U.S. patent application number 15/157075 was filed with the patent office on 2016-12-22 for halogenated compounds for cancer imaging and treatment and methods for their use.
The applicant listed for this patent is British Columbia Cancer Agency Branch, THE UNIVERSITY OF BRITISH COLUMBIA. Invention is credited to Raymond John Andersen, Carmen Adriana Banuelos, Javier GARCIA FERNANDEZ, Yusuke IMAMURA, Jian KUNZHONG, Nasrin R. MAWJI, Marianne Dorothy SADAR, Amy (Hsing Chen) Tien, Jun WANG.
Application Number | 20160367707 15/157075 |
Document ID | / |
Family ID | 52627643 |
Filed Date | 2016-12-22 |
United States Patent
Application |
20160367707 |
Kind Code |
A1 |
Andersen; Raymond John ; et
al. |
December 22, 2016 |
HALOGENATED COMPOUNDS FOR CANCER IMAGING AND TREATMENT AND METHODS
FOR THEIR USE
Abstract
Compounds having a structure of Formula I: ##STR00001## or a
pharmaceutically acceptable salt, tautomer or stereoisomer thereof,
wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, X.sup.1,
X.sup.2, X.sup.3 and X.sup.4 are are as defined herein, and wherein
the compound comprises at least one F, Cl, Br, I or .sup.123I
moiety, are provided. Uses of such compounds for imaging
diagnostics in cancer and therapeutics methods for treatment of
subjects in need thereof, including prostate cancer as well as
methods and intermediates for preparing such compounds are also
provided.
Inventors: |
Andersen; Raymond John;
(Vancouver, CA) ; Banuelos; Carmen Adriana;
(Richmond, CA) ; GARCIA FERNANDEZ; Javier;
(Colunga, ES) ; IMAMURA; Yusuke; (Chiba City,
JP) ; KUNZHONG; Jian; (Vancouver, CA) ; MAWJI;
Nasrin R.; (Burnaby, CA) ; SADAR; Marianne
Dorothy; (West Vancouver, CA) ; WANG; Jun;
(Surrey, CA) ; Tien; Amy (Hsing Chen); (Vancouver,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
THE UNIVERSITY OF BRITISH COLUMBIA
British Columbia Cancer Agency Branch |
VANCOUVER
Vancouver |
|
CA
CA |
|
|
Family ID: |
52627643 |
Appl. No.: |
15/157075 |
Filed: |
May 17, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
14481727 |
Sep 9, 2014 |
9375496 |
|
|
15157075 |
|
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61875556 |
Sep 9, 2013 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 17/14 20180101;
A61K 45/06 20130101; A61P 5/28 20180101; A61K 31/225 20130101; C07C
43/23 20130101; A61K 31/275 20130101; A61K 31/225 20130101; A61P
35/00 20180101; A61K 31/4166 20130101; A61P 43/00 20180101; C07B
59/001 20130101; C07C 69/63 20130101; A61P 17/10 20180101; A61P
27/02 20180101; C07B 2200/05 20130101; A61K 31/275 20130101; A61K
51/04 20130101; A61P 15/08 20180101; A61K 31/167 20130101; A61P
21/00 20180101; A61K 31/09 20130101; A61K 31/4166 20130101; A61K
31/167 20130101; A61P 13/08 20180101; A61K 31/09 20130101; A61K
2300/00 20130101; C07C 57/58 20130101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 51/04 20060101
A61K051/04; A61K 31/09 20060101 A61K031/09; A61K 45/06 20060101
A61K045/06; C07B 59/00 20060101 C07B059/00; C07C 43/23 20060101
C07C043/23; C07C 57/58 20060101 C07C057/58 |
Goverment Interests
STATEMENT OF GOVERNMENT INTEREST
[0002] This invention was made in part with government support
under Grant No. 2R01 CA105304 awarded by the National Cancer
Institute. The United States Government has certain rights in this
invention.
Claims
1. A compound having a structure of Formula I: ##STR00068## or a
pharmaceutically acceptable salt, tautomer or stereoisomer thereof,
wherein: R.sup.1 and R.sup.2 are each independently_H or
C.sub.1-C.sub.10 alkyl, or R.sup.1 and R.sup.2, together with the
carbon atom to which they are bound, are taken together to form a
carbocyclic or heterocyclic ring; R.sup.3, R.sup.4 and R.sup.5 are
each independently H, C.sub.1-C.sub.10 alkyl or C.sub.1-C.sub.10
alkylcarbonyl; and X.sup.1, X.sup.2, X.sup.3 and X.sup.4 are each
independently H, F, Cl, Br, I or .sup.123I, wherein at least one of
X.sup.1, X.sup.2, X.sup.3 or X.sup.4 is F, Cl, Br, I or
.sup.123I.
2. The compound of claim 1, wherein the compound has one of the
following structures (Ia), (Ib), (Id), (Ie), (Ig) or (Ih):
##STR00069## ##STR00070##
3. (canceled)
4. The compound of claim 1, wherein three of x.sup.1, X.sup.2,
X.sup.3 and X.sup.4 are H, and the remaining X.sup.1, X.sup.2,
X.sup.3 or X.sup.4 is F, Cl, Br, I or .sup.123I.
5. The compound of claim 1, wherein X.sup.1 is .sup.123I.
6. The compound of claim 1, wherein X.sup.3 is .sup.123I.
7.-11. (canceled)
12. The compound of claim 1, wherein R.sup.1 and R.sup.2 are each H
or methyl.
13.-15. (canceled)
16. The compound of any one of claims 1-15, wherein at least one of
R.sup.3, R.sup.4 or R.sup.5 is H.
17. (canceled)
18. The compound of claim 1, wherein R.sup.3, R.sup.4 and R.sup.5
are each H.
19. The compound of claim 1, wherein at least one of R.sup.3,
R.sup.4 or R.sup.5 is C.sub.1-C.sub.10 alkyl.
20.-22. (canceled)
23. The compound of claim 19, wherein C.sub.1-C.sub.10 alkyl is
methyl, isopropyl, n-butyl or propargyl.
24.-25. (canceled)
26. The compound of claim 1, wherein at least one of R.sup.3,
R.sup.4 or R.sup.5 is C.sub.1-C.sub.10 alkylcarbonyl.
27. (canceled)
28. The compound of claim 1, wherein R.sup.3, R.sup.4 and R.sup.5
are each C.sub.1-C.sub.10 alkylcarbonyl.
29. The compound of claim 26, wherein C.sub.1-C.sub.10
alkylcarbonyl is methyl carbonyl.
30. The compound of claim 1, wherein the compound has one of the
following structures: ##STR00071## ##STR00072## ##STR00073##
##STR00074## ##STR00075## ##STR00076## ##STR00077## ##STR00078##
##STR00079## ##STR00080## ##STR00081## ##STR00082## ##STR00083##
##STR00084## or a pharmaceutically acceptable salt or stereoisomer
thereof.
31. A pharmaceutical composition comprising a compound of claim 1,
and a pharmaceutically acceptable carrier.
32. A method of imaging cancer, the method comprising administering
a compound of claim 1 to a subject and detecting the presence or
absence of cancer by use of SPECT or PET.
33. The method of claim 32, wherein the method identifies the
presence or absence of a tumor.
34. The method of claim 32, wherein the method identifies the
location of a tumor.
35. The method of claim 32, wherein the cancer is prostate
cancer.
36. The method of claim 35, wherein the prostate cancer is
castration resistant prostate cancer or androgen-dependent prostate
cancer.
37. (canceled)
38. The method of claim 32, wherein the method detects the presence
of splice variants, mutants and/or species which comprise the AR
NTD.
39. A method of imaging prostate, the method comprising
administering a compound of claim 1 to a subject and detecting the
prostate by use of SPECT or PET
40.-44. (canceled)
45. A method for modulating androgen receptor (AR) activity, the
method comprising administering to a mammalian cell a compound of
claim 1.
46. (canceled)
47. The method of claim 45, wherein modulating androgen receptor
(AR) activity is for treatment of at least one indication selected
from the group consisting of: prostate cancer, breast cancer,
ovarian cancer, endometrial cancer, salivary gland carcinoma, hair
loss, acne, hirsutism, ovarian cysts, polycystic ovary disease,
precocious puberty, spinal and bulbar muscular atrophy, and age
related macular degeneration.
48. The method of claim 47, wherein the indication is prostate
cancer.
49.-51. (canceled)
52. A method of modulating androgen receptor (AR) activity, the
method comprising administering a compound of claim 1 to a subject
in need thereof.
53. The method of claim 52, wherein modulating androgen receptor
(AR) activity is for the treatment of one or more of the following:
prostate cancer, breast cancer, ovarian cancer, endometrial cancer,
salivary gland carcinoma, hair loss, acne, hirsutism, ovarian
cysts, polycystic ovary disease, precocious puberty, spinal and
bulbar muscular atrophy, and age related macular degeneration.
54. (canceled)
55. A pharmaceutical composition comprising a compound of claim 1,
an additional therapeutic agent and a pharmaceutically acceptable
carrier.
56. (canceled)
57. The pharmaceutical composition of claim 55, wherein the
additional therapeutic agent is enzalutamide, galeterone, ODM-201,
ARN-509, abiraterone, bicalutamide, nilutamide, flutamide,
cyproterone acetate, docetaxel, Bevacizumab (Avastin), OSU-HDAC42,
VITAXIN, sunitumib, ZD-4054, Cabazitaxel (XRP-6258), MDX-010
(Ipilimumab), OGX 427, OGX 011, finasteride, dutasteride,
turosteride, bexlosteride, izonsteride, FCE 28260, SKF 105,111 or
related compounds thereof.
58. The method of claim 52, wherein the compound is selected from:
##STR00085## ##STR00086## ##STR00087## or a pharmaceutically
acceptable salt or stereoisomer thereof.
Description
PRIOIRTY INFORMATION
[0001] This application claims the benefit of U.S. Provisional
Application No. 61/875,556, filed on Sep. 9, 2013, and which is
incorporated herein be reference in its entirety.
BACKGROUND
Technical Field
[0003] This invention generally relates to radiolabeled compounds
and their use in methods for imaging the prostate gland. For
example, in certain embodiments the compounds are useful for
imaging benign prostate diseases such as benign prostate
hyperplasia. In other embodiments, the compounds are useful for
imaging cancerous prostate diseases, such as prostate cancer
tumors. In certain embodiments the invention relates to radioactive
.sup.123I compounds and their use as an imaging tool in prostate
cancer and benign prostate diseases. The disclosed compounds find
utility in any number of imaging applications, including imaging of
androgen receptor (AR) splice variants in prostate cancers,
including all stages and androgen dependent, androgen-sensitive and
castration-resistant prostate cancers (also referred to as hormone
refractory, androgen-independent, androgen deprivation resistant,
androgen ablation resistant, androgen depletion-independent,
castration-recurrent, anti-androgen-recurrent).
Description of the Related Art
[0004] Androgens mediate their effects through the androgen
receptor (AR). Androgens play a role in a wide range of
developmental and physiological responses and are involved in male
sexual differentiation, maintenance of spermatogenesis, and male
gonadotropin regulation (R. K. Ross, G. A. Coetzee, C. L. Pearce,
J. K. Reichardt, P. Bretsky, L. N. Kolonel, B. E. Henderson, E.
Lander, D. Altshuler & G. Daley, Eur Urol 35, 355-361 (1999);
A. A. Thomson, Reproduction 121, 187-195 (2001); N. Tanji, K. Aoki
& M. Yokoyama, Arch Androl 47, 1-7 (2001)). Several lines of
evidence show that androgens are associated with the development of
prostate carcinogenesis. Firstly, androgens induce prostatic
carcinogenesis in rodent models (R. L. Noble, Cancer Res 37,
1929-1933 (1977); R. L. Noble, Oncology 34, 138-141 (1977)) and men
receiving androgens in the form of anabolic steroids have a higher
incidence of prostate cancer (J. T. Roberts & D. M. Essenhigh,
Lancet 2, 742 (1986); J. A. Jackson, J. Waxman & A. M.
Spiekerman, Arch Intern Med 149, 2365-2366 (1989); P. D. Guinan, W.
Sadoughi, H. Alsheik, R. J. Ablin, D. Alrenga & I. M. Bush, Am
J Surg 131, 599-600 (1976)). Secondly, prostate cancer does not
develop if humans or dogs are castrated before puberty (J. D.
Wilson & C. Roehrborn, J Clin Endocrinol Metab 84, 4324-4331
(1999); G. Wilding, Cancer Sury 14, 113-130 (1992)). Castration of
adult males causes involution of the prostate and apoptosis of
prostatic epithelium while eliciting no effect on other male
external genitalia (E. M. Bruckheimer & N. Kyprianou, Cell
Tissue Res 301, 153-162 (2000); J. T. Isaacs, Prostate 5, 545-557
(1984)). This dependency on androgens provides the underlying
rationale for treating prostate cancer with chemical or surgical
castration (androgen ablation).
[0005] Androgens also play a role in female diseases such as
polycystic ovary syndrome as well as cancers. One example is
ovarian cancer where elevated levels of androgens are associated
with an increased risk of developing ovarian cancer (K. J.
Helzlsouer, A. J. Alberg, G. B. Gordon, C. Longcope, T. L. Bush, S.
C. Hoffman & G. W. Comstock, JAMA 274, 1926-1930 (1995); R. J.
Edmondson, J. M. Monaghan & B. R. Davies, Br J Cancer 86,
879-885 (2002)). The AR has been detected in a majority of ovarian
cancers (H. A. Risch, J Natl Cancer Inst 90, 1774-1786 (1998); B.
R. Rao & B. J. Slotman, Endocr Rev 12, 14-26 (1991); G. M.
Clinton & W. Hua, Crit Rev Oncol Hematol 25, 1-9 (1997)),
whereas estrogen receptor-alpha (ERa) and the progesterone receptor
are detected in less than 50% of ovarian tumors.
[0006] The only effective treatment available for advanced prostate
cancer is the withdrawal of androgens which are essential for the
survival of prostate epithelial cells. Androgen ablation therapy
causes a temporary reduction in tumor burden concomitant with a
decrease in serum prostate-specific antigen (PSA). Unfortunately
prostate cancer can eventually grow again in the absence of
testicular androgens (castration-resistant disease) (Huber et al
1987 Scand J Urol Nephrol. 104, 33-39). Castration-resistant
prostate cancer is biochemically characterized before the onset of
symptoms by a rising titre of serum PSA (Miller et al 1992 J. Urol.
147, 956-961). Once the disease becomes castration-resistant most
patients succumb to their disease within two years.
[0007] The AR has distinct functional domains that include the
carboxy-terminal ligand-binding domain (LBD), a DNA-binding domain
(DBD) comprising two zinc finger motifs, and an N-terminus domain
(NTD) that contains one or more transcriptional activation domains.
Binding of androgen (ligand) to the LBD of the AR results in its
activation such that the receptor can effectively bind to its
specific DNA consensus site, termed the androgen response element
(ARE), on the promoter and enhancer regions of "normally" androgen
regulated genes, such as PSA, to initiate transcription. The AR can
be activated in the absence of androgen by stimulation of the
cAMP-dependent protein kinase (PKA) pathway, with interleukin-6
(IL-6) and by various growth factors (Culig et al 1994 Cancer Res.
54, 5474-5478; Nazareth et al 1996 J. Biol. Chem. 271, 19900-19907;
Sadar 1999 J. Biol. Chem. 274, 7777-7783; Ueda et al 2002 A J.
Biol. Chem. 277, 7076-7085; and Ueda et al 2002 B J. Biol. Chem.
277, 38087-38094). The mechanism of ligand-independent
transformation of the AR has been shown to involve: 1) increased
nuclear AR protein suggesting nuclear translocation; 2) increased
AR/ARE complex formation; and 3) the AR-NTD (Sadar 1999 J. Biol.
Chem. 274, 7777-7783; Ueda et al 2002 A J. Biol. Chem. 277,
7076-7085; and Ueda et al 2002 B J. Biol. Chem. 277, 38087-38094).
The AR may be activated in the absence of testicular androgens by
alternative signal transduction pathways in castration-resistant
disease, which is consistent with the finding that nuclear AR
protein is present in secondary prostate cancer tumors (Kim et al
2002 Am. J. Pathol. 160, 219-226; and van der Kwast et al 1991
Inter. J. Cancer 48, 189-193).
[0008] Available inhibitors of the AR include nonsteroidal
antiandrogens such as bicalutamide (Casodex.TM.), nilutamide,
flutamide, enzulutamide and investigational drug ARN-509 and
steroidal antiandrogens, such as cyproterone acetate. These
antiandrogens target the LBD of the AR and predominantly fail
presumably due to poor affinity and mutations that lead to
activation of the AR by these same antiandrogens (Taplin, M.
E.,
[0009] Bubley, G. J., Kom Y. J., Small E. J., Uptonm M.,
Rajeshkumarm B., Balkm S. P., Cancer Res., 59, 2511-2515 (1999)).
These antiandrogens would also have no effect on the recently
discovered AR splice variants that lack the ligand-binding domain
(LBD) to result in a constitutively active receptor which promotes
progression of castration recurrent prostate cancer (Dehm S M,
Schmidt L J, Heemers H V, Vessella R L, Tindall D J., Cancer Res
68, 5469-77, 2008; Guo Z, Yang X, Sun F, Jiang R, Linn D E, Chen H,
Chen H, Kong X, Melamed J, Tepper C G, Kung H J, Brodie A M,
Edwards J, Qiu Y., Cancer Res. 69, 2305-13, 2009; Hu et al 2009
Cancer Res. 69, 16-22; Sun et al 2010 J Clin Invest. 2010 120,
2715-30).
[0010] Conventional therapy has concentrated on androgen-dependent
activation of the AR through its C-terminal domain. Studies
developing antagonists to the AR have concentrated on the
C-terminus and specifically: 1) the allosteric pocket and AF-2
activity (Estebanez-Perpina et al 2007, PNAS 104, 16074-16079); 2)
in silico "drug repurposing" procedure for identification of
nonsteroidal antagonists (Bisson et al 2007, PNAS 104,
11927-11932); and coactivator or corepressor interactions (Chang et
al 2005, Mol Endocrinology 19, 2478-2490; Hur et al 2004, PLoS Biol
2, E274; Estebanez-Perpina et al 2005, JBC 280, 8060-8068; He et al
2004, Mol Cell 16, 425-438).
[0011] The AR-NTD is also a target for drug development (e.g. WO
2000/001813), since the NTD contains Activation-Function-1 (AF-1)
which is the essential region required for AR transcriptional
activity (Jenster et al 1991. Mol Endocrinol. 5, 1396-404). The
AR-NTD importantly plays a role in activation of the AR in the
absence of androgens (Sadar, M.D. 1999 1 Biol. Chem. 274,
7777-7783; Sadar MD et al 1999 Endocr Relat Cancer. 6, 487-502;
Ueda et al 2002 J. Biol. Chem. 277, 7076-7085; Ueda 2002 J. Biol.
Chem. 277, 38087-38094; Blaszczyk et al 2004 Clin Cancer Res. 10,
1860-9; Dehm et al 2006 J. Biol Chem. 28, 27882-93; Gregory et al
2004 J Biol Chem. 279, 7119-30). The AR-NTD is important in
hormonal progression of prostate cancer as shown by application of
decoy molecules (Quayle et al 2007, Proc Natl Acad Sci U S A.
104,1331-1336).
[0012] While the crystal structure has been resolved for the AR
C-terminus LBD, this has not been the case for the NTD due to its
high flexibility and intrinsic disorder in solution (Reid et al
2002 J. Biol. Chem. 277, 20079-20086) thereby hampering virtual
docking drug discovery approaches. Compounds that modulate AR
include the bis-phenol compounds disclosed in published PCT Nos: WO
2010/000066, WO 2011/082487; WO 2011/082488; WO 2012/145330; WO
2012/139039; WO 2012/145328; WO 2013/028572 and WO
2013/028791,which are hereby incorporated by reference in their
entireties, to the British Columbia Cancer Agency Branch and The
University of British Columbia.
[0013] In addition to compounds which modulate AR, compounds and
methods for imaging the prostate are useful research, diagnostic
and prognostic tools. Such compounds are useful in many
applications, including imaging of benign and/or malignant prostate
cells and tissue. In this regard, positron emission tomography
(PET) is an often used imaging technique for non-invasive
identification of pathological state and tumors. In PET imaging,
the distribution of a radioisotope (e.g., .sup.18F) in the body can
be determined. Thus incorporating .sup.18F into compounds which
concentrate in tumor sites (see e.g., WO 2013/028791) offers
potential for diagnosis, staging, and monitoring treatment of
cancers. However, improved methods for imaging are needed, for
example methods which employ .sup.123I and single photon emission
coupled tomography (SPECT) techniques have potential to improve
methods for imaging AR-rich tissues such as the benign prostate,
and in particular prostate cancers and AR splice variants in
castrate recurrent prostate cancers.
[0014] While significant advances have been made in this field,
there remains a need for improved imaging agents In particular,
methods and compounds suitable for imaging benign and malignant
prostate tissues and cells are needed. The present invention
fulfills these needs, and provides other related advantages.
BRIEF SUMMARY
[0015] Some embodiments of the compounds described herein may be
used for diagnostic purposes to investigate diseases of the
prostate, including cancer. In particular embodiments, the
compounds are useful for imaging diagnostics in cancer. In some
embodiments, such imaging allows for the detection and/or location
of cancer sites (e.g., tumor sites). Furthermore, these compounds
may be used individually or as part of a kit for such purposes.
[0016] The present disclosure is based in part on the surprising
discovery that the compounds described herein, may be used to
modulate AR activity either in vivo or in vitro for both research
and therapeutic uses. Accordingly, embodiments of the compounds are
useful for imaging the prostate. The imaging may be for any number
of diagnostic purposes. For example, in certain embodiments the
compounds are useful for imaging benign prostate cancer diseases.
In other embodiments, the compounds find utility for imaging of
certain cancers, including prostate cancer since certain
embodiments of the compounds localize in prostate tumor sites.
Other imaging agents are androgen mimics; however, in one
embodiment, the compounds are useful for imaging AR splice variants
or any AR species (ie., those mutated in other domains or regions).
The AR may be mammalian. For example, the AR may be human. The
prostate cancer may be castration-resistant prostate cancer. The
prostate cancer may be androgen-dependent prostate cancer.
[0017] In accordance with one embodiment, there is provided a
compound having a structure of Formula I:
##STR00002##
or a pharmaceutically acceptable salt, tautomer or stereoisomer
thereof, wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5,
X.sup.1, X.sup.2, X.sup.3 and X.sup.4 are are as defined herein,
and wherein the compound comprises at least one F, Cl, Br, I or
.sup.123I moiety, are provided.
[0018] In other embodiments pharmaceutical compositions comprising
a compound of structure (I) are provided. Methods employing such
pharmaceutical compositions for imaging cancer are also provided.
Methods for modulating AR activity employing the present compounds
and pharmaceutical compositions are also provided.
[0019] These and other aspects of the invention will be apparent
upon reference to the following detailed description. To this end,
various references are set forth herein which describe in more
detail certain background information, procedures, compounds and/or
compositions, and are each hereby incorporated by reference in
their entirety.
BRIEF DESCRIPTION OF THE DRAWINGS
[0020] In the figures, identical reference numbers identify similar
elements. The sizes and relative positions of elements in the
figures are not necessarily drawn to scale and some of these
elements are arbitrarily enlarged and positioned to improve figure
legibility. Further, the particular shapes of the elements as drawn
are not intended to convey any information regarding the actual
shape of the particular elements, and have been solely selected for
ease of recognition in the figures.
[0021] FIGS. 1A and 1B are graphs showing dose response of a
representative compound (8d) of the invention.
[0022] FIGS. 2A-2D shows specificity of a representative compound
(8d) relative to comparative compounds.
[0023] FIGS. 3A-3C show the characterization data for compound
iii-I.
[0024] FIGS. 4A-4C show the characterization data for compound
iii-Br.
[0025] FIGS. 5A and 5B show the characterization data for compound
iii-Cl.
[0026] FIGS. 6A and 6B show the characterization data for compound
iv-I.
[0027] FIGS. 7A-7C show the characterization data for compound
iv-Br.
[0028] FIGS. 8A-8C show the characterization data for compound
8d.
[0029] FIGS. 9A-9C show the characterization data for compound
9d.
[0030] FIGS. 10A-10C show the characterization data for compound
10d.
[0031] FIG. 11 shows the characterization data for compound
v-F.
[0032] FIGS. 12A and 12B show the characterization data for
compound iv-F.
[0033] FIGS. 13A-13C show the characterization data for compound
11d.
[0034] FIGS. 14A-14E show competitive ligand-binding assay of 8d
and representative ligands from recombinant ligand binding
domains.
[0035] FIG. 15 shows binding experiment of 1d.
[0036] FIG. 16 shows cell viability and proliferation assay of
8d.
DETAILED DESCRIPTION
I. Definitions
[0037] In the following description, certain specific details are
set forth in order to provide a thorough understanding of various
embodiments. However, one skilled in the art will understand that
the invention may be practiced without these details. In other
instances, well-known structures have not been shown or described
in detail to avoid unnecessarily obscuring descriptions of the
embodiments. Unless the context requires otherwise, throughout the
specification and claims which follow, the word "comprise" and
variations thereof, such as, "comprises" and "comprising" are to be
construed in an open, inclusive sense, that is, as "including, but
not limited to." Further, headings provided herein are for
convenience only and do not interpret the scope or meaning of the
claimed invention.
[0038] Reference throughout this specification to "one embodiment"
or "an embodiment" means that a particular feature, structure or
characteristic described in connection with the embodiment is
included in at least one embodiment. Thus, the appearances of the
phrases "in one embodiment" or "in an embodiment" in various places
throughout this specification are not necessarily all referring to
the same embodiment. Furthermore, the particular features,
structures, or characteristics may be combined in any suitable
manner in one or more embodiments. Also, as used in this
specification and the appended claims, the singular forms "a,"
"an," and "the" include plural referents unless the content clearly
dictates otherwise. It should also be noted that the term "or" is
generally employed in its sense including "and/or" unless the
content clearly dictates otherwise.
[0039] The terms below, as used herein, have the following
meanings, unless indicated otherwise:
[0040] "Amino" refers to the --NH.sub.2radical.
[0041] "Cyano" refers to the --CN radical.
[0042] "Halo" or "halogen" refers to bromo, chloro, fluoro or
iodo.
[0043] "Hydroxy" or "hydroxyl" refers to the --OH radical.
[0044] "Imino" refers to the .dbd.NH substituent.
[0045] "Nitro" refers to the --NO.sub.2 radical.
[0046] "Oxo" refers to the .dbd.O substituent.
[0047] "Thioxo" refers to the .dbd.S substituent.
[0048] "Alkyl" refers to a straight or branched hydrocarbon chain
radical which is saturated or unsaturated (i.e., contains one or
more double and/or triple bonds), having from one to twelve carbon
atoms, and which is attached to the rest of the molecule by a
single bond. Alkyls comprising any number of carbon atoms from 1 to
12 are included. An alkyl comprising up to 12 carbon atoms is a
C.sub.1-C.sub.12 alkyl, an alkyl comprising up to 10 carbon atoms
is a C.sub.1-C.sub.10 alkyl, an alkyl comprising up to 6 carbon
atoms is a C.sub.1-C.sub.6 alkyl and an alkyl comprising up to 5
carbon atoms is a C.sub.1-C.sub.5 alkyl. A C.sub.1-C.sub.5 alkyl
includes C.sub.5 alkyls, C.sub.4 alkyls, C.sub.3 alkyls, C.sub.2
alkyls and C.sub.1 alkyl (i.e., methyl) and includes, for example,
and without limitation, saturated C.sub.1-C.sub.5 alkyl,
C.sub.2-C.sub.5 alkenyl and C.sub.2-C.sub.5 alkynyl. Non-limiting
examples of saturated C.sub.1-C.sub.5 alkyl include methyl, ethyl,
n-propyl, i-propyl, sec-propyl, n-butyl, i-butyl, sec-butyl,
t-butyl and n-pentyl. Non-limiting examples of C.sub.2-C.sub.5
alkenyl include vinyl, allyl, isopropenyl, 1-propene-2-yl,
1-butene-1-yl, 1-butene-2-yl, 1-butene-3-yl, 2-butene-1-yl,
2-butene-2-yl, penteneyl and the like. Non-limiting examples of
C.sub.2-C.sub.5 alkynyl include ethynyl, propynyl, butynyl,
pentynyland the like. A C.sub.1-C.sub.6 alkyl includes all moieties
described above for C.sub.1-C.sub.5 alkyls but also includes
C.sub.6 alkyls. A C.sub.1-C.sub.10 alkyl includes all moieties
described above for C.sub.1-C.sub.5 alkyls and C.sub.1-C.sub.6
alkyls, but also includes C.sub.7, C.sub.8, C.sub.9 and C.sub.10
alkyls. Similarily, a C.sub.1-C.sub.12 alkyl includes all the
foregoing moieties, but also includes C.sub.11 and C.sub.12 alkyls.
Unless stated otherwise specifically in the specification, an alkyl
group may be optionally substituted. "Alkylene" or "alkylene chain"
refers to a straight or branched divalent hydrocarbon chain linking
the rest of the molecule to a radical group, consisting solely of
carbon and hydrogen, which is saturated or unsaturated (i.e.,
contains one or more double and/or triple bonds), and having from
one to twelve carbon atoms, e.g., methylene, ethylene, propylene,
n-butylene, ethenylene, propenylene, n-butenylene, propynylene,
n-butynylene, and the like. The alkylene chain is attached to the
rest of the molecule through a single or double bond and to the
radical group through a single or double bond. The points of
attachment of the alkylene chain to the rest of the molecule and to
the radical group can be through one carbon or any two carbons
within the chain. Unless stated otherwise specifically in the
specification, an alkylene chain may be optionally substituted.
[0049] "Alkoxy" refers to a radical of the formula --OR.sub.a where
R.sub.a is an alkyl radical as defined above containing one to
twelve carbon atoms. Unless stated otherwise specifically in the
specification, an alkoxy group may be optionally substituted.
[0050] "Alkylamino" refers to a radical of the formula --NHR.sub.a
or --NR.sub.aR.sub.a where each R.sub.a is, independently, an alkyl
radical as defined above containing one to twelve carbon atoms.
Unless stated otherwise specifically in the specification, an
alkylamino group may be optionally substituted.
[0051] "Alkylcarbonyl" refers to the --C(.dbd.O)Ra moiety, wherein
Ra is an alkyl radical as defined above. A non-limiting example of
an alkyl carbonyl is the methyl carbonyl ("acetal") moiety. Unless
stated otherwise specifically in the specification, an alkyl
carbonyl group may be optionally substituted.
[0052] "Aryl" refers to a hydrocarbon ring system radical
comprising hydrogen, 6 to 18 carbon atoms and at least one aromatic
ring. For purposes of this invention, the aryl radical may be a
monocyclic, bicyclic, tricyclic or tetracyclic ring system, which
may include fused or bridged ring systems. Aryl radicals include,
but are not limited to, aryl radicals derived from aceanthrylene,
acenaphthylene, acephenanthrylene, anthracene, azulene, benzene,
chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane,
indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene,
and triphenylene. Unless stated otherwise specifically in the
specification, the term "aryl" is meant to include aryl radicals
that are optionally substituted.
[0053] "Aralkyl" refers to a radical of the formula
--R.sub.b-R.sub.c where R.sub.b is an alkylene chain as defined
above and R.sub.c is one or more aryl radicals as defined above,
for example, benzyl, diphenylmethyl and the like. Unless stated
otherwise specifically in the specification, an aralkyl group may
be optionally substituted.
[0054] "Carbocyclyl" or "carbocyclic ring" refers to a rings
structure, wherein the the atoms which form the ring are each
carbon. Carbocyclic rings may comprise from 3 to 18 carbon atoms in
the ring. Carbocyclic rings include aryls and cycloalkyls as
defined herein. Unless stated otherwise specifically in the
specification, a carbocyclyl group may be optionally
substituted.
[0055] "Cycloalkyl" refers to a stable non-aromatic monocyclic or
polycyclic hydrocarbon radical consisting solely of carbon and
hydrogen atoms, which may include fused or bridged ring systems,
having from three to fifteen carbon atoms, preferably having from
three to ten carbon atoms, and which is saturated or unsaturated
and attached to the rest of the molecule by a single bond.
Monocyclic radicals include, for example, cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic
radicals include, for example, adamantyl, norbornyl, decalinyl,
7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise
stated specifically in the specification, a cycloalkyl group may be
optionally substituted.
[0056] "Cycloalkylalkyl" refers to a radical of the formula
--R.sub.bR.sub.d where R.sub.b is an alkylene chain as defined
above and R.sub.d is a cycloalkyl radical as defined above. Unless
stated otherwise specifically in the specification, a
cycloalkylalkyl group may be optionally substituted. "Haloalkyl"
refers to an alkyl radical, as defined above, that is substituted
by one or more halo radicals, as defined above, e.g.,
trifluoromethyl, difluoromethyl, trichloromethyl,
2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl,
1,2-dibromoethyl, and the like. Unless stated otherwise
specifically in the specification, a haloalkyl group may be
optionally substituted. "Heterocyclyl" or "heterocyclic ring"
refers to a stable 3- to 18-membered non-aromatic ring radical
which consists of two to twelve carbon atoms and from one to six
heteroatoms selected from the group consisting of nitrogen, oxygen
and sulfur. Heterocyclycl or heterocyclic rings include heteroaryls
as defined below. Unless stated otherwise specifically in the
specification, the heterocyclyl radical may be a monocyclic,
bicyclic, tricyclic or tetracyclic ring system, which may include
fused or bridged ring systems; and the nitrogen, carbon or sulfur
atoms in the heterocyclyl radical may be optionally oxidized; the
nitrogen atom may be optionally quaternized; and the heterocyclyl
radical may be partially or fully saturated. Examples of such
heterocyclyl radicals include, but are not limited to, dioxolanyl,
thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl,
imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,
octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,
2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl,
piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl,
quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl,
tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl,
1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated
otherwise specifically in the specification, Unless stated
otherwise specifically in the specification, a heterocyclyl group
may be optionally substituted.
[0057] "N-heterocyclyl" refers to a heterocyclyl radical as defined
above containing at least one nitrogen and where the point of
attachment of the heterocyclyl radical to the rest of the molecule
is through a nitrogen atom in the heterocyclyl radical. Unless
stated otherwise specifically in the specification, a
N-heterocyclyl group may be optionally substituted.
[0058] "Heterocyclylalkyl" refers to a radical of the formula
--R.sub.bR.sub.e where R.sub.b is an alkylene chain as defined
above and R.sub.e is a heterocyclyl radical as defined above, and
if the heterocyclyl is a nitrogen-containing heterocyclyl, the
heterocyclyl may be attached to the alkyl radical at the nitrogen
atom. Unless stated otherwise specifically in the specification, a
heterocyclylalkyl group may be optionally substituted.
[0059] "Heteroaryl" refers to a 5- to 14-membered ring system
radical comprising hydrogen atoms, one to thirteen carbon atoms,
one to six heteroatoms selected from the group consisting of
nitrogen, oxygen and sulfur, and at least one aromatic ring. For
purposes of this invention, the heteroaryl radical may be a
monocyclic, bicyclic, tricyclic or tetracyclic ring system, which
may include fused or bridged ring systems; and the nitrogen, carbon
or sulfur atoms in the heteroaryl radical may be optionally
oxidized; the nitrogen atom may be optionally quaternized. Examples
include, but are not limited to, azepinyl, acridinyl,
benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl,
benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl,
benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl,
benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl,
benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl
(benzothiophenyl), benzotriazolyl,
benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,
dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl,
isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl,
isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl,
isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl,
oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl,
1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl,
phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl,
pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl,
pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl,
isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl,
triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e. thienyl).
Unless stated otherwise specifically in the specification, a
heteroaryl group may be optionally substituted.
[0060] "N-heteroaryl" refers to a heteroaryl radical as defined
above containing at least one nitrogen and where the point of
attachment of the heteroaryl radical to the rest of the molecule is
through a nitrogen atom in the heteroaryl radical. Unless stated
otherwise specifically in the specification, an N-heteroaryl group
may be optionally substituted. "Heteroarylalkyl" refers to a
radical of the formula --R.sub.bR.sub.f where R.sub.b is an
alkylene chain as defined above and R.sub.f is a heteroaryl radical
as defined above. Unless stated otherwise specifically in the
specification, a heteroarylalkyl group may be optionally
substituted.
[0061] ".sup.123I" refers to the radioactive isotope of iodine
having atomic mass 123. The compounds of structure (I) comprise at
least one .sup.123I moiety. Throughout the present application,
where structures depict a .sup.123I moiety at a certain position it
is meant that the I moiety at this position is enriched for
.sup.123I. In other words, the compounds contain more than the
natural abundance of .sup.123I at the indicated position(s). It is
not required that the compounds comprise 100% .sup.123I at the
indicated positions, provided .sup.123I is present in more than the
natural abundance. Typically the .sup.123I isotope is enriched to
greater than 50%, greater than 60%, greater than 70%, greater than,
80% or greater than 90%, relative to .sup.127I.
[0062] "Thioalkyl" refers to a radical of the formula --SR.sub.a
where R.sub.a is an alkyl radical as defined above containing one
to twelve carbon atoms. Unless stated otherwise specifically in the
specification, a thioalkyl group may be optionally substituted.
[0063] The term "substituted" used herein means any of the above
groups (i.e., alkyl, alkylene, alkoxy, alkylamino, alkylcarbonyl,
thioalkyl, aryl, aralkyl, carbocyclyl, cycloalkyl, cycloalkylalkyl,
haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl,
heteroaryl, N-heteroaryl and/or heteroarylalkyl) wherein at least
one hydrogen atom is replaced by a bond to a non-hydrogen atoms
such as, but not limited to: a halogen atom such as F, Cl, Br, and
I; an oxygen atom in groups such as hydroxyl groups, alkoxy groups,
and ester groups; a sulfur atom in groups such as thiol groups,
thioalkyl groups, sulfone groups, sulfonyl groups, and sulfoxide
groups; a nitrogen atom in groups such as amines, amides,
alkylamines, dialkylamines, arylamines, alkylarylamines,
diarylamines, N-oxides, imides, and enamines; a silicon atom in
groups such as trialkylsilyl groups, dialkylarylsilyl groups,
alkyldiarylsilyl groups, and triarylsilyl groups; and other
heteroatoms in various other groups. "Substituted" also means any
of the above groups in which one or more hydrogen atoms are
replaced by a higher-order bond (e.g., a double- or triple-bond) to
a heteroatom such as oxygen in oxo, carbonyl, carboxyl, and ester
groups; and nitrogen in groups such as imines, oximes, hydrazones,
and nitriles. For example, "substituted" includes any of the above
groups in which one or more hydrogen atoms are replaced with
--NR.sub.gR.sub.h, --NR.sub.gC(.dbd.O)R.sub.h,
--NR.sub.gC(.dbd.O)NR.sub.gR.sub.h, --NR.sub.gC(.dbd.O)OR.sub.h,
--NR.sub.gSO.sub.2R.sub.h, --OC(.dbd.O)N R.sub.gR.sub.h,
--OR.sub.g, --SR.sub.g, --SO.sub.2R.sub.g, --SO.sub.2R.sub.g,
--OSO.sub.2R.sub.g, --SO.sub.2OR.sub.g, .dbd.NSO.sub.2R.sub.g, and
--SO.sub.2NR.sub.gR.sub.h. "Substituted" also means any of the
above groups in which one or more hydrogen atoms are replaced with
--C(.dbd.O)R.sub.g, --C(.dbd.O)OR.sub.g,
--C(.dbd.O)NR.sub.gR.sub.h, --CH.sub.2SO.sub.2R.sub.g,
--CH.sub.2SO.sub.2NR.sub.gR.sub.h. In the foregoing, R.sub.g and
R.sub.h are the same or different and independently hydrogen,
alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl,
cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl,
heterocyclylalkyl, heteroaryl, N-heteroaryl and/or heteroarylalkyl.
"Substituted" further means any of the above groups in which one or
more hydrogen atoms are replaced by a bond to an amino, cyano,
hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkoxy,
alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl,
haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl,
heteroaryl, N-heteroaryl and/or heteroarylalkyl group. In addition,
each of the foregoing substituents may also be optionally
substituted with one or more of the above substituents.
[0064] As used herein, the symbol "" (hereinafter may be referred
to as "a point of attachment bond") denotes a bond that is a point
of attachment between two chemical entities, one of which is
depicted as being attached to the point of attachment bond and the
other of which is not depicted as being attached to the point of
attachment bond. For example, "" indicates that the chemical entity
"XY" is bonded to another chemical entity via the point of
attachment bond. Furthermore, the specific point of attachment to
the non-depicted chemical entity may be specified by inference. For
example, the compound CH.sub.3--R.sup.3, wherein R.sup.3 is H or ""
infers that when R.sup.3 is "XY", the point of attachment bond is
the same bond as the bond by which R.sup.3 is depicted as being
bonded to CH.sub.3.
[0065] "Fused" refers to any ring structure described herein which
is fused to an existing ring structure in the compounds of the
invention. When the fused ring is a heterocyclyl ring or a
heteroaryl ring, any carbon atom on the existing ring structure
which becomes part of the fused heterocyclyl ring or the fused
heteroaryl ring may be replaced with a nitrogen atom.
[0066] The invention disclosed herein is also meant to encompass
the in vivo metabolic products of the disclosed compounds. Such
products may result from, for example, the oxidation, reduction,
hydrolysis, amidation, esterification, and the like of the
administered compound, primarily due to enzymatic processes.
Accordingly, the invention includes compounds produced by a process
comprising administering a compound of this invention to a mammal
for a period of time sufficient to yield a metabolic product
thereof. Such products are typically identified by administering a
radiolabelled compound of the invention in a detectable dose to an
animal, such as rat, mouse, guinea pig, monkey, or to human,
allowing sufficient time for metabolism to occur, and isolating its
conversion products from the urine, blood or other biological
samples.
[0067] "Stable compound" and "stable structure" are meant to
indicate a compound that is sufficiently robust to survive
isolation to a useful degree of purity from a reaction mixture, and
formulation into an efficacious therapeutic agent.
[0068] "Mammal" includes humans and both domestic animals such as
laboratory animals and household pets (e.g., cats, dogs, swine,
cattle, sheep, goats, horses, rabbits), and non-domestic animals
such as wildlife and the like.
[0069] "Optional" or "optionally" means that the subsequently
described event of circumstances may or may not occur, and that the
description includes instances where said event or circumstance
occurs and instances in which it does not. For example, "optionally
substituted aryl" means that the aryl radical may or may not be
substituted and that the description includes both substituted aryl
radicals and aryl radicals having no substitution.
[0070] "Pharmaceutically acceptable carrier, diluent or excipient"
includes without limitation any adjuvant, carrier, excipient,
glidant, sweetening agent, diluent, preservative, dye/colorant,
flavor enhancer, surfactant, wetting agent, dispersing agent,
suspending agent, stabilizer, isotonic agent, solvent, or
emulsifier which has been approved by the United States Food and
Drug Administration as being acceptable for use in humans or
domestic animals.
[0071] "Pharmaceutically acceptable salt" includes both acid and
base addition salts.
[0072] "Pharmaceutically acceptable acid addition salt" refers to
those salts which retain the biological effectiveness and
properties of the free bases, which are not biologically or
otherwise undesirable, and which are formed with inorganic acids
such as, but are not limited to, hydrochloric acid, hydrobromic
acid, sulfuric acid, nitric acid, phosphoric acid and the like, and
organic acids such as, but not limited to, acetic acid,
2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid,
aspartic acid, benzenesulfonic acid, benzoic acid,
4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid,
capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic
acid, citric acid, cyclamic acid, dodecylsulfuric acid,
ethane-1,2-disulfonic acid, ethanesulfonic acid,
2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric
acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic
acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid,
glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric
acid, lactic acid, lactobionic acid, lauric acid, maleic acid,
malic acid, malonic acid, mandelic acid, methanesulfonic acid,
mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic
acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid,
orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic
acid, pyroglutamic acid, pyruvic acid, salicylic acid,
4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid,
tartaric acid, thiocyanic acid, p-toluenesulfonic acid,
trifluoroacetic acid, undecylenic acid, and the like.
[0073] "Pharmaceutically acceptable base addition salt" refers to
those salts which retain the biological effectiveness and
properties of the free acids, which are not biologically or
otherwise undesirable. These salts are prepared from addition of an
inorganic base or an organic base to the free acid. Salts derived
from inorganic bases include, but are not limited to, the sodium,
potassium, lithium, ammonium, calcium, magnesium, iron, zinc,
copper, manganese, aluminum salts and the like. Preferred inorganic
salts are the ammonium, sodium, potassium, calcium, and magnesium
salts. Salts derived from organic bases include, but are not
limited to, salts of primary, secondary, and tertiary amines,
substituted amines including naturally occurring substituted
amines, cyclic amines and basic ion exchange resins, such as
ammonia, isopropylamine, trimethylamine, diethylamine,
triethylamine, tripropylamine, diethanolamine, ethanolamine,
deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol,
dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,
hydrabamine, choline, betaine, benethamine, benzathine,
ethylenediamine, glucosamine, methylglucamine, theobromine,
triethanolamine, tromethamine, purines, piperazine, piperidine,
N-ethylpiperidine, polyamine resins and the like. Particularly
preferred organic bases are isopropylamine, diethylamine,
ethanolamine, trimethylamine, dicyclohexylamine, choline and
caffeine.
[0074] Often crystallizations produce a solvate of the compound of
the invention. As used herein, the term "solvate" refers to an
aggregate that comprises one or more molecules of a compound of the
invention with one or more molecules of solvent. The solvent may be
water, in which case the solvate may be a hydrate. Alternatively,
the solvent may be an organic solvent. Thus, the compounds of the
present invention may exist as a hydrate, including a monohydrate,
dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate and
the like, as well as the corresponding solvated forms. The compound
of the invention may be true solvates, while in other cases, the
compound of the invention may merely retain adventitious water or
be a mixture of water plus some adventitious solvent.
[0075] A "pharmaceutical composition" refers to a formulation of a
compound of the invention and a medium generally accepted in the
art for the delivery of the biologically active compound to
mammals, e.g., humans. Such a medium includes all pharmaceutically
acceptable carriers, diluents or excipients therefor.
[0076] The compounds of the invention, or their pharmaceutically
acceptable salts may contain one or more asymmetric centers and may
thus give rise to enantiomers, diastereomers, and other
stereoisomeric forms that may be defined, in terms of absolute
stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino
acids. The present invention is meant to include all such possible
isomers, as well as their racemic and optically pure forms whether
or not they are specifically depicted herein. Optically active (+)
and (-), (R)- and (S)-, or (D)- and (L)- isomers may be prepared
using chiral synthons or chiral reagents, or resolved using
conventional techniques, for example, chromatography and fractional
crystallization. Conventional techniques for the
preparation/isolation of individual enantiomers include chiral
synthesis from a suitable optically pure precursor or resolution of
the racemate (or the racemate of a salt or derivative) using, for
example, chiral high pressure liquid chromatography (HPLC). When
the compounds described herein contain olefinic double bonds or
other centres of geometric asymmetry, and unless specified
otherwise, it is intended that the compounds include both E and Z
geometric isomers. Likewise, all tautomeric forms are also intended
to be included.
[0077] A "stereoisomer" refers to a compound made up of the same
atoms bonded by the same bonds but having different
three-dimensional structures, which are not interchangeable. The
present invention contemplates various stereoisomers and mixtures
thereof and includes "enantiomers", which refers to two
stereoisomers whose molecules are nonsuperimposeable mirror images
of one another.
[0078] A "tautomer" refers to a proton shift from one atom of a
molecule to another atom of the same molecule. The present
invention includes tautomers of any said compounds.
[0079] The chemical naming protocol and structure diagrams used
herein are a modified form of the I.U.P.A.C. nomenclature system,
using the ACD/Name Version 9.07 software program and/or ChemDraw
Ultra Version 11.0.1 software naming program (CambridgeSoft). For
complex chemical names employed herein, a substituent group is
named before the group to which it attaches. For example,
cyclopropylethyl comprises an ethyl backbone with cyclopropyl
substituent. Except as described below, all bonds are identified in
the chemical structure diagrams herein, except for some carbon
atoms, which are assumed to be bonded to sufficient hydrogen atoms
to complete the valency.
[0080] II. Compounds and Methods
[0081] As noted above, the presently disclosed compounds find
utility in a number of medical imaging application, including
imaging of the prostate. Many currently available imaging agents
tend to accumulate in the bladder, which decreases their
effectiveness as imaging tools specifically for the prostate. While
not wishing to be bound by theory, the present applicants believe
the disclosed compounds are unexpectedly effective for imaging of
the prostate due to their ability to accumulate in the prostate,
rather than the bladder, allowing the prostate gland to be seen.
Accordingly, the compounds may be used in methods for imaging the
prostate, for example to image benign prostate diseases. In other
embodiments, the compounds may be used in methods to image
cancerous prostate diseases, such as tumors of the prostate.
[0082] Androgen ablation therapy causes a temporary reduction in
prostate cancer tumor burden, but the malignancy will begin to grow
again in the absence of testicular androgens to form castrate
resistant prostate cancer (CRPC). A rising titer of serum
prostate-specific antigen (PSA) after androgen ablation therapy
indicates biochemical failure, the emergence of CRPC, and
re-initiation of an androgen receptor (AR) transcription program.
Most patients succumb to CRPC within two years of biochemical
failure.
[0083] AR is a transcription factor and a validated target for
prostate cancer therapy. Current therapies include androgen
ablation and administration of antiandrogens. Most CRPC is
suspected to be AR-dependent. AR has distinct functional domains
that include the C-terminus ligand-binding domain (LBD), a
DNA-binding domain (DBD), and an amino-terminal domain (NTD). AR
NTD contains the activation function-1 (AF-1) that contributes most
of the activity to the AR. Recently, splice variants of the AR that
lack the LBD have been reported in prostate cancer cell lines (VCaP
and 22Rv1), and in CRPC tissues. To date more than 20 splice
variants of AR have been detected. Splice variants V7 and V567es
are clinically relevant with levels of expression correlated to
poor survival and CRPC. AR V567es is solely expressed in 20% of
metastases. Abiraterone resistance is associated with expression of
AR splice variants. Enzalutamide also increases levels of
expression of these constitutively active AR splice variants. These
splice variants lack LBD and thereby would not be inhibited by
current therapies that target the AR LBD such as antiandrogens or
androgen ablation therapy. A single patient with advanced prostate
cancer can have many lesions throughout the body and skeleton and
each tumor can have differing levels of expression of AR.
[0084] Biopsy of metastatic tumors in a patient to determine AR
species is not widely accessible nor feasible to sample tumours in
a patient that may have multiple metastases. Thus it is essential
to develop approaches to detect the expression of all AR species
for the molecular classification of tumors based on the level and
extent of expression of AR splice variants, or other AR species
that cannot be detected using an imaging agent that interacts with
the LBD, to identify patients with potentially aggressive disease
and poor prognosis, or to identify patients that will not respond
to hormone therapies that target the AR LBD. Accordingly, certain
embodiments of the present invention provide a AR NTD-targeted
molecular imaging probe (e.g., compound of formula I) which can be
used to monitor response to therapy and provide insight into the
role of AR in resistance mechanisms.
[0085] One current approach to image AR in prostate cancer uses
positron emission tomography (PET) with
16.beta.[.sup.18F]-fluoro-5.alpha. dihydrotestosterone
(.sup.18F-FDHT) that binds to AR LBD. Unfortunately this imaging
agent cannot detect splice variants lacking LBD. In some
embodiments, the invention employs sequential imaging with
.sup.18F-FDHT to detect full-length AR and gamma radiation emitting
probes to specifically detect the AR NTD which would be the sum of
both full-length AR and variant AR. In other embodiments, the
invention employs sequential imaging with two different PET imaging
agents to detects only full-length AR and another to specifically
detect the AR NTD which would be the sum of both full-length AR and
variant AR. Together these data reveal patients with tumors that
express variant AR (NTD of variant plus full-length AR detected
with NTD isotope minus full-length AR detected with .sup.18F-FDHT).
By using sequential imaging, a discordant distribution or
discordant level of uptake between .sup.18F-FDHT and a radiolabeled
compound of this invention (i.e., compound of structure (I))
indicates the presence of overexpression of splice variants lacking
the LBD.
[0086] Accordingly, certain embodiments of the present invention
are directed to compounds that bind to the AR NTD and are useful
for imaging of tumors with splice variants using SPECT and/or
methods of modulating AR NTD activity. Other embodiments are
directed to compound and methods useful for imaging and/or treating
benign prostate conditions or diseases. In one embodiment, the
present disclosure provides a compound having a structure of
Formula I:
##STR00003##
or a pharmaceutically acceptable salt, tautomer or stereoisomer
thereof, wherein:
[0087] R.sup.1 and R.sup.2 are each independentlyH or
C.sub.1-C.sub.10 alkyl, or R.sup.1 and R.sup.2, together with the
carbon atom to which they are bound, are taken together to form a
carbocyclic or heterocyclic ring;
[0088] R.sup.3, R.sup.4 and R.sup.5 are each independently H,
C.sub.1-C.sub.10 alkyl or C.sub.1-C.sub.10 alkylcarbonyl; and
[0089] X.sup.1, X.sup.2, X.sup.3 and X.sup.4 are each independently
H, F, Cl, Br, I or .sup.123I, wherein at least one of X.sup.1,
X.sup.2, X.sup.3 or X.sup.4 is F, Cl, Br, I or .sup.123I.
[0090] In various embodiments, differerent stereoisomers of the
compound of structure (I) are provided, for example in some
embodiments the compound has one of the following structures (Ia),
(Ib), (Ic) or (Id):
##STR00004##
[0091] In still other embodiments, the compound has one of the
following structures (Ie), (If), (Ig) or (Ih):
##STR00005##
[0092] The compounds contain at least one F, Cl. Br, I or .sup.123I
substitution for use in the imaging and treatment methods described
herein. In some embodiments, the compounds comprise one F, Cl. Br,
I or .sup.123I substitution, for example in certain other
embodiments, three of X.sup.1, X.sup.2, X.sup.3 and X.sup.4 are H,
and the remaining X.sup.1, X.sup.2, X.sup.3 or X.sup.4 is F, Cl.
Br, I or .sup.123I. In some embodiments, the compounds comprise two
F, Cl. Br, I or .sup.123I substitutions (i.e., two of X.sup.1,
X.sup.2, X.sup.3 and X.sup.4 are H, and the other two of X.sup.1,
X.sup.2, X.sup.3 or X.sup.4 are F, Cl. Br, I or .sup.123I). In
other embodiments, the compounds comprise three F, Cl. Br, I or
.sup.123I substitutions (i.e., one of X.sup.1, X.sup.2, X.sup.3 and
X.sup.4 is H, and the remaining X.sup.1, X.sup.2, X.sup.3 or
X.sup.4 is F, Cl. Br, I or .sup.123I) and in other embodiments the
compounds comprise four F, Cl. Br, I or .sup.123I substitutions
(i.e., each of X.sup.1, X.sup.2, X.sup.3 and X.sup.4 are F, Cl. Br,
I or .sup.1231).
[0093] Favorable imaging and/or AR NTD modulating results are
obtained by substitution with F, Cl. Br, I or .sup.123I at any of
the "X" positions. In some of the foregoing embodiments, X.sup.1 is
.sup.123I. In other of the X.sup.3 is .sup.123.sub.I.
[0094] In various embodiments of any of the foregoing, at least one
of R.sup.1 or R.sup.2 is H. For example, in some embodiments
R.sup.1 and R.sup.2 are each H.
[0095] In other embodiments of the foregoing, at least one of
R.sup.1 or R.sup.2 is C.sub.1-C.sub.10 alkyl. For example, in some
embodiments R.sup.1 and R.sup.2 are each C.sub.1-C.sub.10 alkyl. In
some of these embodiments C.sub.1-C.sub.10 alkyl is
C.sub.1-C.sub.10 saturated alky such as methyl.
[0096] In other embodiments, Each R.sup.1 may independently be
C.sub.1-C.sub.5 alkyl. Each R.sup.1 may independently be
C.sub.1-C.sub.4 alkyl. Each R.sup.1 may independently be
C.sub.1-C.sub.3 alkyl. Each R.sup.1 may independently be
C.sub.1-C.sub.2 alkyl. Each R.sup.1 may independently be methyl.
Each R.sup.1 may independently be C.sub.2 alkyl. Each R.sup.1 may
independently be C.sub.3 alkyl. Each R.sup.1 may independently be
C.sub.4 alkyl. Each R.sup.1 may independently be C.sub.5 alkyl.
[0097] In other embodiments, Each R.sup.2 may independently be
C.sub.1-C.sub.5 alkyl. Each R.sup.2 may independently be
C.sub.1-C.sub.4 alkyl. Each R.sup.2 may independently be
C.sub.1-C.sub.3 alkyl. Each R.sup.2 may independently be
C.sub.1-C.sub.2 alkyl. Each R.sup.2 may independently be methyl.
Each R.sup.2 may independently be C.sub.2 alkyl. Each R.sup.2 may
independently be C.sub.3 alkyl. Each R.sup.2 may independently be
C.sub.4 alkyl. Each R.sup.2 may independently be C.sub.5 alkyl.
[0098] In certain of the foregoing embodiments, at least one of
R.sup.3, R.sup.4 or R.sup.5 is H. In certain embodiments, two of
R.sup.3, R.sup.4 and R.sup.5 are H. In other embodiments, R.sup.3,
R.sup.4 and R.sup.5 are each H.
[0099] In still other embodiments of the foregoing compounds of
structure (I), at least one of R.sup.3, R.sup.4 or R.sup.5 is
C.sub.1-C.sub.10 alkyl. For example, in some embodiments two of
R.sup.3, R.sup.4 and R.sup.5 are C.sub.1-C.sub.10 alkyl. In other
embodiments, R.sup.3, R.sup.4 and R.sup.5 are each C.sub.1-C.sub.10
alkyl. In certain of the foregoing embodiments, C.sub.1-C.sub.10
alkyl is saturated C.sub.1-C.sub.10 alkyl. For example, in some
embodiments the saturated C.sub.1-C.sub.10 alkyl is methyl,
isopropyl or n-butyl. In some different embodiments, the
C.sub.1-C.sub.10 alkyl is unsaturated C.sub.1-C.sub.10 alkyl, for
example propargyl.
[0100] In other embodiments, Each R.sup.3 may independently be
C.sub.1-C.sub.5 alkyl. Each R.sup.3 may independently be
C.sub.1-C.sub.4 alkyl. Each R.sup.3 may independently be
C.sub.1-C.sub.3 alkyl. Each R.sup.3 may independently be
C.sub.1-C.sub.2 alkyl. Each R.sup.3 may independently be methyl.
Each R.sup.3 may independently be C.sub.2 alkyl. Each R.sup.3 may
independently be C.sub.3 alkyl. Each R.sup.3 may independently be
C.sub.4 alkyl. Each R.sup.3 may independently be C.sub.5 alkyl.
[0101] In other embodiments, Each R.sup.4 may independently be
C.sub.1-C.sub.5 alkyl. Each R.sup.4 may independently be
C.sub.1-C.sub.4 alkyl. Each R.sup.4 may independently be
C.sub.1-C.sub.3 alkyl. Each R.sup.4 may independently be
C.sub.1-C.sub.2 alkyl. Each R.sup.4 may independently be methyl.
Each R.sup.4 may independently be C.sub.2 alkyl. Each R.sup.4 may
independently be C.sub.3 alkyl. Each R.sup.4 may independently be
C.sub.4 alkyl. Each R.sup.4 may independently be C.sub.5 alkyl.
[0102] In other embodiments, Each R.sup.5 may independently be
C.sub.1-C.sub.5 alkyl. Each R.sup.5 may independently be
C.sub.1-C.sub.4 alkyl. Each R.sup.5 may independently be
C.sub.1-C.sub.3 alkyl. Each R.sup.5 may independently be
C.sub.1-C.sub.2 alkyl. Each R.sup.5 may independently be methyl.
Each R.sup.5 may independently be C.sub.2 alkyl. Each R.sup.5 may
independently be C.sub.3 alkyl. Each R.sup.5 may independently be
C.sub.4 alkyl. Each R.sup.5 may independently be C.sub.5 alkyl.
[0103] In still other embodiments of some of the foregoing
embodiments of the compound of structure (I), at least one of
R.sup.3, R.sup.4 or R.sup.5 is C.sub.1-C.sub.10 alkylcarbonyl. In
some of these embodiments, two of R.sup.3, R.sup.4 and R.sup.5 are
C.sub.1-C.sub.10 alkylcarbonyl. In other of these embodiments,
R.sup.3, R.sup.4 and R.sup.5 are each C.sub.1-C.sub.10
alkylcarbonyl. In some more specific embodiments, the
C.sub.1-C.sub.10 alkylcarbonyl is methyl carbonyl (acetal).
[0104] In other embodiments, Each R.sup.3 may independently be
C.sub.1-C.sub.5 alkylcarbonyl. Each R.sup.3 may independently be
C.sub.1-C.sub.4 alkylcarbonyl. Each R.sup.3 may independently be
C.sub.1-C.sub.3 alkylcarbonyl. Each R.sup.3 may independently be
C.sub.1-C.sub.2 alkylcarbonyl. Each R.sup.3 may independently be
methylcarbonyl. Each R.sup.3 may independently be C.sub.2
alkylcarbonyl. Each R.sup.3 may independently be C.sub.3
alkylcarbonyl. Each R.sup.3 may independently be C.sub.4
alkylcarbonyl. Each R.sup.3 may independently be C.sub.5
alkylcarbonyl.
[0105] In other embodiments, Each R.sup.4 may independently be
C.sub.1-C.sub.5 alkylcarbonyl. Each R.sup.4 may independently be
C.sub.1-C.sub.4 alkylcarbonyl. Each R.sup.4 may independently be
C.sub.1-C.sub.3 alkylcarbonyl. Each R.sup.4 may independently be
C.sub.1-C.sub.2 alkylcarbonyl. Each R.sup.4 may independently be
methylcarbonyl. Each R.sup.4 may independently be C.sub.2
alkylcarbonyl. Each R.sup.4 may independently be C.sub.3
alkylcarbonyl. Each R.sup.4 may independently be C.sub.4
alkylcarbonyl. Each R.sup.4 may independently be C.sub.5
alkylcarbonyl.
[0106] In other embodiments, Each R.sup.5 may independently be
C.sub.1-C.sub.5 alkylcarbonyl. Each R.sup.5 may independently be
C.sub.1-C.sub.4 alkylcarbonyl. Each R.sup.5 may independently be
C.sub.1-C.sub.3 alkylcarbonyl. Each R.sup.5 may independently be
C.sub.1-C.sub.2 alkylcarbonyl. Each R.sup.5 may independently be
methylcarbonyl. Each R.sup.5 may independently be C.sub.2
alkylcarbonyl. Each R.sup.5 may independently be C.sub.3
alkylcarbonyl. Each R.sup.5 may independently be C.sub.4
alkylcarbonyl. Each R.sup.5 may independently be C.sub.5
alkylcarbonyl.
[0107] In some more specific embodiments of the compound of
structure (I), the compound has one of the following structures
from Table 1, or a pharmaceutically acceptable salt thereof:
TABLE-US-00001 TABLE 1 Representative .sup.123I Compounds No.
Structure Name 1 ##STR00006## 3-(4-(2-(4-(3-chloro-2-
hydroxypropoxy)-3- .sup.123iodophenyl)propan-2-
yl)phenoxy)propane-1,2-diol 1a ##STR00007##
(R)-3-(4-(2-(4-((R)-3-chloro-2- hydroxypropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propane-1,2-diol 1b
##STR00008## (S)-3-(4-(2-(4-((S)-3-chloro-2- hydroxypropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propane-1,2-diol 1c
##STR00009## (S)-3-(4-(2-(4-((R)-3-chloro-2- hydroxypropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propane-1,2-diol 1d
##STR00010## (R)-3-(4-(2-(4-((S)-3-chloro-2- hydroxypropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propane-1,2-diol 2
##STR00011## 3-(4-(2-(4-(3-chloro-2-
hydroxypropoxy)phenyl)propan-2-
yl)-2-.sup.123iodophenoxy)propane-1,2- diol 2a ##STR00012##
(R)-3-(4-(2-(4-((R)-3-chloro-2- hydroxypropoxy)phenyl)propan-2-
yl)-2-.sup.123iodophenoxy)propane-1,2- diol 2b ##STR00013##
(S)-3-(4-(2-(4-((S)-3-chloro-2- hydroxypropoxy)phenyl)propan-2-
yl)-2-.sup.123iodophenoxy)propane-1,2- diol 2c ##STR00014##
(S)-3-(4-(2-(4-((R)-3-chloro-2- hydroxypropoxy)phenyl)propan-2-
yl)-2-.sup.123iodophenoxy)propane-1,2- diol 2d ##STR00015##
(R)-3-(4-(2-(4-((S)-3-chloro-2- hydroxypropoxy)phenyl)propan-2-
yl)-2-.sup.123iodophenoxy)propane-1,2- diol 3 ##STR00016##
3-(4-(2-(4-(2-acetoxy-3- chloropropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propane-1,2-diyl diacetate
3a ##STR00017## (S)-3-(4-(2-(4-((R)-2-acetoxy-3- chloropropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propane-1,2-diyl diacetate
3b ##STR00018## (R)-3-(4-(2-(4-((S)-2-acetoxy-3- chloropropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propane-1,2-diyl diacetate
3c ##STR00019## (R)-3-(4-(2-(4-((R)-2-acetoxy-3- chloropropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propane-1,2-diyl diacetate
3d ##STR00020## (S)-3-(4-(2-(4-((S)-2-acetoxy-3- chloropropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propane-1,2-diyl diacetate
4 ##STR00021## 1-chloro-3-(4-(2-(4-(2-hydroxy-3-
methoxypropoxy)phenyl)propan-2-
yl)-2-.sup.123iodophenoxy)propan-2-ol 4a ##STR00022##
(R)-1-chloro-3-(4-(2-(4-((R)-2- hydroxy-3- methoxy propoxy
)phenyl)propan-2- yl)-2-.sup.123iodophenoxy)propan-2-ol 4b
##STR00023## (S)-1-chloro-3-(4-(2-(4-((S)-2- hydroxy-3-
methoxypropoxy)phenyl)propan-2-
yl)-2-.sup.123iodophenoxy)propan-2-ol 4c ##STR00024##
(R)-1-chloro-3-(4-(2-(4-((S)-2- hydroxy-3-
methoxypropoxy)phenyl)propan-2-
yl)-2-.sup.123iodophenoxy)propan-2-ol 4d ##STR00025##
(S)-1-chloro-3-(4-(2-(4-((R)-2- hydroxy-3-
methoxypropoxy)phenyl)propan-2-
yl)-2-.sup.123iodophenoxy)propan-2-ol 5 ##STR00026##
1-(4-(2-(4-(2-acetoxy-3- chloropropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)-3-methoxypropan-2-yl
acetate 5a ##STR00027## (R)-1-(4-(2-(4-((R)-2-acetoxy-3-
chloropropoxy)-3- .sup.123iodophenyl)propan-2-
yl)phenoxy)-3-methoxypropan-2-yl acetate 5b ##STR00028##
(S)-1-(4-(2-(4-((S)-2-acetoxy-3- chloropropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)-3-methoxypropan-2-yl
acetate 5c ##STR00029## (S)-1-(4-(2-(4-((R)-2-acetoxy-3-
chloropropoxy)-3- .sup.123iodophenyl)propan-2-
yl)phenoxy)-3-methoxypropan-2-yl acetate 5d ##STR00030##
(R)-1-(4-(2-(4-((S)-2-acetoxy-3- chloropropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)-3-methoxypropan-2-yl
acetate 6 ##STR00031## 1-butoxy-3-(4-(2-(4-(3-chloro-2-
hydroxypropoxy)-3- .sup.123iodophenyl)propan-2-
yl)phenoxy)propan-2-ol 6a ##STR00032##
(S)-1-butoxy-3-(4-(2-(4-((R)-3- chloro-2-hydroxypropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propan-2-ol 6b ##STR00033##
(S)-1-butoxy-3-(4-(2-(4-((S)-3- chloro-2-hydroxypropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propan-2-ol 6c ##STR00034##
(S)-1-butoxy-3-(4-(2-(4-((R)-3- chloro-2-hydroxypropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propan-2-ol 6d ##STR00035##
(R)-1-butoxy-3-(4-(2-(4-((S-3- chloro-2-hydroxypropoxy)-3-
.sup.123iodophenyl)propan-2- yl)phenoxy)propan-2-ol 7 ##STR00036##
1-(4-(2-(4-(2-acetoxy-3- butoxypropoxy)phenyl)propan-2-
yl)-2-.sup.123iodophenoxy)-3- chloropropan-2-yl acetate 7a
##STR00037## (R)-1-(4-(2-(4-((R)-2-acetoxy-3-
butoxypropoxy)phenyl)propan-2- yl)-2-.sup.123iodophenoxy)-3-
chloropropan-2-yl acetate 7b ##STR00038##
(S)-1-(4-(2-(4-((S)-2-acetoxy-3 butoxypropoxy)phenyl)propan-
yl)-2-.sup.123iodophenoxy)-3- chloropropan-2-yl acetate 7c
##STR00039## (R)-1-(4-(2-(4-((S)-2-acetoxy-3-
butoxypropoxy)phenyl)propan-2- yl)-2-.sup.123iodophenoxy)-3-
chloropropan-2-yl acetate 7d ##STR00040##
(S)-1-(4-(2-(4-((R)-2-acetoxy-3- butoxypropoxy)phenyl)propan-2-
yl)-2-.sup.123iodophenoxy)-3- chloropropan-2-yl acetate 8
##STR00041## 3-(4-(2-(4-(3-chloro-2- hydroxypropoxy)-3-
iodophenyl)propan-2- yl)phenoxy)propane-1,2-diol 8a ##STR00042##
(R)-3-(4-(2-(4-((R)-3-chloro-2- hydroxypropoxy)-3-
iodophenyl)propan-2- yl)phenoxy)propane-1,2-diol 8b ##STR00043##
(S)-3-(4-(2-(4-((S)-3-chloro-2- hydroxypropoxy)-3-
iodophenyl)propan-2- yl)phenoxy)propane-1,2-diol 8c ##STR00044##
(S)-3-(4-(2-(4-((R)-3-chloro-2- hydroxypropoxy)-3-
iodophenyl)propan-2- yl)phenoxy)propane-1,2-diol 8d ##STR00045##
(R)-3-(4-(2-(4-((S)-3-chloro-2- hydroxypropoxy)-3-
iodophenyl)propan-2- yl)phenoxy)propane-1,2-diol 9 ##STR00046##
3-(4-(2-(3-bromo-4-(3-chloro-2- hydroxypropoxy)phenyl)propan-2-
yl)phenoxy)propane-1,2-diol 9a ##STR00047##
(R)-3-(4-(2-(3-bromo-4-((R)-3- chloro-2-
hydroxypropoxy)phenyl)propan-2- yl)phenoxy)propane-1,2-diol 9b
##STR00048## (S)-3-(4-(2-(3-bromo-4-((S)-3- chloro-2-
hydroxypropoxy)phenyl)propan-2 yl)phenoxy)propane-1,2-diol 9c
##STR00049## (S)-3-(4-(2-(3-bromo-4-((R)-3- chloro-2-
hydroxypropoxy)phenyl)propan-2- yl)phenoxy)propane-1,2-diol 9d
##STR00050## (R)-3-(4-(2-(3-bromo-4-((S)-3- chloro-2-
hydroxypropoxy)phenyl)propan-2- yl)phenoxy)propane-1,2-diol 10
##STR00051## 3-(4-(2-(3-chloro-4-(3-chloro-2-
hydroxypropoxy)phenyl)propan-2- yl)phenoxy)propane-1,2-diol 10a
##STR00052## (R)-3-(4-(2-(3-chloro-4-((R)-3- chloro-2-
hydroxypropoxy)phenyl)propan-2- yl)phenoxy)propane-1,2-diol 10b
##STR00053## (S)-3-(4-(2-(3-chloro-4-((S)-3- chloro-2-
hydroxypropoxy)phenyl)propan-2- yl)phenoxy)propane-1,2-diol 10c
##STR00054## (S)-3-(4-(2-(3-chloro-4-((R)-3- chloro-2-
hydroxypropoxy)phenyl)propan-2- yl)phenoxy)propane-1,2-diol 10d
##STR00055## (R)-3-(4-(2-(3-chloro-4-((S)-3- chloro-2-
hydroxypropoxy)phenyl)propan-2- yl)phenoxy)propane-1,2-diol 11
##STR00056## 3-(4-(2-(4-(3-chloro-2- hydroxypropoxy)-3-
fluorophenyl)propan-2- yl)phenoxy)propane-1,2-diol 11a ##STR00057##
(R)-3-(4-(2-(4-((R)-3-chloro-2- hydroxypropoxy)-3-
fluorophenyl)propan-2- yl)phenoxy)propane-1,2-diol 11b ##STR00058##
(S)-3-(4-(2-(4-((S)-3-chloro-2- hydroxypropoxy)-3-
fluorophenyl)propan-2- yl)phenoxy)propane-1,2-diol 11c ##STR00059##
(S)-3-(4-(2-(4-((R)-3-chloro-2- hydroxypropoxy)-3-
fluorophenyl)propan-2- yl)phenoxy)propane-1,2-diol 11d ##STR00060##
(R)-3-(4-(2-(4-((S)-3-chloro-2- hydroxypropoxy)-3-
fluorophenyl)propan-2- yl)phenoxy)propane-1,2-diol
[0108] In some embodiments, compounds of structure I which result
in unstable structures and/or unsatisfied valences are not included
within the scope of the invention.
[0109] In another embodiment, the present disclosure provides a
pharmaceutical composition comprising any of the foregoing
compounds of structure (I) and a pharmaceutically acceptable
carrier.
[0110] Compounds as described herein may be in the free form or in
the form of a salt thereof. In some embodiments, compounds as
described herein may be in the form of a pharmaceutically
acceptable salt, which are known in the art (Berge et al., J.
Pharm. Sci. 1977, 66, 1). Pharmaceutically acceptable salt as used
herein includes, for example, salts that have the desired
pharmacological activity of the parent compound (salts which retain
the biological effectiveness and/or properties of the parent
compound and which are not biologically and/or otherwise
undesirable). Compounds as described herein having one or more
functional groups capable of forming a salt may be, for example,
formed as a pharmaceutically acceptable salt. Compounds containing
one or more basic functional groups may be capable of forming a
pharmaceutically acceptable salt with, for example, a
pharmaceutically acceptable organic or inorganic acid.
Pharmaceutically acceptable salts may be derived from, for example,
and without limitation, acetic acid, adipic acid, alginic acid,
aspartic acid, ascorbic acid, benzoic acid, benzenesulfonic acid,
butyric acid, cinnamic acid, citric acid, camphoric acid,
camphorsulfonic acid, cyclopentanepropionic acid, diethylacetic
acid, digluconic acid, dodecylsulfonic acid, ethanesulfonic acid,
formic acid, fumaric acid, glucoheptanoic acid, gluconic acid,
glycerophosphoric acid, glycolic acid, hemisulfonic acid, heptanoic
acid, hexanoic acid, hydrochloric acid, hydrobromic acid, hydriodic
acid, 2-hydroxyethanesulfonic acid, isonicotinic acid, lactic acid,
malic acid, maleic acid, malonic acid, mandelic acid,
methanesulfonic acid, 2-napthalenesulfonic acid,
naphthalenedisulphonic acid, p-toluenesulfonic acid, nicotinic
acid, nitric acid, oxalic acid, pamoic acid, pectinic acid,
3-phenylpropionic acid, phosphoric acid, picric acid, pimelic acid,
pivalic acid, propionic acid, pyruvic acid, salicylic acid,
succinic acid, sulfuric acid, sulfamic acid, tartaric acid,
thiocyanic acid or undecanoic acid. Compounds containing one or
more acidic functional groups may be capable of forming
pharmaceutically acceptable salts with a pharmaceutically
acceptable base, for example, and without limitation, inorganic
bases based on alkaline metals or alkaline earth metals or organic
bases such as primary amine compounds, secondary amine compounds,
tertiary amine compounds, quaternary amine compounds, substituted
amines, naturally occurring substituted amines, cyclic amines or
basic ion-exchange resins. Pharmaceutically acceptable salts may be
derived from, for example, and without limitation, a hydroxide,
carbonate, or bicarbonate of a pharmaceutically acceptable metal
cation such as ammonium, sodium, potassium, lithium, calcium,
magnesium, iron, zinc, copper, manganese or aluminum, ammonia,
benzathine, meglumine, methylamine, dimethylamine, trimethylamine,
ethylamine, diethylamine, triethylamine, isopropylamine,
tripropylamine, tributylamine, ethanolamine, diethanolamine,
2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine,
lysine, arginine, histidine, caffeine, hydrabamine, choline,
betaine, ethylenediamine, glucosamine, glucamine, methylglucamine,
theobromine, purines, piperazine, piperidine, procaine,
N-ethylpiperidine, theobromine, tetramethylammonium compounds,
tetraethylammonium compounds, pyridine, N,N-dimethylaniline,
N-methylpiperidine, morpholine, N-methylmorpholine,
N-ethylmorpholine, dicyclohexylamine, dibenzylamine,
N,N-dibenzylphenethylamine, 1-ephenamine,
N,N'-dibenzylethylenediamine or polyamine resins. In some
embodiments, compounds as described herein may contain both acidic
and basic groups and may be in the form of inner salts or
zwitterions, for example, and without limitation, betaines. Salts
as described herein may be prepared by conventional processes known
to a person skilled in the art, for example, and without
limitation, by reacting the free form with an organic acid or
inorganic acid or base, or by anion exchange or cation exchange
from other salts. Those skilled in the art will appreciate that
preparation of salts may occur in situ during isolation and
purification of the compounds or preparation of salts may occur by
separately reacting an isolated and purified compound.
[0111] In some embodiments, compounds and all different forms
thereof (e.g. free forms, salts, polymorphs, isomeric forms) as
described herein may be in the solvent addition form, for example,
solvates. Solvates contain either stoichiometric or
non-stoichiometric amounts of a solvent in physical association the
compound or salt thereof. The solvent may be, for example, and
without limitation, a pharmaceutically acceptable solvent. For
example, hydrates are formed when the solvent is water or
alcoholates are formed when the solvent is an alcohol.
[0112] In some embodiments, compounds and all different forms
thereof (e.g. free forms, salts, solvates, isomeric forms) as
described herein may include crystalline and amorphous forms, for
example, polymorphs, pseudopolymorphs, conformational polymorphs,
amorphous forms, or a combination thereof. Polymorphs include
different crystal packing arrangements of the same elemental
composition of a compound. Polymorphs usually have different X-ray
diffraction patterns, infrared spectra, melting points, density,
hardness, crystal shape, optical and electrical properties,
stability and/or solubility. Those skilled in the art will
appreciate that various factors including recrystallization
solvent, rate of crystallization and storage temperature may cause
a single crystal form to dominate.
[0113] In some embodiments, compounds and all different forms
thereof (e.g. free forms, salts, solvates, polymorphs) as described
herein include isomers such as geometrical isomers, optical isomers
based on asymmetric carbon, stereoisomers, tautomers, individual
enantiomers, individual diastereomers, racemates, diastereomeric
mixtures and combinations thereof, and are not limited by the
description of the formula illustrated for the sake of
convenience.
[0114] In some embodiments, pharmaceutical compositions in
accordance with this invention may comprise a salt of such a
compound, preferably a pharmaceutically or physiologically
acceptable salt. Pharmaceutical preparations will typically
comprise one or more carriers, excipients or diluents acceptable
for the mode of administration of the preparation, be it by
injection, inhalation, topical administration, lavage, or other
modes suitable for the selected treatment. Suitable carriers,
excipients or diluents are those known in the art for use in such
modes of administration.
[0115] Suitable pharmaceutical compositions may be formulated by
means known in the art and their mode of administration and dose
determined by the skilled practitioner. For parenteral
administration, a compound may be dissolved in sterile water or
saline or a pharmaceutically acceptable vehicle used for
administration of non-water soluble compounds such as those used
for vitamin K. For enteral administration, the compound may be
administered in a tablet, capsule or dissolved in liquid form. The
tablet or capsule may be enteric coated, or in a formulation for
sustained release. Many suitable formulations are known, including,
polymeric or protein microparticles encapsulating a compound to be
released, ointments, pastes, gels, hydrogels, or solutions which
can be used topically or locally to administer a compound. A
sustained release patch or implant may be employed to provide
release over a prolonged period of time. Many techniques known to
one of skill in the art are described in Remington: the Science
& Practice of Pharmacy by Alfonso Gennaro, 20.sup.th ed.,
Lippencott Williams & Wilkins, (2000). Formulations for
parenteral administration may, for example, contain excipients,
polyalkylene glycols such as polyethylene glycol, oils of vegetable
origin, or hydrogenated naphthalenes. Biocompatible, biodegradable
lactide polymer, lactide/glycolide copolymer, or
polyoxyethylene-polyoxypropylene copolymers may be used to control
the release of the compounds. Other potentially useful parenteral
delivery systems for modulatory compounds include ethylene-vinyl
acetate copolymer particles, osmotic pumps, implantable infusion
systems, and liposomes. Formulations for inhalation may contain
excipients, for example, lactose, or may be aqueous solutions
containing, for example, polyoxyethylene-9-lauryl ether,
glycocholate and deoxycholate, or may be oily solutions for
administration in the form of nasal drops, or as a gel.
[0116] Compounds or pharmaceutical compositions in accordance with
this invention or for use in this invention may be administered by
means of a medical device or appliance such as an implant, graft,
prosthesis, stent, etc. Also, implants may be devised which are
intended to contain and release such compounds or compositions. An
example would be an implant made of a polymeric material adapted to
release the compound over a period of time.
[0117] An "effective amount" of a pharmaceutical composition
according to the invention includes a therapeutically effective
amount or a prophylactically effective amount. A "therapeutically
effective amount" refers to an amount effective, at dosages and for
periods of time necessary, to achieve the desired therapeutic
result, such as reduced tumor size, increased life span or
increased life expectancy. A therapeutically effective amount of a
compound may vary according to factors such as the disease state,
age, sex, and weight of the subject, and the ability of the
compound to elicit a desired response in the subject. Dosage
regimens may be adjusted to provide the optimum therapeutic
response. A therapeutically effective amount is also one in which
any toxic or detrimental effects of the compound are outweighed by
the therapeutically beneficial effects. A "prophylactically
effective amount" refers to an amount effective, at dosages and for
periods of time necessary, to achieve the desired prophylactic
result, such as smaller tumors, increased life span, increased life
expectancy or prevention of the progression of prostate cancer to
lethal CRPC. Typically, a prophylactic dose is used in subjects
prior to or at an earlier stage of disease, so that a
prophylactically effective amount may be less than a
therapeutically effective amount.
[0118] It is to be noted that dosage values may vary with the exact
imaging protocol. For any particular subject, specific dosage
regimens may be adjusted over time according to the individual need
and the professional judgement of the person administering or
supervising the administration of the compositions. Dosage ranges
set forth herein are exemplary only and do not limit the dosage
ranges that may be selected by medical practitioners. The amount of
active compound(s) in the composition may vary according to factors
such as the disease state, age, sex, and weight of the subject.
Dosage regimens may be adjusted to provide the optimum imaging
result. For example, a single bolus may be administered, several
divided doses may be administered over time or the dose may be
proportionally reduced or increased as indicated by the imaging
results. It may be advantageous to formulate parenteral
compositions in dosage unit form for ease of administration and
uniformity of dosage.
[0119] In general, compounds of the invention should be used
without causing substantial toxicity. Toxicity of the compounds of
the invention can be determined using standard techniques, for
example, by testing in cell cultures or experimental animals and
determining the therapeutic index, i.e., the ratio between the LD50
(the dose lethal to 50% of the population) and the LD100 (the dose
lethal to 100% of the population). In some circumstances, such as
in severe disease conditions, substantial excesses of the
compositions may be administered for therapeutic effects. Some
compounds of this invention may be toxic at some concentrations.
Titration studies may be used to determine toxic and non-toxic
concentrations. Toxicity may be evaluated by examining a particular
compound's or composition's specificity across cell lines using PC3
or DU145 cells as possible negative controls since these cells do
not express functional AR. Animal studies may be used to provide an
indication if the compound has any effects on other tissues.
Systemic therapy that targets the AR will not likely cause major
problems to other tissues since antiandrogens and androgen
insensitivity syndrome are not fatal.
[0120] Compounds as described herein may be administered to a
subject. As used herein, a "subject" may be a human, non-human
primate, mammal, rat, mouse, cow, horse, pig, sheep, goat, dog, cat
and the like. The subject may be suspected of having or at risk for
having a cancer, such as prostate cancer, breast cancer, ovarian
cancer, salivary gland carcinoma, or endometrial cancer, or
suspected of having or at risk for having acne, hirsutism,
alopecia, benign prostatic hyperplasia, ovarian cysts, polycystic
ovary disease, precocious puberty, spinal and bulbar muscular
atrophy, or age-related macular degeneration. Diagnostic methods
for various cancers, such as prostate cancer, breast cancer,
ovarian cancer, salivary gland carcinoma, or endometrial cancer,
and diagnostic methods for acne, hirsutism, alopecia, benign
prostatic hyperplasia, ovarian cysts, polycystic ovary disease,
precocious puberty, spinal and bulbar muscular atrophy, or
age-related macular degeneration and the clinical delineation of
cancer, such as prostate cancer, breast cancer, ovarian cancer,
salivary gland carcinoma, or endometrial cancer, diagnoses and the
clinical delineation of acne, hirsutism, alopecia, benign prostatic
hyperplasia, ovarian cysts, polycystic ovary disease, precocious
puberty, spinal and bulbar muscular atrophy, or age-related macular
degeneration are known to those of ordinary skill in the art.
[0121] Compounds for use in the present invention may be obtained
from medical sources or modified using known methodologies from
naturally occurring compounds. In addition, methods of preparing or
synthesizing compounds of the present invention will be understood
by a person of skill in the art having reference to known chemical
synthesis principles. For example, Auzou et al 1974 European
Journal of Medicinal Chemistry 9(5), 548-554 describes suitable
synthetic procedures that may be considered and suitably adapted
for preparing compounds of any one of the compounds of structure
(I) as set out above. Other references that may be helpful include:
Debasish Das, Jyh-Fu Lee and Soofin Cheng "Sulfonic acid
functionalized mesoporous MCM-41 silica as a convenient catalyst
for Bisphenol-A synthesis" Chemical Communications, (2001)
2178-2179; U.S. Pat. No. 2,571,217 Davis, Orris L.; Knight, Horace
S.; Skinner, John R. (Shell Development Co.) "Halohydrin ethers of
phenols." (1951); and Rokicki, G.; Pawlicki, J.; Kuran, W.
"Reactions of 4-chloromethyl-1,3-dioxolan-2-one with phenols as a
new route to polyols and cyclic carbonates." Journal fuer
Praktische Chemie (Leipzig) (1985) 327, 718-722.
[0122] For example, exemplary compounds of the present invention
may be prepared with reference to the following General Reaction
Scheme I, wherein R.sup.1 and R.sup.2 are as described above and
L.sup.1 and L.sup.2 are independently a leacing group:
##STR00061##
[0123] Referring to General Reaction Scheme I, bisphenol compounds
of structure A can be purchased from commercial sources or prepared
according to methods well-known to those of ordinary skill in the
art. Compounds of structure A can be reacted with compounds of
structure B under basic conditions (e.g., NaH) to yield compounds
of structure C. In this regard, particularily useful leaving groups
(L.sup.1) include p-toulenesulfonates ("tosyl"), which can be
prepared by reaction of the corresponding alcohol with tosyl
chloride. Further, various stereoisomers of compound B can be used
depending on the desired stereochemistry of the final product.
Various stereoisomer of B can be purchased or prepared according to
methods known in the art. The radioactive iodine moiety (.sup.123I)
can be installed by reaction of C with an appropriate iodinating
reagent, for example Na.sup.123I and a suitable oxidant (e.g.,
NaClO) to yield D. It should be noted that, although General
Reaction Scheme I depicts iodination at only one position, other
compounds of structure (I) with .sup.123I at different positions
and/or multiple .sup.123I substitutions can be prepared according
to analogous methods known to those of oridinary skill in the
art.
[0124] Reaction of D with epoxide E under basic conditions (e.g.,
NaH) yields compounds of structure F. Again, tosyl leaving groups
have been fond to be particularly useful as the L.sup.2 moiety, and
various stereoisomers of compound F can be used depending on the
desired stereochemistry of the final product. Finally, reaction of
F with an appropriate reagent, such as CeCl3.7H.sub.2O yields G.
Other compounds of structure (I) wherein R.sup.3, R.sup.4 and/or
R.sup.5 are moieties other than H can be prepared by further
modification of compound G. For example alkylation with common
alkylating reagents (e.g., methyl idodide) and/or acylation with
with common acylating reagents (e.g., acetyl chloride) yields
compounds of structure (I) wherein R.sup.3, R.sup.4 and/or R.sup.5
are C.sub.1-C.sub.10 alkyl or C.sub.1-C.sub.10 alkylcarbonyl,
respectively.
[0125] One skilled in the art will recognize that variations to the
order of the steps and reagents discussed in reference to the above
General Synthetic Scheme I are possible. For example, epoxidation
may precede dioxalone formation. Further, .sup.123I atoms may be
introduced via any number of reagents, and iodination is not
limited to those methods depicted or described above. Methods for
such iodination are well known in the art. Methodologies for
preparation of specific exemplary compounds of structure I are
described in more detail in the following examples.
[0126] In addition, protecting group strategies may be employed for
preparation of the compounds disclosed herein. Such strategies are
well known to those of skill in the art.
[0127] Exemplary protecting groups and related strategies are
disclosed in Greene's Protective Groups in Organic Synthesis,
Wiley-Interscience; 4 edition (Oct. 30, 2006), which is hereby
incorporated by reference in its entirety. In certain embodiments,
a protecting group is used to mask an alcohol moiety while
performing other chemical transformations. After removal of the
protecting group, the free hydroxyl is obtained. Such protecting
groups and strategies are well known in the art.
[0128] The present compounds find particular utility in methods for
imaging the prostate. In some embodiments, a method for imaging
benign conditions of the prostate (e.g., benign prostatic
hyperplasia), comprising administering any of the foregoing
pharmaceutical compositions to a subject and detecting the
prostate, is provided. Accordingly, in another embodiment, the
present disclosure provides a method of imaging cancer, the method
comprising administering the foregoing pharmaceutical composition
to a subject and detecting the presence or absence of cancer by use
of SPECT.
[0129] In certain embodiments, the method identifies the presence
or absence of a tumor. For example, some embodiments the method
identifies the location of a tumor. In certain embodiments, the
cancer is prostate cancer, for example, castration resistant
prostate cancer. In other embodiments, the prostate cancer is
androgen-dependent prostate cancer. In some embodiments, the
subject is a mammal such as a human.
[0130] In some other embodiments, the method is useful for
detecting the presence of AR splice variants or other AR species
that cannot be detected by imaging agents that interact with the AR
LBD (i.e., mutations, truncations). Without wishing to be bound by
any particular theory, since the present compounds bind to the AR
N-terminal domain (NTD), even mutants or variants which lack the AR
LBD can be imaged employing the present compounds. Thus, the
present methods may be useful for detecting AR species, including
mutants and variants, which lack the LBD or have LBD mutations, but
do comprise the AR NTD. In other embodiments the method detects the
presence or overexpression of AR splice variants lacking the ligand
binding domain. For example, the method may include sequential
imaging with .sup.18F-FDHT and a compound of the invention and a
discordant distribution or discordant level of uptake between
.sup.18F-FDHT and the compound of the invention indicates the
presence or overexpression of splice variants lacking the ligan
binding domain.
[0131] In other embodiments, the compounds of the invention are
used in single photon emission computed tomography methods to
monitor a patient's response to therapy. In other embodiments, the
methods comprise use of a compound of the invention to detect the
AR NTD.
[0132] In another embodiment, the present disclosure provides the
use of any one of the foregoing compounds of Formula (I) for
imaging cancer. For example in some embodiments, the imaging is in
a human patient.
[0133] In another embodiment, the present disclosure provides the
use of any one of the foregoing compounds of Formula (I) for
imaging the prostate. For example in some embodiments, the imaging
is in a human patient.
[0134] In accordance with another embodiment, there is provided a
use of the compounds of Formula (I) as described anywhere herein
for preparation of a medicament for imaging the prostate. The
imaging may be for imaging of benign postate conditions of for
imaging cancer (e.g., tumors), for example prostate cancer. The
imaging may be by SPECT.
[0135] The imaging may be in a mammalian cell. The imaging may be
in a mammal. The mammal may be a human.
[0136] Alternatively, the compounds may be administred to a mammal
for imaging purposes. The administering and imaging may be to a
mammal in need of diagnosis of at least one indication selected
from the group consisting of: prostate cancer, breast cancer,
ovarian cancer, endometrial cancer, salivary gland carcinoma,
benign prostatic hyperplasia, hair loss, acne, hirsutism, ovarian
cysts, polycystic ovary disease, precocious puberty, spinal and
bulbar muscular atrophy (e.g., Kennedy's disease), and age-related
macular degeneration. The mammalian cell may be a human cell. The
imaging may be for imaging AR splice variants, mutants or other AR
species which comprise the AR NTD.
[0137] In some embodiments, the compounds as described herein or
pharmaceutically acceptable acceptable salts thereof may be used
for imaging and diagnosis of at least one indication selected from
the group consisting of: prostate cancer, breast cancer, ovarian
cancer, endometrial cancer, salivary gland carcinoma, benign
prostatic hyperplasia, hair loss, acne, hirsutism, ovarian cysts,
polycystic ovary disease, precocious puberty, spinal and bulbar
muscular atrophy, and age-related macular degeneration. In some
embodiments, the compounds as described herein or acceptable salts
thereof above may be used in the preparation of a medicament or a
composition for imaging the prostate, for example for imaging
benign prostate conditions or for imaging prostate cancer in a
subject in need of such imaging (for example for diagnosis and/or
location of prostate tumors).
[0138] Some aspects of this invention, make use of compositions
comprising a compound described herein and a pharmaceutically
acceptable excipients or carrier. In some embodiments, the prostate
cancer is castration-resistant prostate cancer (also referred to as
hormone refractory, androgen-independent, androgen deprivation
resistant, androgen ablation resistant, androgen
depletion-independent, castration-recurrent,
anti-androgen-recurrent). In some embodiments the prostate cancer
is androgen-dependent or androgen-sensitive. In other embodiments,
the imaging is for imaging a benign prostate conditions such as
benign prostatic hyperplasia. Methods of imaging any of the
indications described herein are also provided. Such methods may
include administering a compound as described herein or a
composition of a compound as described herein, or an effective
amount of a compound as described herein or composition of a
compound as described herein to a subject in need thereof.
[0139] In other embodiments, the present disclosure provides a
method for modulating androgen receptor (AR) activity, the method
comprising administering to a mammalian cell one or more of the
present compounds. In some embodiments the modulating of androgen
receptor (AR) activity is in a mammalian cell.
[0140] In certain embodiments, the method for modulating androgen
receptor (AR) activity is for treatment of at least one indication
selected from the group consisting of: prostate cancer, breast
cancer, ovarian cancer, endometrial cancer, salivary gland
carcinoma, hair loss, acne, hirsutism, ovarian cysts, polycystic
ovary disease, precocious puberty, spinal and bulbar muscular
atrophy, and age related macular degeneration. In ceretain
embodiments, the indication is prostate cancer. In certain
embodiments, the prostate cancer is castration resistant prostate
cancer. In other embodiments, the prostate cancer is androgen
dependent prostate cancer. In certain embodiments, the spinal and
bulbar muscular atrophy is Kennedy's disease.
[0141] In another aspect, the present disclosure provides a method
of modulating androgen receptor (AR) activity, the method
comprising administering a pharmaceutical composition comprising a
compound as decribed herein to a subject in need thereof.
[0142] In another aspect, the present disclosure provides a
pharmaceutical composition comprising a compound as described
herein, and an additional therapeutic agent and a pharmaceutically
acceptable carrier. In some embodiments, the additional therapeutic
agent is for treating prostate cancer, breast cancer, ovarian
cancer, endometrial cancer, salivary gland carcinoma, hair loss,
acne, hirsutism, ovarian cysts, polycystic ovary disease,
precocious puberty, spinal and bulbar muscular atrophy or age
related macular degeneration. In other embodiments, the additional
therapeutic agent is enzalutamide, galeterone, ARN-509, ODN-201
abiraterone, bicalutamide, nilutamide, flutamide, cyproterone
acetate, docetaxel, Bevacizumab (Avastin), OSU-HDAC42, VITAXIN,
sunitumib, ZD-4054, Cabazitaxel (XRP-6258), MDX-010 (Ipilimumab),
OGX 427, OGX 011, finasteride, dutasteride, turosteride,
bexlosteride, izonsteride, FCE 28260, SKF 105,111 or related
compounds thereof.
[0143] In an exemplary embodiment for imaging the prostate, a dose
of the disclosed compounds in solution (typically 5 to 10
millicuries or 200 to 400 MBq) is typically injected rapidly into a
saline drip running into a vein, in a patient. Then, the patient is
placed in the SPECT for a series of one or more scans which may
take from 20 minutes to as long as an hour (often, only about one
quarter of the body length may be imaged at a time). Methods for
SPECT scanning are well known in the art.
EXAMPLES
[0144] All non-aqueous reactions were performed in flame-dried
round bottomed flasks. The flasks were fitted with rubber septa and
reactions were conducted under a positive pressure of argon unless
otherwise specified. Stainless steel syringes were used to transfer
air- and moisture-sensitive liquids. Flash column chromatography
was performed as described by Still et al. (Still, W. C.; Kahn, M.;
Mitra, A. J. Org. Chem. 1978, 43, 2923) using 230-400 mesh silica
gel. Thin-layer chromatography was performed using aluminum plates
pre-coated with 0.25 mm 230-400 mesh silica gel impregnated with a
fluorescent indicator (254 nm). Thin-layer chromatography plates
were visualized by exposure to ultraviolet light and a "Seebach"
staining solution (700 mL water, 10.5 g Cerium (IV) sulphate
tetrahydrate, 15.0 g molybdato phosphoric acid, 17.5 g sulphuric
acid) followed by heating (.about.1 min) with a heating gun
(.about.250.degree. C.). Organic solutions were concentrated on
Buchi R-114 rotatory evaporators at reduced pressure (15-30 torr,
house vacuum) at 25-40.degree. C.
[0145] Commercial regents and solvents were used as received. All
solvents used for extraction and chromatography were HPLC grade.
Normal-phase Si gel Sep paks.TM. were purchased from waters, Inc.
Thin-layer chromatography plates were Kieselgel 60F.sub.254. All
synthetic reagents were purchased from Sigma Aldrich and Fisher
Scientific Canada.
Example 1
SYNTHESIS OF
(R)-3-(4-(2-(4-((S)-3-CHLORO-2-HYDROXYPROPOXY)-3-.sup.123IODOPHENYL)PROPA-
N-2-YL)PHENOXY)PROPANE-1,2-DIOL (1d)
##STR00062## ##STR00063##
[0147] Compound i
[0148] p-Toluenesulfonyl chloride (6.5 g, 34.1 mmol) was added
portionwise over a period of 10 min to a solution of
(S)-(+)-1,2-isopropylideneglycerol (3.0 g, 22.7 mmol) and DMAP (30
mg, 0.25 mmol) in anhydrous pyridine (30 mL) in a water bath. The
resulting solution was stirred overnight. The pyridine was removed
under reduced pressure, and the residue was diluted with ethyl
acetate (50 mL), washed subsequently with water (2 .times.40 mL),
cold aqueous 1 M HCl (40 mL), saturated NaHCO.sub.3 (40 mL) and
water (40 mL). The organic layer was dried over Mg.sub.2SO.sub.4,
filtered and concentrated to give a light yellow oil. The crude
product was purified by column chromatography (eluent: 10% ethyl
acetate in hexane to 30% ethyl acetate in hexane) to afford
(R)-2,2-Dimethyl-1,3-dioxolane-4-methanol p-toluenesulfonate i
(5.91 g, 90.9% yield) as a colorless viscous oil.
[0149] Compound ii
[0150] Sodium hydride (60% dispersion in mineral oil, 2.27 g, 56.66
mmol, 2.0 equiv) was added slowly to a stirred solution of
Bisphenol A (12.94 g, 56.66 mmol, 2 equiv) in anhydrous dimethyl
formamide (60 mL), at room temperature, and the contents were
stirred under an atmosphere of argon for 20 min. Compound i (8.53
g, 28.33 mmol, 1.0 equiv) was added, and the mixture was allowed to
react at 50-60.degree. C. for 16 h. Next, the reaction was quenched
by the addition of a saturated solution of ammonium chloride (10
mL), and the mixture was extracted with ethyl acetate (3.times.50
mL). The organic layer was washed with deionized water (3.times.40
mL), dried over anhydrous magnesium sulfate, filtered, and then
concentrated under reduced pressure. The resulting residue was
purified by flash column chromatography on silica gel (eluent: 5%
ethyl acetate in hexane to 10% ethyl acetate in hexane) to provide
the title compound (8.10 g, 83.5%) as a sticky oil.
[0151] Compound iii
[0152] Compound ii (200 mg, 0.58 mmol) was dissolved in 4 mL of
methanol. One equivalent of sodium .sup.123iodide (85 mg, 0.58
mmol) and 1.5 equiv of sodium hydroxide (35 mg, 0.88 mmol) were
added and the solution was cooled to 0.degree. C. Aqueous sodium
hypochlorite (800 mg, 1 equiv, 0.58 mmol of sodium hypochlorite)
was then added dropwise over 2 min at 0-3.degree. C. The pH was
kept to 6-7 by adding 10% HCl. The mixture was extracted with
dichloromethane (2.times.20 mL). The organic layer was washed with
deionized water (2.times.20 mL), was dried over anhydrous magnesium
sulfate, was filtered, and was concentrated under reduced pressure
to provide the title compound as a sticky oil.
[0153] Compound iv
[0154] Sodium hydride (60% dispersion in mineral oil, 41.6 mg, 1.04
mmol, 2.0 equiv) was added slowly to a stirred solution of compound
iii in anhydrous dimethyl formamide (3 mL), at room temperature,
and the contents were stirred under an atmosphere of argon for 10
min. A solution of (2R)-(-)-glycidyl tosylate 98% (142 mg, 0.62
mmol, 1.5 equiv) in anhydrous dimethyl formamide (2 mL) was added
via syringe, and the mixture was allowed to react at 65-70.degree.
C. for 40min. Next, the reaction was quenched by the addition of a
saturated solution of ammonium chloride (1 mL), and the mixture was
extracted with dichloromethane (2.times.20 mL). The organic layer
was washed with deionized water (2.times.20 mL), dried over
anhydrous magnesium sulfate, filtered and then concentrated under
reduced pressure to provide a compound iv.
[0155] Compound 1d
[0156] To a solution of compound iv in acetonitrile (15 mL) was
added CeCl.sub.3.7H.sub.2O (391 mg, 1.05 mmol, 2.5 equiv) and the
mixture was refluxed for 1 h. The resulting white paste was
filtered and washed with dichloromethane, and the clear suspension
was concentrated under reduced pressure. The resulting residue was
purified by flash column chromatography on silica gel (eluent: 25%
ethyl acetate in hexane to 70% ethyl acetate in hexane) to provide
compound 1d (59 mg, 19.6% total yield from compound ii) as a sticky
oil.
Example 2
SYNTHESIS OF
(R)-3-(4-(2-(4-((S)-3-CHLORO-2-HYDROXYPROPOXY)-3-IODOPHENYL)PROPAN-2-YL)P-
HENOXY)PROPANE-1,2-DIOL (8d)
##STR00064## ##STR00065##
[0158] Compound iii-I
[0159] Compound ii (400 mg, 1.17 mmol, 1.0 equiv), synthesized
according to Example 1, was dissolved in 8 mL of methanol. Sodium
iodide (157.4 mg, 1.05 mmol, 0.9 equiv) and sodium hydroxide (70.4
mg, 1.76 mmol, 1.5 equiv) were added and the solution was cooled to
0.degree. C. A 5.4% aqueous sodium hypochlorite (1612.9 mg, 1.17
mmol, 1 equiv) was then added dropwise over 5 min at 0-3.degree. C.
After 30 min, the pH was kept to 6-7 by adding 10% HCl. The mixture
was extracted with ethyl acetate (2.times.30 mL). The organic layer
was washed with deionized water (2.times.30 mL), dried over
anhydrous magnesium sulfate, filtered, and concentrated under
reduced pressure to provide the title compound
(S)-4-(2-(4-((2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)phenyl)propan-2-yl)--
2-iodophenol (460 mg, 84%) as an oil.
[0160] Compound iv-I
[0161] Sodium hydride (60% dispersion in mineral oil, 12.8 mg, 0.32
mmol, 1.5 equiv) was added slowly to a stirred solution of compound
iii-I (100 mg, 0.21 mmol, 1.0 equiv) in anhydrous dimethyl
formamide (2 mL), at room temperature, and the contents were
stirred under an atmosphere of argon for 10 min. A solution of
(2R)-(-)-glycidyl tosylate 98% (73 mg, 0.32 mmol, 1.5 equiv) in
anhydrous dimethyl formamide (1 mL) was added via syringe, and the
mixture was allowed to react at room temperature for 16 h. Next,
the reaction was quenched by the addition of a saturated solution
of ammonium chloride (10 mL), and the mixture was extracted with
ethyl acetate (2.times.20 mL). The organic layer was washed with
deionized water (2.times.20 mL), dried over anhydrous magnesium
sulfate, filtered and concentrated under reduced pressure. The
resulting residue was purified by flash column chromatography on
silica gel (eluent: 20% ethyl acetate in hexane to 40% ethyl
acetate in hexane) to provide the title compound
(S)-4-((4-(2-(3-iodo-4-(((R)-oxiran-2-yl)methoxy)phenyl)propan-2-yl)pheno-
xy)methyl)-2,2-dimethyl-1,3-dioxolane (106 mg, 94.6%) as a cream
foam.
[0162] Compound 8d
[0163] To a solution of compound iv (130 mg, 0.25 mmol, 1.0 equiv)
in acetonitrile (10 mL) was added CeCl.sub.3.7H.sub.2O (235 mg,
0.63 mmol, 2.5 equiv) and the mixture was refluxed for 16 h. The
resulting white paste was filtered and washed with ethyl acetate,
and the clear suspension was concentrated under reduced pressure.
The resulting residue was purified by flash column chromatography
on silica gel (eluent: 25% ethyl acetate in hexane to 70% ethyl
acetate in hexane) to provide the title compound 8d (110 mg, 84.6%)
as a transparent oil.
Example 3
SYNTHESIS OF
(R)-3-(4-(2-(4-((S)-3-CHLORO-2-HYDROXYPROPOXY)-3-FLUOROPHENYL)PROPAN-2-YL-
)PHENOXY)PROPANE-1,2-DIOL (11d)
##STR00066##
[0165] Compound v-F
[0166] Selectfluor.RTM. (736.9 mg, 2.08 mmol, 0.95 equiv) was added
slowly to a stirred solution of Bisphenol A (500 mg, 2.19 mmol, 1.0
equiv) in anhydrous acetonitrile (12 mL), at room temperature, and
the contents were stirred under an atmosphere of argon for 16 h.
Then, the reaction was quenched by the addition of water (10 mL),
and the mixture was extracted with ethyl acetate (2.times.20 mL).
The organic layer was washed with deionized water (2.times.20 mL),
dried over anhydrous magnesium sulfate, filtered, and concentrated
under reduced pressure. The resulting residue was purified by flash
column chromatography on silica gel (eluent: 1% ethyl acetate in
dichloromethane to 5% ethyl acetate in dichloromethane) to provide
the title compound (300 mg, 55.7%).
[0167] Compound iii-F
[0168] Sodium hydride (60% dispersion in mineral oil, 32.4 mg, 0.81
mmol, 1.0 equiv) was added slowly to a stirred solution of compound
v-F (200 mg, 0.81 mmol, 1.0 equiv) in anhydrous dimethyl formamide
(8 mL), at room temperature, and the contents were stirred under an
atmosphere of argon for 20 min. Compound i (232 mg, 0.81 mmol, 1.0
equiv) was added, and the mixture was allowed to react at
50-60.degree. C. for 16 h. Next, the reaction was quenched by the
addition of a saturated solution of ammonium chloride (10 mL), and
the mixture was extracted with ethyl acetate (2.times.20 mL). The
organic layer was washed with deionized water (2.times.20 mL),
dried over anhydrous magnesium sulfate, filtered, and then
concentrated under reduced pressure. The resulting residue was
purified by flash column chromatography on silica gel (eluent: 5%
ethyl acetate in hexane to 10% ethyl acetate in hexane) to provide
the title compound (130 mg, 44.5%) as an oil.
[0169] Compound iv-F
[0170] Sodium hydride (60% dispersion in mineral oil, 20.4 mg, 0.51
mmol, 1.5 equiv) was added slowly to a stirred solution of compound
ii-F (124 mg, 0.34 mmol, 1.0 equiv) in anhydrous dimethyl formamide
(2 mL), at room temperature, and the contents were stirred under an
atmosphere of argon for 10 min. A solution of (2R)-(-)-glycidyl
tosylate 98% (116.4 mg, 0.51 mmol, 1.5 equiv) in anhydrous dimethyl
formamide (1 mL) was added via syringe, and the mixture was allowed
to react at room temperature for 16 h. Next, the reaction was
quenched by the addition of a saturated solution of ammonium
chloride (10 mL), and the mixture was extracted with ethyl acetate
(2.times.20 mL). The organic layer was washed with deionized water
(2.times.20 mL), dried over anhydrous magnesium sulfate, filtered
and concentrated under reduced pressure. The resulting residue was
purified by flash column chromatography on silica gel (eluent: 20%
ethyl acetate in hexane to 40% ethyl acetate in hexane) to provide
the title compound (70 mg, 49.4%) as a clear foam.
[0171] Compound 11d
[0172] To a solution of compound iv-F (70 mg, 0.17 mmol, 1.0 equiv)
in acetonitrile (5 mL) was added CeCl.sub.3.7H.sub.2O (160.2 mg,
0.43 mmol, 2.5 equiv) and the mixture was refluxed for 16 h. The
resulting white paste was filtered and washed with ethyl acetate,
and the clear suspension was concentrated under reduced pressure.
The resulting residue was purified by flash column chromatography
on silica gel (eluent: 25% ethyl acetate in hexane to 70% ethyl
acetate in hexane) to provide the title compound 11d (43 mg, 61.3%)
as a transparent oil.
Example 4 (FIGS. 1 and 2)
Compound Activity
[0173] The PSA-luciferase (6.1 kb) reporter contains functional
AREs to which AR binds in response to androgen to induce luciferase
activity. LNCaP cells were transfected with the PSA(6.1
kb)-luciferase reporter for 24 h, and then treated with indicated
concentration of 8d (also referred as EPI-iodide or iodinated EPI)
with synthetic androgen, R1881 (1 nM) for 24 h. After 24 h of
incubation with R1881, the cells were harvested, and relative
luciferase activities were determined (FIG. 1A). To determine the
IC.sub.50, treatments were normalized to the predicted maximal
activity induction (in the absence of test compounds, vehicle only)
(FIG. 1B). From a representative experiment, it was determined that
the 8d has an IC.sub.50 of 1.17.+-.0.22 .mu.M for inhibition of AR
transcriptional activity.
[0174] To assess specificity for the AR, parallel experiments were
performed in LNCaP cells with endogenous AR and ectopic expression
of other closely related human hormone receptors such as the
progesterone receptor-beta (PR.beta.), glucocorticoid receptor (GR)
and estrogen receptor-.alpha. (ER).
[0175] To measure effect on AR, after the LNCaP cells were
transfected with the PSA(6.1 kb)-luciferase reporter for 24 h, they
were then treated with DMSO, 5 .mu.M MDV3100, 25 .mu.M Z (also
referred to as EPI-002 or EPI), or 1.9 .mu.M 8d with or without 1
nM R1881 for 24 h (FIG. 2A). Compound 8d strongly inhibited
androgen-induced PSA luciferase activity.
[0176] LNCaP cells were cotransfected with the expression plasmids
for full-length human PR.beta. and the relative reporter
(PRE-luciferase) for 24 h, and then treated with DMSO, 5 .mu.M
MDV3100, 25 .mu.M Z, or 1.9 .mu.M 8d with or without 10 nM
progesterone for 24 h (FIG. 2B). Compound 8d had no effect on the
transcriptional activity of closely related PR.beta..
[0177] FIG. 2C shows GRE-luciferase activity where LNCaP cells were
cotransfected with the expression plasmids for full-length human GR
and the relative reporter (GRE-luciferase) for 24 h, and then
treated with DMSO, 5 .mu.M MDV3100, 25 .mu.M Z, or 1.9 .mu.M 8d
with or without 10 nM dexamethasone for 48 h.
[0178] FIG. 2D shows ERE-luciferase activity where LNCaP cells were
cotransfected with the expression plasmids for full-length human
ERa and the relative reporter for 24 h, and then treated with DMSO,
5 .mu.M MDV3100, 25 .mu.M Z, or 1.9 .mu.M 8d with or without 10 nM
E2 (estradiol) for 24 h. For 8d, 1 .mu.g/mL=1.9 .mu.M in FIGS.
2A-2D.
[0179] Under conditions where compound 8d strongly inhibited
AR-driven PSA-luciferase activity (FIG. 2A), PR-b, GR, or ER.alpha.
activity were not inhibited (FIGS. 2B-2D). These data support that
compound 8d has specificity for the AR.
##STR00067##
Example 5 (FIG. 14)
Compound Activity
[0180] Competitive ligand-binding assays to detect the displacement
of fluorescently labeled ligand from recombinant LBDs (ligand
binding domains) of AR, PR,
[0181] GR and estrogen receptor (ER) by R1881, MDV3100, Z (also
referred to as EPI-002 or EPI) or 8d (also referred to as iodinated
EPI) were performed. FIGS. 14A-14E display competitive
ligand-binding curves to indicate whether R1881, Z, antiandrogens
(enzalutamide, hydroxyflutamide, bicalutamide) or 8d can displace 1
nM fluorescently labeled cognant ligand from recombinant LBDs of
steroid hormone receptors by using fluorescent polarization (mP).
Serial dilution was performed for each test compound. Mixtures were
incubated for 5 h before measurement of fluorescent polarization.
The data shows 8d does not bind to LBDs of AR, PR, GR, and ER.
Example 6 (FIG. 15)
Compound Activity
[0182] Covalent binding experiments of 1d (also referred to as
.sup.123I-EPI) to recombinant protein AR activation function-1
(AF1) was evaluated by SDS-PAGE (FIG. 15). After 6 h incubation at
room temperature, 1d bound to the recombinant protein AR AF1. Less
binding of 1d was observed when AR AF1 was pre-incubated with cold
Z which is thought to bind to the same site as 1d. The SDS-PAGE gel
was stained with Coomassie blue for loading control of the amount
of AF 1 protein. The data demonstrates that 1d binds to AF1 in the
AR NTD.
Example 7 (FIG. 16)
Compound Activity
[0183] Effects of 8d (also referred to as I-EPI-002) on
androgen-dependent proliferation of LNCaP cells treated with R1881
were compared with PC3 and DU145 cell viability by alamarBlue Cell
Viability Assay (FIG. 16). 8d had no effect on the viability of PC3
and DU145 prostate cancer cells that do not express functional AR,
at concentrations that reduced AR-dependent proliferation of LNCaP
cells. FIG. 16 shows PC3 at day 3, LNCaP at day 4, and DU145 at day
3.
Example 8
Compound Activity
[0184] LNCaP cells were transiently transfected with PSA (6.1
kb)-luciferase for 24 h prior to pre-treatment with compounds of
the invention (e.g., compounds 9d, 10d, 11d) ranging in
concentration from 62.5 ng/ml to 1.5 ug/ml for 1 hour before the
addition of vehicle, or synthetic androgen, R1881 (1 nM) to induce
luciferase production. After 24 h of incubation with R1881, the
cells were harvested, and relative luciferase activities were
determined. To determine the IC.sub.50, treatments were normalized
to the predicted maximal activity induction (in the absence of test
compounds, vehicle only) (FIG. 1B).
TABLE-US-00002 TABLE 1 IC.sub.50 values for 9d, 10d, and 11d
(.mu.M) Compound Trial 1 Trial 2 Trial 3 Trial 4 Average 9d 2.07
2.74 2.96 3.36 2.78 +/- 0.47 10d 2.53 2.30 2.81 2.84 2.62 +/- 0.22
11d 5.95 3.60 7.33 5.40 5.57 +/- 1.34
[0185] Although various embodiments of the invention are disclosed
herein, many adaptations and modifications may be made within the
scope of the invention in accordance with the common general
knowledge of those skilled in this art. Such modifications include
the substitution of known equivalents for any aspect of the
invention in order to achieve the same result in substantially the
same way. Numeric ranges are inclusive of the numbers defining the
range. The word "comprising" is used herein as an open-ended term,
substantially equivalent to the phrase "including, but not limited
to", and the word "comprises" has a corresponding meaning. As used
herein, the singular forms "a", "an" and "the" include plural
referents unless the context clearly dictates otherwise. Thus, for
example, reference to "a thing" includes more than one such thing.
Citation of references herein is not an admission that such
references are prior art to the present invention. Any priority
document(s) and all publications, including but not limited to
patents and patent applications, cited in this specification are
incorporated herein by reference as if each individual publication
were specifically and individually indicated to be incorporated by
reference herein and as though fully set forth herein. The
invention includes all embodiments and variations substantially as
hereinbefore described and with reference to the examples and
drawings.
* * * * *