U.S. patent application number 14/296267 was filed with the patent office on 2015-04-02 for orally bioavailable caffeic acid related anticancer drugs.
This patent application is currently assigned to The Board of Regents of the University of Texas System. The applicant listed for this patent is The Board of Regents of the University of Texas System. Invention is credited to Charles CONRAD, Izabela FOKT, Timothy MADDEN, Jeffrey MYERS, Waldemar PRIEBE, Slawomir SZYMANSKI.
Application Number | 20150094343 14/296267 |
Document ID | / |
Family ID | 38462023 |
Filed Date | 2015-04-02 |
United States Patent
Application |
20150094343 |
Kind Code |
A1 |
PRIEBE; Waldemar ; et
al. |
April 2, 2015 |
ORALLY BIOAVAILABLE CAFFEIC ACID RELATED ANTICANCER DRUGS
Abstract
The present invention concerns compounds and their use to treat
cell proliferative diseases such as cancer. Compounds of the
present invention display significant potency as inhibitors of
Jak2/STAT3 pathways and downstream targets and inhibit the growth
and survival of cancerous cell lines.
Inventors: |
PRIEBE; Waldemar; (Houston,
TX) ; FOKT; Izabela; (Houston, TX) ;
SZYMANSKI; Slawomir; (Spring, TX) ; MADDEN;
Timothy; (Spring, TX) ; MYERS; Jeffrey;
(Bellaire, TX) ; CONRAD; Charles; (Spring,
TX) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
The Board of Regents of the University of Texas System |
Austin |
TX |
US |
|
|
Assignee: |
The Board of Regents of the
University of Texas System
Austin
TX
|
Family ID: |
38462023 |
Appl. No.: |
14/296267 |
Filed: |
June 4, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11695547 |
Apr 2, 2007 |
8779151 |
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14296267 |
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60744105 |
Mar 31, 2006 |
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Current U.S.
Class: |
514/351 ;
514/357; 514/400; 514/521; 514/532; 514/654 |
Current CPC
Class: |
A61P 1/16 20180101; A61P
17/00 20180101; A61P 21/00 20180101; A61P 13/12 20180101; A61P
25/00 20180101; C07D 213/61 20130101; A61P 7/00 20180101; C07C
233/29 20130101; A61P 15/08 20180101; C07D 213/57 20130101; A61P
15/04 20180101; A61P 19/08 20180101; A61P 13/10 20180101; A61P 1/02
20180101; A61P 43/00 20180101; C07C 2601/02 20170501; A61P 19/00
20180101; C07C 2601/20 20170501; C07C 255/31 20130101; C07C 255/41
20130101; A61P 29/00 20180101; C07D 233/64 20130101; A61P 15/14
20180101; A61P 15/00 20180101; A61P 13/00 20180101; A61P 35/02
20180101; A61P 3/00 20180101; A61P 27/02 20180101; A61P 1/04
20180101; C07D 213/75 20130101; A61P 1/18 20180101; C07C 69/732
20130101; C07C 255/44 20130101; C07D 213/64 20130101; A61P 35/00
20180101; A61P 1/00 20180101; A61P 9/00 20180101; A61P 17/06
20180101; A61P 19/02 20180101; A61P 11/00 20180101 |
Class at
Publication: |
514/351 ;
514/357; 514/521; 514/532; 514/654; 514/400 |
International
Class: |
C07D 233/64 20060101
C07D233/64; C07C 255/44 20060101 C07C255/44; C07D 213/57 20060101
C07D213/57; C07C 69/732 20060101 C07C069/732; C07C 233/29 20060101
C07C233/29; C07D 213/64 20060101 C07D213/64; C07D 213/61 20060101
C07D213/61; C07C 255/41 20060101 C07C255/41 |
Goverment Interests
[0002] This invention was made with Government support under grant
number CA101936 awarded by the National Institutes of Health. The
Government has certain rights in the invention.
Claims
1. A method of treating a cell proliferative disease comprising
administering to a patient an effective amount of a compound
selected from the group consisting of: ##STR00060## wherein R.sub.1
is --H or cyano and R.sub.2 is heteroatom-substituted or
heteroatom-unsubstituted C.sub.3-C.sub.7-cycloalkyl; ##STR00061##
wherein X.sub.1 is halo and R.sub.3 is heteroatom-substituted or
heteroatom-unsubstituted C.sub.3-C.sub.7-cycloalkyl,
C.sub.6-C.sub.10-aryl, or C.sub.7-C.sub.10-aralkyl; ##STR00062##
wherein X.sub.2 is halo and R.sub.4 is hydroxy or
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-acyloxy; ##STR00063## wherein: X.sub.3 is halo or
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl or C.sub.1-C.sub.10-alkoxy, R.sub.5 is --H
or cyano, and R.sub.6 is heteroatom-substituted or
heteroatom-unsubstituted C.sub.1-C.sub.10-alkyl,
C.sub.3-C.sub.7-cycloalkyl, C.sub.1-C.sub.10-acyloxy,
C.sub.6-C.sub.10-aryl, or C.sub.7-C.sub.10-aralkyl; ##STR00064##
wherein: X.sub.4 is halo or heteroatom-substituted or
heteroatom-unsubstituted C.sub.1-C.sub.10-alkyl or
C.sub.1-C.sub.10-alkoxy, R.sub.7 is --H or cyano, and R.sub.8 is
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl, C.sub.3-C.sub.7-cycloalkyl,
C.sub.1-C.sub.10-acyloxy, C.sub.6-C.sub.10-aryl, or
C.sub.7-C.sub.10-aralkyl; ##STR00065## wherein: X.sub.5 is
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl or C.sub.1-C.sub.10-alkoxy, R.sub.9 is --H
or cyano, and R.sub.10 is heteroatom-substituted or
heteroatom-unsubstituted C.sub.1-C.sub.10-alkyl,
C.sub.3-C.sub.7-cycloalkyl, C.sub.1-C.sub.10-acyloxy,
C.sub.6-C.sub.10-aryl, or C.sub.7-C.sub.10-aralkyl; ##STR00066##
wherein: A is --C(O)-- or --S(O.sub.2)--, and X.sub.6 is halo or
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl or C.sub.1-C.sub.10-alkoxy, R.sub.11 is
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl, C.sub.3-C.sub.7-cycloalkyl,
C.sub.1-C.sub.10-acyloxy, C.sub.6-C.sub.10-aryl, or
C.sub.7-C.sub.10-aralkyl; ##STR00067## wherein: R.sub.12 is
cyclododecyl, imidazoyl, or cyclohexenyl, R.sub.13 is --H or
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl, C.sub.3-C.sub.7-cycloalkyl,
C.sub.1-C.sub.10-acyloxy, C.sub.6-C.sub.10-aryl, or
C.sub.7-C.sub.10-aralkyl; and ##STR00068## wherein: X.sub.7 is halo
or heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl or C.sub.1-C.sub.10-alkoxy, R.sub.14 is
##STR00069## or a pharmaceutically acceptable salt of any of the
above formulas.
2. The method of claim 1, wherein R.sub.2 is selected from the
group consisting of cyclopropyl, cyclobutyl, cyclopentyl and
cyclohexyl.
3. The method of claim 1, wherein R.sub.3 is selected from the
group consisting of phenyl, benzyl, cyclopropyl, cyclobutyl,
cyclopentyl and cyclohexyl.
4. The method of claim 1, wherein X.sub.1 or X.sub.2 is selected
from the group consisting of --F, --Cl, --Br and --I.
5. The method of claim 1, wherein R.sub.4 is selected from a group
consisting of hydroxy, acetoxy and 2,2-dimethylpropionyloxy.
6. The method of claim 1, wherein X.sub.3 or X.sub.4 is selected
from the group consisting of methoxy, --F, --Cl, --Br and --I.
7. The method of claim 1, wherein R.sub.6 or R.sub.8 is selected
from the group consisting of methyl and cyclopropyl.
8. The method of claim 1, wherein X.sub.5 is selected from a group
consisting of methyl and acetoxymethyl.
9. The method of claim 1, wherein A is --S(O.sub.2)--.
10. (canceled)
11. The method of claim 1, wherein the compound has the formula:
##STR00070## ##STR00071## ##STR00072## ##STR00073## ##STR00074## or
is a pharmaceutically acceptable salt of any of the above
formulas.
12.-44. (canceled)
45. The method of claim 1, wherein the compound has the formula:
##STR00075## or is a pharmaceutically acceptable salt thereof.
46.-86. (canceled)
87. The method of claim 1, wherein the subject is a mammal.
88. The method of claim 87, wherein the mammal is a human.
89. (canceled)
90. The method of claim 1, wherein the cell proliferative disease
is cancer.
91. The method of claim 90, wherein the cancer is melanoma,
non-small cell lung, small cell lung, lung, hepatocarcinoma,
retinoblastoma, astrocytoma, glioblastoma, leukemia, blood, brain,
skin, eye, tongue, gum, neuroblastoma, head, neck, breast,
pancreatic, renal, bone, testicular, ovarian, mesothelioma,
cervical, gastrointestinal, lymphoma, colon, or bladder.
92. The method of claim 1, wherein the cell proliferative disease
is rheumatoid arthritis, inflammatory bowel disease,
osteoarthritis, leiomyomas, adenomas, lipomas, hemangiomas,
fibromas, vascular occlusion, restenosis, arthrosclerosis, a
pre-neoplastic lesion, carcinoma in situ, oral hairy leukoplakia,
or psoriasis.
93. The method of claim 1, wherein STAT3 activation is reduced in a
cell of the subject.
94. The method of claim 1, wherein c-myc expression is reduced in a
cell of the subject.
95.-97. (canceled)
Description
[0001] The present application is a divisional of U.S. application
Ser. No. 11/695,547, filed Apr. 2, 2007, which claims benefit of
priority to U.S. Provisional Application No. 60/744,105, filed Mar.
31, 2006. The entire text of each of the above referenced
disclosures is specifically incorporated herein by reference.
BACKGROUND OF THE INVENTION
[0003] 1. Field of the Invention
[0004] The present invention relates generally to the treatment of
cell proliferative diseases such as cancer. More particularly, it
concerns caffeic acid and related analogs useful for the treatment
of cell proliferative diseases such as cancer, methods of synthesis
of these compounds, and methods of treatment employing these
compounds.
[0005] 2. Description of Related Art
[0006] The compound AG490 is a kinase inhibitor that inhibits Janus
kinase 2/Signal transducer and activator of transcription-3
(Jak2/STAT3) signaling pathway. AG490 belongs to a group of
compounds defined by the parent natural product caffeic acid and
its natural derivatives like caffeic acid benzyl ester.
##STR00001##
[0007] Targeted inhibition of the Jak2/STAT3 pathway with caffeic
acid analogs such as AG490 inhibits tumor cell growth and increases
sensitivity to apoptotic stimuli; thus, inhibitors of this pathway
likely represent potential therapeutics for cancer therapy
(Catlett-Falcone et al., 1999; Alas and Bonavida, 2003; Burdelya et
al., 2002). AG490 would not be considered a drug-like molecule due
to its instability in biological matrices (blood, tissues, etc) and
a lack of potency (Kondo, et al, 2007; Burdelya et al., 2002;
Meydan et al., 1996; Constantin et al., 1998). Receptor-based or
direct activation of Jak2/STAT3 pathway by such stimulators such as
EGF, scr, and IL-6 (multiple interleukins and cytokines) promoting
survival proliferation and angiogenesis of human tumors (Bharti et
al., Verma et al., Kerr et al.), requires inhibitors more potent
and more stable than AG490 to have potential as anti-cancer
drugs.
[0008] Jak2/STAT3 signaling pathways participate in the progression
of a variety of malignancies. STAT3 is constitutively activated in
pancreatic carcinoma, glioblastoma multiforme, and squamous cell
carcinoma of the head and neck, among others, and its activation
has been shown to affect VEGF expression, angiogenesis, tumor
growth, and metastasis in vivo. As such, STAT3 may be an excellent
target for drug development (Yu and Jove, 2004). No effective
inhibitors are currently available.
[0009] AG490, a caffeic acid analog, is sometimes referred
descriptively as a tyrphostin. U.S. Pat. No. 6,426,366 and U.S.
Patent Publication No. 2003/0013748 describe compounds that have
structural similarity with AG490.
[0010] AG490, however, has limited activity in animal studies and
must be used at high concentrations (.about.50 to 100 .mu.M) to
achieve inhibition of Jak2/STAT3 signaling and anti-tumor effects.
This low potency of AG490 is insufficient to warrant clinical
investigation of this compound for the treatment of cancer
(Burdelya et al., 2002; Meydan et al., 1996; Constantin et al.,
1998). Thus a need exists for therapeutics that exhibit strong
anti-proliferative effects through a similar mechanism at lower
therapeutic concentrations.
SUMMARY OF THE INVENTION
[0011] The present invention overcomes limitations in the art by
providing compounds that display improved pharmacological profiles
(e.g., biostability, bioavailability, enhanced tissue penetration,
improved pharmacokinetics, increased potency) when compared with
AG490 and other compounds that are structurally related to caffeic
acid; these compounds block IL-6 mediated Jak2/STAT3 activation at
low micromolar concentrations and suppress related downstream
antiapoptotic, proangiogenic and proliferation promoting signaling.
The present invention involves compounds that have utility as
antitumor and/or chemotherapeutic drugs, methods of synthesizing
these compounds, and methods of using these compounds to treat
patients with cancer.
[0012] Disclosed herein are a new class of compounds that inhibits
Jak2 and STAT3 phosphorylation and many of its related downstream
targets. It also potently inhibits tumor growth in vitro and in
vivo. Unlike AG490, the compounds of the present invention, for
example the caffeic acid analogs with cycloalkyl substituents (see
below), are highly active against a variety of cancers including
pancreatic tumors, such as Colo357-FG, brain tumors, such as
U87-MG, D54, U251 and cancer stem cell lines of glioblastoma
multiforme, and head and neck tumors, including squamous cell
carcinoma cancer cell lines. These compounds inhibit both IL-6, EGF
stimulated and constituative STAT3 activation; suppressed the
expression of Bcl-2, Bcl-XL, survivin, and Mcl-1; and induced
apoptosis, all at low micromolar concentrations.
[0013] One aspect of the present invention provides compounds
according to the structural formulas shown in Table 1.
TABLE-US-00001 TABLE 1 Examples of Different Types of Caffeic Acid
Analogs: ##STR00002## (I) ##STR00003## (II) ##STR00004## (III)
##STR00005## (IV) ##STR00006## (V) ##STR00007## (VI) ##STR00008##
(VII) ##STR00009## (VIII) ##STR00010## (IX)
[0014] In certain aspects of the invention, the chemical structures
shown in Table 1 may be defined as follows: R.sub.1 is --H or cyano
and R.sub.2 is heteroatom-substituted or heteroatom-unsubstituted
C.sub.3-C.sub.7-cycloalkyl; X.sub.1 is halo and R.sub.3 is
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.7-cycloalkyl, C.sub.6-C.sub.10-aryl, or
C.sub.7-C.sub.10-aralkyl; X.sub.2 is halo and R.sub.4 is hydroxy or
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-acyloxy; X.sub.3 is halo or heteroatom-substituted
or heteroatom-unsubstituted C.sub.1-C.sub.10-alkyl or
C.sub.1-C.sub.10-alkoxy; R.sub.5 is --H or cyano, and R.sub.6 is
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl, C.sub.1-C.sub.7-cycloalkyl,
C.sub.1-C.sub.10-acyloxy, C.sub.6-C.sub.10-aryl, or
C.sub.7-C.sub.10-aralkyl; X.sub.4 is halo or heteroatom-substituted
or heteroatom-unsubstituted C.sub.1-C.sub.10-alkyl or
C.sub.1-C.sub.10-alkoxy; R.sub.7 is --H or cyano; R.sub.8 is
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl, C.sub.1-C.sub.7-cycloalkyl,
C.sub.1-C.sub.10-acyloxy, C.sub.6-C.sub.10-aryl, or
C.sub.7-C.sub.10-aralkyl; X.sub.5 is heteroatom-substituted or
heteroatom-unsubstituted C.sub.1-C.sub.10-alkyl or
C.sub.1-C.sub.10-alkoxy; R.sub.9 is --H or cyano; R.sub.10 is
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl, C.sub.1-C.sub.7-cycloalkyl,
C.sub.1-C.sub.10-acyloxy, C.sub.6-C.sub.10-aryl, or
C.sub.7-C.sub.10-aralkyl; A is --C(O)-- or --S(O.sub.2)--; X.sub.6
is halo or heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl or C.sub.1-C.sub.10-alkoxy; R.sub.11 is
heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl, C.sub.1-C.sub.7-cycloalkyl,
C.sub.1-C.sub.10-acyloxy, C.sub.6-C.sub.10-aryl, or
C.sub.7-C.sub.10-aralkyl; R.sub.12 is cyclododecyl, imidazoyl, or
cyclohexenyl; R.sub.13 is --H or heteroatom-substituted or
heteroatom-unsubstituted C.sub.1-C.sub.10-alkyl,
C.sub.1-C.sub.7-cycloalkyl, C.sub.1-C.sub.10-acyloxy,
C.sub.6-C.sub.10-aryl, or C.sub.7-C.sub.10-aralkyl; X.sub.7 is halo
or heteroatom-substituted or heteroatom-unsubstituted
C.sub.1-C.sub.10-alkyl or C.sub.1-C.sub.10-alkoxy; R.sub.14 is:
##STR00011##
[0015] Other aspects of the invention include pharmaceutically
acceptable salts, hydrates, amine-N-oxides, imine-N-oxides,
tautomers, and optical isomers of the compounds described above and
throughout this application.
[0016] In certain embodiments R.sub.2 may cyclopropyl, cyclobutyl,
cyclopentyl or cyclohexyl. In further embodiments R.sub.3 may be
phenyl, benzyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
In yet further embodiments, X.sub.1 or X.sub.2 may be --F, --Cl,
--Br or --I. In still further embodiments, R.sub.4 may be hydroxy,
acetoxy or 2,2-dimethylpropionyloxy. In still yet further
embodiments, X.sub.3 or X.sub.4 may be methoxy, --F, --Cl, --Br or
--I. In some embodiments, R.sub.6 or R.sub.8 may be methyl or
cyclopropyl. In certain aspects, X.sub.5 may be methyl or
acetoxymethyl.
[0017] The compounds shown in Table 2 are specific examples of
compounds provided by this invention:
TABLE-US-00002 TABLE 2 Additional Examples of Caffeic Acid Analogs:
##STR00012## (WP1332) ##STR00013## (WP1331) ##STR00014## (WP1330)
##STR00015## (WP1329) ##STR00016## (WP1328) ##STR00017## (WP1302)
##STR00018## (WP1293) ##STR00019## (WP1286) ##STR00020## (WP1204)
##STR00021## (WP1285) ##STR00022## (WP1284) ##STR00023## (WP1283)
##STR00024## (WP1282) ##STR00025## WP1280 ##STR00026## (WP1273)
##STR00027## (WP1272) ##STR00028## (WP1246) ##STR00029## (WP1229)
##STR00030## (WP1269) ##STR00031## (WP1271) ##STR00032## (WP1268)
##STR00033## (WP1267) ##STR00034## (WP1203) ##STR00035## (WP1201)
##STR00036## (WP1196) ##STR00037## (WP1180) ##STR00038## (WP1179)
##STR00039## (WP1169) ##STR00040## (WP1168) ##STR00041## (WP1167)
##STR00042## (WP1166) ##STR00043## (WP1164) ##STR00044## (WP1163)
##STR00045## (WP1159) ##STR00046## (WP1145) ##STR00047## (WP1193)
##STR00048## (WP1082)
[0018] Some of the compounds in Table 2 are shown as single
enantiomers or diastereomers. The invention provides for all
possible stereoisomers of any of the compounds shown in Table 2
above, as well as those described throughout the application. In
some embodiments, the compound provided will be a single enantiomer
substantially free from other stereoisomers. In other embodiments,
the compound will be a mixture of different stereo isomers, wherein
each stereo isomer has the same molecular formula. In certain of
these embodiments, the invention provides for a racemic mixture of
a given molecular formula.
[0019] Another aspect of the invention comprises compounds having
the formula:
##STR00049##
wherein A is --C(O)-- or --SO.sub.2--. In certain embodiments
R.sub.1 is cyclododecyl,
##STR00050##
In some of these embodiments, X.sub.1, X.sub.2, X.sub.3, and
X.sub.4, are each independently hydrogen, halo, alkyl, alkoxyl,
acetoxyl, alkylacetoxyl, --OH, trihalomethyl, or --NO.sub.2;
Y.sub.1 is halo, --OH or --NO.sub.2; and R.sub.2 is selected from
the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylaryl,
halo, hydrogen, --OH, --NO.sub.2, thioether, amino, --SH, or
--NH.sub.2; R.sub.3 is:
##STR00051##
wherein Z.sub.3 may be a divalent alkyl; and wherein m.sub.1=1, 2,
3, or 4; and R.sub.4 may be hydrogen, --CN, substituted amine,
--CH.sub.2S-alkyl, alkyl, or --CH.sub.2N.sub.3. In some of these
embodiments, R.sub.5 and R.sub.6 are each independently:
##STR00052##
monosaccharide, monosaccharide derivative, polysaccharide,
polysaccharide derivative, aryl or aralkyl; Z is selected from the
group consisting of --NH, --S--, and --O--, and X.sub.5 and X.sub.6
are each independently selected from the group consisting of
hydrogen, upper alkyl, lower alkyl, cycloalkyl, cycloarylalkyl,
aralkyl, aryl, alkoxyl, hydroxyl, hydroxylalkyl, alkylester,
alkylesteralkyl, alkylacetoxyl, or aryloxyl; with the proviso that
if R.sub.4=--CN, substituted amine, --CH.sub.2S-alkyl, alkyl, or
--CH.sub.2N.sub.3, then R.sub.1 is selected from the group
consisting of: cyclododecyl,
##STR00053##
[0020] In further embodiments, R.sub.3 is:
##STR00054##
where X.sub.5 or X.sub.6 is upper alkyl, hydroxyl, aryl, alkoxyl,
aryloxyl; cycloalkyl, cycloarylalkyl, aralkyl, alkylester,
alkylesteralkyl, alkylacetoxy, or aryloxyl.
[0021] In specific embodiments, Z.sub.3 may be --C.sub.2H.sub.4--.
In some examples, X.sub.1, X.sub.2, X.sub.3, and X.sub.4, are each
independently --F, --Cl, --Br, --CH.sub.3, methoxy or
alkylacetoxyl. In further embodiments, X.sub.5 or X.sub.6 is
independently hydrogen, cyclopropyl, cyclobutyl, --CH.sub.3,
--CH.sub.2OH, cyclopentyl, --CH.sub.2OAc,
--CH.sub.2OC(O)C(CH.sub.3).sub.3, --CH.sub.2C.sub.6H.sub.5,
cyclohexyl or aryl.
[0022] In other embodiments, R.sub.5 is an aralkyl having the
structure:
##STR00055##
[0023] an aryl having the structure:
##STR00056##
wherein m=0, 1, 2, 3, 4, 5, 6, or 7 and where X.sub.5 and X.sub.6
are each independently selected from the group consisting of
hydrogen and alkyl, and where R.sub.7, R.sub.8, R.sub.9, R.sub.10,
and R.sub.11 are each independently selected from the group
consisting of hydrogen, halo, alkyl, alkoxy, --OH, trihalomethyl,
and --NO.sub.2.
[0024] A method of treating a cell proliferative disease comprising
administering to a subject an amount of a first compound effective
to treat the cell proliferative disease, wherein the first compound
is caffeic acid, the benzyl ester of caffeic acid, or one of the
compounds the present invention, such as a compound according to
Table 1 or a compound shown in Table 2.
[0025] Another aspect of the present invention concerns a method of
treating a cell proliferative disease comprising administering a
therapeutically relevant amount of a first compound of the present
invention to a subject. The subject may be a mammal, and the mammal
may be a human. The first compound may be comprised in a
pharmaceutically acceptable excipient, diluent, or vehicle. The
cell proliferative disease may be cancer. The cancer may be
melanoma, non-small cell lung, small cell lung, lung,
hepatocarcinoma, retinoblastoma, astrocytoma, glioblastoma,
leukemia, blood, brain, skin, eye, tongue, gum, neuroblastoma,
head, neck, breast, pancreatic, renal, bone, testicular, ovarian,
mesothelioma, cervical, gastrointestinal, lymphoma, colon, or
bladder.
[0026] The cell proliferative disease may be rheumatoid arthritis,
inflammatory bowel disease, osteoarthritis, leiomyomas, adenomas,
lipomas, hemangiomas, fibromas, vascular occlusion, restenosis,
artherosclerosis, a pre-neoplastic lesion, carcinoma in situ, oral
hairy leukoplakia, or psoriasis and the variant forms of psoriasis
including psoriatic arthritis other skin inflammatory conditions
such as urticaria, excema, atopic dermatitis, granuloma annulare,
angiomas, basal cell carcinoma, squamous cell carcinoma, malignant
melanoma, seborrheic dermatitis, rosacea other hyperactive
autoimmune disorders such as rheumatoid arthritis, chronic active
hepatitis, Hashimoto's thyroiditis, lupus, connective tissue
disorders, mixed connective tissue disorders, and neurologic
inflammatory diseases such as multiple sclerosis, inflammatory
leukoencephalitis.
[0027] In certain embodiments, STAT3 activation is reduced in a
cell of the subject. The expression of c-myc may be reduced in a
cell of the subject. The first compound may be administered in
combination with a therapeutically relevant amount of a second
compound. The second compound may be an anti-cancer compound. The
first compound may be administered in combination with a surgery, a
radiation therapy, or a gene therapy.
[0028] Any embodiment discussed herein with respect to one aspect
of the invention applies to other aspects of the invention as well,
unless specifically noted.
[0029] The term "about" is used to indicate that a value includes
the standard deviation of error for the device or method being
employed to determine the value. The use of the term "or" in the
claims is used to mean "and/or" unless explicitly indicated to
refer to alternatives only or the alternatives are mutually
exclusive, although the disclosure supports a definition that
refers to only alternatives and to "and/or." When used in
conjunction with the word "comprising" or other open language in
the claims, the words "a" and "an" denote "one or more," unless
specifically noted. The terms "comprise," "have" and "include" are
open-ended linking verbs. Any forms or tenses of one or more of
these verbs, such as "comprises," "comprising," "has," "having,"
"includes" and "including," are also open-ended. For example, any
method that "comprises," "has" or "includes" one or more steps is
not limited to possessing only those one or more steps and also
covers other unlisted steps. Similarly, any plant that "comprises,"
"has" or "includes" one or more traits is not limited to possessing
only those one or more traits and covers other unlisted traits.
[0030] The terms "inhibiting," "reducing," or "prevention," or any
variation of these terms, when used in the claims and/or the
specification includes any measurable decrease or complete
inhibition to achieve a desired result.
[0031] The term "effective," as that term is used in the
specification and/or claims, means adequate to accomplish a
desired, expected, or intended result.
[0032] As used herein, predominantly one enantiomer or
substantially free from other optical isomers means that the
compound contains at least 95% of one enantiomer, or more
preferably at least 98% of one enantiomer, or most preferably at
least 99% of one enantiomer.
[0033] Other objects, features and advantages of the present
invention will become apparent from the following detailed
description. It should be understood, however, that the detailed
description and any specific examples provided, while indicating
specific embodiments of the invention, are given by way of
illustration only, since various changes and modifications within
the spirit and scope of the invention will become apparent to those
skilled in the art from this detailed description.
BRIEF DESCRIPTION OF THE DRAWINGS
[0034] The following drawings form part of the present
specification and are included to further demonstrate certain
aspects of the present invention. The invention may be better
understood by reference to one or more of these drawings in
combination with the detailed description of specific embodiments
presented herein.
[0035] FIG. 1: Inhibition of constitutively activated STAT3 in
Colo357-FG cells with WP1066 and WP 1193. Western blots of
phosphorylated STAT3 at Tyr705 and total STAT3.
[0036] FIG. 2: Dose-response curve for WP1193 in the presence of
various cell lines. Values are the means.+-.s.d. (error bars) of
triplicate experiments.
[0037] FIG. 3: Dose-response curve for WP1145 in the presence of
various cell lines.
[0038] FIG. 4: Dose-response curve for WP1163 in the presence of
the U87 cell line.
[0039] FIG. 5: Dose-response curve for WP1164 in the presence of
the U87 cell line.
[0040] FIG. 6: Dose-response curve for WP1164 in the presence of
the Colo357-FG cell line.
[0041] FIG. 7: Dose-response curve for WP1166 in the presence of
the U87 cell line.
[0042] FIG. 8: Dose-response curve for WP1167 in the presence of
the U87 cell line.
[0043] FIG. 9: Dose-response curve for WP1168 in the presence of
the U87 cell line.
[0044] FIG. 10: Dose-response curve for WP1169 in the presence of
the U87 cell line.
[0045] FIG. 11: Dose-response curve for WP1229 in the presence of
the U87 and FG cell lines.
[0046] FIG. 12: Dose-response curve for WP1229 in the presence of
the U87 cell line.
[0047] FIG. 13: Dose-response curve for WP1146 in the presence of
the U87 cell line. Values are the means.+-.s.d. (error bars) of
triplicate experiments.
[0048] FIG. 14: Dose-response curve for WP1267 in the presence of
the Colo357-FG cell line. Values are the means.+-.s.d. (error bars)
of triplicate experiments.
[0049] FIG. 15: Dose-response curve for WP1267 in the presence of
the U87 cell line. Values are the means.+-.s.d. (error bars) of
triplicate experiments.
[0050] FIG. 16: Dose-response curve for WP1268 in the presence of
the Colo357-FG cell line. Values are the means.+-.s.d. (error bars)
of triplicate experiments.
[0051] FIG. 17: Dose-response curve for WP1268 in the presence of
the U87 cell line. Values are the means.+-.s.d. (error bars) of
triplicate experiments.
[0052] FIG. 18: Dose-response curve for WP1269 in the presence of
the U87 cell line. Values are the means.+-.s.d. (error bars) of
triplicate experiments.
[0053] FIG. 19: Dose-response curve for WP1269 in the presence of
the Colo357-FG cell line. Values are the means.+-.s.d. (error bars)
of triplicate experiments.
[0054] FIG. 20: Dose-response curve for WP1282 in the presence of
the Colo357-FG cell line.
[0055] FIG. 21: Dose-response curve for WP1282 in the presence of
the U87 cell line.
[0056] FIG. 22: Dose-response curve for WP1283 in the presence of
the Colo357-FG cell line.
[0057] FIG. 23: Dose-response curve for WP1283 in the presence of
the U87 cell line.
[0058] FIG. 24: Dose-response curve for WP1284 in the presence of
the U87 cell line.
[0059] FIG. 25: Dose-response curve for WP1284 in the presence of
the Colo357-FG cell line.
[0060] FIG. 26: Dose-response curve for WP1285 in the presence of
the U87 cell line.
[0061] FIG. 27: Dose-response curve for WP1285 in the presence of
the Colo357-FG cell line.
[0062] FIG. 28: Dose-response curve for WP1286 in the presence of
the U87 cell line.
[0063] FIG. 29: Dose-response curve for WP1286 in the presence of
the Colo357-FG cell line.
[0064] FIG. 30: Dose-response curve for WP1293 in the presence of
the Colo357-FG cell line.
[0065] FIG. 31: Dose-response curve for WP1293 in the presence of
the U87 cell line.
[0066] FIG. 32: Dose-response curve for WP1302 in the presence of
the U87 cell line.
[0067] FIG. 33: Dose-response curve for WP1302 in the presence of
the Colo357-FG cell line.
[0068] FIG. 34: Compound WP1193 more potently than WP1066 inhibits
STAT3 phosphorylation. The graph shows survival (% of control) vs.
concentration in micromolar units.
DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS
I. The Present Invention
[0069] Previous studies have demonstrated that cytokine pathways
that activate transcription factors (e.g., NF-kB, STAT3) are
unregulated or activated by genetic lesions or autocrine/paracrine
mechanisms in multiple tumor types (Hallek et al., 1998; Hideshima
et al., 2002). These pathways contribute to the tumorigenicity and
progression of cancer. In the present invention, new compounds were
synthesized, and in vitro screening revealed that these compounds
can block IL-6 mediated STAT3 activation at low concentrations
(.about.1 .mu.M). As compared to AG490, these compounds are
significantly more potent (20 to 50-fold) in tumor cells in
inhibiting Jak2/STAT3 signaling in IL-6 treated cells. These
compounds can also induce apoptosis in wide range of solid and
hematological tumor cells at concentrations that parallel their
Jak2/STAT3 pathway downregulatory activity. The present invention
discloses compounds that inactivate genes and signaling pathways
important for tumor cell survival and progression, and these
compounds may be used alone or in combination with other agents for
the treatment of cancer.
II. Chemical Definitions
[0070] As used herein, the term "amino" means --NH.sub.2; the term
"nitro" means --NO.sub.2; the term "halo" designates --F, --Cl,
--Br or --I; the term "mercapto" means --SH; the term "cyano" means
--CN; the term "silyl" means --SiH.sub.3, and the term "hydroxy"
means --OH.
[0071] An "alkane" refers to an acyclic branched or unbranched
hydrocarbon, in many cases having the general formula
C.sub.nH.sub.2n+2. An "alkyl" refers to a univalent group derived
from an alkane by removal of a hydrogen atom from any carbon atom
thus having the formula --C.sub.nH.sub.2n+1 in many cases. Alkyl
groups, either straight-chained or branched chained, may be
substituted with additional acyclic alkyl, cycloalkyl, or cyclic
alkyl groups. The alkyl group may be heteroatom-substituted or
heteroatom-unsubstituted, see below. Preferably, the alkyl group
has 1 to 12 carbons. More preferably, it is a lower alkyl having 1
to 7 carbons, more preferably 1 to 4 carbons. An upper alkyl has 8
or more carbon atoms. A "divalent alkyl" refers to a divalent group
derived from an alkane by removal of two hydrogen atoms from either
the same carbon atom (e.g. methylene, ethylidene, propylidene) or
from different carbon atoms (e.g. --C.sub.2H.sub.4--).
[0072] A "cycloalkane" refers to a saturated monocyclic
hydrocarbons with or without side chains.
[0073] A "cycloalkyl" refers to a univalent group derived from
cycloalkane by removal of a hydrogen atom from a ring carbon
atom.
[0074] The term "heteroatom-substituted," when used to modify a
class of organic radicals (e.g. alkyl, aryl, acyl, etc.), means
that one, or more than one, hydrogen atom of that radical has been
replaced by a heteroatom, or a heteroatom containing group.
Examples of heteroatoms and heteroatom containing groups include:
hydroxy, cyano, alkoxy, .dbd.O, .dbd.S, --NO.sub.2,
--N(CH.sub.3).sub.2, amino, or --SH. Specific
heteroatom-substituted organic radicals are defined more fully
below.
[0075] The term "heteroatom-unsubstituted," when used to modify a
class of organic radicals (e.g. alkyl, aryl, acyl, etc.) means that
none of the hydrogen atoms of that radical have been replaced with
a heteroatom or a heteroatom containing group. Substitution of a
hydrogen atom with a carbon atom, or a group consisting of only
carbon and hydrogen atoms, is not sufficient to make a group
heteroatom-substituted. For example, the group
--C.sub.6H.sub.4C.ident.CH is an example of a
heteroatom-unsubstituted aryl group, while --C.sub.6H.sub.4F is an
example of a heteroatom-substituted aryl group. Specific
heteroatom-unsubstituted organic radicals are defined more fully
below.
[0076] The term "heteroatom-unsubstituted C.sub.n-alkyl" refers to
an alkyl, further having a total of n carbon atoms, all of which
are nonaromatic, 3 or more hydrogen atoms, and no heteroatoms. For
example, a heteroatom-unsubstituted C.sub.1-C.sub.10-alkyl has 1 to
10 carbon atoms. The groups, --CH.sub.3, cyclopropylmethyl,
--CH.sub.2CH.sub.3, --CH.sub.2CH.sub.2CH.sub.3,
--CH(CH.sub.3).sub.2, --CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
--CH(CH.sub.3)CH.sub.2CH.sub.3, --CH.sub.2CH(CH.sub.3).sub.2,
--C(CH.sub.3).sub.3, and --CH.sub.2C(CH.sub.3).sub.3 are all
examples of heteroatom-unsubstituted alkyl groups.
[0077] The term "heteroatom-substituted C.sub.n-alkyl" refers to an
alkyl, further having a total of n carbon atoms, all of which are
nonaromatic, 0, 1, or more than one hydrogen atom, at least one
heteroatom, wherein each heteroatom is independently selected from
the group consisting of N, O, F, Cl, Br, I, Si, P, and S. For
example, a heteroatom-substituted C.sub.1-C.sub.10-alkyl has 1 to
10 carbon atoms. The following groups are all examples of
heteroatom-substituted alkyl groups: trifluoromethyl, --CH.sub.2F,
--CH.sub.2Cl, --CH.sub.2Br, --CH.sub.2OH, --CH.sub.2OCH.sub.3,
--CH.sub.2OCH.sub.2CH.sub.3, --CH.sub.2OCH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2OCH(CH.sub.3).sub.2, --CH.sub.2OCH.sub.2CF.sub.3,
--CH.sub.2OCOCH.sub.3, --CH.sub.2NH.sub.2, --CH.sub.2NHCH.sub.3,
--CH.sub.2N(CH.sub.3).sub.2, --CH.sub.2NHCH.sub.2CH.sub.3,
--CH.sub.2N(CH.sub.3)CH.sub.2CH.sub.3,
--CH.sub.2NHCH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2NHCH(CH.sub.3).sub.2, --CH.sub.2OCH(CH.sub.2).sub.2,
--CH.sub.2NHCH(CH.sub.2).sub.2,
--CH.sub.2CH.sub.2NHCH(CH.sub.2).sub.2,
--CH.sub.2N(CH.sub.2CH.sub.3).sub.2, --CH.sub.2CH.sub.2F,
--CH.sub.2CH.sub.2Cl, --CH.sub.2CH.sub.2Br, --CH.sub.2CH.sub.2I,
--CH.sub.2CH.sub.2OH, CH.sub.2CH.sub.2OCOCH.sub.3,
--CH.sub.2CH.sub.2NH.sub.2, --CH.sub.2CH.sub.2N(CH.sub.3).sub.2,
--CH.sub.2CH.sub.2NHCH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2N(CH.sub.3)CH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2NHCH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2NHCH(CH.sub.3).sub.2,
--CH.sub.2CH.sub.2N(CH.sub.2CH.sub.3).sub.2,
--CH.sub.2CH.sub.2NHCO.sub.2C(CH.sub.3).sub.3, and
--CH.sub.2Si(CH.sub.3).sub.3.
[0078] The term "heteroatom-unsubstituted C.sub.n-cycloalkyl"
refers to a cycloalkyl, further having a total of n carbon atoms,
all of which are nonaromatic, 3 or more hydrogen atoms, and no
heteroatoms. For example, a heteroatom-unsubstituted
C.sub.1-C.sub.10-cycloalkyl has 1 to 10 carbon atoms. The groups
--CH(CH.sub.2).sub.2 (cyclopropyl), cyclobutyl, cyclopentyl, and
cyclohexyl, are all examples of heteroatom-unsubstituted cycloalkyl
groups.
[0079] The term "heteroatom-substituted C.sub.n-cycloalkyl" refers
to a cycloalkyl, further having a total of n carbon atoms, all of
which are nonaromatic, 0, 1, or more than one hydrogen atom, at
least one heteroatom, wherein each heteroatom is independently
selected from the group consisting of N, O, F, Cl, Br, I, Si, P,
and S. For example, a heteroatom-substituted
C.sub.1-C.sub.10-cycloalkyl has 1 to 10 carbon atoms.
[0080] The term "heteroatom-unsubstituted C.sub.n-alkenyl" refers
to a radical, having a linear or branched, cyclic or acyclic
structure, further having at least one nonaromatic carbon-carbon
double bond, but no carbon-carbon triple bonds, a total of n carbon
atoms, three or more hydrogen atoms, and no heteroatoms. For
example, a heteroatom-unsubstituted C.sub.2-C.sub.10-alkenyl has 2
to 10 carbon atoms. Heteroatom-unsubstituted alkenyl groups
include: --CH.dbd.CH.sub.2, --CH.dbd.CHCH.sub.3,
--CH.dbd.CHCH.sub.2CH.sub.3, --CH.dbd.CHCH.sub.2CH.sub.2CH.sub.3,
--CH.dbd.CHCH(CH.sub.3).sub.2, --CH.dbd.CHCH(CH.sub.2).sub.2,
--CH.sub.2CH.dbd.CH.sub.2, --CH.sub.2CH.dbd.CHCH.sub.3,
--CH.sub.2CH.dbd.CHCH.sub.2CH.sub.3,
--CH.sub.2CH.dbd.CHCH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2CH.dbd.CHCH(CH.sub.3).sub.2,
--CH.sub.2CH.dbd.CHCH(CH.sub.2).sub.2, and
--CH.dbd.CH--C.sub.6H.sub.5.
[0081] The term "heteroatom-substituted C.sub.n-alkenyl" refers to
a radical, having a single nonaromatic carbon atom as the point of
attachment and at least one nonaromatic carbon-carbon double bond,
but no carbon-carbon triple bonds, further having a linear or
branched, cyclic or acyclic structure, further having a total of n
carbon atoms, 0, 1, or more than one hydrogen atom, and at least
one heteroatom, wherein each heteroatom is independently selected
from the group consisting of N, O, F, Cl, Br, I, Si, P, and S. For
example, a heteroatom-substituted C.sub.2-C.sub.10-alkenyl has 2 to
10 carbon atoms. The groups, --CH.dbd.CHF, --CH.dbd.CHCl and
--CH.dbd.CHBr, are examples of heteroatom-substituted alkenyl
groups.
[0082] The term "heteroatom-unsubstituted C.sub.n-alkynyl" refers
to a radical, having a linear or branched, cyclic or acyclic
structure, further having at least one carbon-carbon triple bond, a
total of n carbon atoms, at least one hydrogen atom, and no
heteroatoms. For example, a heteroatom-unsubstituted
C.sub.2-C.sub.10-alkynyl has 2 to 10 carbon atoms. The groups,
--C.ident.CH, --C.ident.CCH.sub.3, and --C.ident.CC.sub.6H.sub.5
are examples of heteroatom-unsubstituted alkynyl groups.
[0083] The term "heteroatom-substituted C.sub.n-alkynyl" refers to
a radical, having a single nonaromatic carbon atom as the point of
attachment and at least one carbon-carbon triple bond, further
having a linear or branched, cyclic or acyclic structure, and
having a total of n carbon atoms, 0, 1, or more than one hydrogen
atom, and at least one heteroatom, wherein each heteroatom is
independently selected from the group consisting of N, O, F, Cl,
Br, I, Si, P, and S. For example, a heteroatom-substituted
C.sub.2-C.sub.10-alkynyl has 2 to 10 carbon atoms. The group,
--C.ident.CSi(CH.sub.3).sub.3, is an example of a
heteroatom-substituted alkynyl group.
[0084] The term "heteroatom-unsubstituted C.sub.n-aryl" refers to a
radical, having a single carbon atom as a point of attachment,
wherein the carbon atom is part of an aromatic ring structure
containing only carbon atoms, further having a total of n carbon
atoms, 5 or more hydrogen atoms, and no heteroatoms. For example, a
heteroatom-unsubstituted C.sub.6-C.sub.10-aryl has 6 to 10 carbon
atoms. Examples of heteroatom-unsubstituted aryl groups include
phenyl, methylphenyl, (dimethyl)phenyl,
C.sub.6H.sub.4--CH.sub.2CH.sub.3,
C.sub.6H.sub.4--CH.sub.2CH.sub.2CH.sub.3,
--C.sub.6H.sub.4--CH(CH.sub.3).sub.2,
--C.sub.6H.sub.4--CH(CH.sub.2).sub.2,
--C.sub.6H.sub.3(CH.sub.3)CH.sub.2CH.sub.3,
--C.sub.6H.sub.4--CH--CH.sub.2,
--C.sub.6H.sub.4--CH.dbd.CHCH.sub.3, --C.sub.6H.sub.4C.ident.CH,
--C.sub.6H.sub.4C.ident.CCH.sub.3, naphthyl, quinolyl, indolyl, and
the radical derived from biphenyl. The term
"heteroatom-unsubstituted aryl" includes carbocyclic aryl groups,
biaryl groups, and radicals derived from polycyclic fused
hydrocarbons (PAHs).
[0085] The term "heteroatom-substituted C.sub.n-aryl" refers to a
radical, refers to a radical, having either a single aromatic
carbon atom or a single aromatic heteroatom as the point of
attachment, further having a total of n carbon atoms, at least one
hydrogen atom, and at least one heteroatom, further wherein each
heteroatom is independently selected from the group consisting of
N, O, F, Cl, Br, I, Si, P, and S. For example, a
heteroatom-unsubstituted C.sub.1-C.sub.10-heteroaryl has 1 to 10
carbon atoms. The term "heteroatom-substituted aryl" includes
heteroaryl and heterocyclic aryl groups. It also includes those
groups derived from the compounds: pyrrole, furan, thiophene,
imidazole, oxazole, isoxazole, thiazole, isothiazole, triazole,
pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, and the like.
Further examples of heteroatom-substituted aryl groups include the
groups: --C.sub.6H.sub.4F, --C.sub.6H.sub.4Cl, --C.sub.6H.sub.4Br,
--C.sub.6H.sub.4I, --C.sub.6H.sub.4OH, --C.sub.6H.sub.4OCH.sub.3,
--C.sub.6H.sub.4OCH.sub.2CH.sub.3, --C.sub.6H.sub.4OCOCH.sub.3,
--C.sub.6H.sub.4OC.sub.6H.sub.5, C.sub.6H.sub.4NH.sub.2,
--C.sub.6H.sub.4NHCH.sub.3, --C.sub.6H.sub.4NHCH.sub.2CH.sub.3,
--C.sub.6H.sub.4CH.sub.2 Cl, --C.sub.6H.sub.4CH.sub.2Br,
--C.sub.6H.sub.4CH.sub.2OH, --C.sub.6H.sub.4CH.sub.2OCOCH.sub.3,
--C.sub.6H.sub.4CH.sub.2NH.sub.2,
--C.sub.6H.sub.4N(CH.sub.3).sub.2, C.sub.6H.sub.4CH.sub.2 CH.sub.2
Cl, --C.sub.6H.sub.4CH.sub.2CH.sub.2OH,
--C.sub.6H.sub.4CH.sub.2CH.sub.2OCOCH.sub.3,
--C.sub.6H.sub.4CH.sub.2CH.sub.2NH.sub.2,
C.sub.6H.sub.4CH.sub.2CH.dbd.CH.sub.2, C.sub.6H.sub.4CF.sub.3,
--C.sub.6H.sub.4CN, C.sub.6H.sub.4C.ident.CSi(CH.sub.3).sub.3,
C.sub.6H.sub.4COH, --C.sub.6H.sub.4 COCH.sub.3,
--C.sub.6H.sub.4COCH.sub.2CH.sub.3,
--C.sub.6H.sub.4COCH.sub.2CF.sub.3,
--C.sub.6H.sub.4COC.sub.6H.sub.5, --C.sub.6H.sub.4CO.sub.2H,
--C.sub.6H.sub.4CO.sub.2CH.sub.3, --C.sub.6H.sub.4CONH.sub.2,
--C.sub.6H.sub.4CONHCH.sub.3, --C.sub.6H.sub.4CON(CH.sub.3).sub.2,
furanyl, thienyl, pyridyl, pyrrolyl, pyrimidyl, pyrazinyl, and
imidazoyl.
[0086] The term "heteroatom-unsubstituted C.sub.n-aralkyl" refers
to a radical, having a single saturated carbon atom as the point of
attachment, further having a total of n carbon atoms, wherein at
least 6 of the carbon atoms form an aromatic ring structure
containing only carbon atoms, 7 or more hydrogen atoms, and no
heteroatoms. For example, a heteroatom-unsubstituted
C.sub.7-C.sub.10-aralkyl has 7 to 10 carbon atoms. An "aralkyl"
includes an alkyl heteroatom-substituted with an aryl group.
Examples of heteroatom-unsubstituted aralkyls include phenylmethyl
(benzyl) and phenylethyl.
[0087] The term "heteroatom-substituted C.sub.n-aralkyl" refers to
a radical, having a single saturated carbon atom as the point of
attachment, further having a total of n carbon atoms, 0, 1, or more
than one hydrogen atom, and at least one heteroatom, wherein at
least one of the carbon atoms is incorporated an aromatic ring
structures, further wherein each heteroatom is independently
selected from the group consisting of N, O, F, Cl, Br, I, Si, P,
and S. For example, a heteroatom-substituted
C.sub.2-C.sub.10-heteroaralkyl has 2 to 10 carbon atoms.
[0088] The term "heteroatom-unsubstituted C.sub.n-acyl" refers to a
radical, having a single carbon atom of a carbonyl group as the
point of attachment, further having a linear or branched, cyclic or
acyclic structure, further having a total of n carbon atoms, 1 or
more hydrogen atoms, a total of one oxygen atom, and no additional
heteroatoms. For example, a heteroatom-unsubstituted
C.sub.1-C.sub.10-acyl has 1 to 10 carbon atoms. The groups, --COH,
--COCH.sub.3, --COCH.sub.2CH.sub.3, --COCH.sub.2CH.sub.2CH.sub.3,
COCH(CH.sub.3).sub.2, --COCH(CH.sub.2).sub.2, COC.sub.6H.sub.5,
--COC.sub.6H.sub.4CH.sub.3, --COC.sub.6H.sub.4CH.sub.2CH.sub.3,
--COC.sub.6H.sub.4CH.sub.2CH.sub.2CH.sub.3,
--COC.sub.6H.sub.4CH(CH.sub.3).sub.2,
--COC.sub.6H.sub.4CH(CH.sub.2).sub.2, and
--COC.sub.6H.sub.3(CH.sub.3).sub.2, are examples of
heteroatom-unsubstituted acyl groups.
[0089] The term "heteroatom-substituted C.sub.n-acyl" refers to a
radical, having a single carbon atom as the point of attachment,
the carbon atom being part of a carbonyl group, further having a
linear or branched, cyclic or acyclic structure, further having a
total of n carbon atoms, 0, 1, or more than one hydrogen atom, at
least one additional heteroatom in addition to the oxygen of the
carbonyl group, wherein each additional heteroatom is independently
selected from the group consisting of N, O, F, Cl, Br, I, Si, P,
and S. For example, a heteroatom-substituted C.sub.1-C.sub.10-acyl
has 1 to 10 carbon atoms. The term heteroatom-substituted acyl
includes carbamoyl, thiocarboxylate, and thiocarboxylic acid
groups. The groups, --COCH.sub.2CF.sub.3, --CO.sub.2H,
--CO.sub.2CH.sub.3, --CO.sub.2CH.sub.2CH.sub.3,
--CO.sub.2CH.sub.2CH.sub.2CH.sub.3, --CO.sub.2CH(CH.sub.3).sub.2,
--CO.sub.2CH(CH.sub.2).sub.2, --CONH.sub.2, --CONHCH.sub.3,
--CONHCH.sub.2CH.sub.3, --CONHCH.sub.2CH.sub.2CH.sub.3,
--CONHCH(CH.sub.3).sub.2, --CONHCH(CH.sub.2).sub.2,
--CON(CH.sub.3).sub.2, --CON(CH.sub.2CH.sub.3)CH.sub.3,
--CON(CH.sub.2CH.sub.3).sub.2 and --CONHCH.sub.2CF.sub.3, are
examples heteroatom-substituted acyl groups.
[0090] The term "heteroatom-unsubstituted C.sub.n-alkoxy" refers to
a group, having the structure --OR, in which R is a
heteroatom-unsubstituted C.sub.n-alkyl, as that term is defined
above. Heteroatom-unsubstituted alkoxy groups include: --OCH.sub.3,
--OCH.sub.2CH.sub.3, --OCH.sub.2CH.sub.2CH.sub.3,
--OCH(CH.sub.3).sub.2, and --OCH(CH.sub.2).sub.2.
[0091] The term "heteroatom-substituted C.sub.n-alkoxy" refers to a
group, having the structure --OR, in which R is a
heteroatom-substituted C.sub.n-alkyl, as that term is defined
above. For example, --OCH.sub.2CF.sub.3 is a heteroatom-substituted
alkoxy group.
[0092] The term "heteroatom-unsubstituted C.sub.n-alkenyloxy"
refers to a group, having the structure --OR, in which R is a
heteroatom-unsubstituted C.sub.n-alkenyl, as that term is defined
above.
[0093] The term "heteroatom-substituted C.sub.n-alkenyloxy" refers
to a group, having the structure --OR, in which R is a
heteroatom-substituted C.sub.n-alkenyl, as that term is defined
above.
[0094] The term "heteroatom-unsubstituted C.sub.n-alkynyloxy"
refers to a group, having the structure --OR, in which R is a
heteroatom-unsubstituted C.sub.n-alkynyl, as that term is defined
above.
[0095] The term "heteroatom-substituted C.sub.n-alkynyloxy" refers
to a group, having the structure --OR, in which R is a
heteroatom-substituted C.sub.n-alkynyl, as that term is defined
above.
[0096] The term "heteroatom-unsubstituted C.sub.n-aryloxy" refers
to a group, having the structure --OAr, in which Ar is a
heteroatom-unsubstituted C.sub.n-aryl, as that term is defined
above. An example of a heteroatom-unsubstituted aryloxy group is
--OC.sub.6H.sub.5.
[0097] The term "heteroatom-substituted C.sub.n-aryloxy" refers to
a group, having the structure --OAr, in which Ar is a
heteroatom-substituted C.sub.n-aryl, as that term is defined
above.
[0098] The term "heteroatom-unsubstituted C.sub.n-aralkyloxy"
refers to a group, having the structure --OAr, in which Ar is a
heteroatom-unsubstituted C.sub.n-aralkyl, as that term is defined
above.
[0099] The term "heteroatom-substituted C.sub.n-aralkyloxy" refers
to a group, having the structure --OAr, in which Ar is a
heteroatom-substituted C.sub.n-aralkyl, as that term is defined
above.
[0100] The term "heteroatom-unsubstituted C.sub.n-acyloxy" refers
to a group, having the structure --OAc, in which Ac is a
heteroatom-unsubstituted C.sub.n-acyl, as that term is defined
above. A heteroatom-unsubstituted acyloxy group includes
alkylcarbonyloxy and arylcarbonyloxy groups. For example,
--OCOCH.sub.3 is an example of a heteroatom-unsubstituted acyloxy
group.
[0101] The term "heteroatom-substituted C.sub.n-acyloxy" refers to
a group, having the structure --OAc, in which Ac is a
heteroatom-substituted C.sub.n-acyl, as that term is defined above.
A heteroatom-substituted acyloxy group includes alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl,
aminocarbonyl, and alkylthiocarbonyl groups.
[0102] The term "heteroatom-unsubstituted C.sub.n-alkylamino"
refers to a radical, having a single nitrogen atom as the point of
attachment, further having one or two saturated carbon atoms
attached to the nitrogen atom, further having a linear or branched,
cyclic or acyclic structure, containing a total of n carbon atoms,
all of which are nonaromatic, 4 or more hydrogen atoms, a total of
1 nitrogen atom, and no additional heteroatoms. For example, a
heteroatom-unsubstituted C.sub.1-C.sub.10-alkylamino has 1 to 10
carbon atoms. The term "heteroatom-unsubstituted
C.sub.n-alkylamino" includes groups, having the structure --NHR, in
which R is a heteroatom-unsubstituted C.sub.n-alkyl, as that term
is defined above. A heteroatom-unsubstituted alkylamino group would
include --NHCH.sub.3, --NHCH.sub.2CH.sub.3,
--NHCH.sub.2CH.sub.2CH.sub.3, --NHCH(CH.sub.3).sub.2,
--NHCH(CH.sub.2).sub.2, --NHCH.sub.2CH.sub.2CH.sub.2CH.sub.3,
--NHCH(CH.sub.3)CH.sub.2CH.sub.3, --NHCH.sub.2CH(CH.sub.3).sub.2,
--NHC(CH.sub.3).sub.3, --N(CH.sub.3).sub.2,
--N(CH.sub.3)CH.sub.2CH.sub.3, --N(CH.sub.2CH.sub.3).sub.2,
N-pyrrolidinyl, and N-piperidinyl.
[0103] The term "heteroatom-substituted C.sub.n-alkylamino" refers
to a radical, having a single nitrogen atom as the point of
attachment, further having one or two saturated carbon atoms
attached to the nitrogen atom, no carbon-carbon double or triple
bonds, further having a linear or branched, cyclic or acyclic
structure, further having a total of n carbon atoms, all of which
are nonaromatic, 0, 1, or more than one hydrogen atom, and at least
one additional heteroatom, that is, in addition to the nitrogen
atom at the point of attachment, wherein each additional heteroatom
is independently selected from the group consisting of N, O, F, Cl,
Br, I, Si, P, and S. For example, a heteroatom-substituted
C.sub.1-C.sub.10-alkylamino has 1 to 10 carbon atoms. The term
"heteroatom-substituted C.sub.n-alkylamino" includes groups, having
the structure --NHR, in which R is a heteroatom-substituted
C.sub.n-alkyl, as that term is defined above.
[0104] The term "heteroatom-unsubstituted C.sub.n-alkenylamino"
refers to a radical, having a single nitrogen atom as the point of
attachment, further having one or two carbon atoms attached to the
nitrogen atom, further having a linear or branched, cyclic or
acyclic structure, containing at least one nonaromatic
carbon-carbon double bond, a total of n carbon atoms, 4 or more
hydrogen atoms, a total of one nitrogen atom, and no additional
heteroatoms. For example, a heteroatom-unsubstituted
C.sub.2-C.sub.10-alkenylamino has 2 to 10 carbon atoms. The term
"heteroatom-unsubstituted C.sub.n-alkenylamino" includes groups,
having the structure --NHR, in which R is a
heteroatom-unsubstituted C.sub.n-alkenyl, as that term is defined
above. Examples of heteroatom-unsubstituted C.sub.n-alkenylamino
groups also include dialkenylamino and alkyl(alkenyl)amino
groups.
[0105] The term "heteroatom-substituted C.sub.n-alkenylamino"
refers to a radical, having a single nitrogen atom as the point of
attachment and at least one nonaromatic carbon-carbon double bond,
but no carbon-carbon triple bonds, further having one or two carbon
atoms attached to the nitrogen atom, further having a linear or
branched, cyclic or acyclic structure, further having a total of n
carbon atoms, 0, 1, or more than one hydrogen atom, and at least
one additional heteroatom, that is, in addition to the nitrogen
atom at the point of attachment, wherein each additional heteroatom
is independently selected from the group consisting of N, O, F, Cl,
Br, I, Si, P, and S. For example, a heteroatom-substituted
C.sub.2-C.sub.10-alkenylamino has 2 to 10 carbon atoms. The term
"heteroatom-substituted C.sub.n-alkenylamino" includes groups,
having the structure --NHR, in which R is a heteroatom-substituted
C.sub.n-alkenyl, as that term is defined above.
[0106] The term "heteroatom-unsubstituted C.sub.n-alkynylamino"
refers to a radical, having a single nitrogen atom as the point of
attachment, further having one or two carbon atoms attached to the
nitrogen atom, further having a linear or branched, cyclic or
acyclic structure, containing at least one carbon-carbon triple
bond, a total of n carbon atoms, at least one hydrogen atoms, a
total of one nitrogen atom, and no additional heteroatoms. For
example, a heteroatom-unsubstituted C.sub.2-C.sub.10-alkynylamino
has 2 to 10 carbon atoms. The term "heteroatom-unsubstituted
C.sub.n-alkynylamino" includes groups, having the structure --NHR,
in which R is a heteroatom-unsubstituted C.sub.n-alkynyl, as that
term is defined above. An alkynylamino group includes
dialkynylamino and alkyl(alkynyl)amino groups.
[0107] The term "heteroatom-substituted C.sub.n-alkynylamino"
refers to a radical, having a single nitrogen atom as the point of
attachment, further having one or two carbon atoms attached to the
nitrogen atom, further having at least one nonaromatic
carbon-carbon triple bond, further having a linear or branched,
cyclic or acyclic structure, and further having a total of n carbon
atoms, 0, 1, or more than one hydrogen atom, and at least one
additional heteroatom, that is, in addition to the nitrogen atom at
the point of attachment, wherein each additional heteroatom is
independently selected from the group consisting of N, O, F, Cl,
Br, I, Si, P, and S. For example, a heteroatom-substituted
C.sub.2-C.sub.10-alkynylamino has 2 to 10 carbon atoms. The term
"heteroatom-substituted C.sub.n-alkynylamino" includes groups,
having the structure --NHR, in which R is a heteroatom-substituted
C.sub.n-alkynyl, as that term is defined above.
[0108] The term "heteroatom-unsubstituted C.sub.n-arylamino" refers
to a radical, having a single nitrogen atom as the point of
attachment, further having at least one aromatic ring structure
attached to the nitrogen atom, wherein the aromatic ring structure
contains only carbon atoms, further having a total of n carbon
atoms, 6 or more hydrogen atoms, a total of one nitrogen atom, and
no additional heteroatoms. For example, a heteroatom-unsubstituted
C.sub.6-C.sub.10-arylamino has 6 to 10 carbon atoms. The term
"heteroatom-unsubstituted C.sub.n-arylamino" includes groups,
having the structure --NHR, in which R is a
heteroatom-unsubstituted C.sub.n-aryl, as that term is defined
above. A heteroatom-unsubstituted arylamino group includes
diarylamino and alkyl(aryl)amino groups.
[0109] The term "heteroatom-substituted C.sub.n-arylamino" refers
to a radical, having a single nitrogen atom as the point of
attachment, further having a total of n carbon atoms, at least one
hydrogen atom, at least one additional heteroatoms, that is, in
addition to the nitrogen atom at the point of attachment, wherein
at least one of the carbon atoms is incorporated into one or more
aromatic ring structures, further wherein each additional
heteroatom is independently selected from the group consisting of
N, O, F, Cl, Br, I, Si, P, and S. For example, a
heteroatom-substituted C.sub.6-C.sub.10-arylamino has 6 to 10
carbon atoms. The term "heteroatom-substituted C.sub.n-arylamino"
includes groups, having the structure --NHR, in which R is a
heteroatom-substituted C.sub.n-aryl, as that term is defined above.
A heteroatom-substituted arylamino group includes heteroarylamino
groups.
[0110] The term "heteroatom-unsubstituted C.sub.n-aralkylamino"
refers to a radical, having a single nitrogen atom as the point of
attachment, further having one or two saturated carbon atoms
attached to the nitrogen atom, further having a total of n carbon
atoms, wherein at least 6 of the carbon atoms form an aromatic ring
structure containing only carbon atoms, 8 or more hydrogen atoms, a
total of one nitrogen atom, and no additional heteroatoms. For
example, a heteroatom-unsubstituted C.sub.7-C.sub.10-aralkylamino
has 7 to 10 carbon atoms. The term "heteroatom-unsubstituted
C.sub.n-aralkylamino" includes groups, having the structure --NHR,
in which R is a heteroatom-unsubstituted C.sub.n-aralkyl, as that
term is defined above. An aralkylamino group includes
diaralkylamino groups.
[0111] The term "heteroatom-substituted C.sub.n-aralkylamino"
refers to a radical, having a single nitrogen atom as the point of
attachment, further having at least one or two saturated carbon
atoms attached to the nitrogen atom, further having a total of n
carbon atoms, 0, 1, or more than one hydrogen atom, at least one
additional heteroatom, that is, in addition to the nitrogen atom at
the point of attachment, wherein at least one of the carbon atom
incorporated into an aromatic ring, further wherein each heteroatom
is independently selected from the group consisting of N, O, F, Cl,
Br, I, Si, P, and S. For example, a heteroatom-substituted
C.sub.7-C.sub.10-aralkylamino has 7 to 10 carbon atoms. The term
"heteroatom-substituted C.sub.n-aralkylamino" includes groups,
having the structure --NHR, in which R is a heteroatom-substituted
C.sub.n-aralkyl, as that term is defined above. The term
"heteroatom-substituted aralkylamino" includes the term
"heteroaralkylamino."
[0112] The term "heteroatom-unsubstituted C.sub.n-amido" refers to
a radical, having a single nitrogen atom as the point of
attachment, further having a carbonyl group attached via its carbon
atom to the nitrogen atom, further having a linear or branched,
cyclic or acyclic structure, further having a total of n carbon
atoms, 1 or more hydrogen atoms, a total of one oxygen atom, a
total of one nitrogen atom, and no additional heteroatoms. For
example, a heteroatom-unsubstituted C.sub.1-C.sub.10-amido has 1 to
10 carbon atoms. The term "heteroatom-unsubstituted C.sub.n-amido"
includes groups, having the structure --NHR, in which R is a
heteroatom-unsubstituted C.sub.n-acyl, as that term is defined
above. The term amido includes N-alkyl-amido, N-aryl-amido,
N-aralkyl-amido, acylamino, alkylcarbonylamino, arylcarbonylamino,
and ureido groups. The group, --NHCOCH.sub.3, is an example of a
heteroatom-unsubstituted amido group.
[0113] The term "heteroatom-substituted C.sub.n-amido" refers to a
radical, having a single nitrogen atom as the point of attachment,
further having a carbonyl group attached via its carbon atom to the
nitrogen atom, further having a linear or branched, cyclic or
acyclic structure, further having a total of n aromatic or
nonaromatic carbon atoms, 0, 1, or more than one hydrogen atom, at
least one additional heteroatom in addition to the oxygen of the
carbonyl group, wherein each additional heteroatom is independently
selected from the group consisting of N, O, F, Cl, Br, I, Si, P,
and S. For example, a heteroatom-substituted C.sub.1-C.sub.10-amido
has 1 to 10 carbon atoms. The term "heteroatom-substituted
C.sub.n-amido" includes groups, having the structure --NHR, in
which R is a heteroatom-unsubstituted C.sub.n-acyl, as that term is
defined above. The group, --NHCO.sub.2CH.sub.3, is an example of a
heteroatom-substituted amido group.
[0114] The term "heteroatom-unsubstituted C.sub.n-sulfonamido"
refers to a radical, having a single nitrogen atom as the point of
attachment, further having a sulfonyl group attached via its sulfur
atom to the nitrogen atom, further having a linear or branched,
cyclic or acyclic structure, further having a total of n carbon
atoms, 1 or more hydrogen atoms, a total of one oxygen atom, a
total of one nitrogen atom, and no additional heteroatoms. For
example, a heteroatom-unsubstituted C.sub.1-C.sub.10-amido has 1 to
10 carbon atoms. The term amido includes N-alkyl-sulfonamido,
N-aryl-sulfonamido, N-aralkyl-sulfonamido, sulfonylamino,
alkylsulfonamino, and arylsulfonamino groups. The group,
--NHS(O).sub.2CH.sub.3, is an example of a heteroatom-unsubstituted
sulfonamido group.
[0115] The term "heteroatom-substituted C.sub.n-sulfonamido" refers
to a radical, having a single nitrogen atom as the point of
attachment, further having a sulfonyl group attached via its sulfur
atom to the nitrogen atom, further having a linear or branched,
cyclic or acyclic structure, further having a total of n aromatic
or nonaromatic carbon atoms, 0, 1, or more than one hydrogen atom,
at least one additional heteroatom in addition to the sulfur and
oxygen atoms of the sulfonyl group, wherein each additional
heteroatom is independently selected from the group consisting of
N, O, F, Cl, Br, I, Si, P, and S. For example, a
heteroatom-substituted C.sub.1-C.sub.10-sulfonamido has 1 to 10
carbon atoms. The group, --NHS(O).sub.2OCH.sub.3, is an example of
a heteroatom-substituted sulfonamido group.
[0116] The term "heteroatom-unsubstituted C.sub.n-alkylthio" refers
to a group, having the structure --SR, in which R is a
heteroatom-unsubstituted C.sub.n-alkyl, as that term is defined
above. The group, --SCH.sub.3, is an example of a
heteroatom-unsubstituted alkylthio group.
[0117] The term "heteroatom-substituted C.sub.n-alkylthio" refers
to a group, having the structure --SR, in which R is a
heteroatom-substituted C.sub.n-alkyl, as that term is defined
above.
[0118] The term "heteroatom-unsubstituted C.sub.n-alkenylthio"
refers to a group, having the structure --SR, in which R is a
heteroatom-unsubstituted C.sub.n-alkenyl, as that term is defined
above.
[0119] The term "heteroatom-substituted C.sub.n-alkenylthio" refers
to a group, having the structure --SR, in which R is a
heteroatom-substituted C.sub.n-alkenyl, as that term is defined
above.
[0120] The term "heteroatom-unsubstituted C.sub.n-alkynylthio"
refers to a group, having the structure --SR, in which R is a
heteroatom-unsubstituted C.sub.n-alkynyl, as that term is defined
above.
[0121] The term "heteroatom-substituted C.sub.n-alkynylthio" refers
to a group, having the structure --SR, in which R is a
heteroatom-substituted C.sub.n-alkynyl, as that term is defined
above.
[0122] The term "heteroatom-unsubstituted C.sub.n-arylthio" refers
to a group, having the structure --SAr, in which Ar is a
heteroatom-unsubstituted C.sub.n-aryl, as that term is defined
above. The group, --SC.sub.6H.sub.5, is an example of a
heteroatom-unsubstituted arylthio group.
[0123] The term "heteroatom-substituted C.sub.n-arylthio" refers to
a group, having the structure --SAr, in which Ar is a
heteroatom-substituted C.sub.n-aryl, as that term is defined
above.
[0124] The term "heteroatom-unsubstituted C.sub.n-aralkylthio"
refers to a group, having the structure --SAr, in which Ar is a
heteroatom-unsubstituted C.sub.n-aralkyl, as that term is defined
above. The group, --SCH.sub.2C.sub.6H.sub.5, is an example of a
heteroatom-unsubstituted aralkyl group.
[0125] The term "heteroatom-substituted C.sub.n-aralkylthio" refers
to a group, having the structure --SAr, in which Ar is a
heteroatom-substituted C.sub.n-aralkyl, as that term is defined
above.
[0126] The term "heteroatom-unsubstituted C.sub.n-acylthio" refers
to a group, having the structure --SAc, in which Ac is a
heteroatom-unsubstituted C.sub.n-acyl, as that term is defined
above. The group, --SCOCH.sub.3, is an example of a
heteroatom-unsubstituted acylthio group.
[0127] The term "heteroatom-substituted C.sub.n-acylthio" refers to
a group, having the structure --SAc, in which Ac is a
heteroatom-substituted C.sub.n-acyl, as that term is defined
above.
[0128] The term "heteroatom-unsubstituted C.sub.n-alkylsilyl"
refers to a radical, having a single silicon atom as the point of
attachment, further having one, two, or three saturated carbon
atoms attached to the silicon atom, further having a linear or
branched, cyclic or acyclic structure, containing a total of n
carbon atoms, all of which are nonaromatic, 5 or more hydrogen
atoms, a total of 1 silicon atom, and no additional heteroatoms.
For example, a heteroatom-unsubstituted C.sub.1-C.sub.10-alkylsilyl
has 1 to 10 carbon atoms. An alkylsilyl group includes dialkylamino
groups. The groups, --Si(CH.sub.3).sub.3 and
--Si(CH.sub.3).sub.2C(CH.sub.3).sub.3, are examples of
heteroatom-unsubstituted alkylsilyl groups.
[0129] The term "heteroatom-substituted C.sub.n-alkylsilyl" refers
to a radical, having a single silicon atom as the point of
attachment, further having at least one, two, or three saturated
carbon atoms attached to the silicon atom, no carbon-carbon double
or triple bonds, further having a linear or branched, cyclic or
acyclic structure, further having a total of n carbon atoms, all of
which are nonaromatic, 0, 1, or more than one hydrogen atom, and at
least one additional heteroatom, that is, in addition to the
silicon atom at the point of attachment, wherein each additional
heteroatom is independently selected from the group consisting of
N, O, F, Cl, Br, I, Si, P, and S. For example, a
heteroatom-substituted C.sub.1-C.sub.10-alkylsilyl has 1 to 10
carbon atoms.
[0130] The term "pharmaceutically acceptable salts," as used
herein, refers to salts of compounds of this invention that are
substantially non-toxic to living organisms. Typical
pharmaceutically acceptable salts include those salts prepared by
reaction of a compound of this invention with an inorganic or
organic acid, or an organic base, depending on the substituents
present on the compounds of the invention.
[0131] Examples of inorganic acids which may be used to prepare
pharmaceutically acceptable salts include: hydrochloric acid,
phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid,
phosphorous acid and the like. Examples of organic acids which may
be used to prepare pharmaceutically acceptable salts include:
aliphatic mono- and dicarboxylic acids, such as oxalic acid,
carbonic acid, citric acid, succinic acid,
phenyl-heteroatom-substituted alkanoic acids, aliphatic and
aromatic sulfuric acids and the like. Pharmaceutically acceptable
salts prepared from inorganic or organic acids thus include
hydrochloride, hydrobromide, nitrate, sulfate, pyrosulfate,
bisulfate, sulfite, bisulfate, phosphate, monohydrogenphosphate,
dihydrogenphosphate, metaphosphate, pyrophosphate, hydroiodide,
hydrofluoride, acetate, propionate, formate, oxalate, citrate,
lactate, p-toluenesulfonate, methanesulfonate, maleate, and the
like. Other suitable salts are known to one of ordinary skill in
the art.
[0132] Suitable pharmaceutically acceptable salts may also be
formed by reacting the agents of the invention with an organic base
such as methylamine, ethylamine, ethanolamine, lysine, ornithine
and the like. Other suitable salts are known to one of ordinary
skill in the art.
[0133] Pharmaceutically acceptable salts include the salts formed
between carboxylate or sulfonate groups found on some of the
compounds of this invention and inorganic cations, such as sodium,
potassium, ammonium, or calcium, or such organic cations as
isopropylammonium, trimethylammonium, tetramethylammonium, and
imidazolium.
[0134] It should be recognized that the particular anion or cation
forming a part of any salt of this invention is not critical, so
long as the salt, as a whole, is pharmacologically acceptable and
as long as the anion or cation does not contribute undesired
qualities or effects. Further, additional pharmaceutically
acceptable salts are known to those skilled in the art, and may be
used within the scope of the invention. Additional examples of
pharmaceutically acceptable salts and their methods of preparation
and use are presented in Pharmaceutical Salts: Properties,
Selection and Use--A Handbook (2002), which is incorporated herein
by reference.
[0135] As used herein, the term "patient" is intended to include
living organisms in which certain conditions as described herein
can occur. Examples include humans, monkeys, cows, sheep, goats,
dogs, cats, mice, rats, and transgenic species thereof. In a
preferred embodiment, the patient is a primate. In an even more
preferred embodiment, the primate is a human. Other examples of
subjects include experimental animals such as mice, rats, dogs,
cats, goats, sheep, pigs, and cows. The experimental animal can be
an animal model for a disorder, e.g., a transgenic mouse with an
Alzheimer's-type neuropathology. A patient can be a human suffering
from a neurodegenerative disease, such as Alzheimer's disease, or
Parkinson's disease.
[0136] As used herein, the term "IC.sub.50" refers to an inhibitory
dose which is 50% of the maximum response obtained.
[0137] As used herein, the term "water soluble" means that the
compound dissolves in water at least to the extent of 0.010
mole/liter or is classified as soluble according to literature
precedence.
[0138] As used herein, "predominantly one enantiomer" means that
the compound contains at least 85% of one enantiomer, or more
preferably at least 90% of one enantiomer, or even more preferably
at least 95% of one enantiomer, or most preferably at least 99% of
one enantiomer. Similarly, the phrase "substantially free from
other optical isomers" means that the composition contains at most
5% of another enantiomer or diastereomer, more preferably 2% of
another enantiomer or diastereomer, and most preferably 1% of
another enantiomer or diastereomer.
[0139] As used herein the specification, "a" or "an" may mean one
or more. As used herein in the claim(s), when used in conjunction
with the word "comprising" or "having," the words "a" or "an" may
mean one or more than one. As used herein "another" may mean at
least a second or more.
[0140] Other abbreviations used herein are as follows: DMSO,
dimethyl sulfoxide; iNOS, inducible nitric oxide synthase; COX-2,
cyclooxygenase-2; NGF, nerve growth factor; IBMX,
isobutylmethylxanthine; FBS, fetal bovine serum; GPDH, glycerol
3-phosphate dehydrogenase; RXR, retinoid X receptor; TGF-.beta.,
transforming growth factor-.beta.; IFN-.gamma., interferon-.gamma.;
LPS, bacterial endotoxic lipopolysaccharide; TNF-.alpha., tumor
necrosis factor-.alpha.; IL-1.beta., interleukin-1.beta.; GAPDH,
glyceraldehyde-3-phosphate dehydrogenase; MTT,
3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide; TCA,
trichloroacetic acid; HO-1, inducible heme oxygenase.
III. Caffeic Acid, its Derivatives and their Analogs
[0141] The present invention provides caffeic-like compounds for
the treatment of cell proliferative diseases such as cancer. The
compounds of the present invention are shown above, in the summary
of the invention, the claims, as well as the examples below.
IV Cell Proliferative Diseases
[0142] The term "cell proliferative diseases" refers to disorders
resulting from abnormally increased and/or uncontrolled growth of
cell(s) in a multicellular organism that results in harm (e.g.,
discomfort or decreased life expectancy) to the multicellular
organism. Cell proliferative diseases can occur in animals or
humans. Cancer is an example of a cell proliferative disease, and
certain embodiments of the present invention are directed towards
the treatment of cancer.
[0143] In certain embodiments, compounds and methods of the present
invention may be used to treat a wide variety of cancerous states
including, for example, melanoma, non-small cell lung, small cell
lung, lung, hepatocarcinoma, retinoblastoma, astrocytoma,
glioblastoma, leukemia, blood, brain, skin, eye, tongue, gum,
neuroblastoma, head, neck, breast, pancreatic, renal, bone,
testicular, ovarian, mesothelioma, cervical, gastrointestinal,
lymphoma, colon, and/or bladder. The cancer may comprise a tumor
made of cancer cells. These cancerous states may include cells that
are cancerous, pre-cancerous, and/or malignant.
[0144] It is also anticipated that compounds of the present
invention may also be used to treat cell proliferative diseases
other than cancer. Other cell proliferative diseases that may be
treated in certain embodiments of the present invention include,
for example, rheumatiod arthritis, inflammatory bowel disease,
osteoarthritis, leiomyomas, adenomas, lipomas, hemangiomas,
fibromas, vascular occlusion, restenosis, artherosclerosis,
pre-neoplastic lesions (e.g., adenomatous hyperplasia, prostatic
intraepithelial neoplasia), carcinoma in situ, oral hairy
leukoplakia and/or psoriasis and the variant forms of psoriasis
including psoriatic arthritis other skin inflammatory conditions
such as urticaria, excema, atopic dermatitis, granuloma annulare,
angiomas, basal cell carcinoma, squamous cell carcinoma, malignant
melanoma, seborrheic dermatitis, rosacea other hyperactive
autoimmune disorders such as rheumatoid arthritis, chronic active
hepatitis, Hashimoto's thyroiditis, lupus, connective tissue
disorders, mixed connective tissue disorders, and neurologic
inflammatory diseases such as multiple sclerosis, inflammatory
leukoencephalitis.
[0145] The compounds of the present invention may function by
selectively inhibiting STAT3 phosphorylation as has been described
in Iwamaru et al., (2006), which is incorporated herein by
reference. In those studies, WP1066, an inhibitor structurally
related to AG490 but significantly more potent and active, against
human malignant glioma U87-MG and U373-MG cells in vitro and in
vivo. IC.sub.50 values for WP1066 were shown to be 5.6 .mu.M in
U87-MG cells and 3.7 .mu.M in U373-MG cells, which represents
18-fold and eightfold increases in potency, respectively, over that
of AG490. WP1066 activated Bax, suppressed the expression of c-myc,
Bcl-X.sub.L and Mcl-1, and induced apoptosis. Systemic
intraperitoneal administration of WP1066 in mice significantly
(P<0.001) inhibited the growth of subcutaneous malignant glioma
xenografts during the 30-day follow-up period. Immunohistochemical
analysis of the excised tumors revealed that phosphorylated STAT3
levels in the WP1066 treatment group remained inhibited at 3 weeks
after the final WP1066 injection, whereas tumors from the control
group expressed high levels of phosphorylated STAT3. Compounds of
the present invention may even be more potent than WP1066. For
example, as shown in FIG. 34, compound WP1193 was shown to inhibit
STAT3 phosphorylation potently than WP1066 inhibits. Given the
structural similarity between the compounds of the present
invention and the compounds tested in Iwamaru et al., 2006, the
invention contemplates that the compounds of the present invention
will be effective to treat a variety of cancer types, including:
human malignant gliomas, which are the most common malignancies in
the brain, and astrocytic tumors of World Health Organization
grades II-IV (2002). The invention further contemplates that the
compounds of the present invention will be useful to treat those
tumors in which the STAT3 is constitutively activated, such as
those cancers described in Yu and Jove (2004), which is
incorporated herein by reference. These tumors include solid
tumors, such as breast cancer, head and neck cancer, melanoma,
ovarian cancer, lung cancer, pancreatic cancer and prostate cancer,
and blood tumors, such as multiple myeloma, leukemias (e.g.
HTLV-I-dependent, acute myelogenous leukemia, large granular
lymphocyte leukemia), and lymphomas (e.g. EBV-related/Burkitt's,
mycosis fungoides, cutaneous T-cell lymphoma, non-Hodgkin's
lymphoma and anaplastic large-cell lymphoma). The compounds of the
present invention are also expected to be able to penetrate the
blood brain barrier, based on their structural similarity to
WP1066.
[0146] Additionally, compounds of the present invention may be used
to treat diseases other than hyperproliferative diseases. For
example, certain WP compounds may be useful for the treatment of
hypertrophy and ischemia (U.S. Pat. No. 6,433,018) as well as
hepatitis B infection (U.S. Pat. No. 6,420,338). Thus compounds of
the present invention may also be useful for the treatment of other
diseases including hypertrophy, ischemia, and a viral infection
(e.g., hepatitis B infection), psoriasis and the variant forms of
psoriasis including psoriatic arthritis other skin inflammatory
conditions such as urticaria, excema, atopic dermatitis, granuloma
annulare, angiomas, basal cell carcinoma, squamous cell carcinoma,
malignant melanoma, seborrheic dermatitis, rosacea other
hyperactive autoimmune disorders such as rheumatoid arthritis,
chronic active hepatitis, Hashimoto's thyroiditis, lupus,
connective tissue disorders, mixed connective tissue disorders, and
neurologic inflammatory diseases such as multiple sclerosis,
inflammatory leukoencephalitis.
V. Pharmaceutical Compositions
[0147] The anti-tumor compounds of this invention can be
administered to kill certain cells involved in a cell proliferative
disease, such as tumor cells, by any method that allows contact of
the active ingredient with the agent's site of action in the tumor.
They can be administered by any conventional methods available for
use in conjunction with pharmaceuticals, either as individual
therapeutically active ingredients or in a combination of
therapeutically active ingredients. They can be administered alone
but are generally administered with a pharmaceutically acceptable
carrier selected on the basis of the selected route of
administration and standard pharmaceutical practice.
[0148] Aqueous compositions of the present invention will have an
effective amount of the compounds to kill or slow the growth of
cancer cells. Such compositions will generally be dissolved or
dispersed in a pharmaceutically acceptable carrier or aqueous
medium.
[0149] The terms "AG compounds" and "WP compounds" refer to
specific examples of the present invention.
[0150] The phrases "pharmaceutically or pharmacologically
acceptable" refer to molecular entities and compositions that do
not produce an adverse, allergic or other untoward reaction when
administered to an animal, or human, as appropriate. As used
herein, "pharmaceutically acceptable carrier" includes any and all
solvents, dispersion media, coatings, antibacterial and antifungal
agents, isotonic and absorption delaying agents and the like. The
use of such media and agents for pharmaceutical active substances
is well known in the art. Except insofar as any conventional media
or agent is incompatible with the active ingredients, its use in
the therapeutic compositions is contemplated. Supplementary active
ingredients, such as other anti-cancer agents, can also be
incorporated into the compositions.
[0151] In addition to the compounds formulated for parenteral
administration, such as intravenous or intramuscular injection,
other pharmaceutically acceptable forms include, e.g., tablets or
other solids for oral administration; time release capsules; and
any other form currently used, including cremes, lotions,
mouthwashes, inhalants, lipid carriers, liposomes and the like.
[0152] A. Parenteral Administration
[0153] The active compounds will often be formulated for parenteral
administration, e.g., formulated for injection via the intravenous,
intramuscular, subcutaneous, or even intraperitoneal routes. The
preparation of an aqueous composition that contains an
anthracycline of the present invention as an active ingredient will
be known to those of skill in the art in light of the present
disclosure. Typically, such compositions can be prepared as
injectables, either as liquid solutions or suspensions; solid forms
suitable for using to prepare solutions or suspensions upon the
addition of a liquid prior to injection can also be prepared; and
the preparations can also be emulsified.
[0154] Solutions of the active compounds as free base or
pharmacologically acceptable salts can be prepared in water
suitably mixed with a surfactant, such as hydroxypropylcellulose.
Dispersions can also be prepared in glycerol, liquid polyethylene
glycols, and mixtures thereof and in oils. Under ordinary
conditions of storage and use, these preparations contain a
preservative to prevent the growth of microorganisms.
[0155] In some forms, it will be desirable to formulate the
compounds in salt form, generally to improve the solubility and
bioavailability and to provide an active drug form more readily
assimilated. As used herein, the term "pharmaceutically acceptable
salt" refers to compounds which are formed from acidifying a
substituted anthracycline solution with suitable physiologically
tolerated acids. Suitable physiologically tolerated acids are
organic and inorganic acids, such as hydrochloric acid, sulfuric
acid, phosphoric acid, acetic acid, citric acid, oxalic acid,
malonic acid, salicylic acid, maleic acid, methane sulfonic acid,
isothionic acid, lactic acid, gluconic acid, glucuronic acid,
amidosulfuric acid, benzoic acid, tartaric acid and pamoaic acid.
Typically, such salt forms of the active compound will be provided
or mixed prior to use.
[0156] The pharmaceutical forms suitable for injectable use include
sterile aqueous solutions or dispersions; formulations including
sesame oil, peanut oil or aqueous propylene glycol; and sterile
powders for the extemporaneous preparation of sterile injectable
solutions or dispersions. In all cases the form must be sterile and
must be fluid to the extent that easy syringability exists. It must
be stable under the conditions of manufacture and storage and must
be preserved against the contaminating action of microorganisms,
such as bacteria and fungi.
[0157] The active compounds may be formulated into a composition in
a neutral or salt form. Pharmaceutically acceptable salts, include
the acid addition salts and which are formed with inorganic acids
such as, for example, hydrochloric or phosphoric acids, or such
organic acids as acetic, oxalic, tartaric, mandelic, and the
like.
[0158] The compounds of the present invention may also be
formulated into a composition comprising liposomes or any other
lipid carrier. Liposomes include: multivesicular liposomes,
multilamellar liposomes, and unilamellar liposomes.
[0159] The carrier can also be a solvent or dispersion medium
containing, for example, water, ethanol, polyol (for example,
glycerol, propylene glycol, and liquid polyethylene glycol, and the
like), suitable mixtures thereof, and vegetable oils. The proper
fluidity can be maintained, for example, by the use of a coating,
such as lecithin, by the maintenance of the required particle size
in the case of dispersion and by the use of surfactants. The
prevention of the action of microorganisms can be brought about by
various antibacterial ad antifungal agents, for example, parabens,
chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In
many cases, it will be preferable to include isotonic agents, for
example, sugars or sodium chloride. Prolonged absorption of the
injectable compositions can be brought about by the use in the
compositions of agents delaying absorption, for example, aluminum
monostearate and gelatin.
[0160] Sterile injectable solutions are prepared by incorporating
the active compounds in the required amount in the appropriate
solvent with various of the other ingredients enumerated above, as
required, followed by filtered sterilization. Generally,
dispersions are prepared by incorporating the various sterilized
active ingredients into a sterile vehicle which contains the basic
dispersion medium and the required other ingredients from those
enumerated above. In the case of sterile powders for the
preparation of sterile injectable solutions, the preferred methods
of preparation are vacuum-drying and freeze-drying techniques which
yield a powder of the active ingredient plus any additional desired
ingredient from a previously sterile-filtered solution thereof.
[0161] In certain cases, the therapeutic formulations of the
invention could also be prepared in forms suitable for topical
administration, such as in creams and lotions. These forms may be
used for treating skin-associated diseases, such as various
sarcomas.
[0162] Upon formulation, solutions will be administered in a manner
compatible with the dosage formulation and in such amount as is
therapeutically effective. The formulations are easily administered
in a variety of dosage forms, such as the type of injectable
solutions described above, with even drug release capsules and the
like being employable.
[0163] For parenteral administration in an aqueous solution, for
example, the solution should be suitably buffered if necessary and
the liquid diluent first rendered isotonic with sufficient saline
or glucose. These particular aqueous solutions are especially
suitable for intravenous, intramuscular, subcutaneous and
intraperitoneal administration. In this connection, sterile aqueous
media which can be employed will be known to those of skill in the
art in light of the present disclosure. For example, one dosage
could be dissolved in 1 mL of isotonic NaCl solution and either
added to 1000 mL of hypodermoclysis fluid or injected at the
proposed site of infusion, (see for example, "Remington's
Pharmaceutical Sciences" 15th Edition, pages 1035-1038 and
1570-1580). Some variation in dosage will necessarily occur
depending on the condition of the subject being treated. The person
responsible for administration will, in any event, determine the
appropriate dose for the individual subject.
[0164] B. Oral Administration
[0165] In certain embodiments, active compounds may be administered
orally. This is contemplated for agents which are generally
resistant, or have been rendered resistant, to proteolysis by
digestive enzymes. Such compounds are contemplated to include all
those compounds, or drugs, that are available in tablet form from
the manufacturer and derivatives and analogues thereof.
[0166] For oral administration, the active compounds may be
administered, for example, with an inert diluent or with an
assimilable edible carrier, or they may be enclosed in hard or soft
shell gelatin capsule, or compressed into tablets, or incorporated
directly with the food of the diet. For oral therapeutic
administration, the active compounds may be incorporated with
excipients and used in the form of ingestible tablets, buccal
tables, troches, capsules, elixirs, suspensions, syrups, wafers,
and the like. Such compositions and preparations should contain at
least 0.1% of active compound. The percentage of the compositions
and preparations may, of course, be varied and may conveniently be
between about 2 to about 60% of the weight of the unit. The amount
of active compounds in such therapeutically useful compositions is
such that a suitable dosage will be obtained.
[0167] The tablets, troches, pills, capsules and the like may also
contain the following: a binder, as gum tragacanth, acacia,
cornstarch, or gelatin; excipients, such as dicalcium phosphate; a
disintegrating agent, such as corn starch, potato starch, alginic
acid and the like; a lubricant, such as magnesium stearate; and a
sweetening agent, such as sucrose, lactose or saccharin may be
added or a flavoring agent, such as peppermint, oil of wintergreen,
or cherry flavoring. When the dosage unit form is a capsule, it may
contain, in addition to materials of the above type, a liquid
carrier. Various other materials may be present as coatings or to
otherwise modify the physical form of the dosage unit. For
instance, tablets, pills, or capsules may be coated with shellac,
sugar or both. A syrup of elixir may contain the active compounds
sucrose as a sweetening agent methyl and propylparabens as
preservatives, a dye and flavoring, such as cherry or orange
flavor. Of course, any material used in preparing any dosage unit
form should be pharmaceutically pure and substantially non-toxic in
the amounts employed. In addition, the active compounds may be
incorporated into sustained-release preparation and
formulations.
[0168] Upon formulation, the compounds will be administered in a
manner compatible with the dosage formulation and in such amount as
is therapeutically effective. The formulations are easily
administered in a variety of dosage forms, such as those described
below in specific examples.
VI. Therapies
[0169] One of the major challenges in oncology today is the
effective treatment of a given tumor. Tumors are often resistant to
traditional therapies. Thus, a great deal of effort is being
directed at finding efficacious treatment of cancer. One way of
achieving this is by combining new drugs with the traditional
therapies. In the context of the present invention, it is
contemplated that therapies using the compounds could be used in
combination with surgery, chemotherapy, radiotherapy, and/or a gene
therapy.
[0170] "Effective amounts" or a "therapeutically relevant amount"
are those amounts of a compound sufficient to produce a therapeutic
benefit (e.g., effective to reproducibly inhibit decrease, reduce,
inhibit or otherwise abrogate the growth of a cancer cell). An
effective amount, in the context of treating a subject, is
sufficient to produce a therapeutic benefit. The term "therapeutic
benefit" as used herein refers to anything that promotes or
enhances the well-being of the subject with respect to the medical
treatment of the subject's cell proliferative disease. A list of
nonexhaustive examples of this includes extension of the patients
life by any period of time; decrease or delay in the neoplastic
development of the disease; decrease in hyperproliferation;
reduction in tumor growth; delay of metastases; reduction in the
proliferation rate of a cancer cell, tumor cell, or any other
hyperproliferative cell; induction of apoptosis in any treated cell
or in any cell affected by a treated cell; and/or a decrease in
pain to the subject that can be attributed to the patient's
condition.
[0171] The following examples are included to demonstrate preferred
embodiments of the invention. It should be appreciated by those of
skill in the art that the techniques disclosed in the examples
which follow represent techniques discovered by the inventor to
function well in the practice of the invention, and thus can be
considered to constitute preferred modes for its practice. However,
those of skill in the art should, in light of the present
disclosure, appreciate that many changes can be made in the
specific embodiments which are disclosed and still obtain a like or
similar result without departing from the spirit and scope of the
invention.
Example 1
General Method for Synthesis of Compounds
[0172] The following scheme shows a method for preparation of a
specific caffeic acid analog. By using analogs of the starting
materials indicated, a person of skill in the art may use this
method to make other caffeic acid analogs.
##STR00057##
Preparation of 2-methyl-1-oxido-6-substituted pyridines
[0173] Hydrogen peroxide (1 ml of 35%, w/v, 10 mmol) was added to a
solution of the pyridine compound (10 mmol) in glacial acetic acid
(6 ml), and the mixture was heated at 70-80.degree. C. with
stirring for 3 hr. Additional aliquot of 35% (w/v) H.sub.2O.sub.2
was added and the reaction was to allowed to proceed for 9 hr at
70-80.degree. C. The volume of the reaction mixture was reduced in
vacuum, water (2 ml) was added, the mixture was concentrated in
vacuum, and the residue was made alkaline using dry
Na.sub.2CO.sub.3. Chloroform (5 ml) was added, this mixture was
allowed to stand at 25.degree. C. for 5 min., and the insoluble
Na.sub.2CO.sub.3 and NaOAc were removed by filtration. Drying the
filtrate (sodium sulfate) and removal of the solvent in vacuum gave
the target product. Yield depend on substituent in position 2 55%
to 85%. Products without further purification were used in the next
step.
General Method for the Preparation of
2-(acetoxymethyl)-6-substituted Pyridines
[0174] A solution of the 2-methyl-1-oxido-3-substituted pyridine (6
mmol) in acetic anhydride (4.32 g, 42 mmol, 4 ml) was refluxed for
1 hr. Ethanol (3 ml) was added to the reaction mixture, and the
reaction was allowed to proceed at reflux for 10 min. The reaction
mixture was cooled in an ice-water bath poured onto water (10 ml),
and neutralized with 10% aqueous NaHCO.sub.3. Extraction with ether
(2.times.25 ml), washing the extract with brine solution (10 ml),
drying the organic fraction (sodium sulfate), and removal of the
solvent in vacuum gave a residue. Purification of the residue by
silica gel column chromatography using hexanes/ethyl acetate
(70/30, v/v) as eluent afforded the respective product as an oil,
which was subsequently used for the preparation of
2-hydroxymethyl-6-substituted-pyridines.
General Method for the Preparation of
2-hydroxymethyl-6-substituted-pyridine
[0175] A mixture of a 2-(acetoxymethyl)-6-substituted pyridine (5
mmol), 1N NaOH (6 ml) and MeOH (12 ml) was stirred at 25.degree. C.
for 1.5 hr. The reaction mixture was poured onto water (30 ml)
Extraction with ethyl acetate (2.times.50 ml), washing the EtOAc
extract with brine (10 ml), drying the ethyl acetate fraction
(Na.sub.2SO.sub.4), and removal of the solvent in vacuum gave the
residue. Purification of the residue by silica gel column
chromatography using hexanes/ethyl acetate (60/40, v/v) as eluent
afforded the respective product as an oil, which was subsequently
used for the preparation of
6-substituted-2-pyridinecarboxyaldehydes.
General Method for the Preparation of
6-substituted-2-pyridinecarboxyaldehydes
[0176] A solution of anhydrous H.sub.3PO.sub.4 in DMSO (1.5 ml of
1.0M) was added to a solution of the
6-substituted-2-(hydroxymethyl)-pyridine (3 mmol) and
N,N-dicyclohexylcarbodiimide (1.86 g, 9 mmol) in DMSO (7 ml), and
the reaction was allowed to proceed with stirring at 25.degree. C.
for 1.5 hr. The precipitated dicyclohexylourea was filtered, the
filtered solid was washed with ether (15 ml), and the water wash
was extracted with ether (2.times.30 ml). The combined organic
solutions were washed with brine (10 ml), the organic fraction was
dried (sodium sulfate), and the solvent was removed in vacuum. The
residue obtained was purified by silica gel column chromatography
using ether/hexanes (40/60, v/v), as eluent to afford the
respective product.
Preparation of the Ligand:
(S)-2-amino-3-methyl-1,1-diphenylbutan-1-ol
[0177] A solution of phenylmagnesium bromide (3.0 M, 600 mL), 1.7
mol) in diethyl ether was stirred at 0.degree. C. and diluted with
THF (300 mL), followed by portionwise addition of L-valine methyl
ester hydrochloride (50 g, 0.298 mol) while keeping the temperature
below 10.degree. C. After stirring for 3 hr at rt, the reaction
mixture was poured slowly into ice-cold ammonium chloride solution.
Diethyl ether (500 mL) and ethyl acetate (500 mL) were added to the
mixture. After separation of the phases, the aqueous phase was
re-extracted with tert-butylmethyl ether (1 L). The combined
organic phases were stirred at 0.degree. C. and acidified slowly
with 35% hydrochloric acid (about 40 mL) and water. The
hydrochloride precipitate thus formed was filtered off and rinsed
with tert-butylmethyl ether. The mixture was then taken up in
dichloromethane (1 L) and water (1 L) and basified at 0.degree. C.
with 35% sodium hydroxide (about 50 mL). After separation of the
phases the aqueous phase was re-extracted with dichloromethane (1
L). The combined organic phases were washed with water and then
with brine, dried over sodium sulfate and concentrated. After
crystallization from iso-propyl ether
(S)-2-amino-3-methyl-1,1-diphenylbutan-1-ol was obtained (61 g,
87%).
Synthesis of Enantiomeric Amines
[0178] A solution of (S)-2-amino-3-methyl-1,1-diphenylbutan-1-ol
(47 mmol, 12 g) in THF (80 mL) was stirred at temperature below
30.degree. C. followed by slow addition of borane-tetrahydrofuran
solution (1M, 95 mL). The temperature was allowed to rise to room
temperature over 2 h, The reaction mixture was then stirred at
0.degree. C. and the solution of pure
anti-cyclopropyl(phenyl)methamine-benzyl oxime (19 mmol, 5 g), in
THF (10 mL) was added. After stirring the mixture for 20 h at room
temperature, the reaction mixture was cooled to 0.degree. C. and
treated with hydrochloric acid (2N, 100 mL). The mixture was
stirred for 16 h then basified at 0.degree. C. by addition of 35%
sodium hydroxide (100 mL) followed by extraction with ethyl
acetate. Extract was washed with water and brine, dried over sodium
sulfate and evaporated to dryness. Amine was purified by LC
(BIOTAGE SP1 purification system), using chloroform:methanol
(gradient up to 25% of methanol) to give 2.1 g of
(S)-cyclopropyl(phenyl)methanamine, yield 72%.
Preparation of ethyl cyanoacetate
[0179] The mixture of cyanoacetic acid (1 mmol), ethyl alcohol (1
ml) and p-toluenesulfonic acid (0.1 mmol) were refluxed with
toluene (20 ml) for 12 hr. The reaction mixture was washed with
sat. sodium bicarbonate, then with water until neutral. Organic
solution was dried over sodium sulfate. The drying agent and
solvents were removed, and crude ester was distilled under reduced
pressure 97-98.degree. C./21.3 hPa (16 mmHg). Yield 72%.
Preparation of N-(Phenylalkyl)cinnamides (General Procedure)
[0180] The mixture of (S)-cyclopropyl(phenyl)methanamine (4.12 g,
28 mmol) and ethyl cyanoacetate (9.4 g, 84 mmol) in toluene (10 mL)
was prepared and stirred under reflux for 4 hr. Progress of the
reaction was monitored by TLC method. After reaction was completed
the solvent was evaporated to dryness. Product was purified using
LC (Biotage SP1, purification system) to give 4.08 g (68%) of
N--((S)-cyclopropyl(phenyl)methyl)-2-isocyanoacetamide as an
intermediate.
[0181] A mixture of
N--((S)-cyclopropyl(phenyl)methyl)-2-isocyanoacetamide (1 mmol),
6-substituted-2-pyridinecarboxyaldehyde (.mu.mol), and piperidine
(catalytic, 1 drop) in acetonitrile (50 ml) was prepared and
refluxed for 24 hr. Solvent was evaporated to dryness and product
was purified using LC (Biotage SP1, purification system) to give
WP1140 (WP1193) as a white powder (50%).
Synthesis of WP1204
##STR00058##
[0182] Synthesis of N--((S)-1-phenylethyl)acrylamide
[0183] S-(.alpha.)-Methylbenzylamine (6 mL) was dissolved in dry
dichloromethane (10 mL), cooled down to 0.degree. C. and then the
acroyl chloride (4 mL) was added dropwise. The reaction mixture was
stirred at room temperature for 15 min, then solvent was evaporated
to dryness. Crude product was purified by column chromatography
using hexanes, hexanes:ethyl acetate 9:1 to give with a good yield
a pure N--((S)-1-phenylethyl)acrylamide.
Synthesis of
(E)-3-(6-bromopyridin-2-yl)-N--((S)-1-phenylethyl)acrylamide
[0184] To the solution of triphenylphosphine (0.496 mmol) in DMF
(10 mL), a palladium acetate (0.245 mmol) was added at room
temperature under argon atmosphere. The mixture was stirred for 5
min then 2,6-dibromopyridine (12.7 mmol) followed by
N--((S)-1-phenylethyl)acrylamide (14.0 mmol) and triethylamine
(35.9 mmol) were added and the reaction mixture was stirred at
140.degree. C. for 7 hr. The majority of the solvent was removed
under diminished pressure. Diluted HCl was added and the product
was extracted with ethyl acetate. Organic layers were combined,
washed with brine, dried over sodium sulfate. Drying agent and
solvent was removed and product was purified by column
chromatography using hexanes and hexanes:ethyl acetate 2:1 as
eluents, to give pure product with a good yield.
Example 2
NMR Chemical Shifts of Selected Compounds
[0185] The following compounds were prepared according to the
procedure indicated above.
WP1082
(2E)-N-benzyl-2-cyano-3-(cyclohex-3-enyl)acrylamide
[0186] .sup.1HNMR (CDCl.sub.3, .delta.) ppm 7.78 (d, 1H, J=10.5 Hz,
H-3), 7.40-7.32 (m, 5H, Haromat. from benzyl), 6.54 (bs, 1H, NH),
5.79 (m, 1H, H-3'), 5.71 (dddd, 1H, J=11.8 Hz, J=6.4 Hz, J=4.3 Hz,
J=2.0 Hz, H-4'), 3.01-2.93 (m, 1H, H-1'), 4.57 (d, 2H, J=5.7 Hz,
CH.sub.2 from benzyl), 2.23-2.14 (m, 3H, 2', 2', 6'), 2.05-2.00 (m,
1H, 6'), 1.86-1.83 (m, 1H, 6'), 1.66-1.60 (m, 2H, 5', 5')
WP1193
(E)-3-(6-bromopyridin-2-yl)-2-cyano-N--((S)-cyclopropyl(phenyl)methyl)acry-
lamide
[0187] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.22 (s, 1H, H-3),
7.68 (dd, 1H, J=8.9 Hz, J=6.3 Hz, H-4'), 7.60 (d, 1H, J=6.3 Hz,
H-5'), 7.59 (d, 1H, J=8.5 Hz, H-3'), 7.43-7.28 (m, 5H, phenyl),
7.02 (d, 1H, J=7.5 Hz, NH), 4.53 (dd, 1H, J=J=8.5 Hz, H-1''),
1.37-1.23 (m, 1H, H-2''), 0.74-0.64 (m, 2H, CH.sub.2-cyclopropyl),
0.55-0.42 (m, 2H, CH.sub.2-cyclopropyl)
WP1145
(E)-3-(6-bromopyridin-2-yl)-2-cyano-N--((S)-1,2-diphenylethyl)acrylamide
[0188] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.13 (s, 1H, H-3),
7.66 (dd, 1H, J=7.9 Hz, J=7.3 Hz, H-4'), 7.58 (dd, 1H, J=7.9 Hz,
J=1.1 Hz, H-5'), 7.56 (dd, 1H, J=7.3 Hz, J=1 Hz, H-3'), 7.39-7.11
(m, 10H, phenyl), 6.93 (d, 1H, J=7.3 Hz, NH), 5.38 (ddd, 1H,
J=J=7.7 Hz, J=14.4 Hz, H-1''), 3.25 (dd, H, J=14.3 Hz, J=6.4 Hz,
H-2''), 3.17 (dd, H, J=14.3 Hz, J=6.4 Hz, H-2'')
WP1159
(E)-3-(6-bromopyridin-2-yl)-2-cyano-N-(2-phenoxyethyl)acrylamide
[0189] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.22 (s, 1H, H-3),
7.67 (dd, 1H, J=J=7.5 Hz, H-4'), 7.61 (dd, 1H, J=7.5 Hz, J=1.5 Hz,
H-5'), 7.58 (dd, 1H, J=7.5 Hz, J=1.5 Hz, H-3'), 7.32-7.27 (m, 3H,
phenyl), 7.04 (bs, 1H, NH), 7.00-6.91 (m, 2H, phenyl), 4.14 (t, 2H,
J=5.1 Hz, H-2''), 3.85 (dd, 2H, J=10.7 Hz, J=5.1 Hz, H-1'')
WP1163
(E)-3-(6-bromopyridin-2-yl)-2-cyano-N--((R)-cyclopropyl(phenyl)methyl)acry-
lamide
[0190] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.19 (s, 1H, H-3),
7.66 (dd, 1H, J=9.0 Hz, J=6.4 Hz, H-4'), 7.575 (d, 1H, J=6.4 Hz,
H-5'), 7.57 (d, 1H, J=9.0 Hz, H-3'), 7.41-7.26 (m, 5H, phenyl),
7.01 (d, 1H, J=7.5 Hz, NH), 4.50 (dd, 1H, J=J=8.3 Hz, H-1''),
1.35-1.23 (m, 1H, H-2''), 0.72-0.62 (m, 2H, CH.sub.2-cyclopropyl),
0.54-0.40 (m, 2H, CH.sub.2-cyclopropyl)
WP1164
(E)-3-(6-bromopyridin-2-yl)-2-cyano-N--((S)-cyclobutyl(phenyl)methyl)acryl-
amide
[0191] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.18 (s, 1H, H-3),
7.65 (dd, 1H, J=J=7.5 Hz, H-4'), 7.57 (d, 1H, J=7.5 Hz, H-5'), 7.56
(d, 1H, J=7.5 Hz, H-3'), 7.36-7.24 (m, 5H, phenyl), 6.74 (d, 1H,
J=8.0 Hz, NH), 5.05 (dd, 1H, J=J=9.0 Hz, H-1''), 2.78 (dd, 1H,
J=J=9.0 Hz, H-2''), 2.16-1.75 (m, 6H, CH.sub.2 from cyclobutyl)
WP1166
(E)-3-(6-bromopyridin-2-yl)-2-cyano-N--((S)-cyclohexyl(phenyl)methyl)acryl-
amide
[0192] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.19 (s, 1H, H-3),
7.68 (dd, 1H, J=8.0 Hz, J=7.3 Hz, H-4'), 7.59 (dd, 1H, J=8.0 Hz,
J=1.1 Hz, H-5'), 7.58 (dd, 1H, J=7.3 Hz, J=1.1 Hz, H-3'), 7.40-7.29
(m, 5H, phenyl), 6.90 (d, 1H, J=9.2 Hz, NH), 4.87 (dd, 1H, J=J=8.5
Hz, H-1''), 1.93-0.9 (m, 10H, H-cyclohexyl).
WP1167
(E)-3-(6-bromopyridin-2-yl)-2-cyano-N-((1S,2R)-2,3-dihydro-2-hydroxy-1H-in-
den-1-yl)acrylamide
[0193] .sup.1HNMR (DMSO-d6, .delta.) ppm: 8.25 (s, 1H, H-3), 8.24
(d, 1H, J=8.2 Hz, NH), 7.96 (dd, 1H, J=7.6 Hz, Hz, H-4'), 7.89 (dd,
1H, J=7.6 Hz, J=1.0 Hz, H-5'), 7.82 (dd, 1H, J=7.6 Hz, J=1.1 Hz,
H-3'), 7.27-7.21 (m, 4H, H aromat from inden), 5.38 (d, 1H, J=4.5
Hz, OH), 5.34 (dd, 1H, J=8.5 Hz, J=5.2 Hz, H-1''), 4.53 (ddd, 1H,
J=8.2 Hz, J=4.7 Hz, J=1.3 Hz, H-2''), 3.13 (dd, 1H, J=16.4 Hz,
J=5.3 Hz, H-3''), 2.87 (d, 1H, J=16.4 Hz, H-3'')
WP1168
(E)-3-(6-bromopyridin-2-yl)-2-cyano-N-((1R,2S)-2,3-dihydro-2-hydroxy-1H-in-
den-1-yl)acrylamide
[0194] .sup.1HNMR (DMSO-d6, .delta.) ppm: 8.25 (s, 1H, H-3), 8.24
(d, 1H, J=8.2 Hz, NH), 7.96 (dd, 1H, J=7.6 Hz, Hz, H-4'), 7.90 (dd,
1H, J=7.6 Hz, J=1.0 Hz, H-5'), 7.82 (dd, 1H, J=7.6 Hz, J=1.0 Hz,
H-3'), 7.29-7.21 (m, 4H, H aromat from inden), 5.38 (d, 1H, J=4.5
Hz, OH), 5.33 (dd, 1H, J=8.4 Hz, J=5.2 Hz, H-1''), 4.54 (dd, 1H,
J=4.7 Hz, J=1.8 Hz, H-2''), 3.13 (dd, 1H, J=16.4 Hz, J=5.2 Hz,
H-3''), 2.87 (dd, 1H, J=16.4 Hz, J=1.3 Hz, H-3'')
WP1169
(E)-N-benzhydryl-3-(6-bromopyridin-2-yl)-2-cyanoacrylamide
[0195] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.24 (s, 1H, H-3),
7.67 (dd, 1H, J=J=7.6 Hz, H-4'), 7.60 (d, 1H, J=7.6 Hz, H-5'), 7.59
(d, 1H, J=7.6 Hz, H-3'), 7.40-7.27 (m, 10H, phenyl), 7.16 (d, 1H,
J=7.5 Hz, NH), 6.37 (d, 1H, J=8.0 Hz, H-1'')
WP1179
(E)-2-cyano-3-(6-methylpyridin-2-yl)-N--((S)-1-phenylethyl)acrylamide)
[0196] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.29 (s, 1H, H-3),
7.70 (dd, 1H, J=J=7.7 Hz, H-4'),), 7.44-7.26 (m, 7H, H-3', H-5' and
phenyl), 6.81 (d, 1H, J=7.2 Hz, NH), 5.28 (dt, 1H, J=14.4 Hz, J=6.9
Hz, H-1''), 2.66 (s, 3H, Me from methylpyridin), 1.63 (d, 3H, J=6.9
Hz, Me)
WP1180
(E)-2-cyano-3-(6-methylpyridin-2-yl)-N--((R)-1-phenylethyl)acrylamide
[0197] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.29 (s, 1H, H-3),
7.70 (dd, 1H, J=J=7.8 Hz, H-4'),), 7.44-7.26 (m, 7H, H-3', H-5' and
phenyl), 6.81 (d, 1H, J=7.2 Hz, NH), 5.16 (dt, 1H, J=14.2 Hz, J=7.1
Hz, H-1''), 2.66 (s, 3H, Me from methylpyridin), 1.63 (d, 3H, J=7.1
Hz, Me)
WP1196
(E)-2-cyano-3-1H-imidazol-2-yl)-N--((S)-1-phenylethyl)acrylamide
[0198] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 11.25 (bs, 1H, NH),
8.39 (s, 1H, H-3), 7.44-7.26 (m, 7H, H-2', H-3' and phenyl), 7.00
(bs, 1H, NH), 6.76 (d, 1H, J=6.6 Hz, NHCO), 5.20 (dt, 1H, J=14.0
Hz, J=7.1 Hz, H-1''), 1.63 (d, 3H, J=7.1 Hz, Me)
WP1203
(E)-2-cyano-3-cyclododecyl-N--((S)-1-phenylethyl)acrylamide
[0199] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 7.33-7.20 (m, 6H, H-3
and phenyl), 6.33 (d, 1H, J=7.0 Hz, NH), 5.16 (dt, 1H, J=14.4 Hz,
J=7.0 Hz, H-1''), 2.76 (ddd, 2H, J=J=7.0 Hz, J=3.5 Hz, H-2' or
H-12'), 2.54 (dd, 2H, J=J=7.6 Hz, H-2' or H-12'), 1.69-1.37 (m,
22H, H-1', Me and cyclododecyl).
WP1225
(E)-3-(6-chloropyridin-2-yl)-2-cyano-N--((R)-cyclopropyl(phenyl)methyl)acr-
ylamide
[0200] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.25 (s, 1H, H-3),
7.79 (dd, 1H, J=J=7.9 Hz, H-4'), 7.57 (d, 1H, J=7.9 Hz, H-5'), 7.45
(d, 1H, J=7.9 Hz, H-3'), 7.43-7.31 (m, 5H, phenyl), 7.0 (d, 1H,
J=7.5 Hz, NH), 4.54 (dd, 1H, J=J=8.5 Hz, H-1''), 1.37-1.26 (m, 1H,
H-2''), 0.75-0.65 (m, 2H, CH.sub.2-cyclopropyl), 0.58-0.44 (m, 2H,
CH.sub.2-cyclopropyl)
WP1246
(6-((E)-2-((S)-cyclopropyl(phenyl)methylcarbamoyl)-2-cyanovinyl)pyridin-2--
yl)methyl acetate
[0201] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.28 (s, 1H, H-3),
7.83 (dd, 1H, J=J=7.8 Hz, H-4'), 7.52 (d, 1H, J=7.8 Hz, H-5'), 7.45
(d, 1H, J=7.8 Hz, H-3'), 7.43-7.29 (m, 5H, phenyl), 7.02 (d, 1H,
J=7.7 Hz, NH), 5.34 (s, 2H, CH.sub.2Ac), 4.53 (dd, 1H, J=J=8.4 Hz,
H-1''), 2.22 (s, 3H, Ac), 1.37-1.26 (m, 1H, H-2''), 0.71-0.67 (m,
2H, CH.sub.2-cyclopropyl), 0.53-0.45 (m, 2H,
CH.sub.2-cyclopropyl)
WP1267
(E)-3-(6-bromopyridin-2-yl)-2-cyano-N--((R)-cyclobutyl(phenyl)methyl)acryl-
amide
[0202] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.20 (s, 1H, H-3),
7.68 (dd, 1H, J=8.0 Hz, J=7.1 Hz, H-4'), 7.59 (dd, 1H, J=8.0 Hz,
J=1.0 Hz, H-5'), 7.58 (d, 1H, J=7.5 Hz, J=1.0 Hz, H-3'), 7.38-7.26
(m, 5H, phenyl), 6.75 (d, 1H, J=8.5 Hz, NH), 5.08 (dd, 1H, J=J=9.8
Hz, H-1''), 2.85-2.76 (m, 1H, H-2''), 2.16-1.78 (m, 6H, CH.sub.2
from cyclobutyl)
WP1268
(E)-3-(6-bromopyridin-2-yl)-2-cyano-N--((S)-1-hydroxy-3-phenylpropan-2-yl)-
acrylamide
[0203] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.22 (s, 1H, H-3),
7.71 (dd, 1H, J=J=7.65 Hz, H-4'), 7.66 (d, 1H, J=7.65 Hz, H-5'),
7.63 (d, 1H, J=7.65 Hz, H-3'), 7.40-7.28 (m, 5H, phenyl), 6.85 (d,
1H, J=7.1 Hz, NH), 4.4-4.37 (m, 1H, H-1''), 3.83 (dd, 1H, J=11 Hz,
J=3.6 Hz, CH.sub.2Ph), 3.75 (dd, 1H, J=11 Hz, J=4.9 Hz,
CH.sub.2Ph), 3.08 (dd, 1H, J=13.75 Hz, J=7.15 Hz, CH.sub.2OH), 3.01
(dd, 1H, J=13.75 Hz, J=7.15 Hz, CH.sub.2OH), 2.22 (bs, 1H, OH)
WP1269
(E)-3-(6-bromopyridin-2-yl)-2-cyano-N--((R)-2-hydroxy-1-phenylethyl)acryla-
mide
[0204] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.26 (s, 1H, H-3),
7.71 (dd, 1H, J=J=7.65 Hz, H-4'), 7.66 (d, 1H, J=7.65 Hz, H-5'),
7.64 (d, 1H, J=7.65 Hz, H-3'), 7.45-7.38 (m, 5H, phenyl), 7.27 (bs,
1H, NH), 5.29-5.26 (m, 1H, H-1''), 4.05-4.04 (m, 2H, CH.sub.2OH),
2.06 (bs, 1H, OH)
WP1271
(E)-2-cyano-3-(2-fluoropyridin-3-yl)-N--((S)-1-phenylethyl)acrylamide
[0205] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.61 (ddd, 1H, J=9.4
Hz, J=7.8 Hz, J=1.5 Hz, H-6'), 8.51 (s, 1H, H-3), 8.31 (m, 1H,
H-4'), 7.42-7.26 (m, 6H, H-5' and phenyl), 6.54 (d, 1H, J=7.2 Hz,
NH), 5.25 (dt, 1H, J=14.1 Hz, J=6.8 Hz, H-1''), 1.62 (d, 3H, J=6.9
Hz, Me)
WP1272
(E)-2-cyano-N--((S)-cyclopropyl(phenyl)methyl)-3-(2-fluoropyridin-3-yl)acr-
ylamide
[0206] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.63 (ddd, 1H, J=9.4
Hz, J=7.9 Hz, J=1.5 Hz, H-6'), 8.51 (s, 1H, H-3), 8.37 (m, 1H,
H-4'), 7.41-7.27 (m, 6H, H-5' and phenyl), 6.77 (d, 1H, J=7.1 Hz,
NH), 4.51 (dd, 1H, J=J=8.3 Hz, H-1''), 1.35-1.23 (m, 1H, H-2''),
0.73-0.41 (m, 4H, CH.sub.2, from cyclopropyl)
WP1280
(E)-2-cyano-3-(3-fluoropyridin-4-yl)-N--((S)-1-phenylethyl)acrylamide
[0207] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.65 (d, 1H, J=1.5 Hz,
H-2'), 8.59 (d, 1H, J=5.1 Hz, H-6'), 8.52 (d, 1H, J=0.6 Hz, H-3),
7.95 (dd, 1H, J=J=5.4 Hz, H-5'), 7.42-7.29 (m, 5H, phenyl), 6.59
(d, 1H, J=6.6 Hz, NH), 5.25 (dd, 1H, J=J=7.2 Hz, H-1''), 1.62 (d,
3H, J=6.9 Hz, Me)
WP1282
(E)-3-(6-bromopyridin-2-yl)-2-cyano-N--((R)-cyclopentyl(phenyl)methyl)acry-
lamide
[0208] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.16 (s, 1H, H-3),
7.65 (dd, 1H, J=J=8.0 Hz, H-4'), 7.58 (d, 1H, J=8.0 Hz, H-5'), 7.55
(d, 1H, J=8.00 Hz, H-3'), 7.36-7.28 (m, 5H, phenyl), 6.09 (d, 1H,
J=8.0 Hz, NH), 4.86 (dt, 1H, J=J=9.5 Hz, H-1''), 2.37 (ddd, 1H,
J=J=10.2 Hz, J=5.1 Hz, H-2''), 1.92-1.18 (m, 8H, CH.sub.2 from
cyclopentyl)
WP1283
(E)-3-(3-bromopyridin-4-yl)-2-cyano-N--((S)-1-phenylethyl)acrylamide
[0209] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.87 (s, 1H, H-2'),
8.67 (d, 1H, J=5.1 Hz, H-6'), 8.53 (s, 1H, H-3), 7.79 (dd, 1H,
J=J=5.1 Hz, H-5'), 7.41-7.29 (m, 5H, phenyl), 6.61 (d, 1H, J=7.2
Hz, NH), 5.25 (dt, 1H, J=14.2 Hz, J=7.2 Hz, H-1''), 1.62 (d, 3H,
J=6.9 Hz, Me)
WP1284
(E)-3-(5-bromopyridin-3-yl)-2-cyano-N--((S)-1-phenylethyl)acrylamide
[0210] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.86 (d, 1H, J=2.1 Hz,
H-6'), 8.79 (d, 1H, J=2.1 Hz, H-2'), 8.46 (dd, 1H, J=J=2.1 Hz,
H-4'), 8.26 (s, 1H, H-3), 7.42-7.29 (m, 5H, phenyl), 6.55 (d, 1H,
J=8.1 Hz, NH), 5.25 (dt, 1H, J=14.1 Hz, J=6.9 Hz, H-1''), 1.62 (d,
3H, J=6.9 Hz, Me)
WP1285
(E)-3-(2-bromopyridin-3-yl)-2-cyano-N--((S)-1-phenylethyl)acrylamide
[0211] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.59 (d, 1H, J=3.0 Hz,
H-3), 8.47 (dd, 1H, J=4.8 Hz, J=1.8 Hz, H-6'), 8.28 (ddd, 1H, J=7.5
Hz, J=1.8 Hz, J=0.6 Hz, H-4'), 7.41 (ddd, 1H, J=8.1 Hz, J=4.8 Hz,
J=0.6 Hz, H-5'), 7.39-7.29 (m, 5H, phenyl), 6.55 (d, 1H, J=7.5 Hz,
NH), 5.25 (dt, 1H, J=14.1 Hz, J=7.2 Hz, H-1''), 1.62 (d, 3H, J=7.2
Hz, Me)
WP1286
(E)-3-(6-bromopyridin-3-yl)-2-cyano-N--((S)-1-phenylethyl)acrylamide
[0212] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.66 (d, 1H, J=2.1 Hz,
H-2'), 8.27 (s, 1H, H-3), 8.23 (dd, 1H, J=8.4 Hz, J=2.4 Hz, H-4'),
7.63 (d, 1H, J=7.8 Hz, H-5'), 7.35-7.26 (m, 5H, phenyl), 6.56 (d,
1H, J=6.9 Hz, NH), 5.24 (dt, 1H, J=14.1 Hz, J=7.2 Hz, H-1''), 1.61
(d, 3H, J=7.2 Hz, Me)
WP1293
(R)-2-((E)-3-(6-bromopyridin-2-yl)-2-cyanoacrylamido)-2-phenylethyl
acetate
[0213] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.19 (s, 1H, H-3),
7.67 (dd, 1H, J=7.5 Hz, H-4'), 7.59 (dd, 1H, J=7.5 Hz, J=1.2 Hz,
H-5'), 7.58 (dd, 1H, J=7.5 Hz, J=1.2 Hz, H-3'), 7.41-7.21 (m, 5H,
phenyl), 7.22 (d, 1H, J=7.8 Hz, NH), 5.40 (dt, 1H, J=11.4 Hz, J=6.0
Hz, H-1''), 4.43 (d, 2H, J=6.3 Hz, CH.sub.2OAc), 2.08 (s, 3H,
OAc)
WP1302
(R)-2-((E)-3-(6-bromopyridin-2-yl)-2-cyanoacrylamido)-2-phenylethyl
pivalate
[0214] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 8.19 (s, 1H, H-3),
7.67 (dd, 1H, J=8.7 Hz, J=6.3 Hz, H-4'), 7.59 (d, 1H, J=6.3 Hz,
H-5'), 7.58 (d, 1H, J=8.7 Hz, H-3'), 7.38-7.30 (m, 5H, phenyl),
7.26 (d, 1H, J=6.5 Hz, NH), 5.43 (dt, 1H, J=11.4 Hz, J=5.4 Hz,
H-1''), 4.43 (d, 2H, J=5.4 Hz, CH.sub.2O), 2.08 (s, 9H,
CH.sub.3)
Synthesis of WP1201
((S)--N-((E)-2-(6-bromopyridin-2-yl)vinylsulfonyl)-1-phenylethanamine)
##STR00059##
[0216] 2-Chloroethanesulfonyl chloride (5.7 mmol, 930 mg) was
dissolved in dichloromethane (10 mL). Obtained solution was cooled
down to -78.degree. C. Triethylamine (TEA), (5.7 mmol, 0.8 mL) was
added, and the reaction mixture was stirred at -78.degree. C. for
30 min, then for 45 min at 0.degree. C. The reaction mixture was
cooled to -78.degree. C. and the mixture of (S)-methylbenzylamine
(5.7 mmol, 690 mg) and TEA (5.7 mmol, 0.8 mL) in dichloromethane
(10 mL) was added. Stirring at -78.degree. C. was continued for
additional 20 min, then cooling bath was removed and the reaction
mixture was stirred for 20 min at rt. Solvents were evaporated to
dryness, and crude product was purified by LC chromatography (ISCO
LC purification system) using hexanes:ethyl acetate gradient up to
50% ethyl acetate as eluent, to give 0.78 g of sulfonamide (A)
(yield 65%).
[0217] Triphenylphosphine (1.1 mmol, 28 mg) was dissolved in DMF (3
mL). Palladium (II) acetate (0.53 mmol, 113.5 mg) was added and the
mixture was stirred under Ar for 5 min. Sulfonamide (A) (2.8 mmol,
600 mg), 2.6-dibromopyridine (2.8 mmol, 640 mg), TEA (1 mL) and DMF
(3 mL) were added and the mixture was heated under reflux for 2.5
hr. The reaction mixture was cooled down, then diluted with water
(50 mL). Obtained mixture was extracted with ethyl acetate
(3.times.50 mL). Combined extracts were dried over sodium sulfate.
Product was purified by LC (BIOTAGE LC purification system) using
hexanes:ethyl acetate gradient up to 50% ethyl acetate as eluent,
to give 460 mg of WP1201 (yield 45%)
[0218] .sup.1HNMR (CDCl.sub.3, .delta.) ppm: 7.55 (dd, 1H, J=8.0
Hz, J=7.4 Hz, H-4'),), 7.46 (dd, 1H, J=8.0 Hz, J=0.9 Hz, H-5')
7.33-7.20 (m, 5H, phenyl), 7.17 (dd, 1H, J=7.4 Hz, J=0.9 Hz, H-3'),
7.17 (d, 1H, J=14.9 Hz, H-3), 7.09 (d, 1H, J=14.9 Hz, H-2), 4.80
(d, 1H, J=7.1 Hz, NH), 4.62 (dt, 1H, J=13.8 Hz, J=6.9 Hz, H-1''),
1.59 (d, 3H, J=6.9 Hz, Me)
Example 3
General Assay Methods
Cell Cultures
[0219] Glioblastoma U87 and pancreatic cancer cell lines, AsPc-1,
Panc-1, Colo357-FG and Colo357-L3.6 were maintained in DMEM with
10% fetal bovine serum (FBS), 100 mg/ml streptomycin, and 100 IU/ml
penicillin in 5% CO.sub.2 at 37.degree. C.
[0220] Tumor cell lines, were maintained in DMEM with 10% fetal
bovine serum (FBS), 100 microg/ml streptomycin, and 100 IU/ml
penicillin in 5% CO.sub.2 at 37.degree. C.
[0221] AsPc-1: A human pancreatic tumor cell line established from
the ascites of a patient with histopathologically confirmed
adenocarcinoma of the head of the pancreas. See Chen et al.
(1982).
[0222] Panc-1: An epithelioid cell line started from a human
pancreatic carcinoma of ductal cell origin. See Lieber et al.
(1975).
[0223] Colo357 was derived from a metastasis of a pancreatic
adenocarcinoma. See Morgan et al. (1980).
[0224] Colo357-FG and Colo357-L3: Colo357-FG, a fast-growing
variant produced regional lymph node metastasis in 58% of nude mice
after subcutaneous implantation and growth. It also produced
hepatic metastasis in 64% and pulmonary metastasis in 43% of nude
mice after intrasplenic implantation of tumor cells. See Vezeridis
et al. (1990).
[0225] Colo357-L3.5 established by sequential passages of a human
pancreatic cancer cell line through the nude mouse liver. See
Vezeridis et al. (1992).
[0226] WM793 human melanoma tumor cell lines were used from
different stages of progression and their biological and molecular
analyses. See Satyamoorthy et al. (1997).
Cytotoxicity Assay
[0227] For the cytotoxicity assays, 1,500 tumor cells were plated
into 96-well flat-bottom tissue culture plates in complete medium.
After 20 hours fresh media containing different concentrations of
WP1066 was added. Cell number was counted after 72 hours by using
MTS assay (Promega CellTiter AQ Non-Radioactive Cell Proliferation
Assay kit, Madison, Wis., USA) by measuring absorbance at 490 nm
with a 96-well plate reader. Data are presented as relative
inhibition of proliferation plus SD of eight measurements. The
number of cells in the presence of DMSO was taken as 100%.
Apoptosis Assay
[0228] 6.times.10.sup.5 cultured pancreatic cancer cells were
plated on 100-mm dishes for 24 h before treatment. Following
treatment with different times and concentrations of WP1066 or DMSO
(solvent) cells were stained with Annexin V-FITC. Fluorescence was
quantified on a Becton Dickinson (San Jose, Calif.) FACScan for at
least 10,000 events.
Western Blot Analysis
[0229] Cultured pancreatic cancer cells treated with different
times and concentrations of WP compounds or DMSO (solvent) were
lysed in lysis buffer and equal amounts of protein extracts were
fractionated by sodium dodecyl sulfate polyacrylamide gel
electrophoresis and transferred to Hybond-P membranes. Membranes
were immuno-blotted using phospho-specific STAT3, total STAT3,
Bcl-xL, survivin, PARD, caspase-8 and GADPH antibodies. The primary
antibodies were visualized with goat anti-rabbit or goat anti-mouse
peroxidase-conjugated antibodies using an enhanced
chemiluminescence (ECF) system and a Molecular Dynamics Storm
PhosphorImager.
[0230] All of the compositions disclosed and claimed herein can be
made and executed without undue experimentation in light of the
present disclosure. While the compositions and methods of this
invention have been described in terms of preferred embodiments, it
will be apparent to those of skill in the art that variations may
be applied to the compositions and in the steps or in the sequence
of steps of the method described herein without departing from the
concept, spirit and scope of the invention. More specifically, it
will be apparent that certain agents which are both chemically and
physiologically related may be substituted for the agents described
herein while the same or similar results would be achieved. All
such similar substitutes and modifications apparent to those
skilled in the art are deemed to be within the spirit, scope and
concept of the invention as defined by the appended claims.
REFERENCES
[0231] The following references, to the extent that they provide
exemplary procedural or other details supplementary to those set
forth herein, are specifically incorporated herein by reference.
[0232] U.S. Pat. No. 6,433,018 [0233] U.S. Pat. No. 6,420,338
[0234] U.S. Pat. No. 6,426,366 [0235] Arbel et al., Am. J. Obstet.
Gynecol., 188(5):1283-90, 2003. [0236] Bharti et al., J. Immunol.,
171(7):3863-3871, 2003. [0237] Burdelya et al., Mol. Cancer. Ther.,
1(11):893-9, 2002. [0238] Catlett-Falcone et al., Immunity,
10(1):105-15, 1999. [0239] Chen et al., In Vitro, 18(1):24-34,
1982. [0240] Constantin et al., Eur. J. Immunol., 28(11):3523-9,
1998. [0241] Hallek et al., Blood, 91(1):3-21, 1998. [0242]
Hideshima et al., J. Biol. Chem., 277(19):16639-47, 2002. [0243]
Iwamarul et al., Oncogene, 1-10, 2006. [0244] Jernberg-Wiklund et
al., Int. J. Cancer, 51(1):116-23, 1992. [0245] Kerr et al., FEBS
Lett., 546(1):1-5, 2003. [0246] Kuehl et al., Curr. Top Microbiol.
Immunol., 224:277-82, 1997. [0247] Lieber et al., Int. J. Cancer,
15(5):741-747, 1975. [0248] Meydan et al., Nature, 379(6566):645-8,
1996. [0249] Morgan et al., Int. J. Cancer, 25(5):591-598, 1980.
[0250] Remington's Pharmaceutical Sciences" 15th Edition, pages
1035-1038 and 1570-1580. [0251] Satyamoorthy et al., Melanoma Res.,
7(Suppl. 2):S35-S42, 1997. Alas and Bonavida, Clin. Cancer Res.,
9(1):316-26, 2003. [0252] Selvanayagam et al., Blood, 71(1):30-5,
1988. [0253] Verma et al., Cancer Metastasis Rev., 22(4):423-34,
2003. [0254] Vezeridis et al., Cancer, 69(8):2060-2063, 1992.
[0255] Vezeridis et al., J. Surg. Res., 48(1):51-55, 1990. [0256]
Wermuth and Stahl, In: Pharmaceutical Salts: Properties, Selection
and Use--A Handbook, Verlag Helvetica Chimica Acta, 2002. [0257]
World Health Organization grades II-IV, Kleihues et al. (Eds.),
2002. [0258] Yu and Jove, Nature Rev. Cancer, 4(2):97-105,
2004.
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