U.S. patent application number 15/169725 was filed with the patent office on 2017-01-05 for novel cannabinoid receptor 2 (cb2) inverse agonists and therapeutic potential for multiple myeloma and osteoporosis bone diseases.
This patent application is currently assigned to University of Pittsburgh - Of the Commonwealth System of Higher Education. The applicant listed for this patent is University of Pittsburgh - Of the Commonwealth System of Higher Education. Invention is credited to Rentian FENG, Xiang-qun XIE, Peng YANG.
Application Number | 20170001949 15/169725 |
Document ID | / |
Family ID | 48695317 |
Filed Date | 2017-01-05 |
United States Patent
Application |
20170001949 |
Kind Code |
A1 |
XIE; Xiang-qun ; et
al. |
January 5, 2017 |
NOVEL CANNABINOID RECEPTOR 2 (CB2) INVERSE AGONISTS AND THERAPEUTIC
POTENTIAL FOR MULTIPLE MYELOMA AND OSTEOPOROSIS BONE DISEASES
Abstract
Cannabinoid receptor-2 inverse antagonists include compounds
represented by Formula IV, or a pharmaceutically acceptable salt
thereof: ##STR00001## wherein: R.sup.1 and R.sup.2 are
independently H, alkyl, or alkenyl; R.sup.3 is alkyl, alkenyl,
aryl, aralkyl, aralkenyl, heterocyclyl, heterocyclylalkyl,
heteroaryl, heteroarylalkyl; R.sup.4 and R.sup.5 are independently
a bond, alkylenyl, or alkenylenyl; each R.sup.6 and R.sup.7 is
independently selected from the group consisting of OH, F, Cl, Br,
I, (C.sub.1-C.sub.6)alkyl, alkoxy, amino, COOH, CONH.sub.2,
SO.sub.3H, PO.sub.3H.sub.2, CN, SH, NO.sub.2 and CF.sub.3; and p
and q are independently 0, 1, 2, 3, 4, or 5. Such compounds may be
used to treat osteoporosis or multiple myeloma.
Inventors: |
XIE; Xiang-qun; (Sewickley,
PA) ; YANG; Peng; (Pittsburgh, PA) ; FENG;
Rentian; (Pittsburgh, PA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
University of Pittsburgh - Of the Commonwealth System of Higher
Education |
Pittsburgh |
PA |
US |
|
|
Assignee: |
University of Pittsburgh - Of the
Commonwealth System of Higher Education
Pittsburgh
PA
|
Family ID: |
48695317 |
Appl. No.: |
15/169725 |
Filed: |
May 31, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14305941 |
Jun 16, 2014 |
9376380 |
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15169725 |
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13715603 |
Dec 14, 2012 |
8772541 |
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14305941 |
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61576041 |
Dec 15, 2011 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07C 233/11 20130101;
C07C 233/63 20130101; C07C 2603/74 20170501; C07D 211/06 20130101;
C07C 233/36 20130101; C07C 233/40 20130101; C07C 233/78 20130101;
C07C 311/20 20130101; C07C 2601/14 20170501; C07C 311/18 20130101;
C07C 233/22 20130101; C07C 233/13 20130101; C07C 311/05 20130101;
C07D 213/56 20130101; C07C 69/78 20130101; C07C 311/29 20130101;
C07D 235/12 20130101; C07D 203/08 20130101; C07C 311/48 20130101;
C07D 295/135 20130101 |
International
Class: |
C07C 233/40 20060101
C07C233/40; C07D 295/135 20060101 C07D295/135 |
Claims
1. A compound according to Formula I ##STR00116## wherein: D and D'
are independently --H, --OH, --OR.sup.a, (C.sub.1-C.sub.6)alkyl or
##STR00117## R.sup.a is H, straight or branched chain
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-; A, B and Q are
each independently (C.sub.1-C.sub.6)alkylene,
(C.sub.2-C.sub.6)alkenylene or (C.sub.2-C.sub.6)alkynylene; e, f
and g independently are integers between 0 and 6 inclusive; and V,
W, X, Y, and Z are each independently a bond, --C(R''').sub.2--,
--CR'''--, --NR'''--, --N--, --O--, --C(O)--, or --S--; wherein: no
two adjacent members of V, W, X, Y, and Z are simultaneously --O--,
--S--, or --NR'''--; R''' is H, --OH, --OR.sup.a, halogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)haloalkoxy, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, --NH.sub.2,
--NH(C.sub.1-C.sub.6)alkyl, --N[(C.sub.1-C.sub.6)alkyl].sub.2,
--CN, (C.sub.3-C.sub.8)heteroaryl,
(C.sub.3-C.sub.8)heterocycloalkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)heteroaryl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkenylene-, or
(C.sub.1-C.sub.6)alkyl-(C.sub.3-C.sub.8)arylene; l, m, n, p and q
independently are integers between 0 and 2 inclusive, wherein at
least one of l, m, n, or p is not 0; represents the option of
having one or more double bonds; wherein any alkyl, alkylene,
alkenylene, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl is
optionally substituted with halogen, oxo, --COOH, --CN, --NO.sub.2,
--OH, --NR.sup.dR.sup.e, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.6)haloalkoxy,
(C.sub.1-C.sub.6)haloalkyl, (C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heteroaryl, (C.sub.3-C.sub.8)heterocycloalkyl, or
(C.sub.3-C.sub.8)aryloxy; and wherein R.sup.d and R.sup.e are each
independently H, straight or branched (C.sub.1-C.sub.6)alkyl,
optionally substituted (C.sub.3-C.sub.8)aryl, optionally
substituted (C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-, and
H.sub.2N(C.sub.1-C.sub.6)alkylene-, or a pharmaceutically
acceptable salt thereof; with the proviso that when each
##STR00118## is independently a phenyl, B.sub.f and Q.sub.g are
both methylene and e is 0, D is not 4-dimethylaminophenyl.
2. The compound according to claim 1, wherein when D is
##STR00119## each of V, W, X, Y and Z is C(R''') and represents
alternating double bonds.
3. The compound according to claim 2, wherein R''' is
(C.sub.3-C.sub.8)heterocycloalkyl.
4. The compound according to claim 3, wherein R''' is oxetanyl,
tetrahydrofuranyl, tetrahydropyranyl, oxepanyl,
tetrahydrothiophenyl, tetrahydrothiopyranyl, 1,3-dioxanyl,
oxazolidinyl, azetidinyl, pyrrolidinyl, piperidinyl, azepinyl,
piperazinyl, morpholinyl, tetrahydrothiopyranyl-1-oxide,
tetrahydrothiopyranyl-1,1-dioxide, pyrrolidinonyl, piperidinonyl,
azepinonyl, piperazidinonyl, oxazidilinonyl, azetidinonyl, or
morpholinonyl.
5. The compound according to claim 1, selected from the following
table: TABLE-US-00008 ##STR00120## ##STR00121## ##STR00122##
##STR00123## ##STR00124## ##STR00125## ##STR00126## ##STR00127##
##STR00128## ##STR00129## ##STR00130## ##STR00131## ##STR00132##
##STR00133## ##STR00134## ##STR00135## ##STR00136## ##STR00137##
##STR00138## ##STR00139## ##STR00140## ##STR00141## ##STR00142##
##STR00143##
6. (canceled)
7. A compound according to Formula I' ##STR00144## wherein: D and
D' are independently --H, --OH, --OR.sup.a, (C.sub.1-C.sub.6)alkyl
or ##STR00145## R.sup.a', R.sup.a'', and R.sup.a''' are
independently selected from the group consisting of H, straight or
branched chain (C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.5)cycloalkyl,
(C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-; A, B and Q are
each independently (C.sub.1-C.sub.6)alkylene,
(C.sub.2-C.sub.6)alkenylene or (C.sub.2-C.sub.6)alkynylene; e, f
and g independently are integers between 0 and 6 inclusive; V, W,
X, Y, and Z are each independently a bond, --C(R''').sub.2--,
--CR'''--, --NR'''--, --N--, --O--, --C(O)--, or --S--; wherein: no
two adjacent members of V, W, X, Y, and Z are simultaneously --O--,
--S--, or --NR'''--; R''' is H, --OH, --OR.sup.a, halogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)haloalkoxy, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, --NH.sub.2,
--NH(C.sub.1-C.sub.6)alkyl, --N[(C.sub.1-C.sub.6)alkyl].sub.2,
--CN, (C.sub.3-C.sub.8)heteroaryl,
(C.sub.3-C.sub.8)heterocycloalkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)heteroaryl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkenylene-, or
(C.sub.1-C.sub.6)alkyl-(C.sub.3-C.sub.8)arylene; l, m, n, p and q
independently are integers between 0 and 2 inclusive, wherein at
least one of l, m, n, or p is not 0; and represents the option of
having one or more double bonds; wherein any alkyl, alkylene,
alkenylene, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl is
optionally substituted with halogen, oxo, --COOH, --CN, --NO.sub.2,
--OH, --NR.sup.dR.sup.e, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.6)haloalkoxy,
(C.sub.1-C.sub.6)haloalkyl, (C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heteroaryl, (C.sub.3-C.sub.8)heterocycloalkyl, or
(C.sub.3-C.sub.8)aryloxy; and wherein R.sup.d and R.sup.e are each
independently H, straight or branched (C.sub.1-C.sub.6)alkyl,
optionally substituted (C.sub.3-C.sub.8)aryl, optionally
substituted (C.sub.3-C.sub.4)aryl(C.sub.1-C.sub.6)alkylene-, and
H.sub.2N(C.sub.1-C.sub.6)alkylene-, or a pharmaceutically
acceptable salt thereof.
8. The compound according to claim 7, wherein: D is H; D' is
phenyl; B and Q are independently (C.sub.1-C.sub.6)alkylene; e is 0
and each of f and g is 1; and each of R.sup.a', R.sup.a''', and
R.sup.a''' is independently selected from the group consisting of
H, straight chain (C.sub.1-C.sub.6)alkyl, and branched chain
(C.sub.1-C.sub.6)alkyl.
9. The compound according to claim 7 selected from the group
consisting of: ##STR00146##
10. A compound according to Formula II ##STR00147## wherein: A, B'
and E are each independently a bond, --C(R''').sub.2--, --CR'''--,
--NR'''--, --N--, --O--, --C(O)--, or --S--; wherein no two
adjacent members of A, B' and E are simultaneously --O--, --S--, or
--NR'''--; h, j and k independently are integers between 0 and 2
inclusive, wherein at least one of h, j or k is not 0; represents
the option of having one or more double bonds; D and D'' are each
independently --C(O), --CH.sub.2C(O)--, (C.sub.1-C.sub.6)alkylene,
--C(O)NH--, or --NHC(O)--; represents the option of having a
C.sub.5-C.sub.6 fused ring optionally having one or more double
bonds; R and R.sub.1 are each independently OH, --OR.sup.a,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl,
(C.sub.3-C.sub.8)heteroaryl, (C.sub.3-C.sub.8)heterocycloalkyl,
(C.sub.3-C.sub.8)cycloalkyl, (C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkylene-; and R.sup.a is H,
straight or branched chain (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.8)cycloalkyl, (C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-; wherein any
alkyl, alkylene, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl
is optionally substituted with one or more of halogen, oxo, --COOH,
--CN, --NO.sub.2, --OH, --NR.sup.dR.sup.e, (C.sub.1-C.sub.6)alkoxy,
or (C.sub.3-C.sub.8)aryloxy; and wherein R.sup.d and R.sup.e are
each independently H, straight or branched (C.sub.1-C.sub.6)alkyl,
optionally substituted
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-, and
H.sub.2N(C.sub.1-C.sub.6)alkylene-, or a pharmaceutically
acceptable salt thereof; or a compound according to Formula IV or a
pharmaceutically acceptable salt thereof: ##STR00148## wherein:
R.sup.1 and R.sup.2 are independently H, alkyl, or alkenyl; R.sup.3
is alkyl, alkenyl, aryl, aralkyl, aralkenyl, heterocyclyl,
heterocyclylalkyl, heteroaryl, heteroarylalkyl; R.sup.4 and R.sup.5
are independently a bond, alkylenyl, or alkenylenyl; each R.sup.6
and R.sup.7 are independently OH, F, Cl, Br, I,
(C.sub.1-C.sub.6)alkyl, alkoxy, amino, COOH, CONH.sub.2, SO.sub.3H,
PO.sub.3H.sub.2, CN, SH, NO.sub.2 or CF.sub.3; and p and q are
independently 0, 1, 2, 3, 4, or 5; with the proviso that where
R.sup.1 and R.sup.2 are both H, R.sup.4 and R.sup.5 are both
methylene, p and q are 0, R.sup.3 is not 4-dimethylaminophenyl.
11. The compound according to claim 10, wherein the compound is a
compound of Formula II and is phenyl and subscripts h, j and k are
each 1.
12. The compound according to claim 10, wherein the compound is a
compound of Formula II and subscripts h and k independently are 0
and subscript j is 1.
13. The compound according to claim 10, that is ##STR00149##
14-22. (canceled)
23. A pharmaceutical composition comprising a therapeutically
effective amount of a compound claim 1, or a pharmaceutically
acceptable salt thereof; and a pharmaceutically acceptable
carrier.
24-28. (canceled)
29. A method for modulating the activity of a cannabinoid
receptor-2 (CB2) in a subject, comprising contacting the CB-2
receptor with a compound of claim 1, or a pharmaceutically
acceptable salt thereof.
30-37. (canceled)
38. The compound according to claim 10, selected from the following
table: TABLE-US-00009 ##STR00150## ##STR00151## ##STR00152##
##STR00153## ##STR00154## ##STR00155## ##STR00156## ##STR00157##
##STR00158## ##STR00159## ##STR00160## ##STR00161## ##STR00162##
##STR00163## ##STR00164## ##STR00165## ##STR00166##
##STR00167##
39. (canceled)
40. A pharmaceutical composition comprising a therapeutically
effective amount of a compound of Formula IV according to claim 10
and a pharmaceutically acceptable carrier.
41. A method for treating multiple myeloma or osteoporosis in a
subject by modulating the activity of a cannabinoid receptor-2
(CB2), comprising administering to the subject a pharmaceutical
composition comprising a therapeutically effective amount of a
compound of Formula IV according to claim 10.
42-47. (canceled)
Description
[0001] This application is a divisional of U.S. patent application
Ser. No. 14/305,941, filed Jun. 16, 2014, which is a divisional of
U.S. patent application Ser. No. 13/715,603, filed Dec. 14, 2012,
now U.S. Pat. No. 8,772,541, which claims the benefit to priority
from U.S. Provisional Patent Application No. 61/576,041, filed Dec.
15, 2011. All of the references listed above are incorporated
herein by reference in their entirety.
FIELD
[0002] The present invention relates to compounds by the regulation
of cannabinoid receptors. More specifically, the compounds are
inverse antagonists of cannabinoid receptor subtype-2, otherwise
known as CB2.
BACKGROUND
[0003] Osteoporosis is a major metabolic bone disease that affects
44 million Americans or 55% of the people 50 years of age or older,
among which 10 million individuals already have this disease and
the rest 34 million more at increased high risk for osteoporosis.
The disease causes a significant amount of morbidity and mortality
in patients and is often diagnosed after a fracture occurs.
[0004] The endocannabinoid system plays an important role in
regulating skeletal remodeling and bone mass [2, 3]. These
physiological processes are implicated to play a role in the
development and progression of osteoporosis. In particular,
CB2-deficient mice show a remarkably accelerated age-related bone
loss, supported by human genetic studies that portray polymorphisms
in CNR2 gene (encoding CB2) as important genetic risk factors for
osteoporosis. However, CB2-mediated bone anabolic action as well as
the underlying mechanisms has not been fully explored. The present
application provides compounds that can be used as probes to study
the mechanisms involved in CB2-mediated regulation of osteoporotic
signaling.
[0005] Multiple myeloma (MM), an incurable cancer of plasma cells
is the second most common hematological malignancy in the United
States. The disease disproportionately affects males over females,
and is more common in the African American population than in
Caucasians. The etiology of MM is unknown, and at present, there is
no cure available, although modern treatment regimens have been
able to slow disease progression in many patients, and have
extended survival rates to about 3-5 years post-diagnosis.
[0006] MM patients present various symptoms, including
hypercalcemia, anemia, renal failure, and impaired production of
non-pathological immunoglobulins. Many patients also endure
persistent bone pain, which typically stems from small fractures in
the bones. Indeed, the hallmark pathology of MM is increased bone
destruction and development of osteolytic lesions, which are
mediated by high osteoclast (OCL) activity and make the patient
more susceptible to bone fractures.
[0007] Previous work from the present inventors revealed that
compounds belonging to the chemical genus shown below modulated
cannabinoid receptor-2 activity. Preliminary biological data
illustrates that this class of compounds to selectively modulate
the CB2 receptor.
[0008] The cannabinoid receptor subtypes CB1 (brain) and CB2
(spleen) are important G-protein coupled receptor targets for
developing new therapeutic agents. Since the discovery of the
cannabinoid (CB) receptors, their endogenous ligands, and enzymes
implicated to play a role in cannabinoid receptor and ligand
biology there has been intensive pharmacological research into the
therapeutic potential of cannabinergic ligands.
[0009] Clinical data related to the therapeutic potential of CB
ligands for the treatment of nausea, glaucoma, cancer, stroke,
pain, neuronal disorders, osteoporosis, multiple sclerosis, and
autoimmune disorders has generated active interest in cannabinoid
research. While most of the research efforts have focused on the
development of ligands targeted to the CB1 receptor, biological
data indicates that CB1 ligands exert undesirable psychotropic side
effects. These side effects have caused public concern. However,
work to design novel CB2 ligands that do not confer psychotropic
side effects associated with modulation of CB1 activity has been
limited, largely due to a lack of information about the three
dimensional structures of the CB receptors and ligand binding
sites.
[0010] The present inventors have used structure-activity
relationship (SAR), studies to explore and define the chemical
space of CB2 receptor that is involved in ligand binding
interactions. Early studies used to define this chemical space
relied on QSAR/NMR methodologies and in-silico docking experiments
to identify a library of chemically diverse scaffolds as the core
pharmacophore for CB2 receptor ligand design.
SUMMARY
[0011] The present invention uses SAR information to develop a new
class compounds that selectively target and modulate CB2 activity,
particularly with an aim to developing small molecule therapeutics
for treating multiple myeloma and osteoporosis. Compounds according
to the present invention also provide alternate therapeutic
candidates to current medications, such as bisphosphonates,
raloxifene, calcitonin and hormone replacement therapy that are
used to treat osteoporosis and are known to exhibit severe adverse
effects which limits their clinical use.
[0012] According to an embodiment of this invention, therefore, is
provided a compound according to Formula I or a pharmaceutically
acceptable salt thereof.
##STR00002##
[0013] For Formula I compounds D and D' are independently --H,
--OH, --OR.sup.a, (C.sub.1-C.sub.6)alkyl or
##STR00003##
and R.sup.a is H, straight or branched chain
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-.
[0014] In Formula I substituent groups A, B and Q are each
independently (C.sub.1-C.sub.6)alkylene,
(C.sub.2-C.sub.6)alkenylene or (C.sub.2-C.sub.6)alkynylene and
subscripts e, f and g independently are integers between 0 and 6
inclusive.
[0015] V, W, X, Y, and Z are each independently a bond,
--C(R''').sub.2--, --CR'''--, --NR'''--, --N--, --O--, --C(O)--, or
--S--, with the proviso that no two adjacent members of V, W, X, Y,
and Z are simultaneously --O--, --S--, or --NR'''--.
[0016] In Formula I, R''' is H, --OH, --OR.sup.a, halogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)haloalkoxy, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, --NH.sub.2,
--NH(C.sub.1-C.sub.6)alkyl, --N[(C.sub.1-C.sub.6)alkyl].sub.2,
--CN, (C.sub.3-C.sub.8)heteroaryl,
(C.sub.3-C.sub.8)heterocycloalkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)heteroaryl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkenylene-, or
(C.sub.1-C.sub.6)alkyl-(C.sub.3-C.sub.8)arylene and subscripts l,
m, n, p and q independently are integers between 0 and 2 inclusive,
with at least one of l, m, n, or p is not 0.
[0017] represents the option of having one or more double bonds.
For compounds that conform to Formula I, any alkyl, alkylene,
alkenylene, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl is
optionally substituted with halogen, oxo, --COOH, --CN, --NO.sub.2,
--OH, --NR.sup.dR.sup.e, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.6)haloalkoxy,
(C.sub.1-C.sub.6)haloalkyl, (C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heteroaryl, (C.sub.3-C.sub.8)heterocycloalkyl, or
(C.sub.3-C.sub.8)aryloxy; with R.sup.d and R.sup.e each
independently being H, straight or branched (C.sub.1-C.sub.6)alkyl,
optionally substituted (C.sub.3-C.sub.8)aryl, optionally
substituted (C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-, and
H.sub.2N(C.sub.1-C.sub.6)alkylene-.
[0018] When each
##STR00004##
in Formula I is independently a phenyl, B.sub.f and Q.sub.g are
both methylene and e is 0 then D is not 4-dimethylaminophenyl
group.
[0019] Exemplary Formula I compounds are those illustrated in the
table below:
##STR00005## ##STR00006## ##STR00007##
[0020] Another embodiment of the invention is a compound according
to Formula I' or a pharmaceutically acceptable salt thereof:
##STR00008##
[0021] In Formula I', D and D' are independently --H, --OH,
--OR.sup.a, (C.sub.1-C.sub.6)alkyl or
##STR00009##
[0022] Substituents R.sup.a', R.sup.a'' and R.sup.a''' are
independently selected from the group consisting of H, straight or
branched chain (C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, and
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-.
[0023] A, B and Q are each independently (C.sub.1-C.sub.6)alkylene,
(C.sub.2-C.sub.6)alkenylene or (C.sub.2-C.sub.6)alkynylene.
[0024] Subscripts e, f and g independently are integers between 0
and 6 inclusive.
[0025] Ring members V, W, X, Y, and Z are each independently a
bond, --C(R''').sub.2--, --CR'''--, --NR'''--, --N--, --O--,
--C(O)--, or --S--, wherein no two adjacent members of V, W, X, Y,
and Z are simultaneously --O--, --S--, or --NR'''--.
[0026] In Formula I', R''' is H, --OH, --OR.sup.a, halogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)haloalkoxy, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, --NH.sub.2,
--NH(C.sub.1-C.sub.6)alkyl, --N[(C.sub.1-C.sub.6)alkyl].sub.2,
--CN, (C.sub.3-C.sub.8)heteroaryl,
(C.sub.3-C.sub.8)heterocycloalkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)heteroaryl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkenylene-, or
(C.sub.1-C.sub.6)alkyl-(C.sub.3-C.sub.8)arylene.
[0027] Subscripts l, m, n, p and q independently are integers
between 0 and 2 inclusive, wherein at least one of l, m, n, or p is
not 0.
[0028] Also in Formula I', the symbol represents the option of
having one or more double bonds.
[0029] In Formula I' compounds, any alkyl, alkylene, alkenylene,
aryl, heteroaryl, cycloalkyl, or heterocycloalkyl is optionally
substituted with halogen, oxo, --COOH, --CN, --NO.sub.2, --OH,
--NR.sup.dR.sup.e, (C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)haloalkoxy, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.3-C.sub.8)aryl, C.sub.3-C.sub.8)heteroaryl,
(C.sub.3-C.sub.8)heterocycloalkyl, or (C.sub.3-C.sub.8)aryloxy; and
R.sup.d and R.sup.e are each independently H, straight or branched
(C.sub.1-C.sub.6)alkyl, optionally substituted
(C.sub.3-C.sub.8)aryl, optionally substituted
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-, and
H.sub.2N(C.sub.1-C.sub.6)alkylene-.
[0030] According to another embodiment is provided a compound
according to Formula II or a pharmaceutically acceptable salt
thereof.
##STR00010##
[0031] According to Formula II, A, B' and E are each independently
a bond, --C(R''').sub.2--, --CR'''--, --NR'''--, --N--, --O--,
--C(O)--, or --S-- and no two adjacent members of A, B' and E are
simultaneously --O--, --S--, or --NR'''--. Subscripts h, j and k
independently are integers between 0 and 2 inclusive, with at least
one of h, j or k not being 0.
[0032] represents the option of having one or more double bonds and
D and D'' are each independently --C(O), --CH.sub.2C(O)--,
(C.sub.1-C.sub.6)alkylene, --C(O)NH--, or --NHC(O)--.
[0033] For Formula II compounds, represents the option of having a
C.sub.5-C.sub.6 fused ring optionally having one or more double
bonds.
[0034] R and R.sub.1 are each independently OH, --OR.sup.a,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl,
(C.sub.3-C.sub.8)heteroaryl, (C.sub.3-C.sub.8)heterocycloalkyl,
(C.sub.3-C.sub.8)cycloalkyl, (C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkylene- and R.sup.a is H,
straight or branched chain (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.8)cycloalkyl, (C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-.
[0035] For compounds according to Formula II, any alkyl, alkylene,
aryl, heteroaryl, cycloalkyl, or heterocycloalkyl is optionally
substituted with one or more of halogen, oxo, --COOH, --CN,
--NO.sub.2, --OH, --NR.sup.dR.sup.e, (C.sub.1-C.sub.6)alkoxy, or
(C.sub.3-C.sub.8)aryloxy; with R.sup.d and R.sup.e each
independently being H, straight or branched (C.sub.1-C.sub.6)alkyl,
optionally substituted
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-, and
H.sub.2N(C.sub.1-C.sub.6)alkylene-.
[0036] Exemplary Formula II compounds include
##STR00011##
[0037] The invention also provides in another embodiment a compound
according to Formula III' or a pharmaceutically acceptable salt
thereof:
##STR00012##
[0038] In Formula III', X is N, N.sup.+, or --CH--.
[0039] R' is H, straight or branched chain (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.8)cycloalkyl, (C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-.
[0040] Q', R and T are each independently selected from a bond,
--C(O)(C.sub.1-C.sub.6)alkylene-, (C.sub.1-C.sub.6)alkyl,
--S(O).sub.2--, --S(O)--, --S(O).sub.2NHR'',
--O--(C.sub.1-C.sub.6-alkylene)-O--, --OC(O)-- and
--(C.sub.1-C.sub.6-alkylene)-OC(O)--.
[0041] G, H, J, L, and M are each independently a bond,
--C(R''').sub.2--, --CR'''--, --NR'''--, --N--, --O--, --C(O)--,
and --S--, wherein no two adjacent members of G, H, J, L, or M are
simultaneously --O--, --S-- or --NR'''--.
[0042] R''' is H, --OH, --OR.sup.a, halogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)haloalkoxy, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, --NH.sub.2,
--NH(C.sub.1-C.sub.6)alkyl, --N[(C.sub.1-C.sub.6)alkyl].sub.2,
--CN, (C.sub.3-C.sub.8)heteroaryl,
(C.sub.3-C.sub.8)heterocycloalkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)heteroaryl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkylene-, or
(C.sub.1-C.sub.6)alkyl-(C.sub.3-C.sub.8)arylene;
[0043] Subscripts l, m, n, p and q independently are integers
between 0 and 2 inclusive, wherein at least one of l, m, n, or p is
not 0. Subscript o is 0 when X is N or CH, and o is 1 when X is
N.sup.+.
[0044] The symbol represents the option of having one or more
double bonds.
[0045] R.sup.a is H, straight or branched chain
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-.
[0046] In Formula III', any alkyl, alkylene, alkenylene, aryl,
heteroaryl, cycloalkyl, or heterocycloalkyl is optionally
substituted with one or more of halogen, oxo, --COOH, --CN,
--NO.sub.2, --OH, --NR.sup.dR.sup.e, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.6)haloalkoxy,
(C.sub.1-C.sub.6)haloalkyl, (C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heteroaryl, (C.sub.3-C.sub.8)heterocycloalkyl, or
(C.sub.3-C.sub.8)aryloxy; R.sup.d and R.sup.e are each
independently H, straight or branched (C.sub.1-C.sub.6)alkyl,
optionally substituted (C.sub.3-C.sub.8)aryl, optionally
substituted (C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-, or
H.sub.2N(C.sub.1-C.sub.6)alkylene-.
[0047] The present invention also provides a compound according to
Formula III or a pharmaceutically acceptable salt thereof.
##STR00013##
[0048] For Formula III compounds X is N, N.sup.+, or --CH--. R' is
H, straight or branched chain (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.8)cycloalkyl, (C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-.
[0049] Substituent groups Q', R and T are each independently
(C.sub.1-C.sub.6)alkyl, --S(O).sub.2--, --S(O)--,
--S(O).sub.2NHR'', --O--(CH.sub.2).sub.x--O--, --OC(O)-- and
(CH.sub.2).sub.x--OC(O)--, while substituent groups G, H, J, L, and
M each independently being a bond, --C(R''').sub.2--, --CR'''--,
--NR'''--, --N--, --O--, --C(O)--, and --S--. For Formula III
compounds no two adjacent members of G, H, J, L, or M are
simultaneously --O--, --S--, or --NR'''--.
[0050] R''' in Formula III is H, --OH, --OR.sup.a, halogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)haloalkoxy, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, --NH.sub.2,
--NH(C.sub.1-C.sub.6)alkyl, --N[(C.sub.1-C.sub.6)alkyl].sub.2,
--CN, (C.sub.3-C.sub.8)heteroaryl,
(C.sub.3-C.sub.8)heterocycloalkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)heteroaryl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkylene-, or
(C.sub.1-C.sub.6)alkyl-(C.sub.3-C.sub.8)arylene.
[0051] Subscripts l, m, n, p and q independently are integers
between 0 and 2 inclusive, and at least one of l, m, n, or p is not
0.
[0052] in Formula IIII represents the option of having one or more
double bonds. For Formula III compounds, R.sup.a is H, straight or
branched chain (C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-.
[0053] Any alkyl, alkylene, alkenylene, aryl, heteroaryl,
cycloalkyl, or heterocycloalkyl in a Formula IIII compound is
optionally substituted with one or more of halogen, oxo, --COOH,
--CN, --NO.sub.2, --OH, --NR.sup.dR.sup.e, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.6)haloalkoxy,
(C.sub.1-C.sub.6)haloalkyl, (C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heteroaryl, (C.sub.3-C.sub.8)heterocycloalkyl, or
(C.sub.3-C.sub.8)aryloxy; with R.sup.d and R.sup.e each
independently being H, straight or branched (C.sub.1-C.sub.6)alkyl,
optionally substituted (C.sub.3-C.sub.8)aryl, optionally
substituted (C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-, or
H.sub.2N(C.sub.1-C.sub.6)alkylene-.
[0054] According to one embodiment substituent X in Formula III is
N and substituents T and R are each independently --S(O).sub.2--
and Q' is (C.sub.1-C.sub.6)alkyl. Alternatively, substituent X is
--CH-- and each of Q', R and T are independently
--O--(CH.sub.2).sub.x--O--, --OC(O)-- or
(CH.sub.2).sub.x--OC(O)--.
[0055] Exemplary Formula III compounds include
##STR00014##
[0056] The present invention also provides a pharmaceutical
composition comprising a therapeutically effective amount of a
Formulae I, II, or III compound, or a pharmaceutically acceptable
salt thereof; and a pharmaceutically acceptable carrier.
[0057] According to another embodiment is provided a method for
treating multiple myeloma or osteoporosis in a subject by
modulating the activity of a cannabinoid receptor-2 (CB2),
comprising administering to the subject a therapeutically effective
amount of a compound of Formulae I, II or IIII. Also provided is a
method for modulating the activity of a cannabinoid receptor-2
(CB2) by contacting a CB2 receptor with a compound according to
Formulae I, II, or III.
[0058] In one embodiment of the invention is provided a compound
according to Formula IV or a pharmaceutically acceptable salt
thereof:
##STR00015##
[0059] According to Formula IV R.sup.1 and R.sup.2 are
independently H, alkyl, or alkenyl. R.sup.3 is alkyl, alkenyl,
aryl, aralkyl, aralkenyl, heterocyclyl, heterocyclylalkyl,
heteroaryl, or heteroarylalkyl.
[0060] Each of R.sup.4 and R.sup.5 in Formula IV are independently
a bond, alkylenyl, or alkenylenyl. For Formula IV compounds each
R.sup.6 and R.sup.7 are independently OH, F, Cl, Br, I,
(C.sub.1-C.sub.6)alkyl, alkoxy, amino, COOH, CONH.sub.2, SO.sub.3H,
PO.sub.3H.sub.2, CN, SH, NO.sub.2 or CF.sub.3. Subscripts p and q
are independently 0, 1, 2, 3, 4, or 5. According to Formula IV,
when R.sup.1 and R.sup.2 are both H, R.sup.4 and R.sup.5 are both
methylene, and p and q are 0, then R.sup.3 is not
4-dimethylaminophenyl.
[0061] The present invention also provides a pharmaceutical
composition comprising a therapeutically effective amount of a
compound of Formula IV and a pharmaceutically acceptable
carrier.
BRIEF DESCRIPTION OF THE DRAWINGS
[0062] FIGS. 1A-1C. Cannabinoid receptor expression in human cancer
cells. (A) The protein levels of CB1 and CB2 in MM cells (U266,
MM.1S) and non-MM cells (pancreatic cancer panc2.03 9, ovarian
cancer OVCAR3 and SKOV3) were determined by Western blot. Cells in
logarithmic phase were harvested and lysed by RIPA buffer
containing protease cocktail inhibitors. To justify the expression
difference of CB1 and CB2, mouse brain tissue lysate was used for
the specific expression of CB1. (B) CB2 expression in various human
MM cell lines and primary CD138+MM cells. Per the suggestion of the
antibody's manufacturer, the human Jurkat cell lysate was used as
the positive control for CB2 expression. (C) CB2 mRNA level was
measured by RT-PCR in the indicated cell lines, using CB2 specific
primers. GAPDH abundance was employed as the loading controls for
immunoblot and PCR assays.
[0063] FIGS. 2A-2E. Inhibitory effects of CB2 ligands on human MM
cell growth. Human MM cell U266 (3.times.10.sup.4 cells per well in
96-well plate) was exposed to the known CB2 ligands (A) or PAM (B)
at indicated concentrations (0-10 mol/L) for 48 h. Cell
proliferation was measured by [.sup.3H]-thymidine uptake assay as
described in Materials and Methods. (C) U266 cells
(3.times.10.sup.4 per well in 96-well plate) were treated with PAM
at indicated doses for 48 h. Cell viability and the viable cell
number per well were determined using trypan blue exclusion assay.
(D) Human chemoresistant myeloma cell lines MM.1R (dexamethasone),
RPMI-8226/LR5 (melphalan) and their respective parent cells MM.1S
and RPMI-8226 were exposed to PAM for 48 h. Cell proliferation was
measured by .sup.3H-thymidine incorporation. The data presented are
the mean.+-.SD of at least 3 independent measurements. (E)
[S.sup.35]-GTP.gamma.S binding assay of different ligands on human
CB2 receptor expressed in CHO cells.
[0064] FIGS. 3A-3D. Inhibitory activity of PAM (compound (1)) in
CB2 knockdown or agonist treated cells. (A) CB2 gene silencing in
MM1.S cells confirmed by Western blot with CB2 antibody. (B) Human
MM1.S cells expressing shRNA vector or expressing the specific
shRNA against human CB2 were treated with vehicle control (white
column), or the indicated concentrations of PAM (black column) for
48 h. Cell proliferation was measured by [.sup.3H]TdR uptake. (C)
and (D) Human U266 cells (4.times.10.sup.5 cells/well, 24 well
plate) were treated with PAM in the absence or presence of CB2
agonist Win55212-2, or CP55940 for 48 h. The cell viability and the
viable cells/well were determined using trypan blue exclusion
assay. (E) [S.sup.35] GTP .gamma.S binding assay to differentiate
the CB2 receptor modulatory activity of ligands.
[0065] FIG. 4. Effects of XIE35 and compound (1) on RANKL-induced
osteoclast formation in bone marrow mononuclear cells (MNC). Upper
panel: Murine bone marrow MNCs (1.times.10.sup.5 per well) were
isolated by separation on Ficoll-Hypaque gradients as described
previously and cultured in the presence of rmRANKL (12.5 ng/mL)
plus rhM-CSF (10 ng/ml), and drug vehicle, XIE35 (1 .mu.M) or
compound (1) (1 .mu.M) was added to appropriate wells. Half-medium
change was performed every other day using drug-containing media
where appropriate. After 5 days, the cells were fixed and stained
for TRAP activity using a TRAP-staining kit (Sigma-Aldrich)
according to the manufacturer's instructions. Lower panel: Human
bone marrow MNCs (1.times.10.sup.5 per well) were treated with
rhRANKL (50 ng/ml) plus rhM-CSF (10 ng/ml), and the respective
compounds as described above. After three weeks, differentiation
into OCLs was assessed by staining with monoclonal antibody 23c6
using a Vectastatin-ABC-AP kit (Vector Laboratories). The antibody
23c6, which recognizes CD51/61 dimer constituting the OCL
vitronectin receptor, was kindly provided by the Bone Biology
Center of our university. Images were obtained with an Olympus IX70
microscope. Arrows designate the typical multinucleated osteoclasts
with 3 or more nuclei.
[0066] FIGS. 5A-5B. Anti-osteoclastogenesis activity of exemplary
compounds. (A) Compounds 61, 84 and 93 inhibit RANKL-induced
osteoclastogenesis in a dose-dependent manner. RAW 264.7 cells
(3.times.10.sup.3 cells/well) were treated with or without RANKL
(15 ng/mL), followed by addition of the indicated concentrations of
61, 84 and 93 for 5 days and stained for TRAP expression. The data
are the mean of three experiments carried out in triplicate. The
bar indicates the SD. (B) Photographs of cells in the test of
compound 93 (original magnification 100.times.).
DETAILED DESCRIPTION
Definitions
[0067] For the purposes of this disclosure and unless otherwise
specified, "a" or "an" means "one or more."
[0068] "CB" is an abbreviation for "cannabinoid receptor."
[0069] "CB1" is an abbreviation for "cannabinoid receptor
subtype-1."
[0070] "CB2" is an abbreviation for "cannabinoid receptor
subtype-2."
[0071] Generally, reference to a certain element such as hydrogen
or H is meant to include all isotopes of that element. For example,
if an R group is defined to include hydrogen or H, it also includes
deuterium and tritium. Hence, isotopically labeled compounds are
within the scope of the invention.
[0072] In general, "substituted" refers to an organic group as
defined below (e.g., an alkyl group) in which one or more bonds to
a hydrogen atom contained therein are replaced by a bond to
non-hydrogen or non-carbon atoms. Substituted groups also include
groups in which one or more bonds to a carbon(s) or hydrogen(s)
atom are replaced by one or more bonds, including double or triple
bonds, to a heteroatom. Thus, a substituted group will be
substituted with one or more substituents, unless otherwise
specified. In some embodiments, a substituted group is substituted
with 1, 2, 3, 4, 5, or 6 substituents. Examples of substituent
groups include: halogens (i.e., F, Cl, Br, and I); hydroxyls;
alkoxy, alkenoxy, alkynoxy, aryloxy, aralkyloxy, heterocyclyloxy,
and heterocyclylalkoxy groups; carbonyls (oxo); carboxyls; esters;
urethanes; oximes; hydroxylamines; alkoxyamines; aralkoxyamines;
thiols; sulfides; sulfoxides; sulfones; sulfonyls; sulfonamides;
amines; N-oxides; hydrazines; hydrazides; hydrazones; azides;
amides; ureas; amidines; guanidines; enamines; imides; isocyanates;
isothiocyanates; cyanates; thiocyanates; imines; nitro groups;
nitriles (i.e., CN); and the like.
[0073] Substituted ring groups such as substituted cycloalkyl,
aryl, heterocyclyl and heteroaryl groups also include rings and
fused ring systems in which a bond to a hydrogen atom is replaced
with a bond to a carbon atom. Therefore, substituted cycloalkyl,
aryl, heterocyclyl and heteroaryl groups may also be substituted
with substituted or unsubstituted alkyl, alkenyl, and alkynyl
groups as defined below.
[0074] Alkyl groups include straight chain and branched alkyl
groups having from 1 to about 20 carbon atoms, and typically from 1
to 12 carbons or, in some embodiments, from 1 to 8, 1 to 6, or 1 to
4 carbon atoms. Alkyl groups further include cycloalkyl groups as
defined below. Examples of straight chain alkyl groups include
those with from 1 to 8 carbon atoms such as methyl, ethyl,
n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups.
Examples of branched alkyl groups include, but are not limited to,
isopropyl, iso-butyl, sec-butyl, tert-butyl, neopentyl, isopentyl,
and 2,2-dimethylpropyl groups. Representative substituted alkyl
groups may be substituted one or more times with substituents such
as those listed above.
[0075] Cycloalkyl groups are cyclic alkyl groups such as, but not
limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, and cyclooctyl groups. In some embodiments, the
cycloalkyl group has 3 to 8 ring members, whereas in other
embodiments the number of ring carbon atoms range from 3 to 5, 3 to
6, or 3 to 7. Cycloalkyl groups further include mono-, bicyclic and
polycyclic ring systems, such as, for example bridged cycloalkyl
groups as described below, and fused rings, such as, but not
limited to, decalinyl, and the like. In some embodiments,
polycyclic cycloalkyl groups have three rings. Substituted
cycloalkyl groups may be substituted one or more times with,
non-hydrogen and non-carbon groups as defined above. However,
substituted cycloalkyl groups also include rings that are
substituted with straight or branched chain alkyl groups as defined
above. Representative substituted cycloalkyl groups may be
mono-substituted or substituted more than once, such as, but not
limited to, 2,2-, 2,3-, 2,4-2,5- or 2,6-disubstituted cyclohexyl
groups, which may be substituted with substituents such as those
listed above.
[0076] Bridged cycloalkyl groups are cycloalkyl groups in which two
or more hydrogen atoms are replaced by an alkylene bridge, wherein
the bridge can contain 2 to 6 carbon atoms if two hydrogen atoms
are located on the same carbon atom, or 1 to 5 carbon atoms, if the
two hydrogen atoms are located on adjacent carbon atoms, or 2 to 4
carbon atoms if the two hydrogen atoms are located on carbon atoms
separated by 1 or 2 carbon atoms. Bridged cycloalkyl groups can be
bicyclic, such as, for example bicyclo[2.1.1]hexane, or tricyclic,
such as, for example, adamantyl. Representative bridged cycloalkyl
groups include bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl,
bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[3.2.2]nonyl,
bicyclo[3.3.1]nonyl, bicyclo[3.3.2]decanyl, adamantyl,
noradamantyl, bornyl, or norbornyl groups. Substituted bridged
cycloalkyl groups may be substituted one or more times with
non-hydrogen and non-carbon groups as defined above. Representative
substituted bridged cycloalkyl groups may be mono-substituted or
substituted more than once, such as, but not limited to, mono-, di-
or tri-substituted adamantyl groups, which may be substituted with
substituents such as those listed above.
[0077] Cycloalkylalkyl groups are alkyl groups as defined above in
which a hydrogen or carbon bond of an alkyl group is replaced with
a bond to a cycloalkyl group as defined above. In some embodiments,
cycloalkylalkyl groups have from 4 to 20 carbon atoms, 4 to 16
carbon atoms, and typically 4 to 10 carbon atoms. Substituted
cycloalkylalkyl groups may be substituted at the alkyl, the
cycloalkyl or both the alkyl and cycloalkyl portions of the group.
Representative substituted cycloalkylalkyl groups may be
mono-substituted or substituted more than once, such as, but not
limited to, mono-, di- or tri-substituted with substituents such as
those listed above.
[0078] Alkenyl groups include straight and branched chain and
cycloalkyl groups as defined above, except that at least one double
bond exists between two carbon atoms. Thus, alkenyl groups have
from 2 to about 20 carbon atoms, and typically from 2 to 12 carbons
or, in some embodiments, from 2 to 8, 2 to 6, or 2 to 4 carbon
atoms. In some embodiments, alkenyl groups include cycloalkenyl
groups having from 4 to 20 carbon atoms, 5 to 20 carbon atoms, 5 to
10 carbon atoms, or even 5, 6, 7, or 8 carbon atoms. Examples
include, but are not limited to vinyl, allyl,
--CH.dbd.CH(CH.sub.3), --CH.dbd.C(CH.sub.3).sub.2,
--C(CH.sub.3).dbd.CH.sub.2, --C(CH.sub.3).dbd.CH(CH.sub.3),
--C(CH.sub.2CH.sub.3).dbd.CH.sub.2, cyclohexenyl, cyclopentenyl,
cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl, among
others. Representative substituted alkenyl groups may be
mono-substituted or substituted more than once, such as, but not
limited to, mono-, di- or tri-substituted with substituents such as
those listed above.
[0079] Cycloalkenylalkyl groups are alkyl groups as defined above
in which a hydrogen or carbon bond of the alkyl group is replaced
with a bond to a cycloalkenyl group as defined above. Substituted
cycloalkylalkenyl groups may be substituted at the alkyl, the
cycloalkenyl or both the alkyl and cycloalkenyl portions of the
group. Representative substituted cycloalkenylalkyl groups may be
substituted one or more times with substituents such as those
listed above.
[0080] Alkynyl groups include straight and branched chain alkyl
groups, except that at least one triple bond exists between two
carbon atoms. Thus, alkynyl groups have from 2 to about 20 carbon
atoms, and typically from 2 to 12 carbons or, in some embodiments,
from 2 to 8, 2 to 6, or 2 to 4 carbon atoms. Examples include, but
are not limited to --C.ident.CH, --C.ident.C(CH.sub.3),
--C.ident.C(CH.sub.2CH.sub.3), --CH.sub.2C.ident.CH,
--CH.sub.2C.ident.C(CH.sub.3), and
--CH.sub.2C.ident.C(CH.sub.2CH.sub.3), among others. Representative
substituted alkynyl groups may be mono-substituted or substituted
more than once, such as, but not limited to, mono-, di- or
tri-substituted with substituents such as those listed above.
[0081] Aryl groups are cyclic aromatic hydrocarbons that do not
contain heteroatoms. Aryl groups include monocyclic, bicyclic and
polycyclic ring systems. Thus, aryl groups include, but are not
limited to, phenyl, azulenyl, heptalenyl, biphenylenyl, indacenyl,
fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl,
chrysenyl, biphenyl, anthracenyl, indenyl, indanyl, pentalenyl, and
naphthyl groups. In some embodiments, aryl groups contain 6-14
carbons, and in others from 6 to 12 or even 6-10 carbon atoms in
the ring portions of the groups. Although the phrase "aryl groups"
includes groups containing fused rings, such as fused
aromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl,
and the like), it does not include aryl groups that have other
groups, such as alkyl or halo groups, bonded to one of the ring
members. Rather, groups such as tolyl are referred to as
substituted aryl groups. Representative substituted aryl groups may
be mono-substituted or substituted more than once. For example,
monosubstituted aryl groups include, but are not limited to, 2-,
3-, 4-, 5-, or 6-substituted phenyl or naphthyl groups, which may
be substituted with substituents such as those listed above.
[0082] Aralkyl groups are alkyl groups as defined above in which a
hydrogen or carbon bond of an alkyl group is replaced with a bond
to an aryl group as defined above. In some embodiments, aralkyl
groups contain 7 to 20 carbon atoms, 7 to 14 carbon atoms or 7 to
10 carbon atoms. Substituted aralkyl groups may be substituted at
the alkyl, the aryl or both the alkyl and aryl portions of the
group. Representative aralkyl groups include but are not limited to
benzyl and phenethyl groups and fused (cycloalkylaryl)alkyl groups
such as 4-ethyl-indanyl. Representative substituted aralkyl groups
may be substituted one or more times with substituents such as
those listed above.
[0083] Heterocyclyl groups include aromatic (also referred to as
heteroaryl) and non-aromatic ring compounds containing 3 or more
ring members, of which one or more is a heteroatom such as, but not
limited to, N, O, and S. In some embodiments, heterocyclyl groups
include 3 to 20 ring members, whereas other such groups have 3 to
6, 3 to 10, 3 to 12, or 3 to 15 ring members. Heterocyclyl groups
encompass unsaturated, partially saturated and saturated ring
systems, such as, for example, imidazolyl, imidazolinyl and
imidazolidinyl groups. The phrase "heterocyclyl group" includes
fused ring species including those comprising fused aromatic and
non-aromatic groups, such as, for example, benzotriazolyl,
2,3-dihydrobenzo[1,4]dioxinyl, and benzo[1,3]dioxolyl. The phrase
also includes bridged polycyclic ring systems containing a
heteroatom such as, but not limited to, quinuclidyl. However, the
phrase does not include heterocyclyl groups that have other groups,
such as alkyl, oxo or halo groups, bonded to one of the ring
members. Rather, these are referred to as "substituted heterocyclyl
groups". Heterocyclyl groups include, but are not limited to,
aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl,
pyrazolidinyl, thiazolidinyl, tetrahydrothiophenyl,
tetrahydrofuranyl, dioxolyl, furanyl, thiophenyl, pyrrolyl,
pyrrolinyl, imidazolyl, imidazolinyl, pyrazolyl, pyrazolinyl,
triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl,
thiazolinyl, isothiazolyl, thiadiazolyl, oxadiazolyl, piperidyl,
piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl,
tetrahydrothiopyranyl, oxathiane, dioxyl, dithianyl, pyranyl,
pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl,
dihydropyridyl, dihydrodithiinyl, dihydrodithionyl,
homopiperazinyl, quinuclidyl, indolyl, indolinyl, isoindolyl,
azaindolyl (pyrrolopyridyl), indazolyl, indolizinyl,
benzotriazolyl, benzimidazolyl, benzofuranyl, benzothiophenyl,
benzthiazolyl, benzoxadiazolyl, benzoxazinyl, benzodithiinyl,
benzoxathiinyl, benzothiazinyl, benzoxazolyl, benzothiazolyl,
benzothiadiazolyl, benzo[1,3]dioxolyl, pyrazolopyridyl,
imidazopyridyl (azabenzimidazolyl), triazolopyridyl,
isoxazolopyridyl, purinyl, xanthinyl, adeninyl, guaninyl,
quinolinyl, isoquinolinyl, quinolizinyl, quinoxalinyl,
quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl, pteridinyl,
thianaphthalenyl, dihydrobenzothiazinyl, dihydrobenzofuranyl,
dihydroindolyl, dihydrobenzodioxinyl, tetrahydroindolyl,
tetrahydroindazolyl, tetrahydrobenzimidazolyl,
tetrahydrobenzotriazolyl, tetrahydropyrrolopyridyl,
tetrahydropyrazolopyridyl, tetrahydroimidazopyridyl,
tetrahydrotriazolopyridyl, and tetrahydroquinolinyl groups.
Representative substituted heterocyclyl groups may be
mono-substituted or substituted more than once, such as, but not
limited to, pyridyl or morpholinyl groups, which are 2-, 3-, 4-,
5-, or 6-substituted, or disubstituted with various substituents
such as those listed above.
[0084] Heteroaryl groups are aromatic ring compounds containing 5
or more ring members, of which, one or more is a heteroatom such
as, but not limited to, N, O, and S. Heteroaryl groups include, but
are not limited to, groups such as pyrrolyl, pyrazolyl, triazolyl,
tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl, pyridazinyl,
pyrimidinyl, pyrazinyl, thiophenyl, benzothiophenyl, furanyl,
benzofuranyl, indolyl, azaindolyl (pyrrolopyridyl), indazolyl,
benzimidazolyl, imidazopyridyl (azabenzimidazolyl),
pyrazolopyridyl, triazolopyridyl, benzotriazolyl, benzoxazolyl,
benzothiazolyl, benzothiadiazolyl, imidazopyridyl,
isoxazolopyridyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl,
guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl,
quinoxalinyl, and quinazolinyl groups. Although the phrase
"heteroaryl groups" includes fused ring compounds such as indolyl
and 2,3-dihydro indolyl, the phrase does not include heteroaryl
groups that have other groups bonded to one of the ring members,
such as alkyl groups. Rather, heteroaryl groups with such
substitution are referred to as "substituted heteroaryl groups."
Representative substituted heteroaryl groups may be substituted one
or more times with various substituents such as those listed
above.
[0085] Heterocyclylalkyl groups are alkyl groups as defined above
in which a hydrogen or carbon bond of an alkyl group is replaced
with a bond to a heterocyclyl group as defined above. Substituted
heterocyclylalkyl groups may be substituted at the alkyl, the
heterocyclyl or both the alkyl and heterocyclyl portions of the
group. Representative heterocyclyl alkyl groups include, but are
not limited to, 4-ethyl-morpholinyl, 4-propylmorpholinyl,
furan-2-yl methyl, furan-3-yl methyl, pyridine-3-yl methyl,
tetrahydrofuran-2-yl ethyl, and indol-2-yl propyl. Representative
substituted heterocyclylalkyl groups may be substituted one or more
times with substituents such as those listed above.
[0086] Heteroaralkyl groups are alkyl groups as defined above in
which a hydrogen or carbon bond of an alkyl group is replaced with
a bond to a heteroaryl group as defined above. Substituted
heteroaralkyl groups may be substituted at the alkyl, the
heteroaryl or both the alkyl and heteroaryl portions of the group.
Representative substituted heteroaralkyl groups may be substituted
one or more times with substituents such as those listed above.
[0087] Groups described herein having two or more points of
attachment (i.e., divalent, trivalent, or polyvalent) within the
compound of the invention are designated by use of the suffix,
"ene." For example, divalent alkyl groups are alkylene groups,
divalent aryl groups are arylene groups, divalent heteroaryl groups
are divalent heteroarylene groups, and so forth. Substituted groups
having a single point of attachment to the compound of the
invention are not referred to using the "ene" designation. Thus,
e.g., chloroethyl is not referred to herein as chloroethylene.
[0088] Alkoxy groups are hydroxyl groups (--OH) in which the bond
to the hydrogen atom is replaced by a bond to a carbon atom of a
substituted or unsubstituted alkyl group as defined above. Examples
of linear alkoxy groups include but are not limited to methoxy,
ethoxy, propoxy, butoxy, pentoxy, hexoxy, and the like. Examples of
branched alkoxy groups include but are not limited to isopropoxy,
sec-butoxy, tert-butoxy, isopentoxy, isohexoxy, and the like.
Examples of cycloalkoxy groups include but are not limited to
cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and
the like. Representative substituted alkoxy groups may be
substituted one or more times with substituents such as those
listed above.
[0089] The terms "aryloxy" and "arylalkoxy" refer to, respectively,
a substituted or unsubstituted aryl group bonded to an oxygen atom
and a substituted or unsubstituted aralkyl group bonded to the
oxygen atom at the alkyl. Examples include but are not limited to
phenoxy, naphthyloxy, and benzyloxy. Representative substituted
aryloxy and arylalkoxy groups may be substituted one or more times
with substituents such as those listed above.
[0090] The term "amine" (or "amino") as used herein refers to
--NHR.sup.4 and --NR.sup.5R.sup.6 groups, wherein R.sup.4, R.sup.5
and R.sup.6 are independently hydrogen, or a substituted or
unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl,
heterocyclylalkyl or heterocyclyl group as defined herein. In some
embodiments, the amine is NH.sub.2, methylamino, dimethylamino,
ethylamino, diethylamino, propylamino, isopropylamino, phenylamino,
or benzylamino.
[0091] The term "amide" refers to a --NR'R''C(O)-- group wherein R'
and R'' each independently refer to a hydrogen,
(C.sub.1-C.sub.5)alkyl, or (C.sub.3-C.sub.6)aryl.
[0092] The term `nitrile or cyano" can be used interchangeably and
refer to a --CN group which is bound to a carbon atom of a
heteroaryl ring, aryl ring and a heterocycloalkyl ring.
[0093] The substituent --CO.sub.2H, may be replaced with
bioisosteric replacements such as:
##STR00016##
and the like, wherein R has the same definition as R' and R'' as
defined herein. See, e.g., THE PRACTICE OF MEDICINAL CHEMISTRY
(Academic Press: New York, 1996), at page 203.
[0094] Those of skill in the art will appreciate that compounds of
the invention may exhibit the phenomena of tautomerism,
conformational isomerism, geometric isomerism and/or optical
isomerism. As the formula drawings within the specification and
claims can represent only one of the possible tautomeric,
conformational isomeric, optical isomeric or geometric isomeric
forms, it should be understood that the invention encompasses any
tautomeric, conformational isomeric, optical isomeric and/or
geometric isomeric forms of the compounds having one or more of the
utilities described herein, as well as mixtures of these various
different forms.
[0095] "Tautomers" refers to isomeric forms of a compound that are
in equilibrium with each other. The concentrations of the isomeric
forms will depend on the environment the compound is found in and
may be different depending upon, for example, whether the compound
is a solid or is in an organic or aqueous solution. For example, in
aqueous solution, pyrazoles may exhibit the following isomeric
forms, which are referred to as tautomers of each other:
##STR00017##
[0096] As readily understood by one skilled in the art, a wide
variety of functional groups and other structures may exhibit
tautomerism, and all tautomers of compounds as described herein are
within the scope of the present invention.
[0097] Stereoisomers of compounds, also known as "optical isomers,"
include all chiral, diastereomeric, and racemic forms of a
structure, unless the specific stereochemistry is expressly
indicated. Thus, compounds used in the present invention include
enriched or resolved optical isomers at any or all asymmetric atoms
as are apparent from the depictions. Both racemic and
diastereomeric mixtures, as well as the individual optical isomers
can be isolated or synthesized so as to be substantially free of
their enantiomeric or diastereomeric partners, and these are all
within the scope of the invention.
[0098] "A pharmaceutically acceptable carrier" is a phrase that
denotes a carrier such as but not limited to a diluent, an
excipient, a wetting agent, a buffering agent, a suspending agent,
a lubricating agent, an adjuvant, a vehicle, a delivery system, an
emulsifier, a disintegrant, an absorbent, a preservative, a
surfactant, a colorant, a flavorant, a sweetener, or a mixture of
any two or more thereof. Pharmaceutically acceptable excipients and
carriers are generally known and, hence, are included in the
instant invention. Such materials are described, for example, in
REMINGTON: THE SCIENCE AND PRACTICE OF PHARMACY, 21.sup.st ed., The
University of Philadelphia (2005).
[0099] Pharmaceutical compositions and medicaments may be prepared
by mixing one or more compounds of the invention, prodrugs thereof,
pharmaceutically acceptable salts thereof, stereoisomers thereof,
tautomers thereof, or solvates thereof, with pharmaceutically
acceptable carriers, excipients, binders, diluents or the like to
prevent and treat disorders associated with cannabinoid receptors.
The compounds and compositions of the invention may be used to
prepare formulations and medicaments that prevent or treat a
variety of disorders associated with cannabinoid receptors, and
described herein. For example, disorders and diseases such as
obesity, smoking addiction, cardimetabolic risk factors, and other
disorder and diseases associated with the central nervous system.
Such compositions can be in the form of, for example, granules,
powders, tablets, capsules, syrup, suppositories, injections,
emulsions, elixirs, suspensions or solutions. The instant
compositions can be formulated for various routes of
administration, for example, by oral, parenteral, topical, rectal,
nasal, vaginal administration, or via implanted reservoir.
Parenteral or systemic administration includes, but is not limited
to, subcutaneous, intravenous, intraperitoneally, intramuscular,
intra-articular, intrasynovial, intrasternal, intrathecal,
intralesional and intracranial injections. The following dosage
forms are given by way of example and should not be construed as
limiting the instant invention.
[0100] Pharmaceutically acceptable salts of the invention compounds
are considered within the scope of the present invention. The
compound of the invention has a number of basic nitrogen groups,
and as such, pharmaceutically acceptable salts can be formed with
inorganic acids (such as hydrochloric acid, hydroboric acid, nitric
acid, sulfuric acid, and phosphoric acid), organic acids (e.g.
formic acid, acetic acid, trifluoroacetic acid, fumaric acid,
oxalic acid, tartaric acid, lactic acid, maleic acid, citric acid,
succinic acid, malic acid, methanesulfonic acid, benzenesulfonic
acid, and p-toluenesulfonic acid) or acidic amino acids (such as
aspartic acid and glutamic acid). The compounds of the present
invention may have acidic substituent groups, and in such cases, it
can form salts with metals, such as alkali and earth alkali metals
(e.g. Na.sup.+, Li.sup.+, K.sup.+, Ca.sup.2+, Mg.sup.2+,
Zn.sup.2+), organic amines (e.g. ammonia, trimethylamine,
triethylamine, pyridine, picoline, ethanolamine, diethanolamine,
triethanolamine) or basic amino acids (e.g. arginine, lysine and
ornithine).
[0101] Certain compounds within the scope of the invention are
derivatives referred to as prodrugs. The expression "prodrug"
denotes a derivative of a known direct acting drug, e.g. esters and
amides, which derivative has enhanced delivery characteristics and
therapeutic value as compared to the drug, and is transformed into
the active drug by an enzymatic or chemical process; see Notari, R.
E., "Theory and Practice of Prodrug Kinetics," Methods in
Enzymology 112: 309-23 (1985); Bodor, N., "Novel Approaches in
Prodrug Design," Drugs of the Future 6: 165-82 (1981); and
Bundgaard, H., "Design of Prodrugs: Bioreversible-Derivatives for
Various Functional Groups and Chemical Entities," in DESIGN OF
PRODRUGS (H. Bundgaard, ed.), Elsevier (1985), Goodman and Gilmans,
THE PHARMACOLOGICAL BASIS OF THERAPEUTICS, 8th ed., McGraw-Hill
(1992).
[0102] For oral, buccal, and sublingual administration, powders,
suspensions, granules, tablets, pills, capsules, gelcaps, and
caplets are acceptable as solid dosage forms. These can be
prepared, for example, by mixing one or more compounds of the
instant invention, or pharmaceutically acceptable salts or
tautomers thereof, with at least one additive such as a starch or
other additive. Suitable additives are sucrose, lactose, cellulose
sugar, mannitol, maltitol, dextran, starch, agar, alginates,
chitins, chitosans, pectins, tragacanth gum, gum arabic, gelatins,
collagens, casein, albumin, synthetic or semi-synthetic polymers or
glycerides. Optionally, oral dosage forms can contain other
ingredients to aid in administration, such as an inactive diluent,
or lubricants such as magnesium stearate, or preservatives such as
paraben or sorbic acid, or anti-oxidants such as ascorbic acid,
tocopherol or cysteine, a disintegrating agent, binders,
thickeners, buffers, sweeteners, flavoring agents or perfuming
agents. Tablets and pills may be further treated with suitable
coating materials known in the art.
Compounds
[0103] The compounds represented by Formulae I, I', II, III, III',
and VI (below) are potent modulators of cannabinoid receptor-2
(CB2). As stated above, the endocannabinoid system, particularly
CB2, plays an important role in various physiological processes,
including regulation of skeletal remodeling and bone mass. These
physiological processes are implicated to play a role in the
development and progression of osteoporosis. While the
physiological role of endocannabinoid system in multiple myeloma
(MM), is not clear, cell based studies using Formulae I, I', II,
III, III', and IV compounds have revealed an anti-MM activity.
[0104] In one aspect, compounds represented by Formula I, II, and
III, or their pharmaceutically acceptable salts are provided:
##STR00018##
[0105] For Formula I compounds D is H, --OH, --OR.sup.a,
(C.sub.1-C.sub.6)alkyl or
##STR00019##
In one embodiment, D is
##STR00020##
and is an aromatic phenyl group that can be optionally substituted
by one, two, or three substituent groups. represents the point of
attachment of the ring to the Formula I scaffold. According to
Formula I, D can be an --OR.sup.a, or a (C.sub.1-C.sub.6)alkyl
group. When D is OR.sup.a, however, R.sup.a is selected from the
group consisting of hydrogen, straight or branched chain
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-. According to the
above formulas, A, B and Q in Formula I are each independently
(C.sub.1-C.sub.6)alkylene, (C.sub.2-C.sub.6)alkenylene and
(C.sub.2-C.sub.6)alkynylene, while subscripts e, f and g
independently are integers between 0 and 6 inclusive.
[0106] For compounds that conform to Formula I, substituent groups
V, W, X, Y, or Z are each independently a bond, --C(R''').sub.2--,
--CR'''--, --NR'''--, --N--, --O--, --C(O)--, and --S--. However,
no two adjacent members of V, W, X, Y, or Z in Formula I are
simultaneously --O--, --S--, or --NR'''--. According to Formula I
compounds, V, W, X, Y, and Z are each independently a bond,
--C(R''').sub.2--, --CR'''--, --NR'''--, --N--, --O--, --C(O)--, or
--S--. Compounds according to Formula I encompass, therefore,
species in which one, two or each of the three ring structures
represents a heteroaryl group, a heterocycle group, a cycloalkyl
group or an aryl ring. Exemplary of heteroaryl and heterocycle
rings are oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, oxepanyl,
tetrahydrothiophenyl, tetrahydrothiopyranyl, 1,3-dioxanyl,
oxazolidinyl, azetidinyl, pyrrolidinyl, piperidinyl, azepinyl,
piperazinyl, morpholinyl, tetrahydrothiopyranyl-1-oxide,
tetrahydrothiopyranyl-1, 1-dioxide, pyrrolidinonyl, piperidinonyl,
azepinonyl, piperazidinonyl, oxazidilinonyl, azetidinonyl, or
morpholinonyl. When any of V, W, X, Y, or Z is a --C(R''').sub.2--,
--CR'''--, --NR'''-- group, substituent R''' is H, OH, OR.sup.a,
halogen, (C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)haloalkoxy, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, --NH.sub.2,
--NH(C.sub.1-C.sub.6)alkyl, --N[(C.sub.1-C.sub.6)alkyl].sub.2,
--CN, (C.sub.3-C.sub.8)heteroaryl,
(C.sub.3-C.sub.8)heterocycloalkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)heteroaryl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkenylene-, or
(C.sub.1-C.sub.6)alkyl-(C.sub.3-C.sub.8)arylene.
[0107] Subscripts l, m, n, p and q independently are integers
between 0 and 2 inclusive and at least one of l, m, n, p, or q is
not zero (0). To accommodate for the presence of aromatic and
non-aromatic ring systems, Formula I recites to represent the
option of having one or more double bonds within a ring system.
Further, for compounds that conform to Formula I, any alkyl,
alkylene, alkenylene, aryl, heteroaryl, cycloalkyl, or
heterocycloalkyl is optionally substituted with one or more
halogen, oxo, --COOH, --CN, --NO.sub.2, --OH, --NR.sup.dR.sup.e,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)haloalkoxy, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.3-C.sub.8)aryl, (C.sub.3-C.sub.8)heteroaryl,
(C.sub.3-C.sub.8)heterocycloalkyl, or (C.sub.3-C.sub.8)aryloxy;
with each of R.sup.d and R.sup.e being independently H, straight or
branched (C.sub.1-C.sub.6)alkyl, optionally substituted
(C.sub.3-C.sub.8)aryl, optionally substituted
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-, or
H.sub.2N(C.sub.1-C.sub.6)alkylene-. In one embodiment, the compound
of Formula I is subject to the proviso that where each
##STR00021##
are independently phenyl, B.sub.f and Q.sub.g are both methylene,
and subscript e is 0, D is a 4-dimethylaminophenyl. However, the
methods of treatment described below are not necessarily subject to
this proviso.
[0108] In one aspect, the compounds of Formula I also include the
compounds of Formula IV:
##STR00022##
[0109] For Formula IV compounds, R.sup.1 and R.sup.2 are
independently H, alkyl, or alkenyl and substituent R.sup.3 can be
alkyl, alkenyl, aryl, aralkyl, aralkenyl, heterocyclyl,
heterocyclylalkyl, heteroaryl, or heteroarylalkyl. In one
embodiment of Formula IV, R.sup.4 and R.sup.5 are independently a
bond, alkylenyl, or alkenylenyl and each R.sup.6 and R.sup.7 is
independently H, OH, F, Cl, Br, I, alkoxy, amino, --COOH,
--C(O)NH.sub.2, SO.sub.3H, PO.sub.3H.sub.2, --CN, --SH, --NO.sub.2,
or CF.sub.3 groups. For Formula IV compounds, subscripts p and q
are independently 0, 1, 2, 3, 4, or 5. In some embodiments, the
compounds of Formula IV are subject to the proviso that when
R.sup.1 and R.sup.2 are both H, R.sup.4 and R.sup.5 are both
methylene, and subscripts p and q are 0, R.sup.3 in Formula IA is
not 4-dimethylaminophenyl.
[0110] In some embodiments of Formula IV, R.sup.3 is a substituted
phenyl ring belonging to Formula B, with each R.sup.8 being
independently selected from OH, F, Cl, Br, I,
(C.sub.1-C.sub.6)alkyl, alkoxy, amino, --COOH, --C(O)NH.sub.2,
SO.sub.3H, PO.sub.3H.sub.2, --CN, --SH, --NO.sub.2, or CF.sub.3 and
t is 0, 1, 2, 3, 4 or 5.
[0111] In one embodiment of Formula IV, R.sup.1 and R.sup.2 are
independently a H or a C.sub.1-C.sub.8 alkyl group, substituent
R.sup.3 is alkyl, aryl, or aralkyl, where alkyl has from 1-8
carbons and substituent groups R.sup.4 and R.sup.5 are
independently a bond, C.sub.1-C.sub.8 alkylenyl, or C.sub.1-C.sub.8
alkenylenyl. According to this embodiment, each R.sup.6 and R.sup.7
is independently OH, F, Cl, Br, I, C.sub.1-C.sub.8 alkoxy, or
amino, subscripts p, q, and t are independently 0, 1, or 2 and each
R.sup.8 is independently OH, F, Cl, Br, I, alkoxy, amino.
[0112] For certain Formula IV compounds, R.sup.1 and R.sup.2
independently are H, R.sup.4 and R.sup.5 are both a bond,
methylene, or ethylene and R.sup.3 is
##STR00023##
For these compounds each R.sup.8 is independently OH, F, Cl, Br, I,
methoxy, trifluoromethoxy, NH.sub.2, dimethylamino, or diethylamino
group and p, q, and t are independently 0, 1, or 2.
[0113] Exemplary Formula I compounds without limitation are those
shown below in Table 1:
TABLE-US-00001 TABLE 1 ##STR00024## ##STR00025## ##STR00026##
##STR00027## ##STR00028## ##STR00029## ##STR00030## ##STR00031##
##STR00032## ##STR00033## ##STR00034## ##STR00035## ##STR00036##
##STR00037## ##STR00038## ##STR00039## ##STR00040## ##STR00041##
##STR00042##
[0114] As described generally above, the invention also provides
compounds according to Formula I':
##STR00043##
[0115] In some embodiments, the Formula I' compound is one where D
is H; D' is phenyl; B and Q are independently
(C.sub.1-C.sub.6)alkylene; e is 0 and each of f and g is 1; and
each of R.sup.a', R.sup.a'''. and R.sup.a''' is independently
selected from the group consisting of H, and straight or branched
chain (C.sub.1-C.sub.6)alkyl.
[0116] Exemplary Formula I' compounds include those in the
following table:
TABLE-US-00002 ##STR00044## ##STR00045## ##STR00046## ##STR00047##
##STR00048## ##STR00049##
[0117] In another embodiment, is provided a CB2 ligand that
conforms to Formula II.
##STR00050##
[0118] For Formula II compounds or their pharmaceutically
acceptable salts, substituent groups A, B' and E are each
independently a bond, --C(R''').sub.2--, --CR'''--, --NR'''--,
--N--, --O--, --C(O)--, or --S--, however, no two adjacent members
of A, B' and E can simultaneously be --O--, --S--, or --NR'''--.
While subscripts h, j and k independently are integers between 0
and 2 inclusive, at least one of h, j or k is not 0 in Formula
II.
[0119] In one embodiment, D and D'' are each independently --C(O),
--CH.sub.2C(O)--, (C.sub.1-C.sub.6)alkylene, --C(O)NH--, or
--NHC(O)--. Formula II, furthermore, prescribes compounds that have
a fused ring system, with representing the option of having a
C.sub.5 member or a C.sub.6 member fused ring that optionally has
one or more double bonds.
[0120] For Formula II compounds, R and R.sub.1 are each
independently --OH, --OR.sup.a, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)haloalkyl, (C.sub.2-C.sub.6)alkenyl,
(C.sub.2-C.sub.6)alkynyl, (C.sub.3-C.sub.8)heteroaryl,
(C.sub.3-C.sub.8)heterocycloalkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkylene-.
[0121] When Group R and R.sub.1 are --OR.sup.a, R.sup.a is H,
straight or branched chain (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.8)cycloalkyl, (C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-.
[0122] For compounds conforming to Formula II, any alkyl, alkylene,
aryl, heteroaryl, cycloalkyl, or heterocycloalkyl is optionally
substituted with one or more halogen, oxo, --COOH, --CN,
--NO.sub.2, --OH, --NR.sup.dR.sup.e, (C.sub.1-C.sub.6)alkoxy, or
(C.sub.3-C.sub.8)aryloxy; with each R.sup.d and R.sup.e
independently being H, straight or branched (C.sub.1-C.sub.6)alkyl,
optionally substituted
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-, or
H.sub.2N(C.sub.1-C.sub.6)alkylene-.
[0123] According to one embodiment are provided Formula II
compounds in which represents an optionally substituted phenyl
group, groups A and E are independently --N--, --O--, or --S-- and
represents the option of having one or more double bonds. As
prescribed, therefore, compounds that conform to Formula II include
without limitation analogs of phenylimidazole, 3H-indazole, indole,
benz[d]oxazole, or 2,3-dihydrobenzo[d]oxazole,
benzo[d]thiazole.
[0124] Exemplary of Formula II compounds without limitation are the
following:
##STR00051##
[0125] Also provided by the present invention are modulators of the
CB2 receptor that conform to Formula III.
##STR00052##
[0126] For Formula III compounds X is N, or --CH--, groups Q', R
and T are each independently (C.sub.1-C.sub.6)alkyl,
--S(O).sub.2--, --S(O)--, --S(O).sub.2NHR'',
--O--(CH.sub.2).sub.x--O--, --OC(O)-- or (CH.sub.2).sub.x--OC(O)--,
while groups G, H, J, L, or M are each independently a bond,
--C(R''').sub.2--, --CR'''--, --NR'''--, --N--, --O--, --C(O)--, or
--S--.
[0127] For Formula III compounds, however, no two adjacent members
of G, H, J, L, or M can simultaneously be --O--, --S--, or
--NR'''--. According to one embodiment, R''' is H, OH, OR.sup.a,
halogen, (C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)haloalkoxy, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, --NH.sub.2,
--NH(C.sub.1-C.sub.6)alkyl, --N[(C.sub.1-C.sub.6)alkyl].sub.2,
--CN, (C.sub.3-C.sub.8)heteroaryl,
(C.sub.3-C.sub.8)heterocycloalkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)heteroaryl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkylene-, or
(C.sub.1-C.sub.6)alkyl-(C.sub.3-C.sub.8)arylene. The dashed circle,
represents the option of having one or more double bonds and for
Formula III compounds, subscripts l, m, n, p and q independently
are integers between 0 and 2 inclusive, with at least one of l, m,
n, p, or q being a non-zero (0) integer.
[0128] When R''' is --OR.sup.a, group R.sup.a is hydrogen, straight
or branched chain (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.8)cycloalkyl, (C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-.
[0129] Furthermore, for compounds conforming to Formula III, any
alkyl, alkylene, alkenylene, aryl, heteroaryl, cycloalkyl, or
heterocycloalkyl is optionally substituted with one or more members
selected from the group consisting of halogen, oxo, --COOH, --CN,
--NO.sub.2, --OH, --NR.sup.dR.sup.e, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.6)haloalkoxy,
(C.sub.1-C.sub.6)haloalkyl, (C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heteroaryl, (C.sub.3-C.sub.8)heterocycloalkyl, or
(C.sub.3-C.sub.8)aryloxy; with each R.sup.d and R.sup.e group being
independently H, straight or branched (C.sub.1-C.sub.6)alkyl,
optionally substituted (C.sub.3-C.sub.8)aryl, optionally
substituted (C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-, or
H.sub.2N(C.sub.1-C.sub.6)alkylene-.
[0130] For certain compounds according to Formula III, substituent
X is a nitrogen, substituent groups T and R are each --S(O).sub.2--
and Q' is a (C.sub.1-C.sub.6)alkyl. For other Formula III
compounds, X is --CH-- and each of Q, R and T are independently
--O--(CH.sub.2).sub.x--O--, --OC(O)-- or (CH.sub.2).sub.x--OC(O)--.
Illustrated below, without limitation are two compounds that
conform to Formula III.
##STR00053##
Pharmaceutical Compositions
[0131] Compounds of Formula I, II, III or IV can each be
administered to a patient or subject in need of treatment either
individually, or in combination with other therapeutic agents that
have similar biological activities. For example, Formulae I, II,
III or IV compounds and compositions can be administered as a
single dose or as multiple daily doses by a practicing medical
practitioner. When combination therapy is used, however, the
compound and the other therapeutic agent are administered
separately at different time intervals, or simultaneously.
[0132] Pharmaceutical formulations may include a compound I, II,
III or IV, or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier. In some embodiments, the
composition further contains, in accordance with accepted practices
of pharmaceutical compounding, one or more additional therapeutic
agents, pharmaceutically acceptable excipients, diluents,
adjuvants, stabilizers, emulsifiers, preservatives, colorants,
buffers, flavor imparting agents.
[0133] In one embodiment, the pharmaceutical composition includes a
compound according to Formula I or a pharmaceutically acceptable
salt thereof, and a pharmaceutically acceptable carrier.
##STR00054##
[0134] For Formula I compounds D is H, OH, OR.sup.a,
(C.sub.1-C.sub.6)alkyl and
##STR00055##
In one embodiment, D is
##STR00056##
and is an aromatic phenyl group that can be optionally substituted
by one, two, or three substituent groups. represents the point of
attachment of the ring to the Formula I scaffold. According to
Formula I, D can be an --OR.sup.a, or a (C.sub.1-C.sub.6)alkyl
group. When D is OR.sup.a, R.sup.a is H, straight or branched chain
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.8)cycloalkyl,
(C.sub.3-C.sub.14)aryl,
(C.sub.3-C.sub.14)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.14)heteroaryl-(C.sub.1-C.sub.6)alkylene-, or
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-. Groups A, B and Q
in Formula I are each independently (C.sub.1-C.sub.6)alkylene,
(C.sub.2-C.sub.6)alkenylene or (C.sub.2-C.sub.6)alkynylene, while
subscripts e, f and g independently are integers between 0 and 6
inclusive. In Formula I compounds, V, W, X, Y, and Z are each
independently a bond, --C(R''').sub.2--, --CR'''--, --NR'''--,
--N--, --O--, --C(O)--, or --S--. However, no two adjacent members
of V, W, X, Y, and Z in Formula I are simultaneously --O--, --S--,
or --NR'''--, Compounds according to Formula I encompass,
therefore, species in which one, two or each of the three ring
structures represents a heteroaryl group, a heterocycle group, a
cycloalkyl group or an aryl ring. Exemplary of heteroaryl and
heterocycle rings are oxetanyl, tetrahydrofuranyl,
tetrahydropyranyl, oxepanyl, tetrahydrothiophenyl,
tetrahydrothiopyranyl, 1,3-dioxanyl, oxazolidinyl, azetidinyl,
pyrrolidinyl, piperidinyl, azepinyl, piperazinyl, morpholinyl,
tetrahydrothiopyranyl-1-oxide, tetrahydrothiopyranyl-1,1-dioxide,
pyrrolidinonyl, piperidinonyl, azepinonyl, piperazidinonyl,
oxazidilinonyl, azetidinonyl, and morpholinonyl.
[0135] In one embodiment, the pharmaceutical formulation includes a
compound of Formula I, where any of V, W, X, Y, or Z is a
--C(R''').sub.2--, --CR'''--, --NR'''-- group, substituent R''' is
--H, --OH, --OR.sup.a, halogen, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.6)haloalkoxy,
(C.sub.1-C.sub.6)haloalkyl, (C.sub.2-C.sub.6)alkenyl,
(C.sub.2-C.sub.6)alkynyl, --NH.sub.2, --NH(C.sub.1-C.sub.6)alkyl,
--N[(C.sub.1-C.sub.6)alkyl].sub.2, --CN,
(C.sub.3-C.sub.8)heteroaryl, (C.sub.3-C.sub.8)heterocycloalkyl,
(C.sub.3-C.sub.8)cycloalkyl, (C.sub.3-C.sub.8)aryl,
(C.sub.3-C.sub.8)heterocycloalkyl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)heteroaryl-(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkylene-,
(C.sub.3-C.sub.8)aryl(C.sub.1-C.sub.6)alkenylene-, or
(C.sub.1-C.sub.6)alkyl-(C.sub.3-C.sub.8)arylene; l, m, n, p and q
independently are integers between 0 and 2 inclusive, and at least
one of l, m, n, p, or q is not zero (0). To accommodate for the
presence of aromatic and non-aromatic ring systems, Formula I
recites to represent the option of having one or more double bonds
within a ring system. Further, for compounds that conform to
Formula I, any alkyl, alkylene, alkenylene, aryl, heteroaryl,
cycloalkyl, or heterocycloalkyl is optionally substituted with
halogen, oxo, --COOH, --CN, --NO.sub.2, --OH, --NR.sup.dR.sup.e,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)haloalkoxy, (C.sub.1-C.sub.6)haloalkyl,
(C.sub.3-C.sub.8)aryl, (C.sub.3-C.sub.8)heteroaryl,
(C.sub.3-C.sub.8)heterocycloalkyl and (C.sub.3-C.sub.8)aryloxy;
with each of R.sup.d and R.sup.e being H, straight or branched
(C.sub.1-C.sub.6)alkyl, optionally substituted
(C.sub.3-C.sub.8)aryl, optionally substituted
(C.sub.3-C.sub.14)aryl(C.sub.1-C.sub.6)alkylene-, and
H.sub.2N(C.sub.1-C.sub.6)alkylene-.
[0136] In one embodiment, the pharmaceutical composition comprises
a compound of Formula IV, or a pharmaceutically acceptable salt
thereof, and a pharmaceutically acceptable carrier:
##STR00057##
[0137] In such pharmaceutical compositions including a compound of
Formula IV, R.sup.1 and R.sup.2 are independently H, alkyl, or
alkenyl; R.sup.3 is alkyl, alkenyl, aryl, aralkyl, aralkenyl,
heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl;
R.sup.4 and R.sup.5 are independently a bond, alkylenyl, or
alkenylenyl; each R.sup.6 and R.sup.7 is independently OH, F, Cl,
Br, I, (C.sub.1-C.sub.6)alkyl, alkoxy, amino, --COOH,
--C(O)NH.sub.2, SO.sub.3H, PO.sub.3H.sub.2, --CN, --SH, --NO.sub.2,
or CF.sub.3; and p and q are independently 0, 1, 2, 3, 4, or 5.
[0138] In one embodiment, the pharmaceutical composition comprises
a compound selected from those illustrated in Table 1 or a
pharmaceutically acceptable salt, and a pharmaceutically acceptable
carrier.
Multiple Myeloma and/or Osteoporosis
[0139] It has been determined that the compounds of Formulae I, I',
II, III, III' and IV are CB-2 receptor inverse agonists. A link is
also hereby established between CB-2 receptors and the treatment of
multiple myeloma and osteoporosis. By using two distinct CB2
antibodies that are able to distinguish active and inactive CB2
receptors, the present investors found that the CB2 receptors on
human MM cells are in active form. Based on the high levels of
expression on MM cells and nature of the CB2 receptor on human MM
cells, it is expected that CB2 ligands with antagonistic activities
can be employed to inhibit MM growth. Thus, by administering a
compound of Formula I, I', II, III, III' or IV to a subject, or by
contacting a compound of Formula I, I', II, III, III' or IV with
multiple myeloma cells or cells causing osteoporosis, the activity
of the CB-2 receptors may be modulated such that the cells are
inactivated, their activity is substantially altered, or the
subject is treated.
[0140] In one embodiment, a method for treating multiple myeloma or
osteoporosis in a subject by modulating the activity of a
cannabinoid receptor-2 (CB2), includes administering to the subject
a pharmaceutical composition comprising a therapeutically effective
amount of a compound of Formula IV:
##STR00058##
In such methods utilizing Formula IV, R.sup.1 and R.sup.2 are
independently H, alkyl, or alkenyl; R.sup.3 is alkyl, alkenyl,
aryl, aralkyl, aralkenyl, heterocyclyl, heterocyclylalkyl,
heteroaryl, heteroarylalkyl; R.sup.4 and R.sup.5 are independently
a bond, alkylenyl, or alkenylenyl; each R.sup.6 and R.sup.7 is
independently OH, F, Cl, Br, I, (C.sub.1-C.sub.6)alkyl, alkoxy,
amino, --COOH, --C(O)NH.sub.2, SO.sub.3H, PO.sub.3H.sub.2, --CN,
--SH, --NO.sub.2, CF.sub.3; and p and q are independently 0, 1, 2,
3, 4, or 5. In one embodiment, the treatment is for multiple
myeloma.
[0141] In another aspect, a method for modulating the activity of a
cannabinoid receptor-2 (CB2) in a subject, includes contacting the
CB-2 receptor with a compound of Formula IV:
##STR00059##
In such methods utilizing Formula IV, R.sup.1 and R.sup.2 are
independently H, alkyl, or alkenyl; R.sup.3 is alkyl, alkenyl,
aryl, aralkyl, aralkenyl, heterocyclyl, heterocyclylalkyl,
heteroaryl, heteroarylalkyl; R.sup.4 and R.sup.5 are independently
a bond, alkylenyl, or alkenylenyl; each R.sup.6 and R.sup.7 is
independently OH, F, Cl, Br, I, (C.sub.1-C.sub.6)alkyl, alkoxy,
amino, --COOH, --C(O)NH.sub.2, SO.sub.3H, PO.sub.3H.sub.2, --CN,
--SH, --NO.sub.2, CF.sub.3; and p and q are independently 0, 1, 2,
3, 4, or 5.
[0142] In another aspect, a method is provided including modulating
the activity of a cannabinoid receptor-2 (CB2) in a subject
suffering from osteoporosis. In one embodiment, the modulating
includes administering a CB-2 receptor inverse agonist to the
subject. In one embodiment of the methods, the CB-2 receptor
inverse agonist includes a compound of Formula IV:
##STR00060##
In such methods utilizing Formula IV, R.sup.1 and R.sup.2 are
independently H, alkyl, or alkenyl; R.sup.3 is alkyl, alkenyl,
aryl, aralkyl, aralkenyl, heterocyclyl, heterocyclylalkyl,
heteroaryl, heteroarylalkyl; R.sup.4 and R.sup.5 are independently
a bond, alkylenyl, or alkenylenyl; each R.sup.6 and R.sup.7 is
independently OH, F, Cl, Br, I, (C.sub.1-C.sub.6)alkyl, alkoxy,
amino, --COOH, --C(O)NH.sub.2, SO.sub.3H, PO.sub.3H.sub.2, --CN,
--SH, --NO.sub.2, CF.sub.3; and p and q are independently 0, 1, 2,
3, 4, or 5.
[0143] The compositions may be administered orally, or
parenterally, the term "parenteral" as used herein includes
subcutaneous injections, intravenous, intramuscular, intrasternal
injection or infusion techniques. Suitable oral compositions in
accordance with the invention include without limitation tablets,
troches, lozenges, aqueous or oily suspensions, dispersible powders
or granules, emulsion, hard or soft capsules, syrups or elixirs.
The dosage form may be a single unit dosage form that includes any
of the compounds of Formula I, I', II, III, III' or IV, or a
pharmaceutically salt thereof. Such dosage forms may include a
pharmaceutically acceptable carrier.
[0144] Compositions for parenteral administrations are administered
in a sterile medium. Depending on the vehicle used and
concentration the concentration of the drug in the formulation, the
parenteral formulation can either be a suspension or a solution
containing dissolved drug. Adjuvants such as local anesthetics,
preservatives and buffering agents can also be added to parenteral
compositions.
[0145] The present invention, thus generally described, will be
understood more readily by reference to the following examples,
which are provided by way of illustration and are not intended to
be limiting of the present invention.
Synthesis of Compounds
General Synthetic Methodology
[0146] The preparation of substituted amide compounds used in the
coupling reactions below were prepared via hydrolysis of
corresponding 2-phenylacetonitriles in concentrated H.sub.2SO.sub.4
(Scheme 1). The reactions were typically conducted at about
0.degree. C. for about 12 hours.
##STR00061##
[0147] The compounds described below and shown in Table 1, were
synthesized by the method a or b of Scheme. In Scheme 1a, the
coupling reaction between the amide and aldehyde was performed in
anhydrous dichloroethane (DCE) with the catalyst trimethylsilyl
chloride (TMSCl) at about 70.degree. C. for 3-12 hours. In Scheme
1b, the coupling reaction was performed in anhydrous
dichloromethane (DCM) with the catalyst F.sub.3CSO.sub.3SiMe.sub.3,
at room temperature for about 12 hours.
##STR00062##
Chemistry
[0148] All reagents were purchased from commercial sources and used
without further purification. Analytical thin-layer chromatography
(TLC) was performed on SiO.sub.2 plates on Alumina. Visualization
was accomplished by UV irradiation at 254 nm. Preparative TLC was
conducted using Preparative Silica gel TLC plates (1000 .mu.m, 20
cm.times.20 cm). Flash column chromatography was performed using
Biotage Isolera flash purification system. .sup.1H NMR was recorded
on a Bruker 400 MHz spectrometer. Chemical shifts are reported as 6
values in parts per million (ppm) as referenced to residual
solvent. .sup.1H NMR spectra are tabulated as follows: chemical
shift, multiplicity (s=singlet, bs=broad singlet, d=doublet,
t=triplet, q=quartet, m=multiplet), coupling constant(s), and
number of protons. Flash column chromatography was performed using
SiO.sub.2 60 (particle size 0.040-0.055 mm, 230-400 mesh).
[0149] The chemical purity of the target compounds was >95% as
determined using the following conditions: a Shimadzu HPLC-MS-MS
with a HAMILTON reversed phase column (HxSil, C18, 3 .mu.m,
2.1.times.50 mm (H2)); Eluent A: 5% CH.sub.3CN in H.sub.2O, eluent
B: 90% CH.sub.3CN in H.sub.2O (Table 2); flow rate of 0.2 mL/min;
UV detector, 254 nm.
TABLE-US-00003 TABLE 2 HPLC-MS-MS Eluent Compositions T (min) A (%)
B (%) 0.0 5 95 3.00 5 95 7.00 90 10 12.00 90 10 18.00 5 95 20.00
stop stop
Preparation of 2-Penylacetamide
[0150] Benzyl cyanide (5 g, 42.7 mmol) was added slowly to
concentrated sulfuric acid (20 ml) cooled by water-ice bath. The
solution was stirred overnight. The reaction mixture was poured
into ice water and neutralized with 20% NaOH. The aqueous phase was
extracted by ethyl acetate (3.times.15 mL). The combined organic
layers were washed with water (3.times.10 mL) and brine (3.times.10
mL), dried over anhydrous MgSO.sub.4, filtered, and concentrated
under reduced pressure. The residue was recrystallized from ethyl
acetate and hexane to give the title compound (4.5 g, 78%). .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 7.54 (s, 1H), 7.20-7.32 (m,
5H), 6.87 (s, 1H), 3.38 (s, 2H).
2-(4-Chlorophenyl)acetamide
[0151] The same procedure was followed using 2-(4-chloro)benzyl
cyanide. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.49 (s, 1H),
7.34-7.35 (m, 2H), 7.26-7.27 (m, 2H), 6.92 (s, 1H), 3.34-3.37 (m,
2H).
(4-(Trifluoromethyl)phenyl)acetamide
[0152] The same procedure was followed using
4-trifluoromethylbenzyl cyanide. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.65 (d, J=8.0 Hz, 2H), 7.58 (s, 1H), 7.49
(d, J=8.0 Hz, 2H), 7.00 (s, 1H), 3.51 (s, 2H).
[0153] General Protocol for the Coupling Reaction Between Amide and
Aldehyde--Method 1.
[0154]
N,N'-((4-(dimethylamino)phenyl)methylene)bis(2-phenylacetamide)
(1). To a suspension of 4-(dimethylamino)benzaldehyde (149 mg, 1
mmol) and 2-phenylacetamide (270 mg, 2 mmol) in 2 mL anhydrous DCE,
TMSCl (216 mg, 2 mmol) was added. The mixture was heated at
70.degree. C. for 12 h, then cooled to room temperature and the
crude product precipitated from the solution. The crude product was
recrystallized with methanol and hexane to give the final product
(140 mg, 35%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.89 (d,
J=8.0 Hz, 2H), 7.59 (s, 2H), 7.41 (d, J=8.8 Hz, 2H), 7.21-7.32 (m,
10H), 6.54 (t, J=8.0 Hz, 1H), 3.52 (dd, J=14.0, 15.6 Hz, 4H), 3.06
(s, 6H).
[0155] N,N'-(phenylmethylene)bis(2-phenylacetamide) (2). Yield:
67%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.78 (d, J=7.2 Hz,
2H), 7.21-7.35 (m, 15H), 6.55 (t, J=7.8 Hz, 1H), 3.50 (dd, J=13.8,
20.4 Hz, 4H).
[0156] N,N'-((2-fluorophenyl)methylene)bis(2-phenylacetamide) (3).
Yield: 64%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.87 (d,
J=7.8 Hz, 2H), 7.44 (t, J=7.8 Hz, 1H), 7.36 (q, J=6.6 Hz, 1H),
7.17-7.29 (m, 12H), 6.74 (t, J=7.8 Hz, 1H), 3.48 (dd, J=14.4, 24.0
Hz, 4H).
[0157] N,N'-((4-fluorophenyl)methylene)bis(2-phenylacetamide) (5).
Yield: 72%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.79 (d,
J=8.0 Hz, 2H), 7.16-7.36 (m, 14H), 6.54 (t, J=8.0 Hz, 1H), 3.52
(dd, J=14.4, 15.6 Hz, 4H).
[0158] N,N'-((4-chlorophenyl)methylene)bis(2-phenylacetamide) (6).
Yield: 71%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.83 (d,
J=7.8 Hz, 2H), 7.41 (d, J=7.8 Hz, 2H), 7.21-7.32 (m, 12H), 6.51 (t,
J=7.8 Hz, 1H), 3.50 (dd, J=14.4, 17.4 Hz, 4H).
[0159] N,N'-(p-tolylmethylene)bis(2-phenylacetamide) (8). Yield:
70%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.71 (d, J=8.0 Hz,
2H), 7.13-7.32 (m, 14H), 6.52 (t, J=8.0 Hz, 1 Hz, 1H), 3.51 (dd,
J=14.4, 15.6 Hz, 4H), 2.29 (S, 3H).
[0160] N,N'-((4-methoxyphenyl)methylene)bis(2-phenylacetamide)
(10). Yield: 62%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.70
(d, J=7.6 Hz, 2H), 7.21-7.31 (m, 12H), 6.89 (d, J=8.8 Hz, 2H), 6.51
(t, J=8.0 Hz, 1H), 3.74 (s, 3H), 3.50 (dd, J=14.0, 17.2 Hz,
4H).
[0161]
N,N'-((2-(trifluoromethyl)phenyl)methylene)bis(2-phenylacetamide)
(13). Yield: 70%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.87
(d, J=7.2 Hz, 2H), 7.78 (d, J=7.6 Hz, 1H), 7.68-7.73 (m, 2H), 7.54
(t, J=7.6 Hz, 1H), 7.19-7.30 (m, 10H), 6.83 (t, J=6.8 Hz, 1H), 3.46
(dd, J=14.0, 17.6 Hz, 4H).
[0162]
N,N'-((4-(trifluoromethyl)phenyl)methylene)bis(2-phenylacetamide)
(14). Yield: 75%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.89
(d, J=7.6 Hz, 2H), 7.72 (d, J=7.6 Hz, 2H), 7.51 (d, J=7.6 Hz, 2H),
7.25-7.30 (m, 10H), 6.58 (t, J=7.2 Hz, 1H), 3.53 (s, 4H).
[0163] N,N'-((4-nitrophenyl)methylene)bis(2-phenylacetamide) (15).
Yield: 84%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.94 (d,
J=7.8 Hz, 2H), 8.14 (d, J=8.4 Hz, 2H), 7.50 (d, J=8.4 Hz, 2H),
7.16-7.25 (m, 10H), 6.52 (t, J=7.8 Hz, 1H), 3.47 (s, 4H).
[0164]
N,N'-((4-(diethylamino)phenyl)methylene)bis(2-phenylacetamide)
(17). Yield: 14%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.60
(d, J=8.0 Hz, 2H), 7.21-7.31 (m, 10H), 7.07 (d, J=8.4 Hz, 2H), 6.60
(d, J=8.8 Hz, 2H), 6.43 (t, J=8.0 Hz, 1H), 3.44-3.52 (m, 4H),
3.29-3.34 (m, 4H), 1.06 (t, J=7.6 Hz, 6H). LC-MS (ESI): m/z 430.3
(M+H).sup.+.
[0165]
N,N'-((4-(dibutylamino)phenyl)methylene)bis(2-phenylacetamide)
(18). Yield: 15%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.58
(d, J=8.0 Hz, 2H), 7.20-7.31 (m, 10H), 7.06 (d, J=8.8 Hz, 2H), 6.57
(d, J=8.8 Hz, 2H), 6.41 (t, J=8.0 Hz, 1H), 3.47-3.48 (m, 4H),
3.22-3.26 (m, 4H), 1.43-1.50 (m, 4H), 1.26-1.35 (m, 4H), 0.91 (t,
J=7.6 Hz, 6H). LC-MS (ESI): m/z 486.2 (M+H).sup.+.
[0166]
N,N'-((4-(piperidin-1-yl)phenyl)methylene)bis(2-phenylacetamide)
(19). Yield: 45%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.80
(d, J=8.0 Hz, 2H), 7.15-7.32 (m, 14H), 6.51 (t, J=8.0 Hz, 1H),
3.37-3.52 (m, 8H), 1.61-1.83 (m, 6H). LC-MS (ESI): m/z 442.3
(M+H).sup.+.
[0167] N,N'-(phenylmethylene)bis(2-(4-chlorophenyl)acetamide) (21).
Yield: 52%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.88 (d,
J=7.8 Hz, 2H), 7.25-7.41 (m, 13H), 6.52 (t, J=7.8 Hz, 1H), 3.50 (s,
4H).
[0168]
N,N'-((2-fluorophenyl)methylene)bis(2-(4-chlorophenyl)acetamide)
(22). Yield: 63%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.93
(d, J=7.2 Hz, 2H), 7.45 (t, J=7.2 Hz, 1H), 7.33-7.37 (m, 5H),
7.25-7.26 (m, 4H), 7.18-7.21 (m, 2H), 6.73 (t, J=7.2 Hz, 1H), 3.48
(s, 4H).
[0169]
N,N'-((4-fluorophenyl)methylene)bis(2-(4-chlorophenyl)acetamide)
(23). Yield: 67%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.82
(d, J=7.6 Hz, 2H), 7.34-7.37 (m, 6H), 7.27 (d, J=8.4 Hz, 4H), 7.19
(t, J=8.8 Hz, 2H), 6.51 (t, J=8.0 Hz, 1H), 3.51 (s, 4H).
[0170]
N,N'-((4-chlorophenyl)methylene)bis(2-(4-chlorophenyl)acetamide)
(24). Yield: 40%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.88
(d, J=7.8 Hz, 2H), 7.42 (d, J=8.4 Hz, 2H), 7.31-7.35 (m, 6H), 7.26
(d, J=8.4 Hz, 4H), 6.47 (t, J=7.8 Hz, 1H), 3.50 (s, 4H).
[0171] N,N'-(p-tolylmethylene)bis(2-(4-chlorophenyl)acetamide)
(25). Yield: 68%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.76
(d, J=8.0 Hz, 2H), 7.34 (d, J=8.4 Hz, 4H), 7.28 (d, J=8.4 Hz, 4H),
7.17 (q, J=8.0 Hz, 4H), 6.50 (t, J=7.6 Hz, 1H), 3.51 (s, 4H), 2.29
(s, 3H).
[0172]
N,N'-((4-methoxyphenyl)methylene)bis(2-(4-chlorophenyl)acetamide)
(26).
[0173] Yield: 61%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.73
(d, J=7.6 Hz, 2H), 7.34 (d, J=8.8 Hz, 4H), 7.26 (d, J=8.4 Hz, 4H),
7.21 (d, J=8.8 Hz, 2H), 6.90 (d, J=8.8 Hz, 2H), 6.46 (t, J=8.0 Hz,
1H), 3.74 (s, 3H), 3.49 (s, 4H).
[0174]
N,N'-((2-(trifluoromethyl)phenyl)methylene)bis(2-(4-chlorophenyl)ac-
etamide) (27). Yield: 72%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.04 (d, J=6.0 Hz, 2H), 7.81 (d, J=7.8 Hz, 1H), 7.72 (d,
J=7.8 Hz, 1H), 7.62 (t, J=7.8 Hz, 1H), 7.52 (t, J=7.8 Hz, 1H),
7.27-7.30 (m, 8H), 7.06 (t, J=7.2 Hz, 1H), 3.52 (s, 4H).
[0175]
N,N'-((4-(trifluoromethyl)phenyl)methylene)bis(2-(4-chlorophenyl)ac-
etamide) (28). Yield: 65%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.96 (d, J=7.6 Hz, 2H), 7.73 (d, J=8.4 Hz, 2H), 7.54 (d,
J=8.4 Hz, 2H), 7.35 (d, J=8.4 Hz, 4H), 7.28 (d, J=8.4 Hz, 4H), 6.57
(t, J=7.6 Hz, 1H), 3.54 (s, 4H).
[0176]
N,N'-((4-nitrophenyl)methylene)bis(2-(4-chlorophenyl)acetamide)
(29). Yield: 80%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.02
(d, J=7.2 Hz, 2H), 8.21-8.23 (m, 2H), 7.57 (d, J=7.8 Hz, 2H),
7.33-7.35 (m, 4H), 7.26 (d, J=6.6 Hz, 4H), 6.55 (t, J=7.8 Hz, 1H),
3.53 (S, 4H).
[0177]
N,N'-(phenylmethylene)bis(2-(4-(trifluoromethyl)phenyl)acetamide)
(30). Yield: 73%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.94
(d, J=7.8 Hz, 2H), 7.63 (d, J=7.8 Hz, 4H), 7.48 (d, J=8.4 Hz, 4H),
7.29-7.37 (m, 5H), 6.55 (t, J=7.8 Hz, 1H), 3.63 (s, 4H).
[0178]
N,N'-((2-fluorophenyl)methylene)bis(2-(4-(trifluoromethyl)phenyl)ac-
etamide) (31). Yield: 66%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.99 (d, J=7.2 Hz, 2H), 7.63 (d, J=7.8 Hz, 4H), 7.46 (d,
J=7.8 Hz, 5H), 7.36-7.40 (m, 1H), 7.21 (t, J=7.8 Hz, 2H), 6.74 (t,
J=7.8 Hz, 1H), 3.60 (s, 4H).
[0179]
N,N'-((4-fluorophenyl)methylene)bis(2-(4-(trifluoromethyl)phenyl)ac-
etamide) (32). Yield: 70%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.95 (d, J=8.4 Hz, 2H), 7.63 (d, J=8.4 Hz, 4H), 7.47 d,
J=8.4 Hz, 4H), 7.37 (dd, J=5.4, 8.4 Hz, 2H), 7.19 (t, J=8.4 Hz,
2H), 6.51 (t, J=7.8 Hz, 1H), 3.62 (s, 4H).
[0180]
N,N'-((4-chlorophenyl)methylene)bis(2-(4-(trifluoromethyl)phenyl)ac-
etamide) (33). Yield: 75%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.96 (d, J=7.8 Hz, 2H), 7.63 (d, J=7.2 Hz, 4H), 7.46 (d,
J=7.8 Hz, 4H), 7.43 (dd, J=1.8, 8.4 Hz, 2H), 7.34-7.35 (m, 2H),
6.50 (t, J=7.2 Hz, 1H), 3.62 (s, 4H).
[0181]
N,N'-(p-tolylmethylene)bis(2-(4-(trifluoromethyl)phenyl)acetamide)
(34). Yield: 71%. 1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.87 (d,
J=8.4 Hz, 2H), 7.63 (d, J=7.8 Hz, 4H), 7.47 (d, J=8.4 Hz, 4H), 7.20
(d, J=8.4 Hz, 2H), 7.15 (d, J=7.8 Hz, 2H), 6.49 (t, J=7.8 Hz, 1H),
3.61 (s, 4H), 2.28 (s, 3H).
[0182]
N,N'-((4-methoxyphenyl)methylene)bis(2-(4-(trifluoromethyl)phenyl)a-
cetamide) (35). Yield: 76%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.85 (d, J=7.8 Hz, 2H), 7.63 (d, J=7.8 Hz, 4H), 7.47 (d,
J=7.8 Hz, 4H), 7.24 (d, J=9.0 Hz, 2H), 6.91 (d, J=8.4 Hz, 2H), 6.48
(t, J=7.8 Hz, 1H), 3.74 (s, 3H), 3.61 (s, 4H).
[0183]
N,N'-((4-(trifluoromethyl)phenyl)methylene)bis(2-(4-(trifluoromethy-
l)phenyl)acetamide) (36). Yield: 64%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.08 (d, J=7.2 Hz, 2H), 7.73 (d, J=7.8 Hz,
2H), 7.63 (d, J=7.8 Hz, 4H), 7.56 (d, J=7.8 Hz, 2H), 7.48 (d, J=7.8
Hz, 4H), 6.60 (t, J=7.8 Hz, 1H), 3.35-3.43 (m, 4H).
[0184] Synthetic Method 2.
[0185] N,N'-(2-phenylethane-1,1-diyl)bis(2-phenylacetamide) (37).
To a well stirred suspension of 2-phenylacetamide (540 mg, 4 mmol)
in dry DCM (2 mL) was added the 2-phenylacetaldehyde (240 mg, 2
mmol) and trimethylsilyltrifluoromethane sulonate (22 mg, 0.1
mmol). The mixture was vigorously stirred for 12 h at room
temperature, diluted with toluene (4 mL), and filtered. The
precipitate was washed several times with toluene which was
recrystallized with methanol and hexane to give the final product
(140 mg, 19%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.45 (d,
J=7.6 Hz, 2H), 7.14-7.28 (m, 15H), 5.55 (t, J=7.6 Hz, 1H), 3.39 (s,
4H), 2.93 (d, J=7.2 Hz, 2H). LC-MS (ESI): m/z 373.0
(M+H).sup.+.
[0186] N,N'-(3-phenylpropane-1,1-diyl)bis(2-phenylacetamide) (38).
Yield: 52%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.45 (d,
J=7.6 Hz, 2H), 7.10-7.31 (m, 15H), 5.26 (t, J=7.6 Hz, 1H),
3.38-3.46 (m, 4H), 2.47-2.53 (m, 2H), 1.88-1.94 (m, 2H). LC-MS
(ESI): m/z 387.3 (M+H).sup.+.
[0187] (E)-N,N'-(3-phenylprop-2-ene-1,1-diyl)bis(2-phenylacetamide)
(39). Yield: 18%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.61
(d, J=7.6 Hz, 2H), 7.21-7.37 (m, 15H), 6.41-6.45 (m, 1H), 6.27-6.32
(m, 1H), 6.04-6.09 (m, 1H), 3.44-3.52 (m, 4H). LC-MS (ESI): m/z
385.1 (M+H).sup.+.
[0188] N,N'-((4-isopropoxyphenyl)methylene)bis(2-phenylacetamide)
(12). Yield: 8.1%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.69
(d, J=7.6 Hz, 2H), 7.18-7.13 (m, 12H), 6.87 (d, J=6.8 Hz, 2H), 6.48
(t, J=8.0 Hz, 1H), 4.58-4.61 (m, 1H), 3.45-3.53 (m, 4H), 1.25 (d,
J=6.0 Hz, 6H). LC-MS (ESI): m/z 417.2 (M+H).sup.+.
[0189] N,N'-((4-bromophenyl)methylene)bis(2-phenylacetamide) (7).
Yield: 71%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.82 (d,
J=7.6 Hz, 2H), 7.53-7.56 (m, 2H), 7.21-7.33 (m, 12H), 6.48 (t,
J=7.6 Hz, 1H), 3.46-3.54 (m, 4H). LC-MS (ESI): m/z 437.0
(M+H).sup.+.
[0190] N,N'-((4-ethoxyphenyl)methylene)bis(2-phenylacetamide) (11).
Yield: 30%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.69 (d,
J=8.0 Hz, 2H), 7.18-7.31 (m, 10H), 6.88 (d, J=6.4 Hz, 2H), 6.48 (t,
J=8.0 Hz, 1H), 3.98-4.03 (m, 2H), 3.45-3.52 (m, 4H), 1.31 (t, J=6.8
Hz, 3H). LC-MS (ESI): m/z 403.1 (M+H).sup.+.
[0191] N,N'-(pentane-1,1-diyl)bis(2-phenylacetamide) (40). Yield:
63%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.25 (d, J=8.0 Hz,
2H), 7.19-7.30 (m, 8H), 5.30 (t, J=7.6 Hz, 1H), 3.36-3.44 (m, 4H),
1.56-1.62 (m, 2H), 1.14-1.26 (m, 4H), 0.81 (t, J=7.2 Hz, 3H). LC-MS
(ESI): m/z 339.1 (M+H).sup.+.
[0192] N,N'-(hexane-1,1-diyl)bis(2-phenylacetamide) (41). Yield:
73%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.25 (d, J=8.0 Hz,
2H), 7.19-7.30 (m, 8H), 5.29 (t, J=7.6 Hz, 1H), 3.39-3.44 (m, 4H),
1.57-1.59 (m, 2H), 1.18-1.23 (m, 6H), 0.82 (t, J=6.8 Hz, 3H). LC-MS
(ESI): m/z 353.3 (M+H).sup.+.
[0193] N,N'-((3-fluorophenyl)methylene)bis(2-phenylacetamide) (4).
Yield: 27%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.83 (d,
J=8.0 Hz, 2H), 7.06-7.42 (m, 14H), 6.53 (t, J=7.6 Hz, 1H),
3.47-3.55 (m, 4H). LC-MS (ESI): m/z 377.2 (M+H).sup.+.
[0194] N,N'-((4-isopropylphenyl)methylene)bis(2-phenylacetamide)
(9). Yield: 15%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.73
(d, J=8.0 Hz, 2H), 7.20-7.32 (m, 14H), 6.51 (t, J=8.0 Hz, 1H),
3.46-3.54 (m, 4H), 2.85-2.89 (m, 1H), 1.19 (d, J=6.8 Hz, 6H). LC-MS
(ESI): m/z 401.2 (M+H).sup.+.
[0195] N,N'-(2-Phenylethane-1,1-diyl)bis(2-phenylacetamide) (44).
To a well stirred suspension of 2-phenylacetamide (540 mg, 4 mmol)
in dry dichloromethane (2 mL) was added the 2-phenylacetaldehyde
(240 mg, 2 mmol) and trimethylsilyltrifluoromethanesulfonate (22
mg, 0.1 mmol). The mixture was vigorouslystirred for 12 h at room
temperature, diluted with toluene (4 mL), and filtered. The
precipitate was washed several times with toluene which was
recrystallized with methanol and hexane to give the final product
(560 mg, 76%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.45 (d,
J=7.6 Hz, 2H), 7.14-7.28 (m, 15H), 5.55 (t, J=7.6 Hz, 1H), 3.39 (s,
4H), 2.93 (d, J=7.2 Hz, 2H). HPLC-MS (ESI): m/z 373.2
(M+H).sup.+.
[0196] N,N'-(3-Phenylpropane-1,1-diyl)bis(2-phenylacetamide) (45).
Compound 45 was prepared from 2-phenylacetamide and
3-phenylpropanal using the procedure for compound 44. Yield: 92%.
.sup.1HNMR (400 MHz, DMSO-d.sub.6) .delta. 8.45 (d, J=7.6 Hz, 2H),
7.10-7.31 (m, 15H), 5.26 (t, J=7.6 Hz, 1H), 3.38-3.46 (m, 4H),
2.47-2.53 (m, 2H), 1.88-1.94 (m, 2H). LC-MS (ESI): m/z 387.3
(M+H).sup.+.
[0197] (E)-N,N'-(3-Phenylprop-2-ene-1,1-diyl)bis(2-phenylacetamide)
(46). Compound 46 was prepared from 2-phenylacetamide and
cinnamaldehyde using the procedure for compound 44. Yield: 88%.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.61 (d, J=7.6 Hz, 2H),
7.21-7.37 (m, 15H), 6.41-6.45 (m, 1H), 6.27-6.32 (m, 1H), 6.04-6.09
(m, 1H), 3.44-3.52 (m, 4H). LC-MS (ESI): m/z 385.1 (M+H).sup.+.
[0198]
N,N'-((4-(Diethylamino)phenyl)methylene)bis(3-phenylpropanamide)
(47). Compound 47 was prepared from 3-phenylpropanamide and
4-(diethylamino)benzaldehyde according to the procedure for
compound 1. Yield: 66%. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
8.29-8.30 (m, 2H), 7.17-7.29 (m, 10H), 6.92 (d, J=8.4 Hz, 2H), 6.56
(d, J=8.4 Hz, 2H), 6.46 (t, J=8.0 Hz, 1H), 2.82 (t, J=7.6 Hz, 4H),
2.42-2.47 (m, 4H), 1.07 (t, J=6.8 Hz, 6H). LC-MS (ESI): m/z 458.2
(M+H).sup.+. HRMS (ESI) for C.sub.29H.sub.36N.sub.3O.sub.2
(MH.sup.+): calcd, 458.2802. found, 458.2795.
[0199]
N,N'-((4-(Diethylamino)phenyl)methylene)bis(3-phenylacrylamide)
(48). Compound 48 was prepared from cinnamamide and
4-(diethylamino)benzaldehyde according to the procedure for
compound 1. Yield: 68%. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
8.68-8.70 (m, 2H), 7.38-7.58 (m, 12H), 7.19 (d, J=8.8 Hz, 2H), 6.79
(d, J=16.0 Hz, 2H), 6.66-6.68 (m, 3H), 3.29-3.35 (m, 4H), 1.08 (t,
J=7.2 Hz, 6H). LC-MS (ESI): m/z 454.2 (M+H).sup.+. HRMS (ESI) for
C.sub.29H.sub.32N.sub.3O.sub.2 (MH.sup.+): calcd, 454.2489. found,
454.2487.
[0200] N,N'-((4-(Diethylamino)phenyl)methylene)dibenzamide (49).
Compound 49 was prepared from benzamide and
4-(diethylamino)-benzaldehyde according to the procedure for
compound 1. Yield: 73%. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
9.24 (d, J=7.6 Hz, 1H), 7.88-7.93 (m, 4H), 7.81 (d, J=8.8 Hz, 2H),
7.46-7.65 (m, 9H), 7.20 (m, 1H), 3.70-3.81 (m, 4H), 1.17 (t, J=7.2
Hz, 6H). LC-MS (ESI): m/z 402.2 (M+H).sup.+. HRMS (ESI) for
C.sub.25H.sub.28N.sub.3O.sub.2 (MH.sup.+): calcd, 402.2176. found,
402.2167.
[0201]
N,N'-((4-(diethylamino)phenyl)methylene)bis(2-methylpropanamide)
(50). Compound 50 was prepared from isobutyramide and
4-(diethylamino)benzaldehyde according to the procedure for
compound 1. Yield: 80%. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
7.18 (d, J=8.4 Hz, 2H), 6.70-6.73 (m, 2H), 6.56 (s, 1H), 3.35-3.50
(m, 2H), 2.47-2.54 (m, 4H), 1.13-1.16 (m, 12H). LC-MS (ESI): m/z
334.2 (M+H).sup.+. HRMS (ESI) for C.sub.19H.sub.32N.sub.3O.sub.2
(MH.sup.+): calcd, 334.2489. found 334.2483.
[0202]
N,N'-((4-(diethylamino)phenyl)methylene)bis(2,2-dimethylpropanamide-
) (51). Compound 51 was prepared from pivalamide and
4-(diethylamino)benzaldehyde using method 1. Yield: 77%. .sup.1H
NMR (400 MHz, DMSO) .delta. 7.70 (d, J=8.8 Hz, 2H), 7.03 (d, J=8.8
Hz, 2H), 6.62 (d, J=8.8 Hz, 2H), 6.52 (t, J=8.4 Hz, 1H), 3.30-3.33
(m, 4H), 1.12 (s, 18H), 1.05-1.08 (m, 6H). LC-MS (ESI): m/z 262.2
(M+H).sup.+. HRMS (ESI) for C.sub.21H.sub.36N.sub.3O.sub.2
(MH.sup.+): calcd, 362.2802. found 362.2795.
[0203] N,N'-((4-(diethylamino)phenyl)methylene)dipentanamide (52).
Compound 52 was prepared from pentanamide and
4-(diethylamino)benzaldehyde using method 1. Yield: 69%. .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 7.18 (d, J=8.8 Hz, 2H), 6.71 (d,
J=8.8 Hz, 2H), 6.58 (t, J=8.4 Hz, 1H), 3.33-3.41 (m, 4H), 2.25 (t,
J=7.2 Hz, 4H), 1.58-1.66 (m, 4H), 1.33-1.43 (m, 4H), 1.14 (t, J=7.2
Hz, 6H), 0.95 (t, J=2.8 Hz, 6H). LC-MS (ESI): m/z 362.2
(M+H).sup.+. HRMS (ESI) for C.sub.21H.sub.36N.sub.3O.sub.2
(MH.sup.+): calcd, 362.2802. found 362.2792.
[0204] N,N'-((4-(diethylamino)phenyl)methylene)dihexanamide (53).
Compound 53 was prepared from hexanamide and
4-(diethylamino)benzaldehyde using method 1. Yield: 76%. .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 7.19 (d, J=8.8 Hz, 2H), 6.71 (d,
J=8.8 Hz, 2H), 6.58 (t, J=8.4 Hz, 1H), 3.35-3.41 (m, 4H), 2.19-2.26
(m, 4H), 1.61-1.68 (m, 4H), 1.32-1.35 (m, 8H), 1.14 (t, J=6.8 Hz,
6H), 0.94 (t, J=2.8 Hz, 6H). LC-MS (ESI): m/z 390.3 (M+H).sup.+.
HRMS (ESI) for C.sub.23H.sub.40N.sub.3O.sub.2 (MH.sup.+): calcd,
390.3115. found 390.3108.
[0205] N,N'-((4-(diethylamino)phenyl)methylene)dioctanamide (54).
Compound 54 was prepared from octanamide and
4-(diethylamino)benzaldehyde using method 1. Yield: 68%. .sup.1H
NMR (400 MHz, DMSO) .delta. 8.21 (d, J=8.0 Hz, 2H), 7.07 (d, J=8.8
Hz, 2H), 6.61 (d, J=8.8 Hz, 2H), 6.42 (t, J=8.0 Hz, 1H), 3.29-3.31
(m, 4H), 2.06-2.14 (m, 4H), 1.47-1.50 (m, 4H), 1.08-1.24 (m, 16H),
1.06 (t, J=7.2 Hz, 6H), 0.94 (t, J=7.2 Hz, 6H). LC-MS (ESI): m/z
446.3 (M+H).sup.+. HRMS (ESI) for C.sub.27H.sub.48N.sub.3O.sub.2
(MH.sup.+): calcd, 446.3741. found 446.3734.
[0206] N,N'-((4-(diethylamino)phenyl)methylene)bis(decanamide)
(55). Compound 55 was prepared from decanamide and
4-(diethylamino)benzaldehyde using method 1. Yield: 56%. .sup.1H
NMR (400 MHz, DMSO) .delta. 8.99 (d, J=9.6 Hz, 1H), 8.38 (d, J=8.8
Hz, 1H), 6.99 (d, J=8.8 Hz, 2H), 6.58 (d, J=8.4 Hz, 2H), 5.84 (d,
J=8.4 Hz, 1H), 3.27-3.30 (m, 4H), 2.33 (t, J=7.2 Hz, 4H), 2.14 (t,
J=7.2 Hz, 4H), 1.50-1.55 (m, 4H), 1.24-1.28 (m, 24H), 1.06 (t,
J=7.2 Hz, 6H), 0.87 (t, J=7.2 Hz, 6H). LC-MS (ESI): m/z 502.4
(M+H).sup.+.
##STR00063##
.sup.aReagents and conditions: (i) adamantan-1-amine, methanol,
refluxed, 10 h; (ii) heptan-1-amine, methanol, refluxed, 12 h;
(iii) p-toluidine or 4-chloroaniline, methanol, refluxed, 12 h;
(iv) NaBH.sub.4, methanol, r.t, 12 h; (v) acyl chloride or sulfonyl
chloride, anhydrous DCM, TEA, r.t, 12 h.
[0207] The synthetic routes to obtain compounds 55-99 are outlined
in Scheme 2 above. The commercially available
4-(diethylamino)benzaldehyde was reacted with adamantan-1-amine in
methanol to give A, which, when treated with NaBH.sub.4 gave the
secondary amine B. Finally, the coupling reaction between
intermediate B and selected acyl chloride or sulfonyl chloride
yielded the corresponding compounds 56-66. Taking heptan-1-amine,
p-toluidine or 4-chloroaniline as the starting material, the
synthesis of target compounds 67-99 was accomplished using a
procedure similar to that utilized for preparing compounds 56-66.
The final compounds were purified by flash column
chromatography.
General Procedure for Synthesis of Secondary Amine Building
Blocks.
[0208]
(3s,5s,7s,E)-N-(4-(Diethylamino)benzylidene)adamantan-1-amine (A).
(3s,5s,7s)-adamantan-1-amine hydrochloride (3.75 g, 20 mmol) was
added slowly to a solution of 4-(diethylamino)benzaldehyde and
methanol (50 mL). The mixture was stirred and refluxed for 12 h.
The reaction mixture was cooled to room temperature and the solvent
was removed by evaporation in vacuum to give the crude compound A,
which was used to the next step without further purification.
[0209] (3s,5s,7s)-N-(4-(Diethylamino)benzyl)adamantan-1-amine (B).
The crude compound A was dissolved in methanol (50 mL) and
NaBH.sub.4 (1.14 g, 30 mmol) was added. The mixture was continued
to stir for 12 h at room temperature. The reaction solution was
poured into water and extracted with EA. The combined organic
layers were washed with water and brine, and then dried over
Na.sub.2SO.sub.4. The mixture was filtered and the solvent was
evaporated in vacuum. The residue was purified by flash
chromatography on silica gel to obtain B (5.8 g, 88%). .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.13 (d, J=8.0 Hz, 2H), 6.62 (d,
J=8.0 Hz, 2H), 3.65 (bs, 1H), 3.42-3.48 (m, 2H), 3.28-3.39 (m, 4H),
2.05-2.07 (m, 3H), 1.58-1.71 (m, 12H), 1.07 (t, J=6.8 Hz, 6H).
LC-MS (ESI): m/z 313.2 (M+H).sup.+.
[0210] 4-(((4-Chlorophenyl)amino)methyl)-N,N-diethylaniline (C).
Yield: 78%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.03-7.14
(m, 4H), 6.56-6.62 (m, 4H), 6.19-6.22 (m, 1H), 4.06-4.07 (m, 2H),
3.27-3.34 (m, 4H), 1.06 (t, J=6.8 Hz, 3H). LC-MS (ESI): m/z 289.0
(M+H).sup.+.
Synthesis of Compounds 56-99
[0211]
N-((3s,5s,7s)-Adamantan-1-yl)-N-(4-(diethylamino)benzyl)benzenesulf-
onamide (56). The intermediate B (328 mg, 1.0 mmol) in
dichloromethane (DCM, 5 mL) was chilled in ice with the exclusion
of moisture and them triethylamine (122 mg, 1.2 mmol) was added to
it. The resulting solution was treated dropwise under stirring with
benzenesulfonyl chloride (177 mg, 1.0 mmol) also dissolved in DCM
over 30 min at 0.degree. C. and them left overnight at room
temperature. The reaction solution was poured into water and
extracted with EA. The combined organic layers were washed with
water and brine, and then dried over Na.sub.2SO.sub.4. The mixture
was filtered and the solvent was evaporated in vacuum. The residue
was purified by flash chromatography on silica gel to obtain 56
(400 mg, 85%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.13 (d,
J=8.0 Hz, 2H), 6.62 (d, J=8.0 Hz, 2H), 3.65 (bs, 1H), 3.42-3.48 (m,
2H), 3.28-3.39 (m, 4H), 2.05-2.07 (m, 3H), 1.58-1.71 (m, 12H), 1.07
(t, J=6.8 Hz, 6H). LC-MS (ESI): m/z 453.1 (M+H).sup.+.
[0212]
N-((3s,5s,7s)-Adamantan-1-yl)-N-(4-(diethylamino)benzyl)-4-fluorobe-
nzenesulfonamide (57) was prepared in a manner analogous to that
for compound 56. Yield: 87%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.87-7.89 (m, 2H), 7.38-7.43 (m, 2H), 7.21 (d, J=8.8 Hz,
2H), 6.64 (d, J=8.8 Hz, 2H), 4.59 (s, 2H), 3.29-3.36 (m, 4H),
1.88-1.93 (m, 9H), 1.42-1.51 (m, 6H), 1.07-1.10 (m, 6H). LC-MS
(ESI): m/z 471.0 (M+H).sup.+.
[0213] N-((3s, 5s,
7s)-Adamantan-1-yl)-4-chloro-N-(4-(diethylamino)benzyl)benzenesulfonamide
(58) was prepared in a manner analogous to that for compound 56.
Yield: 92%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.83 (d,
J=8.4 Hz, 2H), 7.64 (d, J=8.8 Hz, 2H), 7.21 (d, J=8.8 Hz, 2H), 6.65
(d, J=8.8 Hz, 2H), 4.60 (s, 2H), 3.29-3.37 (m, 4H), 1.88-1.93 (m,
9H), 1.42-1.51 (m, 6H), 1.07-1.11 (m, 6H). LC-MS (ESI): m/z 487.1
(M+H).sup.+.
[0214]
N-((3r)-Adamantan-1-yl)-N-(4-(diethylamino)benzyl)-4-methoxybenzene-
sulfonamide (59) was prepared in a manner analogous to that for
compound 56. Yield: 89%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.72-7.74 (m, 2H), 7.21 (d, J=8.8 Hz, 2H), 7.07-7.10 (m,
2H), 6.64 (d, J=8.8 Hz, 2H), 4.56 (s, 2H), 3.85 (s, 3H), 3.29-3.33
(m, 4H), 1.87-1.92 (m, 9H), 1.42-1.50 (m, 6H), 1.07-1.10 (m, 6H).
LC-MS (ESI): m/z 483.0 (M+H).sup.+.
[0215] N-((3s, 5s,
7s)-Adamantan-1-yl)-N-(4-(diethylamino)benzyl)-4-methylbenzenesulfonamide
(60) was prepared in a manner analogous to that for compound 56.
Yield: 86%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.70 (d,
J=8.4 Hz, 2H), 7.38 (d, J=8.0 Hz, 2H), 7.23 (d, J=8.8 Hz, 2H), 6.65
(d, J=8.8 Hz, 2H), 4.59 (s, 2H), 3.30-3.42 (m, 4H), 2.40 (s, 3H),
1.87-1.92 (m, 9H), 1.40-1.50 (m, 6H), 1.03-1.12 (m, 6H). LC-MS
(ESI): m/z 467.2 (M+H).sup.+.
[0216] N-((3s, 5s,
7s)-Adamantan-1-yl)-N-(4-(diethylamino)benzyl)-3-methylbenzenesulfonamide
(61) was prepared in a manner analogous to that for compound 56.
Yield: 84%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.58-7.62
(m, 2H), 7.42-7.48 (m, 2H), 7.23 (d, J=8.4 Hz, 2H), 6.65 (d, J=8.4
Hz, 2H), 4.60 (s, 2H), 3.30-3.34 (m, 4H), 2.39 (s, 3H), 1.87-1.92
(m, 9H), 1.41-1.50 (m, 6H), 1.07-1.11 (m, 6H). LC-MS (ESI): m/z
467.1 (M+H).sup.+.
[0217] N-((3s, 5s,
7s)-Adamantan-1-yl)-N-(4-(diethylamino)benzyl)-4-isopropylbenzenesulfonam-
ide (62) was prepared in a manner analogous to that for compound
56. Yield: 71%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.72
(d, J=8.0 Hz, 2H), 7.44 (d, J=8.0 Hz, 2H), 7.20 (d, J=8.8 Hz, 2H),
6.64 (d, J=8.8 Hz, 2H), 4.57 (s, 2H), 3.27-3.29 (m, 4H), 2.97-3.01
(m, 1H), 1.89-1.92 (m, 9H), 1.41-1.51 (m, 6H), 1.19-1.24 (m, 6H),
1.08-1.10 (m, 6H). LC-MS (ESI): m/z 495.2 (M+H).sup.+.
[0218]
N-((3s,5s,7s)-Adamantan-1-yl)-N-(4-(diethylamino)benzyl)cyclohexane-
carboxamide (63) was prepared in a manner analogous to that for
compound 56. Yield: 89%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.03 (d, J=8.8 Hz, 2H), 6.68 (d, J=8.8 Hz, 2H), 4.49 (s, 2H),
3.35-3.40 (m, 4H), 2.36-2.42 (m, 1H), 2.22 (d, J=2.4 Hz, 6H),
2.03-2.07 (m, 3H), 1.54-1.76 (m, 14H), 1.18-1.21 (m, 8H). LC-MS
(ESI): m/z 423.5 (M+H).sup.+.
[0219]
N-((3s,5s,7s)-Adamantan-1-yl)-N-(4-(diethylamino)benzyl)octanamide
(64) was prepared in a manner analogous to that for compound 56.
Yield: 81%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.05 (d,
J=8.4 Hz, 2H), 6.68 (d, J=8.8 Hz, 2H), 4.51 (s, 2H), 3.34-3.39 (m,
4H), 2.25-2.33 (m, 8H), 2.05 (s, 3H), 1.60-1.70 (m, 8H), 1.17-1.31
(m, 14H), 0.88 (t, J=6.8 Hz, 3H). LC-MS (ESI): m/z 439.4
(M+H).sup.+.
[0220] N-((3s, 5s,
7s)-Adamantan-1-yl)-2-(4-chlorophenyl)-N-(4-(diethylamino)benzyl)acetamid-
e (65) was prepared in a manner analogous to that for compound 56.
Yield: 79%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.28 (d,
J=8.8 Hz, 2H), 7.14 (d, J=8.4 Hz, 2H), 7.09 (d, J=8.4 Hz, 2H), 6.71
(d, J=8.8 Hz, 2H), 4.50 (s, 2H), 3.62 (s, 2H), 3.36-3.41 (m, 4H),
2.27 (s, 6H), 2.06-2.07 (m, 3H), 1.60-1.70 (m, 6H), 1.21 (t, J=7.2
Hz, 6H). LC-MS (ESI): m/z 465.2 (M+H).sup.+.
[0221]
N-((3s,5s,7s)-Adamantan-1-yl)-N-(4-(diethylamino)benzyl)-2-phenylac-
etamide (66) was prepared in a manner analogous to that for
compound 56. Yield: 70%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.29-7.32 (m, 2H), 7.22-7.26 (m, 3H), 7.10 (d, J=8.4 Hz, 2H), 6.71
(d, J=8.8 Hz, 2H), 4.50 (s, 2H), 3.67 (s, 2H), 3.36-3.41 (m, 4H),
2.28 (s, 6H), 2.06-2.07 (m, 3H), 1.60-1.70 (m, 6H), 1.21 (t, J=7.2
Hz, 6H). LC-MS (ESI): m/z 431.1 (M+H).sup.+.
[0222]
N-(4-(Diethylamino)benzyl)-N-heptyl-4-methylbenzenesulfonamide (67)
was prepared in a manner analogous to that for compound 56. Yield:
87%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.74 (d, J=8.4 Hz,
2H), 7.29-7.33 (m, 2H), 7.09 (d, J=8.8 Hz, 2H), 6.61 (d, J=8.4 Hz,
2H), 4.23 (s, 2H), 3.33-3.38 (m, 4H), 3.06-3.09 (m, 2H), 2.46 (s,
3H), 1.17-1.39 (m, 16H), 1.10-1.15 (m, 3H). LC-MS (ESI): m/z 430.7
(M+H).sup.+.
[0223]
N-(4-(Diethylamino)benzyl)-N-heptyl-3-methylbenzenesulfonamide (68)
was prepared in a manner analogous to that for compound 56. Yield:
89%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.61 (bs, 2H),
7.49-7.50 (m, 2H), 7.06 (d, J=8.4 Hz, 2H), 6.60 (d, J=8.4 Hz, 2H),
4.14 (s, 2H), 3.28-3.33 (m, 4H), 2.96-3.00 (m, 2H), 2.40 (s, 3H),
1.14-1.24 (m, 4H), 1.04-1.08 (m, 12H), 0.81-0.83 (m, 3H). LC-MS
(ESI): m/z 432.0 (M+H).sup.+.
[0224]
N-(4-(Diethylamino)benzyl)-N-heptyl-4-isopropylbenzenesulfonamide
(69) was prepared in a manner analogous to that for compound 56.
Yield: 81%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.77 (d,
J=8.4 Hz, 2H), 7.37 (d, J=8.0 Hz, 2H), 7.07 (d, J=8.4 Hz, 2H), 6.61
(d, J=8.8 Hz, 2H), 4.24-4.27 (m, 2H), 3.34-3.38 (m, 4H), 2.97-3.32
(m, 3H), 1.10-1.38 (m, 22H), 0.85-0.89 (m, 3H). LC-MS (ESI): m/z
460.2 (M+H).sup.+.
[0225]
N-(4-(Diethylamino)benzyl)-N-heptyl-4-methoxybenzenesulfonamide
(70) was prepared in a manner analogous to that for compound 56.
Yield: 89%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.77-7.80 (m,
2H), 7.08 (d, J=8.4 Hz, 2H), 6.99 (d, J=8.8 Hz, 2H), 6.61 (d, J=8.4
Hz, 2H), 4.22 (s, 2H), 3.90 (s, 3H), 3.33-3.38 (m, 4H), 3.05-3.09
(m, 2H), 1.15-1.40 (m, 16H), 0.83-0.89 (m, 3H). LC-MS (ESI): m/z
448.2 (M+H).sup.+.
[0226]
N-(4-(Diethylamino)benzyl)-N-heptyl-4-isopropoxybenzenesulfonamide
(71) was prepared in a manner analogous to that for compound 56.
Yield: 61%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.76 (d,
J=8.08 Hz, 2H), 7.13 (d, J=8.8 Hz, 2H), 7.06 (d, J=8.4 Hz, 2H),
6.60 (d, J=8.0 Hz, 2H), 4.10 (s, 2H), 3.86 (s, 3H), 3.28-3.34 (m,
4H), 2.93-2.96 (m, 2H), 1.03-1.23 (m, 20H), 0.79-0.83 (m, 3H).
LC-MS (ESI): m/z 475.4 (M+H).sup.+.
[0227]
N-(4-(Diethylamino)benzyl)-4-fluoro-N-heptylbenzenesulfonamide (72)
was prepared in a manner analogous to that for compound 56. Yield:
77%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.83-7.87 (m, 2H),
7.16-7.21 (m, 2H), 7.05 (d, J=8.4 Hz, 2H), 6.60 (d, J=8.4 Hz, 2H),
4.25 (s, 2H), 3.33-3.38 (m, 4H), 3.08-3.13 (m, 2H), 1.02-1.40 (m,
16H), 0.88-0.92 (m, 3H). LC-MS (ESI): m/z 436.0 (M+H).sup.+.
[0228]
4-Chloro-N-(4-(diethylamino)benzyl)-N-heptylbenzenesulfonamide (73)
was prepared in a manner analogous to that for compound 56. Yield:
83%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.77 (d, J=8.4 Hz,
2H), 7.48 (d, J=8.4 Hz, 2H), 7.06 (d, J=8.8 Hz, 2H), 6.60 (d, J=8.4
Hz, 2H), 4.25 (s, 2H), 3.33-3.38 (m, 4H), 3.08-3.12 (m, 2H),
1.04-1.42 (m, 16H), 0.88-0.92 (m, 3H). LC-MS (ESI): m/z 450.7
(M+H).sup.+.
[0229]
N-(4-(Diethylamino)benzyl)-N-heptyl-1-phenylmethanesulfonamide (74)
was prepared in a manner analogous to that for compound 56. Yield:
63%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.35-7.40 (m, 5H),
7.16 (d, J=8.4 Hz, 2H), 6.65 (d, J=8.4 Hz, 2H), 4.05-4.25 (m, 4H),
3.34-3.39 (m, 4H), 2.93-2.96 (m, 2H), 1.03-1.41 (m, 16H), 0.88-0.92
(m, 3H). LC-MS (ESI): m/z 431.2 (M+H).sup.+.
[0230] N-(4-(Diethylamino)benzyl)-N-heptylbutane-1-sulfonamide (75)
was prepared in a manner analogous to that for compound 56. Yield:
60%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.19 (d, J=8.4 Hz,
2H), 6.66 (d, J=8.8 Hz, 2H), 4.31 (s, 2H), 3.40-3.40 (m, 4H),
3.14-3.16 (m, 2H), 2.88-2.92 (m, 2H), 1.77-1.81 (m, 2H), 0.88-1.57
(m, 24H). LC-MS (ESI): m/z 398.0 (M+H).sup.+.
[0231] N-(4-(Diethylamino)benzyl)-N-heptyl-2-phenylacetamide (76)
was prepared in a manner analogous to that for compound 56. Yield:
70%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.25-7.33 (m, 5H),
7.08 (d, J=8.4 Hz, 1H), 6.94 (d, J=8.8 Hz, 1H), 6.65-6.69 (m, 2H),
4.47-4.51 (m, 2H), 3.81-3.82 (m, 2H), 3.31-3.40 (m, 5H), 3.24-3.28
(m, 1H), 1.12-1.29 (m, 16H), 0.88-0.92 (m, 3H). LC-MS (ESI): m/z
396.1 (M+H).sup.+.
[0232]
2-(4-Chlorophenyl)-N-(4-(diethylamino)benzyl)-N-heptylacetamide
(77) was prepared in a manner analogous to that for compound 56.
Yield: 65%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.16-7.35
(m, 4H), 7.17 (d, J=8.4 Hz, 1H), 7.08 (d, J=6.4 Hz, 1H), 6.66-6.71
(m, 2H), 4.51 (s, 2H), 3.80-3.81 (m, 2H), 3.35-3.39 (m, 5H),
3.26-3.28 (m, 1H), 1.14-1.67 (m, 16H), 0.89-0.92 (m, 3H). LC-MS
(ESI): m/z 428.8 (M+H).sup.+.
[0233]
N-(4-(Diethylamino)benzyl)-4-(dimethylamino)-N-heptylbenzamide (78)
was prepared in a manner analogous to that for compound 56. Yield:
80%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.37 (d, J=8.4 Hz,
2H), 7.09 (bs, 2H), 6.64-6.66 (m, 4H), 4.55 (s, 2H), 3.32-3.37 (m,
6H), 2.98 (s, 6H), 1.57-1.61 (m, 2H), 1.14-1.28 (m, 14H), 0.86 (t,
J=7.2 Hz, 6H). LC-MS (ESI): m/z 424.4 (M+H).sup.+.
[0234] N-(4-(Diethylamino)benzyl)-N-heptylcyclohexanecarboxamide
(79) was prepared in a manner analogous to that for compound 56.
Yield: 91%. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 6.97-7.06 (m,
4H), 6.59-6.66 (m, 4H), 4.11-4.46 (m, 2H), 3.14-3.37 (m, 6H),
2.47-2.51 (m, 1H), 1.51-1.82 (m, 10H), 1.01-1.49 (m, 16H),
0.84-0.90 (m, 3H). LC-MS (ESI): m/z 387.1 (M+H).sup.+.
[0235] N-(4-(Diethylamino)benzyl)-N-heptyloctanamide (80) was
prepared in a manner analogous to that for compound 56. Yield: 95%.
.sup.1H NMR (400 MHz, MeOD) .delta. 7.02-7.09 (m, 2H), 6.67-6.87
(m, 2H), 4.49 (s, 2H), 3.24-3.41 (m, 6H), 2.43 (t, J=7.2 Hz, 2H),
1.13-1.68 (m, 26H), 0.89-0.95 (m, 3H). LC-MS (ESI): m/z 402.9
(M+H).sup.+.
[0236]
N-(4-Chlorophenyl)-N-(4-(diethylamino)benzyl)benzenesulfonamide
(81) was prepared in a manner analogous to that for compound 56.
Yield: 73%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.64-7.73
(m, 5H), 7.32-7.34 (m, 2H), 6.96-7.05 (m, 4H), 6.50 (d, J=7.2 Hz,
2H), 4.63 (s, 2H), 3.23-3.26 (m, 4H), 1.02-1.04 (m, 6H). LC-MS
(ESI): m/z 428.9 (M+H).sup.+.
[0237]
N-(4-Chlorophenyl)-N-(4-(diethylamino)benzyl)-4-fluorobenzenesulfon-
amide (82) was prepared in a manner analogous to that for compound
56. Yield: 69%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.68-7.70 (m, 2H), 7.44-7.49 (m, 2H), 7.33 (d, J=8.8 Hz, 2H), 7.06
(d, J=8.4 Hz, 2H), 6.96 (d, J=8.8 Hz, 2H), 6.50 (d, J=8.8 Hz, 2H),
4.62 (s, 2H), 3.22-3.28 (m, 4H), 1.02 (t, J=6.8 Hz, 6H). LC-MS
(ESI): m/z 446.9 (M+H).sup.+.
[0238]
4-Chloro-N-(4-chlorophenyl)-N-(4-(diethylamino)benzyl)benzenesulfon-
amide (83) was prepared in a manner analogous to that for compound
56. Yield: 65%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.70
(d, J=8.8 Hz, 2H), 7.63 (d, J=8.8 Hz, 2H), 7.34 (d, J=8.8 Hz, 2H),
7.07 (d, J=8.8 Hz, 2H), 6.96 (d, J=8.8 Hz, 2H), 6.50 (d, J=8.8 Hz,
2H), 4.62 (s, 2H), 3.22-3.28 (m, 4H), 1.01-1.06 (m, 6H). LC-MS
(ESI): m/z 463.0 (M+H).sup.+.
[0239]
N-(4-Chlorophenyl)-N-(4-(diethylamino)benzyl)-4-methoxybenzenesulfo-
namide (84) was prepared in a manner analogous to that for compound
56. Yield: 55%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.56
(d, J=8.8 Hz, 2H), 7.32 (d, J=8.8 Hz, 2H), 7.13 (d, J=8.8 Hz, 2H),
7.05 (d, J=8.8 Hz, 2H), 6.96 (d, J=8.8 Hz, 2H), 6.50 (d, J=8.8 Hz,
2H), 4.58 (s, 2H), 3.86 (s, 3H), 3.22-3.27 (m, 4H), 1.02 (t, J=6.8
Hz, 6H). LC-MS (ESI): m/z 459.1 (M+H).sup.+.
[0240]
N-(4-Chlorophenyl)-N-(4-(diethylamino)benzyl)-4-methylbenzenesulfon-
amide (85) was prepared in a manner analogous to that for compound
56. Yield: 79%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.31-7.51 (m, 6H), 7.03-7.05 (m, 2H), 6.96 (d, J=8.8 Hz, 2H), 6.49
(d, J=8.8 Hz, 2H), 4.59 (s, 2H), 3.22-3.27 (m, 4H), 2.42 (s, 3H),
1.02 (t, J=6.8 Hz, 6H). LC-MS (ESI): m/z 443.2 (M+H).sup.+.
[0241]
N-(4-Chlorophenyl)-N-(4-(diethylamino)benzyl)-3-methylbenzenesulfon-
amide (86) was prepared in a manner analogous to that for compound
56. Yield: 65%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.38-7.53 (m, 4H), 7.32 (d, J=8.4 Hz, 2H), 7.04 (d, J=8.8 Hz, 2H),
6.96 (d, J=8.8 Hz, 2H), 6.50 (d, J=8.8 Hz, 2H), 4.61 (s, 2H),
3.22-3.28 (m, 4H), 2.40 (s, 3H), 1.02 (t, J=6.8 Hz, 6H). LC-MS
(ESI): m/z 443.0 (M+H).sup.+.
[0242]
N-(4-Chlorophenyl)-N-(4-(diethylamino)benzyl)-4-isopropylbenzenesul-
fonamide (87) was prepared in a manner analogous to that for
compound 56. Yield: 89%. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.56 (d, J=8.4 Hz, 2H), 7.49 (d, J=8.0 Hz, 2H), 7.32 (d,
J=8.8 Hz, 2H), 7.06 (d, J=8.8 Hz, 2H), 6.96 (d, J=8.8 Hz, 2H), 6.50
(d, J=8.8 Hz, 2H), 4.61 (s, 2H), 3.23-3.26 (m, 4H), 2.98-3.05 (m,
1H), 1.25 (d, J=6.8 Hz, 6H), 1.03 (t, J=6.4 Hz, 6H). LC-MS (ESI):
m/z 471.1 (M+H).sup.+.
[0243] N,2-bis(4-Chlorophenyl)-N-(4-(diethylamino)benzyl)acetamide
(88) was prepared in a manner analogous to that for compound 56.
Yield: 79%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.44 (d,
J=8.4 Hz, 2H), 7.32 (d, J=8.4 Hz, 2H), 7.09-7.14 (m, 4H), 6.91 (d,
J=8.8 Hz, 2H), 6.54 (d, J=8.8 Hz, 2H), 4.71 (s, 2H), 3.42 (s, 2H),
3.25-3.31 (m, 4H), 1.05 (t, J=6.4 Hz, 6H). LC-MS (ESI): m/z 442.8
(M+H).sup.+.
[0244]
N-(4-Chlorophenyl)-N-(4-(diethylamino)benzyl)cyclohexanecarboxamide
(89) was prepared in a manner analogous to that for compound 56.
Yield: 68%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.44 (d,
J=8.4 Hz, 2H), 7.11 (d, J=8.4 Hz, 2H), 6.89 (d, J=8.4 Hz, 2H), 6.55
(d, J=8.4 Hz, 2H), 4.65 (s, 2H), 3.26-3.31 (m, 4H), 2.08 (bs, 1H),
1.36-1.63 (m, 7H), 0.93-1.13 (m, 9H). LC-MS (ESI): m/z 399.4
(M+H).sup.+.
[0245] N-(4-(Diethylamino)benzyl)-N-(p-tolyl)benzenesulfonamide
(90) was prepared in a manner analogous to that for compound 56.
Yield: 80%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.60-7.72
(m, 5H), 7.04 (d, J=8.0 Hz, 2H), 6.96 (d, J=8.8 Hz, 2H), 6.87 (d,
J=8.4 Hz, 2H), 6.50 (d, J=8.8 Hz, 2H), 4.60 (s, 2H), 3.23-3.28 (m,
4H), 2.23 (s, 3H), 1.03 (t, J=7.2 Hz, 6H). LC-MS (ESI): m/z 408.9
(M+H).sup.+.
[0246]
N-(4-(Diethylamino)benzyl)-4-fluoro-N-(p-tolyl)benzenesulfonamide
(91) was prepared in a manner analogous to that for compound 56.
Yield: 83%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.66-7.68
(m, 2H), 7.43-7.47 (m, 2H), 7.06 (d, J=8.0 Hz, 2H), 6.96 (d, J=8.8
Hz, 2H), 6.89 (d, J=8.4 Hz, 2H), 6.50 (d, J=8.8 Hz, 2H), 4.60 (s,
2H), 3.22-3.30 (m, 4H), 2.23 (s, 3H), 1.02 (t, J=7.2 Hz, 6H). LC-MS
(ESI): m/z 427.2 (M+H).sup.+.
[0247]
4-Chloro-N-(4-(diethylamino)benzyl)-N-(p-tolyl)benzenesulfonamide
(92) was prepared in a manner analogous to that for compound 56.
Yield: 85%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.67-7.70
(m, 2H), 7.61-7.63 (m, 2H), 7.06 (d, J=8.0 Hz, 2H), 6.96 (d, J=8.8
Hz, 2H), 6.90 (d, J=8.4 Hz, 2H), 6.50 (d, J=8.8 Hz, 2H), 4.60 (s,
2H), 3.22-3.28 (m, 4H), 2.23 (s, 3H), 1.02 (t, J=7.2 Hz, 6H). LC-MS
(ESI): m/z 442.8 (M+H).sup.+.
[0248]
N-(4-(Diethylamino)benzyl)-4-methoxy-N-(p-tolyl)benzenesulfonamide
(93) was prepared in a manner analogous to that for compound 56.
Yield: 91%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.53-7.56
(m, 2H), 7.11-7.13 (m, 2H), 7.04 (d, J=8.4 Hz, 2H), 6.96 (d, J=8.4
Hz, 2H), 6.88 (d, J=8.4 Hz, 2H), 6.49 (d, J=8.8 Hz, 2H), 4.56 (s,
2H), 3.86 (s, 3H), 3.22-3.27 (m, 4H), 2.23 (s, 3H), 1.02 (t, J=7.2
Hz, 6H). LC-MS (ESI): m/z 439.1 (M+H).sup.+.
[0249]
N-(4-(Diethylamino)benzyl)-4-methyl-N-(p-tolyl)benzenesulfonamide
(94) was prepared in a manner analogous to that for compound 56.
Yield: 94%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.50 (d,
J=8.0 Hz, 2H), 7.40 (d, J=8.4 Hz, 2H), 7.04 (d, J=8.0 Hz, 2H), 6.96
(d, J=8.8 Hz, 2H), 6.87 (d, J=8.4 Hz, 2H), 6.49 (d, J=8.8 Hz, 2H),
4.57 (s, 2H), 3.22-3.27 (m, 4H), 2.42 (s, 3H), 2.22 (s, 3H), 1.02
(t, J=7.2 Hz, 6H). LC-MS (ESI): m/z 423.0 (M+H).sup.+.
[0250]
N-(4-(Diethylamino)benzyl)-3-methyl-N-(p-tolyl)benzenesulfonamide
(95) was prepared in a manner analogous to that for compound 56.
Yield: 72%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.37-7.51
(m, 4H), 7.05 (d, J=8.4 Hz, 2H), 6.96 (d, J=8.4 Hz, 2H), 6.86-6.88
(m, 2H), 6.50 (d, J=8.8 Hz, 2H), 4.59 (s, 2H), 3.22-3.28 (m, 4H),
2.39 (s, 3H), 2.23 (s, 3H), 1.03 (t, J=7.2 Hz, 6H). LC-MS (ESI):
m/z 423.4 (M+H).sup.+.
[0251]
N-(4-(Diethylamino)benzyl)-4-isopropyl-N-(p-tolyl)benzenesulfonamid-
e (96) was prepared in a manner analogous to that for compound 56.
Yield: 86%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.54-7.56
(m, 2H), 7.47 (d, J=8.4 Hz, 2H), 7.05 (d, J=8.0 Hz, 2H), 6.96 (d,
J=8.4 Hz, 2H), 6.89 (d, J=8.4 Hz, 2H), 6.49 (d, J=8.8 Hz, 2H), 4.58
(s, 2H), 3.22-3.28 (m, 4H), 2.98-3.05 (m, 1H), 2.23 (s, 3H), 1.25
(d, J=7.2 Hz, 6H), 1.02 (t, J=7.2 Hz, 6H). LC-MS (ESI): m/z 450.9
(M+H).sup.+.
[0252] N-(4-(Diethylamino)benzyl)-2-phenyl-N-(p-tolyl)acetamide
(97) was prepared in a manner analogous to that for compound 56.
Yield: 86%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.17-7.27
(m, 5H), 7.05 (d, J=7.2 Hz, 2H), 6.97 (d, J=8.0 Hz, 2H), 6.91 (d,
J=8.8 Hz, 2H), 6.54 (d, J=8.8 Hz, 2H), 4.69 (s, 2H), 3.35-3.40 (m,
2H), 3.25-3.31 (m, 4H), 2.30 (s, 3H), 1.05 (t, J=7.2 Hz, 6H). LC-MS
(ESI): m/z 387.5 (M+H).sup.+.
[0253]
2-(4-Chlorophenyl)-N-(4-(diethylamino)benzyl)-N-(p-tolyl)acetamide
(98) was prepared in a manner analogous to that for compound 56.
Yield: 86%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.31 (d,
J=8.4 Hz, 2H), 7.18 (d, J=8.0 Hz, 2H), 7.08 (d, J=8.0 Hz, 2H), 6.98
(d, J=8.0 Hz, 2H), 6.91 (d, J=8.4 Hz, 2H), 6.54 (d, J=8.8 Hz, 2H),
4.68 (s, 2H), 3.35-3.38 (m, 2H), 3.25-3.32 (m, 4H), 2.30 (s, 3H),
1.05 (t, J=7.2 Hz, 6H). LC-MS (ESI): m/z 420.7 (M+H).sup.+.
[0254] N-(4-(Diethylamino)benzyl)-N-(p-tolyl)cyclohexanecarboxamide
(99) was prepared in a manner analogous to that for compound 56.
Yield: 84%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.18 (d,
J=8.0 Hz, 2H), 6.94 (d, J=8.0 Hz, 2H), 6.89 (d, J=8.4 Hz, 2H), 6.54
(d, J=8.4 Hz, 2H), 4.63 (s, 2H), 3.26-3.31 (m, 4H), 2.30 (s, 3H),
2.08-2.14 (m, 1H), 1.35-1.62 (m, 8H), 0.86-1.09 (m, 8H). LC-MS
(ESI): m/z 379.5 (M+H).sup.+.
Methods and Uses
[0255] Inventive compounds that conform to Formulae I, I', II, III,
III' and IV are useful for modulating CB2 receptor activity. The
endocannabinoid system that is formed of the CB1 and CB2 receptors
plays an important physiological role glaucoma, cancer, stroke,
pain, neuronal disorders, osteoporosis, multiple sclerosis, and
autoimmune disorders. The present invention focuses on the of small
molecule therapeutic agents that selectively target and modulate
the activity of the CB2 receptor, as well as the use of such
compounds to treat multiple myeloma and osteoporosis.
Multiple Myeloma (MM)
[0256] CB2 signaling plays an important role in B cell maturation
and functions. The observation that CB2 is consistently
overexpressed in malignant B cells and MM cells when compared to
reactive lymphoid tissue or normal purified B lymphocytes, led to
the conclusion that agents capable of selectively targeting CB2
receptor and capable of modulating its activity may play a role in
the treatment of MM.
[0257] For example, FIGS. 1A-1C illustrates higher expression of
cannabinoid receptor 2 in various multiple myeloma cells grown in
culture using Western blot or RT-PCR analysis. These figures also
shows that CB1 is primarily expressed in brain tissue and its
expression in MM cells is negligible.
[0258] Several compounds according to the present invention were
synthesized as described above. Radiometric binding studies using
cultured cells expressing CB2 and [.sup.35S]-GTP .gamma.S binding
assays were used to demonstrate the CB2 binding ability of
compounds in accordance with the invention and to identify whether
the inventive Formulae I, II or III compounds were agonist,
antagonist or inverse agonists of CB2.
Biological Testing Data
1. Cell Culture and Reagents
[0259] Human MM cell lines U266, H929, OPM2, RPMI-8226 and its
subline RPMI 8226/LR5 (resistant to melphalan), MM.1 S and its
subline MM.1 R (resistant to dexamethasone) were cultured as
described earlier. The chemoresistant cell lines were cultured in
the presence of melphalan or dexamethasone, and resistance
phenotype was confirmed by cell proliferation assays. zVAD-fmk was
from Calbiochem (San Diego, Calif., USA). Cannabinoid ligands
SR141716 (CB1 inverse agonist), CP55940 (CB1/CB2 agonist),
Win55212-2 (CB1/CB2 agonist), SR144528 (CB2 inverse agonist) were
provided by NIH-NIDA-NDSP program. The radioligand
[.sup.3H]-CP55940 used for receptor binding assay was obtained from
Perkin-Elmer (Boston, Mass., USA).
2. Biological Assays
[0260] Briefly, the bioassay is carried out using the Perkin Elmer
96-well Top Counter. Competition binding assay was used to evaluate
the CB receptor binding affinity (K.sub.i) of the screened ligands
by displacing [.sup.3H]CP-55940. The procedure is as follows.
[0261] The CB receptor binding affinity (Ki) of the in-silico
screened ligands is determined via displacement of
[.sup.3H]CP-55,940. In competition binding experiments, the tested
compound dilutions are carried out in duplicate in TME buffer (25
mM Tris, 5 mM MgCl.sub.2, 1 mM EDTA) containing 0.1% (w/v) fatty
acid free bovine serum albumin (BSA), pH 7.4. Various
concentrations of the tested compound are added in the same volume
to 3 nM [.sup.3H]CP-55,940. TME buffer and cell membrane
preparations expressing CB receptors (5 .mu.g per well) are added
to a final volume of 200 .mu.L. For the saturation binding
experiments, varying concentrations of [.sup.3H]CP-55,940 (0.05-1.5
nM) with or without 2 .mu.M of unlabeled ligands (CP-55,940) are
incubated with the receptor membrane preparations to determine
K.sub.d and nonspecific binding. After the binding suspensions are
incubated at 30.degree. C. for 1 hr, the reaction is terminated by
rapid filtration through microfiltration plates (Unifilter GF/B
filterplate, PerkinElmer), followed by 5 washes with ice cold TME
buffer containing 0.1% BSA on a Packard Filtermate Harvester
(PerkinElmer). The plates are then dried overnight and 30 .mu.l
MicroScint 0 scintillation cocktail is added to each well of the
dried filter plates. The bound radioactivity is then counted using
a Perkin Elmer 96-well TopCounter. The K.sub.i is calculated by
using nonlinear regression analysis (Prism 5; GraphPad Software
Inc., San Diego, Calif.), with the K.sub.d values for
[.sup.3H]CP-55,940 determined from saturation binding experiments.
This assay is used for determination of the binding affinity
parameters (K.sub.i) of ligand-receptor interactions for the CB
receptor.
[0262] A tritiated thymidine incorporation assay assay was carried
out to investigate the effects of CB2 ligands on cell
proliferation. U266, RPMI-8226 (3.times.10.sub.4 cells/well), MM.1
S cells (6.times.10.sup.4 cells/well), and their resistant sublines
were cultured in 96-well culture plates with or without drugs for
48 hours. DNA synthesis was measured by .sup.3H-thymidine uptake as
described previously.
3. Compounds of the Invention Inhibit Proliferation of Human MM
Cells
[0263] Cell-based cAMP assays were used to identify the mode by
which the CB2 ligands according to the present invention modulate
CB2 receptor. Briefly, cAMP cell-based assays were used to
investigate whether a given Formula I, I', II, III, III' or IV
compound was an agonist, antagonist, or an inverse agonist of MM
cells expressing the CB2 receptor and to measure the anti-MM
activity of the compounds of the present invention. Known CB2
agonists, antagonists and inverse agonist were used as positive
controls. The data illustrates that compounds of of the invention
bind tightly to the CB2 receptors with K.sub.i values in the
nanomolar range. See Table 3.
TABLE-US-00004 TABLE 3A Compounds And Radioligand Binding Data For
Compounds Of Structure ##STR00064## Compd R.sub.1 R.sub.2 MW cLog P
K.sub.i (CB.sub.2), nM.sup.b, c K.sub.i (CB.sub.1), nM.sup.a, d
SI.sup.e 1 H p-(CH.sub.3).sub.2N--C.sub.6H.sub.4 401.50 4.04 777
>20,000 >26 2 H Ph 358.43 3.93 9,930 NT 3 H
o-F--C.sub.6H.sub.4 376.42 4.08 35,330 NT 4 H m-F--C.sub.6H.sub.4
376.42 4.08 12,670 NT 5 H p-F--C.sub.6H.sub.4 376.42 4.08 10,900 NT
6 H p-Cl--C.sub.6H.sub.4 392.88 4.54 3,081 NT 7 H
p-Br--C.sub.6H.sub.4 437.33 4.70 2,226 NT 8 H
p-CH.sub.3--C.sub.6H.sub.4 372.46 4.45 494 109 9 H
p-i-C.sub.3H.sub.7--C.sub.6H.sub.4 400.51 5.18 85 >20,000
>235 10 H p-CH.sub.3O--C.sub.6H.sub.4 388.46 3.78 783 >20,000
>26 11 H p-C.sub.2H.sub.5O--C.sub.6H.sub.4 402.49 4.13 1,500 NT
12 H p-i-C.sub.3H.sub.7O--C.sub.6H.sub.4 416.51 4.55 313 >20,000
>64 13 H o-CF.sub.3--C.sub.6H.sub.4 426.43 4.81 11,780 NT 14 H
p-CF.sub.3--C.sub.6H.sub.4 426.43 4.81 596 >20,000 >34 15 H
p-NO.sub.2--C.sub.6H.sub.4 403.43 3.87 NB NT 16 H
p-H.sub.2N--C.sub.6H.sub.4 373.45 2.51 12,550 NT 17 H
p-(C.sub.2H.sub.5).sub.2N--C.sub.6H.sub.4 429.55 4.76 64 >20,000
>313 18 H p-(C.sub.4H.sub.9).sub.2N--C.sub.6H.sub.4 485.66 6.69
221 >20,000 >90 19 H p-piperidyl-C.sub.6H.sub.4 441.56 4.89
595 >20,000 >34 20 H p-(Benzyl).sub.2N--C.sub.6H.sub.4 553.69
7.33 203 >20,000 >99 21 Cl Ph 427.32 5.14 NB NT 22 Cl
o-F--C.sub.6H.sub.4 445.31 5.29 10,850 NT 23 Cl p-F--C.sub.6H.sub.4
445.31 5.29 NB NT 24 Cl p-Cl--C.sub.6H.sub.4 461.77 5.75 154
>20,000 >130 25 Cl p-CH.sub.3--C.sub.6H.sub.4 441.35 5.66 462
>20,000 >43 26 Cl p-CH.sub.3O--C.sub.6H.sub.4 457.35 4.98 310
>20,000 >65 27 Cl o-CF.sub.3--C.sub.6H.sub.4 495.32 6.02 158
>20,000 >127 28 Cl p-CF.sub.3--C.sub.6H.sub.4 495.32 6.02 101
>20,000 >198 29 Cl p-NO.sub.2--C.sub.6H.sub.4 472.32 5.08 NB
NT 30 CF.sub.3 Ph 494.43 5.69 NB NT 31 CF.sub.3 o-F--C.sub.6H.sub.4
512.42 5.83 NB NT 32 CF.sub.3 p-F--C.sub.6H.sub.4 512.42 5.83 NB NT
33 CF.sub.3 p-Cl--C.sub.6H.sub.4 528.87 6.29 NB NT 34 CF.sub.3
p-CH.sub.3--C.sub.6H.sub.4 508.46 6.20 NB NT 35 CF.sub.3
p-CH.sub.3O--C.sub.6H.sub.4 524.45 5.53 NB NT 36 CF.sub.3
p-CF.sub.3--C.sub.6H.sub.4 562.43 6.57 NB NT 37 H
C.sub.6H.sub.5CH.sub.2 372.46 3.99 NB NT 38 H
C.sub.6H.sub.5CH.sub.2CH.sub.2 386.49 4.44 9,319 NT 39 H
C.sub.6H.sub.5CH.dbd.CH 384.47 4.54 5,683 NT 40 H n-C.sub.4H.sub.9
338.44 3.75 35,970 NT 41 H n-C.sub.5H.sub.11 352.47 4.19 18,200 NT
42.sup.f, g 2.1 NT 43.sup.f, h NT 10.6 44 H C.sub.6H.sub.5CH.sub.2
372.46 3.99 NB NB 45 H C.sub.6H.sub.5CH.sub.2CH.sub.2 386.49 4.44
9,319 NB 46 H C.sub.6H.sub.5CH.dbd.CH 384.47 4.54 5,683 NB
TABLE-US-00005 TABLE 3B Compounds And Radioligand Binding Data For
Compounds Of Structure ##STR00065## K.sub.i (CB.sub.2), K.sub.i
(CB.sub.1), Compd Y MW cLog P nM.sup.b, c nM.sup.a, d SI.sup.e 47
CH.sub.2CH.sub.2 457.61 5.64 231 >20,000 >93 48 CH.dbd.CH
453.58 5.80 167 >20,000 >119 49 bond 401.50 4.80 688
>20,000 >29
TABLE-US-00006 TABLE 3C Compounds And Radioligand Binding Data For
Compounds Of Structure ##STR00066## K.sub.i K.sub.i cLog
(CB.sub.2), (CB.sub.1), Compd R.sup.a' R.sup.a'' R.sup.a''' MW P
nM.sup.b, c nM.sup.a, d SI.sup.e 50 H CH.sub.3 CH.sub.3 333.46 3.57
2636 NB 51 CH.sub.3 CH.sub.3 CH.sub.3 361.52 4.69 3553 NB 52 H H
C.sub.3H.sub.7 361.52 4.27 182 >20,000 >109 53 H H
C.sub.4H.sub.9 389.57 5.16 25 >20,000 >800 54 H H
C.sub.6H.sub.13 445.68 7.9 146 >20,000 >136 55 H H
C.sub.8H.sub.17 501.79 10.0 160 >20,000 >125
TABLE-US-00007 TABLE 3D Compounds And Radioligand Binding Data For
Compounds Of Structure Compd R K.sub.i (CB.sub.2), nM.sup.b, c
K.sub.i (CB.sub.1), nM.sup.a, d SI.sup.e ##STR00067## B H 19950 NT
130 56 ##STR00068## 84 11000 130 57 ##STR00069## 25 4268 170 58
##STR00070## 173 2033 11 59 ##STR00071## 137 7300 53 60
##STR00072## 47 NB 425 61 ##STR00073## 19 8224 432 62 ##STR00074##
457 NT 63 ##STR00075## 35 NB >571 64 ##STR00076## 638 NT 65
##STR00077## 38 NB >526 66 ##STR00078## 60 NB >333
##STR00079## 67 ##STR00080## 2745 NT 68 ##STR00081## 2303 NT 69
##STR00082## 13000 NT 70 ##STR00083## 5193 NT 71 ##STR00084## NB NT
72 ##STR00085## 1101 NT 73 ##STR00086## 6740 NT 74 ##STR00087## 273
NT 75 ##STR00088## 680 NT 76 ##STR00089## 1312 NT 77 ##STR00090##
696 NT 78 ##STR00091## 1280 NT 79 ##STR00092## 212 NT 80
##STR00093## NB NT ##STR00094## G H 6741 NT 81 ##STR00095## 20 1773
88 82 ##STR00096## 73 1126 15 83 ##STR00097## 36 6617 183 84
##STR00098## 14 NB >1,428 85 ##STR00099## 37 137 3.7 86
##STR00100## 2.8 866 309 87 ##STR00101## 222 NT 88 ##STR00102## 136
NB 147 89 ##STR00103## 164 NB 121 ##STR00104## 90 ##STR00105## 3.4
514 151 91 ##STR00106## 5.6 858 153 92 ##STR00107## 3.0 412 137 93
##STR00108## 0.5 1297 2594 94 ##STR00109## 5.4 437 80 95
##STR00110## 5.8 218 37 96 ##STR00111## 4.3 3365 782 97
##STR00112## 72 NB >277 98 ##STR00113## 107 3200 29 99
##STR00114## 222 202 0.9 .sup.a, bBinding affinities of compounds
for CB.sub.1 and CB.sub.2 receptor were evaluated using
[.sup.3H]CP-55,940 radioligand competition binding assay. .sup.cNB
no binding K.sub.i > 20,000 nM. .sup.dNT = not tested. .sup.eSI:
selectivity index for CB.sub.2, calculated as
K.sub.i(CB.sub.1)/K.sub.i(CB.sub.2) ratio. .sup.f The binding
affinities of reference compounds were evaluated in parallel with
compounds 1-99 under the same conditions. .sup.g CB.sub.2 reference
compound SR 144528. .sup.h CB.sub.1 reference compound SR
141716.
[0264] The above compounds in Table 3, however, showed very poor to
no binding activity to the CB1 receptor, thus illustrating the
selectivity for CB2. FIG. 2A shows the dose dependent inhibition of
DNA synthesis in MM cells by the known CB2 selective inverse
agonists (SR144528 and AM630) and a known CB2 selective agonist
(Hu308) while FIG. 2B illustrates dose dependent inhibition of DNA
synthesis in MM cells using a representative compound (1) according
to the present invention.
##STR00115##
[0265] As demonstrated in FIG. 2A, the known inverse agonist had a
modest activity in inhibiting DNA synthesis in MM cells, while the
known agonist Hu308 showed no effect even at a concentration as
high as 10 .mu.M. In contrast, compound (1), showed potent
inhibition of DNA synthesis in MM cells a dose-dependent manner
(ICso: 1.25 .mu.M). c-AMP studies showed this compound to be an
inverse agonist of CB2. See FIG. 2C. It was surprising to discover
that compound (1) selectively inhibited the growth of two human
chemoresistant myeloma cell lines MM.1 R, resistant to
dexamethasone and RPMI-226/LR5 resistant to melphalan with
IC.sub.50 values in the range of about 0.6 to 1.2 .mu.M. However,
no growth inhibitory effect was seen for their respective parent
cells MM.1S and RPMI-8226.
[0266] Since drug resistance is a prevalent problem in multiple
myeloma clinical treatment, the ability of compounds according to
this invention to overcome chemoresistance of MM cells against
conventional drugs such as dexamethasone or melphalan provides an
unexpected advantageous benefit in MM treatment. The present
inventors have hypothesized that the cell inhibitory activity of
the inventive compounds is due to their ability to activate
apoptotic processes. Biological studies by the inventors have shown
that while mitogen-activated protein kinase (MAPK) family members
activated in response to cell stress are crucial for triggering
apoptosis, are not up regulated in MM cells treated with compound
(1).
[0267] Further studies revealed that compound (1) did not have any
effect on H2A.X phosphorylation that is critical for DNA damage and
apoptosis through sustained activation of JNK. However, endoplasmic
reticulum (ER) stress-induced transcription factor, CHOP, was
transiently upregulated, which promoted upregulation of
CHOP-targeted gene death receptor (DR) 5a, but not death receptor-4
(DR4). Compound (1) also altered certain proteins that are known to
play a role in the cell-growth cycle. For instance, contacting
cells with various concentrations of compound (1), showed that
while number of cells in the G.sub.0-G.sub.1 phase does not change,
the number of cells in the S-phase and those in the G2-M phase
decreased in a dose dependent manner. To further explore this
observation, the present inventors investigated the effect of
compound (1) on several proteins involved in cell G2-M phase
transition. In vitro studies indicate that compound (1) prevents
the tyrosine phosphatase Cdc25C from activating cyclin-B bound Cdc2
that is responsible for triggering mitosis and G1 to S phase and G2
to M phase cell transitions.
[0268] Taken together, the biological data indicates that compound
(1) negatively regulates myeloma cell cycle transitions by
modulating the expression and/or activity of various proteins that
are known to be important for cell cycle phase transitions, which
ultimately result in cell growth inhibition and cell death. The
role of CB2 in mediating the inhibition of MM cell growth was
studied using compound (1) as an exemplary compound according to
the present invention. As illustrated by Western blot analysis
(FIGS. 3A and 3B), a stable knockout of CB2, largely abrogated the
anti-MM activity of compound (1), suggesting a direct role for the
cannabinoid receptor 2 in MM cell death. Pretreatment of MMcells
with known agonists Win55212-2 or CP55940 and subsequent contact of
these cells with compound (1) showed that pretreatment with a CB2
specific agonist attenuated compound (1) mediated inhibition of MM
cell growth demonstrating that the CB2 receptor is a mediator of MM
cell death. See FIGS. 3C and 3D. FIG. 3E compares the CB2 receptor
modulatory activity of several known CB2 ligands to PAM (compound
(1)). Surprisingly, the present inventors discovered that PAM is an
inverse CB2 receptor agonist.
[0269] The above results and data illustrated in the figures
illustrate that compounds that conform to Formulae I, I', II, III,
III' and IV are a novel class of candidate therapeutics for
treating multiple myeloma.
Osteoporosis
[0270] In a further embodiment of the invention, Formulae I, I',
II, III, III' and IV compounds are candidate therapeutics for
treating a patient or subject suffering from osteoporosis and in
need of treatment. The present inventors surprisingly found that
compounds according to the present invention suppress osteoclast
activity while enhancing the activity of osteoblasts. The inventive
compounds, therefore, provide a two pronged therapeutic approach to
the treatment of osteoporosis, which differs from conventional
therapeutic regimens that rely on inhibition of osteoclast activity
to treat osteoporosis.
[0271] Using in vitro cell based binding assays, c-AMP regulation
and/or .sup.35S-GTP-.gamma.S binding assays described above, the
present inventors tested the ability of compound (1) to inhibit
osteoclast activity and prevent bone loss either alone or in
combination with the clinically approved drug Zoledronic in mouse
OVX osteoporosis models. Recent data indicate that compound (1)
selectively perturbs CB2 receptor and has exhibits anti-osteoclast
activity on both primary mouse and human bone marrow (BM)
preosteoclasts. Compound (1) was also found to be non-toxic when
administered at concentrations as high as 1 .mu.M.
[0272] FIG. 4 illustrates the anti-osteoclast activity of compound
(1). Briefly, this figure compares the inhibitory effects of
compound (1) and a known CB2 antagonist (XIE-35), from the
inventors laboratory on RANKL-induced osteoclast formation in bone
marrow mononuclear cells. It is clear that XIE-35 exerts a weak
inhibitory effect on RANKL/M-CSF induced osteoclast formation as
shown by the presence of numerous osteoclasts (arrows). In
contrast, compound (1) exerts strong anti-osteoclast effect
completely blocking osteoclast formation suggesting a more
favorable spatial orientation that enhances this compounds
interactions with appropriate amino acid residues in the CB2
receptors ligand binding pocket. As stated above CB2 is highly
expressed on osteoclasts and is believed to play a role in
osteoclast formation, maturation, and modulate processes that are
involved in bone resorption.
[0273] As a first step in evaluating the therapeutic potential of
compounds of the present invention as candidate therapeutics for
treating osteoporosis, the present inventors will conduct studies
directed to measuring the ability of these CB2 ligands inhibit
osteoclast (OCL) formation and prevent bone resorption as further
explained below.
a. Mouse OCL Formation and Bone Resorption Using Marrow Cells and
Preosteoclast Cell Line RAW264.7
[0274] Briefly, bone marrow (BM) cells will be flushed from the
long bones of 4-month-old mice, plated onto petri dishes, and
incubated for 48 h in the presence of M-CSF (100 ng/ml).
Nonadherent erythrocytes will be removed, and the adherent cells
will be washed with PBS and resuspended in culture medium. The
resulting M-CSF-dependent bone marrow macrophages cells will be
plated onto 96-well plates (5.times.10.sup.4 cells per well) in 100
.mu.l .alpha.-MEM containing 10% FBS, antibiotics, rhM-CSF (10
ng/ml), and rmRANKL (15 ng/ml), with or without a Formulae I, II or
III compound and inhibition of osteoclast formation by compounds of
the invention will be determined fixing the cells and staining the
fixed cells with a TRAP staining kit (Sigma-Aldrich) according to
the manufacturer's instructions.
[0275] The positively stained cells that contain three or more
nuclei will be counted as osteoclasts. Mouse OCL formation using
RAW264.7 cells (4.times.10.sup.3/well) will be conducted using the
similar procedures. To perform bone resorption pit assay, murine
M-CSF-dependent marrow cells (2.times.10.sup.5/well) will be seeded
on the dentin slices in 96-well plate and treated as above for
three weeks. The presence of osteoclasts on dentin slices will be
confirmed by TRAP staining and bone resorption lacunae will be
stained with hematoxylin.
b. Human Osteoclast Maturation and Function (Bone Resorption
Assay)
[0276] This assay will measure the effects of our CB2 ligands on
human OCL formation and activity. Briefly, nonadherent BM
mononuclear cells (10.sup.5 cells/well) will be seeded in 96-well
plates in .alpha.-MEM containing 20% horse serum, 10 ng/ml rhM-CSF,
and 50 ng/ml rhRANKL, in the presence or absence of CB2 ligands.
Half-media changes will be carried out twice a week where
appropriate. Differentiation into OCLs will be assessed by staining
with monoclonal antibody 23c6 that recognizes CD51/61 dimer
constituting the OCL vitronectin receptor, using a
Vectastatin-ABC-AP kit.
[0277] OCL number per well, nuclei per OCL, and OCL size will be
counted and measured using an inverted microscope and SPOT
software. For bone resorption, non-adherent marrow cells
(10.sup.5/well) will be seeded on the dentin slices in 96-well
plate and treated as above for four weeks. Bone resorption lacunae
will be stained with hematoxylin. All these tests will be conducted
using previously published protocols from the inventors [28].
c. Exploration of the Cellular Pathways and Molecular Mechanisms by
which the CB2 Ligands Exhibit Inhibitory Effects on OCL.
[0278] To investigate the molecular mechanism for the
anti-osteoclast activity of compounds encompassed by the present
invention, granulocyte macrophage colony forming units (CFU-GM)
cells that are known to be committed to osteoclast (OCL) precursor
cell will be plated at a cell density of 5.times.10.sup.4/well in
96-well culture plates in the presence of a-MEM medium containing
10% FCS and 100 pg/ml GM-CSF.
[0279] Cells will be incubated with an exemplary Formulae I, II or
III compound at one or more dose for 48 h. Control wells containing
cells but no test compound will also be maintained under identical
tissue culture conditions. Cells will be pulsed with [.sup.3H]-TdR
37 kBq/well during the last 8 h of culture, harvested onto
glass-fiber filter mats using an automatic cell harvester, and
counted using a beta plate scintillation counter. Thymidine uptake
will be measured as described in the literature and the data will
be analyzed and presented graphically as counts per minute
(CPM).
[0280] Furthermore, given that compounds that conform to Formulae
I, II or III are selectivity and tight-binding ligands of CB2
receptor, these compounds are valuable chemical probes for
elucidating the signal pathways that are important in the onset and
progression of osteoporosis. Recent studies have demonstrated the
high expression of cannabinoid receptor CB2 on osteoclasts. In view
of the immune monocyte origin of osteoclasts, targeting CB2
receptor has been proposed to regulate the ratio of activated
osteoclast to osteoblast, which is critical for the seizure of bone
loss and pathological fractures in osteoporosis induced by aging or
cancers. See for example, Ofek O, et al. Proc Natl Acad Sci USA.
2006; 103:696-701; Idris A I, et al. Drug News Perspect. 2008;
21:533-40; and Bab I A. Ann NY Acad Sci. 2007; 116:414-22.
[0281] The effect of Formulae I, I', II, III, III' and IV compounds
on differentiation and activity of osteoblasts will also be
investigated. Three kinds of experiments: alkaline phosphatase
(ALP) activity (an osteoblast differentiation marker), expression
of osteoblast related genes (Bsp, Ocn and Runx2) and
transactivation of osteoblast-specific Og2 (mouse osteocalcin gene
2), and mineralization assay will be used for this study.
[0282] To identify specific genes associated with osteoblast
development MC3T3-E1 preosteoblastic cells MC-4 will be cultured in
.alpha.-MEM containing ascorbic acid (50 .mu.g/ml) for 6 days.
Cells will be incubated with an exemplary Formulae I, II or III
compound at one or more dose for 48 h. Control wells containing
cells but no test compound will also be maintained under identical
tissue culture conditions. Total RNA will be isolated and analyzed
by quantitative real-time (RT)-PCR using specific primers for Bsp
(bone sialoprotein), Ocn (osteocalcin), and Runx2 mRNAs, which are
normalized to Gapdh mRNA.
[0283] In a separate experiment, MC-42 cells that are stably
transfected 1.3-kb Og2 (mouse osteocalcin gene 2) promoter driving
expression of a firefly luciferase gene, will be plated in 35-mm
plates and cultured in .alpha.-MEM-containing ascorbic acid (50
.mu.g/mL) for 15 days. Cells will be incubated with an exemplary
compound of the invention at one or more dose for 24 hours.
Following incubation, cells are harvested and assayed for
luciferase and ALP activity. The results were normalized to total
protein content of the sample.
d. Mineralization Assay
[0284] To determine the effect of exemplary Formulae I, I', II,
III, III' and IV compounds on the mineralization potential of cells
in culture, MC-42 cells will grown in culture as described above
for 15 days. Inorganic phosphate will be added to a final
concentration of 5.0 mM in the presence or absence of a fixed
concentrations of a Formulae I, I', II, III, III' and IV compound
for 48 hours. Following incubation, cells will be stained using the
von Kossa method and imaged by direct scanning of the
mineralization dish using the ScanMaker 9800 XL as previously
described.
e. In Vivo Studies
[0285] Based on the results from ex-vivo studies, compounds
identified as having potent CB2 modulatory effects will be
evaluated in the OVX mouse model using 25 .mu.g zoledronic acid
(ZOL, s.c., Novartis) as a positive control. Briefly,
ovariectomized (OVX) C57-BL6 mice (8-wk-old female, Charles River
Lab., Wilmington, Mass.) will be divided into 5 groups, (i) sham
group; (ii) OVX+vehicle; (iii) OVX+ZOL 25 .mu.g; (iv) OVX+CB2
ligand (low dose, ip); and (v) OVX+CB2 ligand (high dose, ip for
each test compound), with ten (10) mice per group. Dosing with
exemplary Formulae I, I', II, III, III' and IV compounds will be
commenced on day six (6) after ovariectomy or sham ovariectomy and
dosing will be continued for four weeks. At the end of the four
week study, tibial bone mineral density will be measured for each
mouse in the test and sham groups by microcomputed tomography
(micro-CT). Additionally, the osteoclast forming capacity will be
evaluated using bone marrow from treated and untreated mice.
Results from the in vivo study will be used to advance certain
potent compounds to more advanced clinical trials.
f. Osteoclast Formation Bioactivity
[0286] A number of compounds according to the invention were
selected as representative candidates to be evaluated against
RANKL-induced osteoclast differentiation on RAW 264.7 cells. RAW
264.7 is a mouse monocytic cell line that is used as a standard
osteoclast differentiation model. As shown in FIGS. 5A-5B, the
inventors tested the effect of these candidates on osteoclast (OCL)
formation using RAW 264.7 cells. Each ligand that was tested
induced a concentration-dependent inhibition of osteoclastogenesis
and all of the tested compounds generally showed strong potency in
suppressing OCL formation at 10 .mu.M, with inhibition rates of
>95%.
[0287] Compounds 84 and 93 also showed good inhibition activity at
low dose of 1 .mu.M (FIGS. 5A-5B). Meanwhile, these results
indicated that the inhibition activities are consistent with the
CB.sub.2 binding affinities. Especially, compound 93 showed the
strongest inhibition activity, with inhibition rates of 46%, 97%,
and 100% at 0.1, 1, and 10 .mu.M, respectively.
[0288] The same techniques demonstrated that compound 17 showed
strong inhibition activity, with inhibition rates of 72%, 79%, and
84% at 0.1, 1, and 10 .mu.M, respectively.
Cytotoxicity Studies Using Normal Human Cells
[0289] The compounds of the invention showed promising inhibition
activity with respect to osteoclastogenesis. To examine whether the
impaired osteoclastogenesis in the presence of PAM compounds is due
to their cell toxicity, the inventors investigated the cytotoxicity
profile of PAM compounds on normal human cells.
[0290] Peripheral blood was drawn in a heparinized syringe from
healthy fasting volunteers who had been without medication for at
least 2 weeks. The peripheral blood mononuclear cell (PBMC)
fraction was obtained by gradient centrifugation over
Ficoll-Hypaque (Amersham), as described by Feng, R. et al. S.
KD5170, a novel mercaptoketone-based histone deacetylase inhibitor,
exerts antimyeloma effects by DNA damage and mitochondrial
signaling, Mol. Cancer Ther. 2008, 7, 1494-1505. PBMC were washed
three times with ice-cold PBS, followed by resuspension at
5.times.10.sub.5/mL in the culture medium supplemented with 10%
inactivated FBS, 2 mM glutamine, 100 U/mL penicillin, and 100
.mu.g/mL streptomycin (Sigma). The compounds in a stock solution
(50 mMin DMSO) were diluted with the culture medium to application
conditions and further used for the treatment of PBMC for 3 days.
The final DMSO concentrations are always 0.02%. After treatment for
72 h, cell viability was determined using trypan blue exclusion
assay. These human cell studies conformed to the guidelines of the
Institutional Review Board of the University of Pittsburgh, Pa.
[0291] After treatment of these normal cells with compounds 9, 12,
17, 84, and 93 for 3 days, the trypan blue exclusion assays
indicated that the cell viability was not significantly affected in
comparing with the vehicle control group. For instance, compound 17
did not show any cytotoxic effects at the concentration (1 .mu.M)
of 79% inhibition of osteoclastogenesis, and only slight effects on
cell viability were observed at high concentration of 10 .mu.M.
[0292] Similarly for compounds 84 and 93, cell viability was not
significantly affected in comparing with the vehicle control group
at 1.25 and 2.5 .mu.M, and only some effects on cell viability were
observed at high concentrations of 5 and 10 .mu.M. Compound 84 did
not show any cytotoxic effects at 1.25 .mu.M (97% inhibition rate
at 1 .mu.M), and only slight effect on cell viability were observed
at high concentration of 5 .mu.M.
[0293] These results show that the compounds possess favorable
therapeutic indexes and the inhibition of human osteoclastogenesis
is not a result of their cytotoxicity.
EQUIVALENTS
[0294] While certain embodiments have been illustrated and
described, it should be understood that changes and modifications
can be made therein in accordance with ordinary skill in the art
without departing from the technology in its broader aspects as
defined in the following claims.
[0295] The present disclosure is not to be limited in terms of the
particular embodiments described in this application. Many
modifications and variations can be made without departing from its
spirit and scope, as will be apparent to those skilled in the art.
Functionally equivalent methods and compositions within the scope
of the disclosure, in addition to those enumerated herein, will be
apparent to those skilled in the art from the foregoing
descriptions. Such modifications and variations are intended to
fall within the scope of the appended claims. The present
disclosure is to be limited only by the terms of the appended
claims, along with the full scope of equivalents to which such
claims are entitled. It is to be understood that this disclosure is
not limited to particular methods, reagents, compounds compositions
or biological systems, which can of course vary. It is also to be
understood that the terminology used herein is for the purpose of
describing particular embodiments only, and is not intended to be
limiting.
[0296] In addition, where features or aspects of the disclosure are
described in terms of Markush groups, those skilled in the art will
recognize that the disclosure is also thereby described in terms of
any individual member or subgroup of members of the Markush
group.
[0297] As will be understood by one skilled in the art, for any and
all purposes, particularly in terms of providing a written
description, all ranges disclosed herein also encompass any and all
possible subranges and combinations of subranges thereof. Any
listed range can be easily recognized as sufficiently describing
and enabling the same range being broken down into at least equal
halves, thirds, quarters, fifths, tenths, etc. As a non-limiting
example, each range discussed herein can be readily broken down
into a lower third, middle third and upper third, etc. As will also
be understood by one skilled in the art all language such as "up
to," "at least," "greater than," "less than," and the like, include
the number recited and refer to ranges which can be subsequently
broken down into subranges as discussed above. Finally, as will be
understood by one skilled in the art, a range includes each
individual member.
[0298] All publications, patent applications, issued patents, and
other documents referred to in this specification are herein
incorporated by reference as if each individual publication, patent
application, issued patent, or other document was specifically and
individually indicated to be incorporated by reference in its
entirety. Definitions that are contained in text incorporated by
reference are excluded to the extent that they contradict
definitions in this disclosure.
[0299] While several, non-limiting examples have been illustrated
and described, it should be understood that changes and
modifications can be made therein in accordance with ordinary skill
in the art without departing from the invention in its broader
aspects as defined in the following claims.
* * * * *