.dbd.O)R.sup.1, Z.sub.n-C(.dbd.O)OR.sup.1, alkyl, allyl,
alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein
said alkyl, ally), alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, or Z.sub.n-Ar or
Z.sub.n-heteroaryl may be substituted or unsubstituted; R.sup.1 is
H, alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or
Z.sub.n-heteroaryl, wherein said alkyl, allyl, alkenyl, alkynyl,
heteroalkyl, heteroallyl, heteroalkenyl, heteroalkynyl, alkoxy,
heteroalkoxy, Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl,
Z.sub.n-Ar or Z.sub.n-heteroaryl may be substituted or
unsubstituted; Z is an alkylene having at least 1 carbon, or an
alkenylene or alkynylene each having at least 2 carbons, wherein
said alkylene, alkenylene, or alkynylene may be substituted or
unsubstituted; and n is zero or any integer.
12. The method of claim 11, wherein said tumor is selected from the
group consisting of breast, cervical, stomach, colon, bladder,
rectal, liver, pancreatic, lung, cervix uteri, corpus uteri, ovary,
prostate, testis, renal, brain/cns, head, neck, throat, anal and
oral cancers, eye or ocular cancer, skin melanoma, Ewing's Sarcoma,
Kaposi's Sarcoma, basal cell carcinoma and squamous cell carcinoma,
small cell lung cancer, mouth/pharynx, esophageal, larynx, kidney
and lymphoma, acute lymphocytic leukemia, and acute myelogenous
leukemia.
13. A method of inhibiting MDM2 expression in a mammal, comprising
administering an amount of a compound effective to inhibit said
expression, said compound having the Formula (I): ##STR00021##
where Ar is aryl or heteroaryl, each of which may be substituted or
unsubstituted; W is H, Z.sub.n-F, Z.sub.n-CI, Z.sub.n-Br,
Z.sub.n-I, Z.sub.n-CF.sub.3, Z.sub.n-NO.sub.2, Z.sub.n-OR.sup.1,
Z.sub.n-NR.sup.1R.sup.2, Z.sub.n-COOR.sup.1, Z.sub.n-SR.sup.1,
Z.sub.n-(C.dbd.O)R.sup.1, Z.sub.n-O(C.dbd.O)R.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O)R.sup.1, Z.sub.n-(C.dbd.O)NR.sup.1, alkyl,
allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein
said alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or
Z.sub.n-heteroaryl may be substituted or unsubstituted; X is
Z.sub.n, Z.sub.n-O, Z.sub.n-S, Z.sub.n-NR.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O), Z.sub.n-C.dbd.O, Z.sub.n-OC(.dbd.O), or
Z.sub.n-C(.dbd.O)O; R.sup.1 and R.sup.2 are independently H, an
amine protecting group, an alcohol protecting group, an acid
protecting group, a sulfur protecting group, alkyl, allyl, alkenyl,
alkynyl, heteroalkyl, heteroallyl, heteroalkenyl, heteroalkynyl,
alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl,
Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein said alkyl, allyl,
alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl may be
substituted or unsubstituted, or R.sup.1 together with R.sup.2 and
N forms a saturated or partially unsaturated heterocycle ring
having 1 or more heteroatoms in said ring, wherein said heterocycle
may be substituted or unsubstituted and wherein said heterocycle
may be fused to an aromatic ring; Z is an alkylene having at least
1 carbon, or an alkenylene or alkynylene each having at least 2
carbons, wherein said alkylene, alkenylene, or alkynylene may be
substituted or unsubstituted; and n is zero or any integer.
14. A method of inhibiting MDM2 expression in a mammal, comprising
administering an amount of a compound effective to inhibit said
expression, said compound having the Formula (II): ##STR00022##
where Ar is aryl or heteroaryl, each of which may be substituted or
unsubstituted; W is H, Z.sub.n-F, Z.sub.n-CI, Z.sub.n-Br,
Z.sub.n-I, Z.sub.n-CF.sub.3, Z.sub.n-NO.sub.2, Z.sub.n-OR.sup.1,
Z.sub.n-NR.sup.1R.sup.2, Z.sub.n-COOR.sup.1, Z.sub.n-SR.sup.1.
Z.sub.n-(C.dbd.O)R.sup.1, Z.sub.n-O(C.dbd.O)R.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O)R.sup.1, Z.sub.n-(C.dbd.O)NR.sup.1, alkyl,
allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein
said alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or
Z.sub.n-heteroaryl may be substituted or unsubstituted; X is
Z.sub.n, Z.sub.n-O, Z.sub.n-S, Z.sub.n-NR.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O), Z.sub.n-C.dbd.O, Z.sub.n-OC(.dbd.O), or
Z.sub.n-C(.dbd.O)O; R.sup.1 and R.sup.2 are independently H,
an-amine protecting group, an alcohol protecting group, an acid
protecting group, a sulfur protecting group, alkyl, ally), alkenyl,
alkynyl, heteroalkyl, heteroallyl, heteroalkenyl, heteroalkynyl,
alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl,
Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein said alkyl. allyl,
alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl may be
substituted or unsubstituted, or R.sup.1 together with R.sup.2 and
N forms a saturated or partially unsaturated heterocycle ring
having 1 or more heteroatoms in said ring, wherein said heterocycle
may be substituted or unsubstituted and wherein said heterocycle
may be fused to an aromatic ring; R.sub.3 is an
electron-withdrawing moiety; Z is an alkylene having at least 1
carbon, or an alkenylene, or alkynylene each having at least 2
carbons, wherein said alkylene, alkenylene, or alkynylene may be
substituted or unsubstituted; and n=zero or any integer.
15. A method of inhibiting MDM2 expression in a mammal, comprising
administering an amount of a compound effective to inhibit said
expression, said compound having the Formula (III): ##STR00023##
where Ar is aryl or heteroaryl, each of which may be substituted or
unsubstituted; R.sup.4 is H, an amine protecting group,
Z.sub.n-OR.sup.1, Z.sub.n-SR.sup.1, Z.sub.n-NR.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O)R.sup.1, Z.sub.n-C.dbd.OR.sup.1,
Z.sub.n-OC(.dbd.O)R.sup.1, Z.sub.n-C(.dbd.O)OR.sup.1, alkyl, allyl,
alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein
said alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, or Z.sub.n-Ar or
Z.sub.n-heteroaryl may be substituted or unsubstituted; R.sup.1 is
H, alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or
Z.sub.n-heteroaryl, wherein said alkyl, allyl, alkenyl, alkynyl,
heteroalkyl, heteroallyl, heteroalkenyl, heteroalkynyl, alkoxy,
heteroalkoxy, Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl,
Z.sub.n-Ar or Z.sub.n-heteroaryl may be substituted or
unsubstituted; Z is an alkylene having at least 1 carbon, or an
alkenylene or alkynylene each having at least 2 carbons, wherein
said alkylene, alkenylene, or alkynylene may be substituted or
unsubstituted; and n is zero or any integer.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional application of U.S.
application Ser. No. 10/517,781 filed Apr. 20, 2005, now pending;
which is a 35 USC .sctn. 371 National Stage application of
International Application No. PCT/US03/18962 filed Jun. 12, 2003;
which claims the benefit under 35 USC .sctn. 119(e) to U.S.
Application Ser. No. 60/444,429 filed Feb. 3, 2003 and U.S.
Application Ser. No. 60/388,255 filed Jun. 13, 2002, now abandoned.
The disclosure of each of the prior applications is considered part
of and is incorporated by reference in the disclosure of this
application.
BACKGROUND OF THE INVENTION
Field of the Invention
[0002] This invention relates to novel boronic chalcone compounds
and uses thereof. The compounds of this invention are particularly
useful for the treatment of tumors and cancers.
[0003] Throughout this application, various publications are
referenced by author and date. Full citations for these
publications may be found listed at the end of the specification
immediately preceding the claims. The disclosures of these
publications are hereby incorporated by reference in their
entireties into this application in order to more fully describe
the state of the art as known to those skilled therein as of the
date of this invention described and claimed herein,
[0004] Breast cancer is expected to account for 203,500 new cancer
cases and 39,600 deaths in 2002 (Jemal, A. et al., CA Cancer J.
Clin. 2002, 52, 23-47). Although major advances have been made in
early detection, prevention, and treatment, the need for more
effective therapy in the fight against late stage breast cancer
continues. Currently there is no curative treatment for women with
metastatic breast cancer once they have failed adjuvant therapies.
New and effective cytotoxic agents with novel mechanisms of action
are therefore urgently needed for the treatment of women with
metastatic breast cancer. Hormone and chemotherapy while of
substantial palliative benefit, has had little impact on overall
survival and the mortality rate from metastatic breast cancer. At
the present time standard treatments for metastatic breast cancer
include paclitaxel in combination with vinca alkaloids, etoposide,
and other regimens that include agents such as anthacyclines,
alkylating agents, antimetabolites, tamoxifen, and aromatase
inhibitors (Chabner B A, Collins J M, Cancer chemotherapy principal
and practice, pp 9-13, and 40-85. B. Lippincott Company,
Philadelphia, 1990). The ultimate conclusion of these numerous
studies over the last 50 years is that although these therapies
provides significant palliative effect in the majority of with
metastatic breast patients, it is likely to be curative. Although
major advances have been made in early detection, prevention, and
treatment of early disease, the need for more effective therapy in
the fight against late stage breast cancer continues.
[0005] Recently the mouse double minute 2 (MDM2) oncogene has been
suggested as a target for breast cancer therapy (Juven-Gershon, T.
and Oren, M. Mol. Med., 1999, 5, 71-83; Momand, J. et al., Nucleic
Acids Res., 1998, 26, 3453-3459). MDM2 is amplified or
overexpressed in human breast cancer, and MDM2 levels are
associated with poor prognosis of human breast cancer. The
oncoprotein MDM2 inhibits the tumor suppressor protein p53 by
binding to the p53 transactivation domain. The p53 gene is
inactivated in human cancer either by mutations or by binding to
oncogenic proteins such as MDM2 (Lane, D. P. and Hall, P. A.,
Trends BioChem. Sci, 1997, 22, 372-374; Oliner, J. D. et al.,
Nature, 1992, 358, 80-83; Lozano, G.; Montes de Oca Luna, R.,
Biochim. Biophys. Acta, 1998, 1377, M55-M59; Wang, H. et al.,
Clinical Cancer Res., 2001, 7, 3613-3624). In breast tumors, over
expression of MDM2 inactivates an otherwise intact p53, disabling
the genome integrity checkpoint and allowing cell cycle progression
of defective cells (Boyd, M. T. et al., J. Biol. Chem., 2000, 275,
31883-31890). Studies comparing MDM2 overexpression and p53
mutation concluded that these are mutually exclusive events,
supporting the notion that the primary impact of MDM2 amplification
in cancer cells is the inactivation of the endogenous wild-type p53
(Wang et al., supra). It has been shown recently that a peptide
homologue of p53 is sufficient to induce p53-dependent death of
cells overexpressing MDM2 (Wasylyk, C., et al., Oncogene, 1999, 18,
1921-1934). This result provides clear evidence that disruption of
the p53/MDM2 complex might be effective in cancer therapy. It has
been shown that MDM2 additionally has a role in tumor growth
p53-independent mechanisms (Baker, S. J., et al., Science, 1990,
249, 912-915; Diller, L, et al., Mol. Cell. Biol., 1990, 10,
5772-5781; Fakharzadeh, S. S., et al., EMBO J., 1991, 10, 1565;
Lundgren, K.; Montes de Oca Luna, R., et al., Genes Dev., 1997, 11,
714-725; Zhang, R. and Wang, H., Curr. Pharm. Des., 2000, 6,
393-416; Chabner, B. A. and Collins, J. M., Cancer chemotherapy
principal and practice; Lippincott Williams & Wilkins
Publishers Philadelphia, 1990; pp 9-13 and 40-85B).
[0006] Chalcones are a class of anticancer agents that have shown
promising therapeutic efficacy for the management of human cancers.
Chalcones, considered as the precursor of flavonoids and
isoflavonoids, are abundant in edible plants. Chemically they
comprise open-chain flavonoids in which the two aromatic rings are
joined by a three-carbon .alpha., .beta.-unsaturated carbonyl
system. For example, chalcones have been observed to inhibit the
proliferation of both established and primary ovarian cancer cells
(De Vincenzo, R., et al., Anticancer Drug Des., 1995, 10, 481-490).
In vivo, chalcones have been demonstrated to be effective as
antitumor agents in skin carcinogenesis (Statomi, Y., Int. J.
Cancer, 1993, 55, 506-514; Yamamoto, S. et al., Carcinogenesis,
1991, 12, 317-323) and chemopreventive agents in several
experimental models (Makita, H., et al., Cancer Res., 1996, 56,
4904-4909; Rui, H., J. Cell. Biochem., 1997, 67, 7-11; Wattenberg,
L. W., et al., Cancer Lett., 1994, 83, 165-169). Recent studies
have shown that these chalcones induce apoptosis in variety of cell
types including breast cancers (Claude-Alain, C., et al.,
Anticancer Res., 2001, 21, 3949-3956; WO 01/117988; WO 96/19209;
U.S. Pat. No. 5,808,137; Maggiolini, M., et al., J. Steroid
BioChem. Mol. Biol., 2002, 82, 315-322; Stall, R., et al.,
Biochemistry, 2001, 40, 336-344; DiCesare, N. and Lakowicz, J. R.,
Tetrahedron. Lett., 2002, 43, 2615-2618). Biochemical experiments
have shown that these compounds could disrupt the MDM2/p53 protein
complex, releasing p53 from both the p53/MDM2 and DNA-bound
p53/MDM2 complexes (Stoll et al., supra).
[0007] Carboxylic chalcones have shown promising therapeutic
efficacy for the management of human cancers (Daskiewicz, J. B., et
al., Tetrahedron Lett. 1999, 40, 7095-7098; Devincenzo, R., et al.,
Anti-Cancer Drug Des. 1995, 10, 481-490). Previous studies (Stoll
et al., supra; Kussie, P. H., et al., Science, 1996, 274, 948-953)
on the binding modes of carboxylic acid analogs of chalcones with
MDM2 revealed that the carboxylic acid group could be placed near
the base of lysine 51 (K51), which is found in a salt bridge
interaction with glutamic acid 25 (E25). It was presumed that the
acid group of the chalcone forms a salt bridge with K51 and
simultaneously breaks the salt bridge with E25 of the MDM2.
However, carboxylic acid analogs of chalcone reported in the
literature (Stoll et al., supra) are equally toxic to both normal
and malignant breast epithelial cells. The toxicity to normal
breast cells may be due to MDM2/p53 independent mechanisms.
Therefore, a chalcone derivative that could strongly and
irreversibly bind to and disrupt MDM2 protein complexes may be
selectively toxic to MDM2 overexpressing breast cancer cells.
[0008] Boronic acids are Lewis acids and isosteres of carboxylic
acid. The pKa's of boronic acids are about 9-10, and therefore at
physiological pH boronic acids remain unionized
(Tongcharoensirikui, P., et al., Abstracts of Papers, 222nd ACS
national meeting, Chicago, Ill., August 26-30; American chemical
society, Washington, D.C., 2001; MEDI-224). Thus, a coordinate
covalent bond (boron-nitrogen) can be formed between a electron
deficient boronic acid moiety and electron donating amino group,
which may strongly enhance binding of boronic-chalcones with the
lysine 51 of MDM2 at neutral pH when compared to the corresponding
carboxylic acid analog of chalcones.
[0009] Boronic chalcone analogs have been previously described.
These compounds have been used as fluorescent probes that may be
useful for detection of fluorides (DiCesare, N. and Lakowicz, J.
R., supra) and saccharides such as glucose that may be applicable
to the design of biosensors for diabetes (DiCesare, N. and
Lakowicz, J. R., supra). However, prior to this invention no
investigations into the anticancer activity of boronic-chalcones on
different cancer cell lines have been reported.
SUMMARY OF THE INVENTION
[0010] Surprisingly, it has now been found that certain novel
chalcones derivatives, in particular boronic chalcone derivatives,
possess antiproliferative activity on cancer cells at micromolar
concentrations. Accordingly, this invention provides the design and
synthesis of novel boronic chalcone derivatives, and pharmaceutical
compositions containing these compounds. Several the compounds
described herein were observed to have high activity in the breast
cancer cell lines tested and has been shown to be 6-9 fold less
toxic to normal MCF-12A cell lines compared to normal breast
epithelial cell lines. The novel boronic chalcone analogs disclosed
herein should overcome the limiting lack of specificity of
carboxylic acid analogs of chalcones.
[0011] The present invention farther investigates the potential
value of MDM2 as a drug target for breast cancer therapy. For
example, a chalcone derivative of this invention that inhibits MDM2
expression or binds to and disrupts the MDM2 protein complex may be
a useful compound for the treatment of breast cancer. While not
wishing to be bound by any theory, it is believed that the boronic
acid analog might form a stronger salt bridge with K51 of MDM2 than
the corresponding carboxylic acid analogs of chalcones and will
selectively inhibit growth of breast cancer cells. Accordingly, a
set of boronic acid-chalcone derivatives were designed and tested
their ability to selectively kill breast cancer over normal breast
epithelial cells.
[0012] In general, one embodiment of the invention relates to
boronic-chalcone compounds of the general Formula I:
##STR00001##
where
[0013] Ar is aryl or heteroaryl, each of which may be substituted
or unsubstituted;
[0014] W is H, Z.sub.n-F, Z.sub.n-CI, Z.sub.n-Br, Z.sub.n-I,
Z.sub.n-CF.sub.3, Z.sub.n-NO.sub.2, Z.sub.n-OR.sup.1,
Z.sub.n-NR.sup.1R.sup.2, Z.sub.n-COOR.sup.1, Z.sub.n-SR.sup.1,
Z.sub.n-(C.dbd.O)R.sup.1, Z.sub.n-O(C.dbd.O)R.sup.1;
Z.sub.n-NR.sup.1(C.dbd.O)R.sup.1, Z.sub.n-(C.dbd.O)NR.sup.1, alkyl,
allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein
said alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or
Z.sub.n-heteroaryl may be substituted or unsubstituted;
[0015] X is Z.sub.n, Z.sub.n-O, Z.sub.n-S, Z.sub.n-NR.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O), Z.sub.n-C.dbd.O, Z.sub.n-OC(.dbd.O), or
Z.sub.n-C(.dbd.O)O;
[0016] R.sup.1 and R.sup.2 are independently H, an amine protecting
group, an alcohol protecting group, an acid protecting group, a
sulfur protecting group, alkyl, allyl, alkenyl, alkynyl,
heteroalkyl, heteroallyl, heteroalkenyl, heteroalkynyl, alkoxy,
heteroalkoxy, Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl,
Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein said alkyl, allyl,
alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl may be
substituted or unsubstituted,
[0017] or R.sup.1 together with R.sup.2 and N forms a saturated or
partially unsaturated heterocycle ring having 1 or more heteroatoms
in said ring, wherein said heterocycle may be substituted or
unsubstituted and wherein said heterocycle may be fused to an
aromatic ring;
[0018] Z is an alkylene having at least 1 carbon, or an alkenylene
or alkynylene each having at least 2 carbons, wherein said
alkylene, alkenylene, or alkynylene may be substituted or
unsubstituted; and
[0019] n is zero or any integer.
[0020] The invention is also directed to pharmaceutically
acceptable prodrugs, pharmaceutically active metabolites, and
pharmaceutically acceptable salts of the compound of Formula I.
Methods of making the compounds of Formula I are also
described.
[0021] In another embodiment, this invention relates to compounds
of the general Formula II:
##STR00002##
where Ar, W, X and n are as defined above and R.sub.3 is an
electron-withdrawing moiety. The invention is also directed to
pharmaceutically acceptable prodrugs, pharmaceutically active
metabolites, and pharmaceutically acceptable salts of the compound
of Formula II. Methods of making the compounds of Formula II are
also described.
[0022] In another embodiment, this invention relates to compounds
of the general Formula III:
##STR00003##
where Ar is as defined above and R.sup.4 is H, an amine protecting
group, Z.sub.n-OR.sup.1, Z.sub.n-SR.sup.1, Z.sub.n-NR.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O)R.sup.1, Z.sub.n-C.dbd.OR.sup.1,
Z.sub.n-OC(.dbd.O)R.sup.1, Z.sub.n-C(.dbd.O)OR.sup.1, alkyl, allyl,
alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein
said alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or
Z.sub.n-heteroaryl may be substituted or unsubstituted, and where
Z.sub.n, R.sup.1 and n are as defined above. The invention is also
directed to pharmaceutically acceptable prodrugs, pharmaceutically
active metabolites, and pharmaceutically acceptable salts of the
compound of Formula III. Methods of making the compounds of Formula
III are also described.
[0023] The invention further relates to a method for treating
proliferative diseases such as cancers. More specifically, one
embodiment of this invention provides a method of treating or
preventing a tumor or cancer in a patient comprising administering
to said patient in need thereof an effective amount of a compound
having the Formula I-III or a pharmaceutically-acceptable salt or
in vivo cleavable prodrug thereof. Other aspects of the invention
include methods for treating cancers mediated by MDM2. Examples of
cancers that may be treated of prevented by the compounds of this
invention include, but are not limited to, breast, colorectal,
cervical, ovarian, brain, acute leukemia, gastric, non-small cell
lung, pancreatic, and renal cancer.
[0024] The invention also features methods of combination therapy,
such as a method for treating cancer, wherein the above methods
further include providing radiation therapy or chemotherapy. The
chemotherapy or radiation therapy may be administered before,
concurrently, or after the administration of a disclosed compound
according to the needs of the patient.
[0025] In a further aspect the present invention provides methods
of inhibiting MDM2 expression in a mammal, comprising administering
an amount of a compound effective to inhibit said expression, said
compound having the Formula I-III or a pharmaceutically acceptable
salt or in vivo cleavable prodrug thereof.
[0026] The invention also relates to pharmaceutical compositions
comprising an effective amount of an agent selected from compounds
of Formulas I-III or a pharmaceutically acceptable prodrug,
pharmaceutically active metabolite, or pharmaceutically acceptable
salt thereof.
[0027] Additional advantages and novel features of this invention
shall be set forth in part in the description that follows, and in
part will become apparent to those skilled in the art upon
examination of the following specification or may be learned by the
practice of the invention. The advantages of the invention may be
realized and attained by means of the instrumentalities,
combinations, compositions, and methods particularly pointed out in
the appended claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0028] The accompanying drawings, which are incorporated herein and
form a part of the specification, illustrate non-limiting
embodiments of the present invention, and together with the
description, serve to explain the principles of the invention.
[0029] FIG. 1 shows a reaction scheme for the synthesis of compound
4.
[0030] FIG. 2 shows a reaction scheme for the synthesis of compound
8.
[0031] FIG. 3 shows a reaction scheme for the synthesis of
compounds 14 and 15.
[0032] FIG. 4 shows a reaction scheme for the synthesis of compound
19.
DETAILED DESCRIPTION OF THE INVENTION
[0033] The invention features novel boronic chalcone compounds
having Formulas I-III, pharmaceutical compositions thereof, and
methods of using such compounds and compositions. The inventive
compounds of the Formulas I-III are useful, for example, for
treating a tumor or cancer in a patient. Such compounds have
particular utility as therapeutic agents for diseases that can be
treated by the inhibition of MDM2 expression.
[0034] The term "tumor" as used herein refers to abnormal growth in
tissue which occurs when cellular proliferation is more rapid than
normal tissue and continues to grow after the stimuli that
initiated the new growth cease. Tumors generally exhibit partial or
complete lack of structural organization and functional
coordination with the normal tissue, and usually form a distinct
mass of tissue which may be benign (benign tumor) or malignant
(carcinoma). Tumors tend to be highly vascularized.
[0035] The term "cancer" is used as a general term herein to
describe malignant tumors or carcinoma. These malignant tumors may
invade surrounding tissues, may metastasize to several sites and
are likely to recur after attempted removal and to cause death of
the patient unless adequately treated. As used herein, the terms
carcinoma and cancer are subsumed under the term tumor.
[0036] In general, one embodiment of the invention relates to novel
boronic chalcone compounds of the general Formula I:
##STR00004##
where
[0037] Ar is aryl or heteroaryl, each of which may be substituted
or unsubstituted;
[0038] W is H, Z.sub.n-F, Z.sub.n-Cl, Z.sub.n-Br, Z.sub.n-I,
Z.sub.n-CF.sub.3, Z.sub.n-NO.sub.2, Z.sub.n-OR.sup.1,
Z.sub.n-NR.sup.1R.sup.2, Z.sub.n-COOR.sup.1, Z.sub.n-SR.sup.1,
Z.sub.n-(C.dbd.O)R.sup.1, Z.sub.n-O(C.dbd.O)R.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O)R.sup.1, Z.sub.n-(C.dbd.O)NR.sup.1, alkyl,
allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein
said alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or
Z.sub.n-heteroaryl may be substituted or unsubstituted;
[0039] X is Z.sub.n, Z.sub.n-O, Z.sub.n-S, Z.sub.n-NR.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O), Z.sub.n-C.dbd.O, Z.sub.n-OC(.dbd.O), or
Z.sub.n-C(.dbd.O)O;
[0040] R.sup.1 and R.sup.2 are independently H, an amine protecting
group, an alcohol protecting group, an acid protecting group, a
sulfur protecting group, alkyl, allyl, alkenyl, alkynyl,
heteroalkyl, heteroallyl, heteroalkenyl, heteroalkynyl, alkoxy,
heteroalkoxy, Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl,
Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein said alkyl, allyl,
alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl may be
substituted or unsubstituted, or R.sup.1 together with R.sup.2 and
N forms a saturated or partially unsaturated heterocycle ring
having 1 or more heteroatoms in said ring, wherein said heterocycle
may be substituted or unsubstituted and wherein said heterocycle
may be fused to an aromatic ring;
[0041] Z is an alkylene having at least 1 carbon, or an alkenylene
or alkynylene each having at least 2 carbons, wherein said
alkylene, alkenylene, or alkynylene may be substituted or
unsubstituted; and
[0042] n is zero or any integer.
[0043] The term "aryl" means a monovalent aromatic hydrocarbon
monocyclic radical of 6 to 10 ring atoms or a polycyclic aromatic
hydrocarbon, optionally substituted independently with one or more
substituents described herein. Examples of aryl groups include, but
are not limited to, phenyl, 1-naphthyl, 2-naphthyl, and derivatives
thereof.
[0044] The term "heteroaryl" means a monovalent monocyclic aromatic
radical of 5 to 10 ring atoms or a polycyclic aromatic radical,
containing one or more ring heteroatoms selected from N, O, or S,
the remaining ring atoms being C. The aromatic radical is
optionally substituted independently with one or more substituents
described herein. Examples Include, but are not limited to, furyl,
thienyl, pyrrolyl, pyrazolyl, pyrimidinyl, imidazolyl, indolyl,
quinolyl, benzopyranyl, thiazolyl, oxazolyl, isoxazolyl,
thiophenyl, 1,3,4-triazolyl, tetrazolyl, pyridinyl, pyrimidinyl,
pyridazinyl, indolyl, and derivatives thereof.
[0045] Alkyl groups include aliphatic (i.e., hydrocarbyl or
hydrocarbon radical structures containing hydrogen and carbon
atoms) with a free valence. Alkyl groups are understood to include
straight chain and branched structures. The alkyl group can be
substituted with one or more substituents which are independently
selected from the substituents described herein. Examples of alkyl
groups include, but are not limited to, methyl, ethyl, n-propyl,
isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl,
isopentyl, tert-pentyl, hexyl, isohexyl, and the like.
[0046] Alkenyl groups are analogous to alkyl groups, but have at
least one double bond (two adjacent sp.sup.2 carbon atoms).
Depending on the placement of a double bond and substituents, if
any, the geometry of the double bond may be entgegen (E), or
zusammen (Z), cis, or trans. Similarly, alkynyl groups are
analogous to alkyl groups, but have at least one triple bond (two
adjacent sp carbon atoms). Unsaturated alkenyl or alkynyl groups
may have one or more double or triple bonds, respectively, or a
mixture thereof; like alkyl groups, unsaturated groups may be
straight chain or branched, and they may be substituted as
described both above for alkyl groups and throughout the disclosure
by example. The alkenyl or alkynyl groups can be substituted with
one or more substituents which are independently selected from the
substituents described herein.
[0047] The term "alkylene" means a linear or branched saturated
divalent hydrocarbon radical of one to twelve carbon atoms, e.g.,
methylene, ethylene, propylene, 2-methylpropylene, pentylene, and
the like.
[0048] The term "alkenylene" refers to a linear or branched
divalent hydrocarbon radical of two to twelve carbons containing at
least one double bond, wherein the alkenylene radical may be
optionally substituted independently with one or more substituents
described herein. Examples include, but are not limited to,
ethenylene, propenylene, and the like.
[0049] The term "alkynylene" to a linear or branched divalent
hydrocarbon radical of two to twelve carbons containing at least
one triple bond, wherein the alkynylene radical may be optionally
substituted independently with one or more substituents described
herein.
[0050] The term "allyl" refers to a radical having the formula
R.sup.1C.dbd.CHCHR.sup.2, wherein R.sup.1 and R.sup.2 are
independently alkyl, alkenyl, alkynyl, cycloalkyl,
heterocycloalkyl, aryl, heteroaryl, or any substituent as defined
herein, wherein the allyl may be optionally substituted
independently with one or more substituents described herein.
[0051] The term "cycloalkyl" refers to saturated or partially
unsaturated cyclic hydrocarbon radical having from three to twelve
carbon atoms, wherein the cycloalkyl may be optionally substituted
independently with one or more substituents described herein. The
term "cycloalkyl" further includes bicyclic and tricyclic
cycloalkyl structures, wherein the bicyclic and tricyclic
structures may include a saturated or partially unsaturated
cycloalkyl fused to a saturated or partially unsaturated cycloalkyl
or heterocycloalkyl ring or an aryl or heteroaryl ring. Examples of
cycloalkyl groups include, but are not limited to, cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like.
[0052] The term "heteroalkyl" refers to saturated linear or
branched-chain monovalent hydrocarbon radical of one to twelve
carbon atoms, wherein at least one of the carbon atoms is replaced
with a heteroatom selected from N, O, or S, and wherein the radical
may be a carbon radical or heteroatom radical (i.e., the heteroatom
may appear in the middle or at the end of the radical). The
heteroalkyl radical may be optionally substituted independently
with one or more substituents described herein. The term
"heteroalkyl" encompasses alkoxy and heteroalkoxy radicals.
[0053] The term "heterocycloalkyl" refers to a saturated or
partially unsaturated cyclic radical of 3 to 8 ring atoms in which
at least one ring atom is a heteroatom selected from nitrogen,
oxygen and sulfur, the remaining ring atoms being C where one or
more ring atoms may be optionally substituted independently with
one or more substituent described below. The radical may be a
carbon radical or heteroatom radical. "Heterocycloalkyl" also
includes radicals where heterocycle radicals are fused with
aromatic or heteroaromatic rings. Examples of heterocycloalkyl
rings include, but are not limited to, piperidyl, quinolyl,
isothiazolyl, piperidinyl, morpholinyl, piperazinyl,
tetrahydrofuryl, tetrahydropyrrolyl, pyrrolidinyl,
octahydroindolyl, octahydrobenzothiofuranyl, and
octahydrobenzofuranyl.
[0054] The term "heteroalkenyl" refers to linear or branched-chain
monovalent hydrocarbon radical of two to twelve carbon atoms,
containing at least one double bond, e.g., ethenyl, propenyl, and
the like, wherein at least one of the carbon atoms is replaced with
a heteroatom selected from N, O, or S, and wherein the radical may
be a carbon radical or heteroatom radical (i.e., the heteroatom may
appear in the middle or at the end of the radical). The
heteroalkenyl radical may be optionally substituted independently
with one or more substituents described herein, and includes
radicals having "cis" and "trans" orientations, or alternatively,
"E" and "Z" orientations.
[0055] The term "heteroalkynyl" refers to a linear or branched
monovalent hydrocarbon radical of two to twelve carbon atoms
containing at least one triple bond. Examples include, but are not
limited to, ethynyl, propynyl, and the like, wherein at least one
of the carbon atoms is replaced with a heteroatom selected from N,
O, or S, and wherein the radical may be a carbon radical or
heteroatom radical (i.e., the heteroatom may appear in the middle
or at the end of the radical). The heteroalkynyl radical may be
optionally substituted independently with one or more substituents
described herein.
[0056] The term "heteroallyl" refers to radicals having the formula
RC.dbd.CHCHR.sup.1, wherein R and R.sup.1 are independently alkyl,
alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl,
or any substituent as defined herein, wherein at least one of the
carbon atoms is replaced with a heteroatom selected from N, O, or
S, and wherein the radical may be a carbon radical or heteroatom
radical (i.e., the heteroatom may appear in the middle or at the
end of the radical). The heteroallyl may be optionally substituted
independently with one or more substituents described herein.
[0057] As used herein, the term "electron withdrawing moiety" is
known in the art, and refers to a group which has a greater
electron withdrawing effect than hydrogen. A variety of
electron-withdrawing groups are known, and include halogens (e.g.,
fluoro, chloro, bromo, and iodo groups), NO.sub.2, NH.sub.2, CN,
SO.sub.2, SO.sub.2Ar, COOH, OAr, COOR.sup.1, OR.sup.1, COR.sup.1,
SH, SR.sup.1, OH, CF.sub.3, Ar, alkenyl, alkynyl or allyl, wherein
said Ar, alkenyl, alkynyl and allyl may be optionally unsubstituted
or substituted with an electron withdrawing moiety.
[0058] The term "amino protecting group" refers to those organic
groups intended to protect nitrogen atoms against undesirable
reactions during synthetic procedures and include, but are not
limited to, benzyl, benzyloxycarbonyl (CBZ), tert-butoxycarbonyl
(Boc), trifluoroacetyl, and the like.
[0059] The term "alcohol protecting group" refers to those organic
groups intended to protect alcohol groups or substituents against
undesirable reactions during synthetic procedures and include, but
are not limited to, (trimethylsilyl)ethoxymethyl (SEM), tert-butyl,
methoxymethyl (MOM), and the like.
[0060] The term "sulfur protecting groups" refers to those organic
groups intended to protect sulfur groups or substituents against
undesirable reactions during synthetic procedures and include, but
are not limited to, benzyl, (trimethylsilyl)ethoxymethyl (SEM),
tert-butyl, trityl and the like.
[0061] The term "acid protecting groups" refers to those organic
groups intended to protect acid groups or substituents against
undesirable reactions during synthetic procedures and include, but
are not limited to, benzyl, (trimethylsilyl)ethoxymethyl (SEM),
methylethyl and tert-butyl esters, and the like.
[0062] In general, the various moieties or functional groups of the
compounds of Formulas I-III may be optionally substituted by one or
more substituents. Examples of substituents suitable for purposes
of this invention include, but are not limited to, F, CI, Br, I,
alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-OR.sup.1,
Z.sub.n-NO.sub.2) Z.sub.n-CN, Z.sub.n--CO.sub.2R.sup.1,
Z.sub.n-(C.dbd.O)R.sup.1, Z.sub.n-O(C.dbd.O)R.sup.1,
Z.sub.n-O-alkyl, Z.sub.n-OAr, Z.sub.n-SH, Z.sub.n-SR.sup.1,
Z.sub.n-SOR.sup.1, Z.sub.n--SO.sub.2R.sup.1, Z.sub.n-S--Ar,
Z.sub.n-SOAr, Z.sub.n-SO.sub.2Ar, Z.sub.n-Ar, Z.sub.n-heteroaryl,
Z.sub.n-(C.dbd.O)NR.sup.1R.sup.2, Z.sub.n-NR.sup.1R.sup.2,
Z.sub.n-NR.sup.1(C.dbd.O)R.sup.1, Z.sub.n-SO.sub.2NR.sup.1R.sup.2,
PO.sub.3H.sub.2, SO.sub.3H.sub.2, amine protecting groups, alcohol
protecting groups, sulfur protecting groups, or acid protecting
groups, where:
[0063] Z is alkylene having from 1 to 4 carbons, or alkenylene or
alkynylene each having from 2 to 4 carbons, wherein said alkylene,
alkenylene, or alkynylene may be substituted or unsubstituted;
[0064] n is zero or any integer,
[0065] R.sup.1 and R.sup.2 are alkyl, allyl, alkenyl, alkynyl,
heteroalkyl, heteroallyl, heteroalkenyl, heteroalkynyl, alkoxy,
heteroalkoxy, Z.sub.n-cycloalkyl, or Z.sub.n-heterocycloalkyl, Ar
or heteroaryl, wherein said alkyl, allyl, alkenyl, alkynyl,
heteroalkyl, heteroallyl, heteroalkenyl, heteroalkynyl, alkoxy,
heteroalkoxy, Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Ar, or
heteroaryl may be substituted or unsubstituted.
[0066] The compounds of this invention may possess one or more
asymmetric centers; such compounds can therefore be produced as
individual (R)- or (S)-stereoisomers or as mixtures thereof. Unless
indicated otherwise, the description or naming of a particular
compound in the specification and claims is intended to include
both individual enantiomers and mixtures, racemic or otherwise,
thereof. Accordingly, this invention also includes racemates and
resolved enantiomers, and diastereomers compounds of the Formulas
I-III. The methods for the determination of stereochemistry and the
separation of stereoisomers are well known in the art (see
discussion in Chapter 4 of "Advanced Organic Chemistry", 4th
edition J. March, John Wiley and Sons, New York, 1992).
[0067] Certain of the compounds of this invention, in
pharmaceutical dosage form, may be used as a method of treating a
cancer or as a prophylactic agent for preventing a disease or
condition from manifesting itself. Accordingly, this invention
further includes compositions including, but not limited to,
solvates, pharmaceutically acceptable prodrugs, pharmaceutically
active metabolites, and pharmaceutically acceptable salts of
compounds of Formulas I-III. The term "solvate" refers to an
aggregate of a molecule with one or more solvent molecules.
[0068] In certain pharmaceutical dosage forms, the pro-drug form of
the compounds according to the present invention may be preferred.
A "prodrug" is a compound that may be converted under physiological
conditions or by solvolysis to the specified compound or to a
pharmaceutically acceptable salt of such compound.
[0069] A "pharmaceutically active metabolite" is a
pharmacologically active product produced through metabolism in the
body of a specified compound or salt thereof. Metabolites of a
compound may be identified using routine techniques known in the
art and their activities determined using tests such as those
described herein.
[0070] Prodrugs and active metabolites of a compound may be
identified using routine techniques known in the art. Various forms
of prodrugs are known in the art. For examples of such prodrug
derivatives, see, for example, a) Design of Prodrugs, edited by H.
Bundgaard, (Elsevier, 1985) and Methods in Enzymology, Vol. 42, p.
309-396, edited by K. Widder, et al. (Academic Press, 1985); b) A
Textbook of Drug Design and Development, edited by
Krogsgaard-Larsen and H. Bundgaard, Chapter 5 "Design and
Application of Prodrug" (H. Bundgaard p. 113-191 (1991)); c) H.
Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992); d) H.
Bundgaard, et al., Journal of Pharmaceutical Sciences, 77:285
(1988); and e) N. Kakeya, et al., Chem. Pharm. Bull., 32: 692
(1984).
[0071] The compounds of this invention, including prodrug forms of
these agents, can be provided in the form of pharmaceutically
acceptable salts. As used herein, the term pharmaceutically
acceptable salts or complexes refers to appropriate salts or
complexes of the active compounds according to the present
invention which retain the desired biological activity of the
parent compound and exhibit limited toxicological effects to normal
cells. Nonlimiting examples of such salts are acid addition salts
formed with inorganic acids (for example, hydrochloric acid,
hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and
the like), and salts formed with organic acids such as acetic acid,
oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic
acid, benzoic acid, tannic acid, pamoic acid, alginic acid, and
polyglutamic acid, among others
[0072] The inventive compounds may be prepared using the reaction
routes and synthesis schemes as described below, employing the
techniques available in the art using starting materials that are
readily available. For example, in one embodiment boronic chalcone
compound (4), which is based upon a chemical structure I:
##STR00005##
can be prepared according to the reaction scheme shown in FIG. 1.
In FIG. 1, aldehyde (1) and ketone (2) undergo a Claisen-Schmidt
aldol condensation upon treatment with potassium hydroxide in
methanol according to standard methods to provide compound (3).
Treatment of compound (3) with pinacol (bromoalkyl)boronate in the
presence of sodium hydride in THF followed by deprotection under
alkaline conditions provides the desired compound (4).
[0073] In another embodiment, this invention relates to compounds
of the general Formula II:
##STR00006##
where Ar, W, X and n are as defined above and R.sub.3 is an
electron-withdrawing moiety. Boronic chalcone compound (8), which
is based upon structure II can prepared according to the general
reaction scheme shown in FIG. 2. In FIG. 2, ketone (5) and aldehyde
(6) undergo a Claisen-Schmidt aldol condensation upon treatment
with potassium hydroxide in methanol according to standard methods
to provide compound (7). Treatment of compound (7) with pinacol
(bromoalkyl)boronate in the presence of sodium hydride in THF,
followed by deprotection under alkaline conditions provides the
desired compound (8).
[0074] FIG. 3 shows another general reaction scheme for the
syntheses of compounds (14) and (15), which are also based upon a
chemical structure II. According to FIG. 3, aldehyde (9) and ketone
(10) undergo a Claisen-Schmidt aldol condensation upon treatment
with potassium hydroxide in methanol according to standard methods
to provide compound (II). Compound (II) is converted to the acid
chloride (12) by treating with thionyl chloride. Treatment of the
add chloride (12) with pinacol
bis(trimethylsilyl)aminoalkylboronate (13) provides compound (14).
Pinacol bis(trimethylsilyl)aminoalkylboronate (13) can be easily
prepared as shown in FIG. 3. Alternatively, treatment of compound
(12) with an alkylamineboronic acid provides compound (15).
[0075] In another embodiment, this invention relates to compounds
of the general Formula III:
##STR00007##
where Ar is as defined above and R.sup.4 is H, an amine protecting
group, Z.sub.n-OR.sup.1, Z.sub.n-SR.sup.1, Z.sub.n--R.sup.1,
Z.sub.n-NR.sup.1(C.dbd.O)R.sup.1, Z.sub.n-C.dbd.OR.sup.1,
Z.sub.n-OC(.dbd.O)R.sup.1, Z.sub.n-C(.dbd.O)OR.sup.1, alkyl, allyl,
alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, Z.sub.n-cycloalkyl,
Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or Z.sub.n-heteroaryl, wherein
said alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,
heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
Z.sub.n-cycloalkyl, Z.sub.n-heterocycloalkyl, Z.sub.n-Ar or
Z.sub.n-heteroaryl may be substituted or unsubstituted, and where
Z.sub.n, R.sup.1 and n are as defined above.
[0076] FIG. 4 shows a general reaction scheme for the synthesis of
chalcone compound (19) based upon structure III. For example,
reacting 1-aryl-2-propynon (16) with lithium chloride in acetic
acid at room temperature provides 3-chloro-1-arylpropenone (17).
Treatment of compound (17) with an arylaminoboronic acid (18) in
the presence of triethylamine provides compound (19).
[0077] It is important to note that the methods of synthesizing the
disclosed compounds are general examples, and one of ordinary skill
may readily determine or provide alternative syntheses for
producing compounds according to the present invention without
engaging in undue experimentation. Using the general and specific
synthetic methodologies described herein, a number of the chemical
compounds as set forth in Table 1 were synthesized.
[0078] The compounds of the present invention are useful for
treating or preventing benign and malignant tumors, including
various cancers such as, cervical, anal and oral cancers, stomach,
colon, bladder, rectal, liver, pancreatic, lung, breast, cervix
uteri, corpus uteri, ovary, prostate, testis, renal, brain/cns
(e.g., gliomas), head and neck, eye or ocular, throat, skin
melanoma, acute lymphocytic leukemia, acute myelogenous leukemia,
Ewing's Sarcoma, Kaposi's Sarcoma, basal cell carcinoma and
squamous cell carcinoma, small cell lung cancer, mouth/pharynx,
esophageal, larynx, kidney and lymphoma, among others. In addition,
conditions such as neurofibromatosis, tuberous sclerosis (each of
which conditions produces benign tumors of the skin), hemangiomas
and lymphangiogenesis, among others, may be treated effectively
with compounds according to the present invention.
[0079] Methods of treating tumors and/or cancer according to the
present invention comprise administering to a patient in need
thereof an effective amount of one or more compounds according to
the present invention or a pharmaceutically acceptable salt or in
vivo cleavable prodrug thereof.
[0080] The invention also features methods of combination therapy,
such as a method for treating cancer, wherein the above methods
further include providing radiation therapy or chemotherapy, for
example, with mitotic inhibitors such as a taxane or a vinca
alkaloid. Examples of mitotic inhibitors include paclitaxel,
docetaxel, vincristine, vinblastine, vinorelbine, and vinflunine.
Other therapeutic combinations include a MEK inhibitor of the
invention and an anticancer agent such as cisplatin, 5-fluorouracil
or 5-fluoro-2-4(1H,3H)-pyrimidinedione (5FU), flutamide, and
gemcitabine. The chemotherapy or radiation therapy may be
administered before, concurrently, or after the administration of a
disclosed compound according to the needs of the patient.
[0081] Numerous biological assays have been used and are accepted
by those skilled in the art to assess the anti-tumor and
anti-cancer activity of compounds according to the present
invention. Any of these methods can be used to evaluate the
activity of the compounds disclosed herein. One common method of
assessing activity is through the use of test panels of cancer cell
lines. These tests evaluate the in vitro anti-cancer activity of
particular compounds in cancer cell lines, and provide predictive
data with respect to the use of tested compounds in vivo. Other
assays include in vivo evaluations of the compound's effect on
human or in an appropriate animal model, for example, using mouse
tumor cells implanted into or grafted onto mice or in other
appropriate animal models.
[0082] In the case of testing the anti-cancer activity of compounds
according to the present invention, an assay based on human breast
cancer MDA-MB-231 (estrogen receptor negative) and wtMCF7 (estrogen
receptor positive) cells may be employed as described in Example 2.
In this assay, cells are seeded onto a 96-well plate and treated
with a compound according to the present invention at a known
concentration. The cell numbers are counted and compared against
controls. Percent inhibition is readily determined from the data
obtained. Other methods known in the art may also be used without
undue experimentation to assay the anti-cancer activity of the
disclosed compounds.
[0083] Therapeutically effective amounts of the compounds of the
invention may also be used to treat diseases mediated by expression
of MDM2. An "effective amount" is intended to mean that amount of
compound that, when administered to a mammal in need of such
treatment, is sufficient to inhibit or attenuate expression of
MDM2. Thus, for example, a therapeutically effective amount of a
compound selected from Formulas I-III, or a salt, active metabolite
or prodrug thereof, is a quantity sufficient to modulate, regulate,
or inhibit expression of MDM2.
[0084] The amount of a given agent that will correspond to such an
amount will vary depending upon factors such as the particular
compound, disease condition and its severity, the identity (e.g.,
weight) of the mammal in need of treatment, but can nevertheless be
routinely determined by one skilled in the art. "Treating" is
intended to mean at least the mitigation of a disease condition in
a mammal, such as a human, and includes, but is not limited to,
preventing the disease condition from occurring in a mammal,
particularly when the mammal is found to be predisposed to having
the disease condition but has not yet been diagnosed as having it,
modulating and/or inhibiting the disease condition; and/or
alleviating the disease condition.
[0085] In order to use a compound of the Formula I-III, or a
pharmaceutically acceptable salt or in vivo cleavable prodrug
thereof, for the therapeutic treatment (including prophylactic
treatment) of mammals including humans, it is normally formulated
in accordance with standard pharmaceutical practice as a
pharmaceutical composition. According to this aspect of the
invention there is provided a pharmaceutical composition that
comprises a compound of the Formula I-III, or a pharmaceutically
acceptable salt or in vivo cleavable prodrug thereof, as defined
hereinbefore in association with a pharmaceutically acceptable
diluent or carrier.
[0086] The compounds of this invention may be incorporated into
formulations for all routes of administration including for
example, oral, topical and parenteral including intravenous,
intramuscular, eye or ocular, intraperitoneal, intrabuccal,
transdermal and in suppository form.
[0087] In the pharmaceutical aspect according to the present
invention, the compound according to the present invention is
formulated preferably in admixture with a pharmaceutically
acceptable carrier, excipient or additive. In general, it is
preferable to administer the pharmaceutical composition in orally
administrable form, but for treatment of a number of conditions, a
number of other formulations may be administered via a topical,
parenteral, intravenous, intramuscular, transdermal, buccal,
subcutaneous, suppository or other route, including an eye or
ocular route. Intravenous and intramuscular formulations are
preferably administered in sterile saline. Of course, one of
ordinary skill in the art may modify the formulations within the
teachings of the specification to provide numerous formulations for
a particular route of administration without rendering the
compositions of the present invention unstable or compromising
their therapeutic activity. In particular, the modification of the
present compounds to render them more soluble in water or other
vehicle, for example, may be easily accomplished by minor
modifications (salt formulation, esterification, etc.) which are
well within the ordinary skill in the art. It is also well within
the skill of those skilled in the art to modify the route of
administration and dosage regimen of a particular compound in order
to manage the pharmacokinetics of the present compounds for maximum
beneficial effect to the patient.
[0088] The compositions of the invention may be in a form suitable
for oral use (for example as tablets, lozenges, hard or soft
capsules, aqueous or oily suspensions, emulsions, dispersible
powders or granules, syrups or elixirs), for topical use (for
example as creams, ointments, gels, or aqueous or oily solutions or
suspensions), for administration by inhalation (for example as a
finely divided powder or a liquid aerosol), for administration by
insufflation (for example as a finely divided powder) or for
parenteral administration (for example as a sterile aqueous or oily
solution for intravenous, subcutaneous, or intramuscular dosing or
as a suppository for rectal dosing). For example, compositions
intended for oral use may contain, for example, one or more
coloring, sweetening, flavoring and/or preservative agents.
[0089] Suitable pharmaceutically-acceptable excipients for a tablet
formulation include, for example, inert diluents such as lactose,
sodium carbonate, calcium phosphate or calcium carbonate,
granulating and disintegrating agents such as corn starch or
algenic acid; binding agents such as starch; lubricating agents
such as magnesium stearate, stearic acid or talc; preservative
agents such as ethyl or propyl p-hydroxybenzoate, and
anti-oxidants, such as ascorbic acid. Tablet formulations may be
uncoated or coated either to modify their disintegration and the
subsequent absorption of the active ingredient within the
gastrointestinal tract, or to improve their stability and/or
appearance, in either case, using conventional coating agents and
procedures well known in the art.
[0090] Compositions for oral use may be in the form of hard gelatin
capsules in which the active ingredient is mixed with an inert
solid diluent, for example, calcium carbonate, calcium phosphate or
kaolin, or as soft gelatin capsules in which the active ingredient
is mixed with water or an oil such as peanut oil, liquid paraffin,
or olive oil.
[0091] Aqueous suspensions generally contain the active ingredient
in finely powdered form together with one or more suspending
agents, such as sodium carboxymethylcellulose, methylcellulose,
hydroxypropylmethylcellulose, sodium alginate,
polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or
wetting agents such as lecithin or condensation products of an
alkylene oxide with fatty acids (for example polyoxethylene
stearate), or condensation products of ethylene oxide with long
chain aliphatic alcohols, for example heptadecaethyleneoxycetanol,
or condensation products of ethylene oxide with partial esters
derived from fatty acids and a hexitol such as polyoxyethylene
sorbitol monooleate, or condensation products of ethylene oxide
with partial esters derived from fatty acids and hexitol
anhydrides, for example polyethylene sorbitan monooleate. The
aqueous suspensions may also contain one or more preservatives
(such as ethyl or propyl p-hydroxybenzoate, anti-oxidants (such as
ascorbic acid), coloring agents, flavoring agents, and/or
sweetening agents (such as sucrose, saccharine, or aspartame).
[0092] Oily suspensions may be formulated by suspending the active
ingredient in a vegetable oil (such as arachis oil, olive oil,
sesame oil or coconut oil) or in a mineral oil (such as liquid
paraffin). The oily suspensions may also contain a thickening agent
such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents
such as those set out above, and flavoring agents may be added to
provide a palatable oral preparation. These compositions may be
preserved by the addition of an anti-oxidant such as ascorbic
acid.
[0093] Dispersible powders and granules suitable for preparation of
an aqueous suspension by the addition of water generally contain
the active ingredient together with a dispersing or wetting agent,
suspending agent and one or more preservatives. Suitable dispersing
or wetting agents and suspending agents are exemplified by those
already mentioned above. Additional excipients such as sweetening,
flavoring and coloring agents, may also be present.
[0094] The pharmaceutical compositions of the invention may also be
in the form of oil-in-water emulsions. The oily phase may be a
vegetable oil, such as olive oil or arachis oil, or a mineral oil,
such as for example liquid paraffin or a mixture of any of these.
Suitable emulsifying agents may be, for example,
naturally-occurring gums such as gum acacia or gum tragacanth,
naturally-occurring phosphatides such as soya bean, lecithin, an
esters or partial esters derived from fatty acids and hexitol
anhydrides (for example sorbitan monooleate) and condensation
products of the said partial esters with ethylene oxide such as
polyoxyethylene sorbitan monooleate. The emulsions may also contain
sweetening, flavoring and preservative agents.
[0095] Syrups and elixirs may be formulated with sweetening agents
such as glycerol, propylene glycol, sorbitol, aspartame or sucrose,
and may also contain a demulcent, preservative, flavoring and/or
coloring agent.
[0096] The pharmaceutical compositions may also be in the form of a
sterile injectable aqueous or oily suspension, which may be
formulated according to known procedures using one or more of the
appropriate dispersing or wetting agents and suspending agents,
which have been mentioned above, A sterile injectable preparation
may also be a sterile injectable solution or suspension in a
non-toxic parenterally-acceptable diluent or solvent, for example a
solution in 1,3-butanediol.
[0097] Suppository formulations may be prepared by mixing the
active ingredient with a suitable non-irritating excipient which is
solid at ordinary temperatures but liquid at the rectal temperature
and will therefore melt in the rectum to release the drug. Suitable
excipients include, for example, cocoa butter and polyethylene
glycols.
[0098] Topical formulations, such as creams, ointments, gels and
aqueous or oily solutions or suspensions, may generally be obtained
by formulating an active ingredient with a conventional, topically
acceptable, vehicle or diluent using conventional procedures well
known in the art.
[0099] Compositions for administration by insufflation may be in
the form of a finely divided powder containing particles of average
diameter of, for example, 30 urn or much less, the powder itself
comprising either active ingredient alone or diluted with one or
more physiologically acceptable carriers such as lactose. The
powder for insufflation is then conveniently retained in a capsule
containing, for example, 1 to 50 mg of active ingredient for use
with a turbo-inhaler device, such as is used for insufflation of
the known agent sodium cromoglycate.
[0100] Compositions for administration by inhalation may be in the
form of a conventional pressurized aerosol arranged to dispense the
active ingredient either as an aerosol containing finely divided
solid or liquid droplets. Conventional aerosol propellants such as
volatile fluorinated hydrocarbons or hydrocarbons may be used and
the aerosol device is conveniently arranged to dispense a metered
quantity of active ingredient.
[0101] For further information on formulations, see Chapter 25.2 in
Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch;
Chairman of Editorial Board), Pergamon Press 1990, which is
specifically incorporated herein by reference.
[0102] The amount of a compound of this invention that is combined
with one or more excipients to produce a single dosage form will
necessarily vary depending upon the host treated and the particular
route of administration. For example, a formulation intended for
oral administration to humans will may contain, for example, from
0.5 mg to 2 g of active agent compounded with an appropriate and
convenient amount of excipients which may vary from about 5 to
about 98 percent by weight of the total composition. Dosage unit
forms will generally contain about 1 mg to about 500 mg of an
active ingredient. For further information on routes of
administration and dosage regimes, see Chapter 25.3 in Volume 5 of
Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of
Editorial Board), Pergamon Press 1990, which is specifically
incorporated herein by reference.
[0103] The size of the dose for therapeutic or prophylactic
purposes of a compound of Formula I-III will naturally vary
according to the nature and severity of the conditions, the age and
sex of the animal or patient and the route of administration,
according to well known principles of medicine.
[0104] In one aspect of this invention, the compounds of this
invention or pharmaceutical salts or prodrugs thereof may be
formulated into pharmaceutical compositions for administration to
animals or humans to treat or prevent solid tumors or cancer.
EXAMPLES
[0105] In order to illustrate the invention, the following examples
are included. However, it is to be understood that these examples
do not limit the invention and are only meant to suggest a method
of practicing the invention. Persons skilled in the art will
recognize that the chemical reactions described may be readily
adapted to prepare a number of other boronic chalcones of the
invention, and alternative methods for preparing the compounds of
this invention are deemed to be within the scope of this invention.
For example, the synthesis of non-exemplified compounds according
to the invention may be successfully performed by modifications
apparent to those skilled in the art, e.g., by appropriately
protecting interfering groups, by utilizing other suitable reagents
known in the art other than those described, and/or by making
routine modifications of reaction conditions. Alternatively, other
reactions disclosed herein or known in the art will be recognized
as having applicability for preparing other compounds of the
invention.
[0106] In the examples below, unless otherwise indicated all
temperatures are set forth in degrees Celsius. Reagents were
purchased from commercial suppliers such as Aldrich Chemical Co.,
Lancaster, TCI or Maybridge, and used without further purification
unless otherwise indicated. The reactions set forth below were done
generally under a positive pressure of nitrogen or argon or with a
drying tube (unless otherwise stated) in anhydrous solvents. The
reaction flasks were typically fitted with rubber septa for the
introduction of substrates and reagents via syringe. Glassware was
oven dried and/or heat dried.
Example 1
General Procedure for the Preparation of Boronic Chalcone
Analogs
[0107] To a solution of an acetophenone in methanol (10 mL/mmol)
was added an aldehyde (1.5 equiv) followed by an aqueous solution
of KOH (50%, 1 mL/mmol of acetophenone). The mixture was heated at
70.degree. C. for 4-6 hours and monitored by TLC. Water (20 mL) was
then added, the methanol was evaporated, and the solution was
extracted with CH.sub.2CI.sub.2. The organic layer was dried and
evaporated to dryness, then purified by recrystallization with
methanol-water to afford the desired chalcone.
Example 2
Compound 3a
[0108] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. (ppm): 7.53 (d,
J=8.4 Hz, 2H), 7.71 (d, J=16 Hz, 1H), 7.79 (d, J=16 Hz, 1H), 7.82
(m, 4H), 8.05 (d, J=8 Hz, 2H); m.p. 268-270.degree. C.; mass
spectrum (EI mode): m/z=377 [M].sup.+. (.sup.10B), 378 [M].sup.+.
(.sup.11B); Yield 86%.
Example 3
Compound 3b
[0109] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. (ppm): 7.59 (d,
J=8 Hz, 1H), 7.69-7.73 (m, 2H), 7.77 (d, J=8 Hz, 2H), 7.83 (d,
J=15.6 Hz, 1H), 7.98 (d, J=2 Hz, 1H), 8.07 (d, J=8 Hz, 2H); m.p.
267-268.degree. C.; mass spectrum (EI mode): m/z=319 [M].sup.+.
(.sup.35CI.sup.35CI.sup.10B), 320 [M].sup.+.
(.sup.35CI.sup.35CI.sup.11B), 321 [M].sup.+.
(.sup.35CI.sup.37CI.sup.10B), 322 [M].sup.+.
(.sup.35CI.sup.37CI.sup.11B), 323 [M].sup.+.
(.sup.37CI.sup.37CI.sup.10B), 324 [M].sup.+.
(.sup.37CI.sup.37CI.sup.11B); Yield 80%.
Example 4
Compound 3c
[0110] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. (ppm): 7.14 (d,
J=8.4 Hz, 1H), 7.66 (d, J=8 Hz, 2H), 7.68 (d, J=15.2 Hz, 1H), 7.72
(d, J=15.2 Hz, 1H), 7.77 (d, J=8.4 Hz, 1H), 7.86 (d, J=2.4 Hz, 1H),
8.05 (d, J=8 Hz, 2H); m.p. 242-243.degree. C.; mass spectrum (EI
mode): m/z=303 [M].sup.+. (.sup.35CI.sup.10B), 304 [M].sup.+.
(.sup.35CI.sup.11B), 305 [M].sup.+. (.sup.37CI.sup.10B), 306
[M].sup.+. (.sup.37CI.sup.11B); Yield 90%.
Example 5
Compound 3d
[0111] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. (ppm): 7.15 (dd,
J=8.4 Hz, 8.8 Hz, 1H), 7.61 (m, 2H), 7.51 (m, 1H), 7.63 (d, J=15.6
Hz, 1H), 7.73 (d, J=15.6 Hz, 1H), 7.77 (m, 1H), 8.05 (d, J=7.8 Hz,
2H); m.p. 264-255.degree. C.; mass spectrum (EI mode): m/z=287
[M].sup.+. (.sup.10B), 288 [M].sup.+. (.sup.11B); Yield 85%.
Example 6
Compound 3e
[0112] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. (ppm): 7.12 (d,
J=8.4 Hz, 1H), 7.66 (d, J=16 Hz, 1H), 7.71 (d, J=16 Hz, 1H), 7.73
(m, 1H), 7.77 (d, J=8 Hz, 2H), 8.01 (d, J=2 Hz, 1H), 8.05 (d, J=8
Hz, 2H); m.p. 163-164.degree. C.; mass spectrum (EI mode): m/z=347
[M].sup.+. (.sup.79Br.sup.10B), 348 [M].sup.+. (.sup.79Br.sup.11B),
349 [M].sup.+. (.sup.81Br.sup.10B), 350 [M].sup.+.
(.sup.81Br.sup.11B); Yield 89%.
Example 7
Compound 3f
[0113] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. (ppm): 4.81 (s,
2H), 7.04 (d, J=8.8 Hz, 2H), 7.62 (d, J=15.6 Hz, 1H), 7.66 (d, J=8
Hz, 2H), 7.8 (d, J=8.4 Hz, 2H), 7.97 (d, J=15.6 Hz, 1H), 8.13 (d,
J=8.8 Hz, 2H); m.p. 281-282.degree. C.; mass spectrum (EI mode):
m/z=408 [M].sup.+. Yield 90%.
Example 8
Compound 3g
[0114] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. (ppm): 4.91 (s,
2H), 7.07 (d, J=9.2 Hz, 2H), 7,58 (d, J=8.8 Hz, 1H), 7.67 (m, 2H),
7.82 (d, J=15.6 Hz, 1H), 7.95 (d, J=2 Hz, 1H), 8.12 (d, J=9.2 Hz,
2H); m.p. 186-187.degree. C.; mass spectrum (EI mode): m/z=350
[M].sup.+. (.sup.35CI.sup.35CI), 352 [M].sup.+.
(.sup.35CI.sup.37CI), 354 [M].sup.+. 37CI.sup.37CI); Yield 95%.
Example 9
Compound 6
[0115] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. (ppm): 6.93 (d,
J=8 Hz, 2H), 7.36 (d, J=8.4 Hz, 2H), 7.52 (d, J=15.6 Hz, 1H), 7.71
(d, J=15.6 Hz, 1H), 7.76 (d, J=8 Hz, 2H), 8.0 (d, J=8.4 Hz, 2H);
m.p. 215-216.degree. C.; mass spectrum (EI mode): m/z=350
[M].sup.+. Yield 98
Example 10
Compound 7
[0116] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. (ppm): 3.41 (s,
2H), 7.01 (d, J=8.8 Hz, 2H), 7.12 (d, J=8.8 Hz, 2H), 7.5 (d, J=8.4
Hz, 2H), 7.65 (d, J=15.6 Hz, 1H), 7.80 (d, J=15.6 Hz, 1H), 8.05 (d,
J=8.4 Hz, 2H); mass spectrum (EI mode): m/z=407 [M].sup.+.
(.sup.10B), 408[M].sup.+. (.sup.11B); Yield 45%.
Example 11
Compound 8
[0117] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. (ppm): 7.67-7.741
(m, 6H), 7.82 (d, J=15.6 Hz, 1H), 7.99 (dd, J=8.2 Hz, 1.6 Hz, 1H),
8.19 (d, J=1.6 Hz, 1H); m.p. 283-285.degree. C.; mass spectrum (EI
mode): m/z-319 [M].sup.+. (.sup.35CI.sup.35CI.sup.10B), 320
[M].sup.+. (.sup.35CI.sup.35CI.sup.11B), 321 [M].sup.+.
(.sup.35CI.sup.37CI.sup.10B), 322 [M].sup.+.
(.sup.35CI.sup.37CI.sup.11B), 323 [M].sup.+.
(.sup.37CI.sup.37CI.sup.10B), 324 [M].sup.+.
(.sup.37CI.sup.37CI.sup.11B); Yield 68%.
Example 12
Compound 9
[0118] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. (ppm): 4.79 (s,
2H), 7.02 (d, J=8.8 Hz, 2H), 7.76 (d, J=15.6 Hz, 1H), 7.83-7.91 (m,
4H), 8.11 (dd, J=8 Hz, 2 Hz, 1H), 8.4 (d, J=2 Hz, 1H); m.p.
209-210.degree. C.; mass spectrum (EI mode): m/z=350 [M].sup.+.
(.sup.35CI.sup.35CI), 352 [M].sup.+. (.sup.35CI.sup.37CI), 354
[M].sup.+. (.sup.37CI.sup.37CI); Yield 78%.
Example 13
Compound 11
[0119] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. (ppm); 6.31 (dd,
J=2.4 Hz, 3.2 Hz, 1H), 7.15 (d, J=2.4 Hz, 1H), 7.25 (d, J=3.2 Hz,
1H), 7.55 (d, J=15.6 Hz, 1H), 7.66 (m, 2H), 7.7 (d, J=15.6 Hz, 1H),
7.79 (m, 2H); m.p. 229-231.degree. C.; mass spectrum (EI mode):
m/z=240 [M].sup.+. (.sup.10B), 241[M].sup.+. (.sup.11B); Yield
60%.
TABLE-US-00001 TABLE 2 Elemental analytical data of the chalcone
analogs Molecular Calculated Found Compound formula C H N C H N 3a
C.sub.15H.sub.12BIO.sub.3 47.67 3.20 -- 47.72 3.22 -- 3b
C.sub.15H.sub.11BCI.sub.2O.sub.3 56.13 3.45 -- 56.23 3.52 -- 3c
C.sub.15H.sub.11BCIFO.sub.3 59.16 3.64 59.40 3.71 3d
C.sub.15H.sub.11BF.sub.2O.sub.3 62.54 3.85 -- 62.42 3.92 -- 3e
C.sub.15H.sub.11BBrFO.sub.3 51.63 3.18 -- 51.52 3.24 -- 3f
C.sub.17H.sub.13IO.sub.4 50.02 3.21 -- 50.16 3.29 -- 3g
C.sub.17H.sub.12CI.sub.2O.sub.4 58.14 3.44 -- 57.98 3.50 -- 6
C.sub.15H.sub.11IO.sub.2 51.45 3.17 -- 51.50 3.22 -- 7
C.sub.16H.sub.14BIO.sub.4 47.10 3.46 -- 47.22 3.51 -- 8
C.sub.15H.sub.11BCI.sub.2O.sub.3 56.13 3.45 -- 56.28 3.50 -- 9
C.sub.17H.sub.12CI.sub.2O.sub.4 58.14 3.44 -- 58.22 3.42 -- 10
C.sub.13H.sub.11NO 79.16 5.62 7.10 79.34 5.68 7.12 11
C.sub.13H.sub.12BNO.sub.3 64.77 5.02 5.81 64.86 5.10 5.89
Example 12
Cytotoxicity of Boronic Chalcone Analogs
[0120] The cytotoxicity of the boronic chalcone analogs and
carboxylic acid chalcones were evaluated and compared by MTT
survival assays of human breast cancer MDA-MB-435, MDA-MB-231,
Wt-MCF7t and MCF-10A cells treated at different concentrations of
the analogs.
[0121] Human breast cancer MDA-MB-231 (estrogen independent) and
wtMCF7 (estrogen dependent) human breast cancer cells were
maintained in DMEM medium supplemented with 5% fetal bovine serum,
2 mM glutamine, and 100 units/mL penicillin/streptomycin.
MDA-MB-435 (estrogen independent) human breast cancer cells were
maintained in IMEM medium supplemented with 5% fetal bovine serum
and 2 mM glutamine. Each cell line used has unique growth
characteristics spanning the spectrum of tumor invasiveness,
differentiation and estrogen dependence. Normal breast epithelial
cell lines, MCF-10A and MCF-12A, were maintained in 5% and 10%
horse serum in DMEM:Ham's F12 media, respectively, supplemented
with 2 mM glutamine, 100 units/mL penicillin/streptomycin, 0.02
.mu.g/mL EGF, 0.01 mg/mL insulin, and 0.1 .mu.g/mL cholera
toxin.
[0122] Cells were incubated at 37.degree. C. in a 5% CO.sub.2
atmosphere. The MTT colorimetric assay was used to determine growth
inhibition (Mosmann, T., J. Immunol. Methods, 1983, 65, 55-63).
Cells were plated in 96-well plates and allowed to attach for 24
hours. Chalcone derivatives based on compounds 20 and 21 below were
dissolved in DMSO at 10 mM concentrations. Cells were expose in
quadruplicate well to chalcone concentrations of 0.5-100
.quadrature.M for 96 hours. After 96 hours the media was aspirated,
and 100 .quadrature.L of 1 mg/mL MTT solution (Sigma Chemical Co.)
diluted in serum free media was added to each well. After 4 hours
of incubation, the MTT solution was removed and 200 .quadrature.L
of 1:1 (v/v) solution of DMSO: ethanol was added to each well to
dissolve formazan crystals. The absorbance at A.sub.540 nm was
determined on a plate reader. IC.sub.50 values were determined from
log plots of percent of control vs. concentration. Each compound
was assayed twice in quadruplicate. Analogs 3g and 9 have been
previously described (Stoll R., et al., supra; Kussie, P. H., et
al., supra) and were included for comparative purposes.
TABLE-US-00002 Compound A B C ##STR00008## 3a I H B(OH).sub.2 3b CI
CI B(OH).sub.2 3c F CI B(OH).sub.2 3d F F B(OH).sub.2 3e F Br
B(OH).sub.2 3f I H OCH.sub.2COOH 3g CI CI OCH.sub.2COOH 6 I H OH 7
I H OCH.sub.2B(OH).sub.2 ##STR00009## 8 CI CI B(OH).sub.2 9 CI CI
OCH.sub.2COOH
[0123] IC.sub.50 values were used to determine growth inhibition in
the presence of chalcone derivatives. Of particular interest are
compounds able to differentially inhibit growth such that human
breast cancer cell lines are inhibited but normal breast epithelial
cells are significantly less inhibited. Compound 3a, 3d, 3e, 7, 8,
10 and 11 are 5-10 fold more toxic to human breast cancer cell
lines compared to normal breast epithelial cell lines (Table 1). In
the presence of these compounds, cell growth in the human breast
cancer cell lines MDA-MB-435, MDA-MB-231, and wt-MCF7 is inhibited,
indicated by the range of low IC.sub.50 values from 3.5 to 23. Cell
growth in the normal breast epithelial cell lines MCF-1QA and
MCF-12A is less inhibited, shown by higher IC50 values ranging from
11 to 75.
TABLE-US-00003 TABLE 1 Chalcones inhibit growth of human breast
cell lines.sup.a MDA- MCF- MB-435 MDA-MB-231 Wt-MCF7 MCF-10A 12A
Compound IC.sub.50 IC.sub.50 IC.sub.50 IC.sub.50 IC.sub.50 3a 10
8.8 7.0 75 63 3b 3.5 9.5 5.0 18 11 3c 16 8.5 6.0 25 22 3d 8.8 8.8
7.8 18 39 3e 8.8 9.5 8.5 17 38 3f 18 44 9 44 38 3g 9 9 13 13 15 6
4.5 8 7 15 30 7 18 11 9.5 38 100 8 4 8 5.5 18 15 9 13 18 15 12 28
10 15 15 9 63 38 11 15 23 19 38 60 .sup.aIC.sub.50 values expressed
in .mu.M; see biology section for details of the MTT assay.
[0124] The foregoing description is considered as illustrative only
of the principles of the invention. Further, since numerous
modifications and changes will be readily apparent to those skilled
in the art, it is not desired to limit the invention to the exact
construction and process shown as described above. Accordingly, all
suitable modifications and equivalents may be resorted to falling
within the scope of the invention as defined by the claims that
follow.
[0125] The words "comprise," "comprising," "include," "including,"
and "includes" when used in this specification and in the following
claims are intended to specify the presence of stated features,
integers, components, or steps, but they do not preclude the
presence or addition of one or more other features, integers,
components, steps, or groups thereof.
* * * * *