U.S. patent application number 12/396849 was filed with the patent office on 2009-11-26 for eponemycin and epoxomicin analogs and uses thereof.
This patent application is currently assigned to EISAI R&D MANAGEMENT CO., LTD.. Invention is credited to Sergei Agoulnik, Kozo Akasaka, Francis Fang, Jean-Christophe Harmange, Lynn Hawkins, Yimin Jiang, Charles Johannes, Xiang-Yi Li, Pamela McGuinness, Erin A. Murphy, Shawn Schiller, Mary Vermeulen, Jiayi Wu.
Application Number | 20090291998 12/396849 |
Document ID | / |
Family ID | 26994981 |
Filed Date | 2009-11-26 |
United States Patent
Application |
20090291998 |
Kind Code |
A1 |
Agoulnik; Sergei ; et
al. |
November 26, 2009 |
EPONEMYCIN AND EPOXOMICIN ANALOGS AND USES THEREOF
Abstract
The present invention provides compounds having formula (1):
wherein R.sub.1-R.sub.6, A, J, D, A, G, Q, w, x, y, and z are as
described generally and in classes and subclasses herein, and
additionally provides pharmaceutical compositions thereof, and
methods for the use thereof in the treatment of cancer and/or
inflammatory disorders, and more generally as proteasome
inhibitors.
Inventors: |
Agoulnik; Sergei;
(Wilmington, MA) ; Akasaka; Kozo; (Belmont,
MA) ; Fang; Francis; (Andover, MA) ; Harmange;
Jean-Christophe; (Andover, MA) ; Hawkins; Lynn;
(Concord, MA) ; Jiang; Yimin; (Londonderry,
NH) ; Johannes; Charles; (Newbury, MA) ; Li;
Xiang-Yi; (Andover, MA) ; McGuinness; Pamela;
(Methuen, MA) ; Murphy; Erin A.; (Atkinson,
NH) ; Schiller; Shawn; (Haverhill, MA) ;
Vermeulen; Mary; (Ipswich, MA) ; Wu; Jiayi;
(Brookline, MA) |
Correspondence
Address: |
MYERS BIGEL SIBLEY & SAJOVEC
PO BOX 37428
RALEIGH
NC
27627
US
|
Assignee: |
EISAI R&D MANAGEMENT CO.,
LTD.
|
Family ID: |
26994981 |
Appl. No.: |
12/396849 |
Filed: |
March 3, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10501120 |
Dec 20, 2004 |
7524883 |
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PCT/US2003/000390 |
Jan 8, 2003 |
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12396849 |
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60346711 |
Jan 8, 2002 |
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60373011 |
Apr 16, 2002 |
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Current U.S.
Class: |
514/414 ;
514/456; 514/466; 514/469; 514/475; 548/465; 549/405; 549/435;
549/469; 549/548 |
Current CPC
Class: |
A61K 38/00 20130101;
C07D 407/12 20130101; C07F 9/65502 20130101; A61P 35/02 20180101;
A61P 37/00 20180101; C07D 405/12 20130101; C07D 303/36 20130101;
A61P 35/00 20180101; C07D 209/48 20130101; A61P 29/00 20180101;
A61P 31/18 20180101; C07F 5/025 20130101; A61P 43/00 20180101; C07K
5/06139 20130101 |
Class at
Publication: |
514/414 ;
549/548; 549/435; 549/469; 548/465; 549/405; 514/475; 514/466;
514/469; 514/456 |
International
Class: |
A61K 31/4035 20060101
A61K031/4035; C07D 303/36 20060101 C07D303/36; C07D 317/06 20060101
C07D317/06; C07D 307/78 20060101 C07D307/78; C07D 209/04 20060101
C07D209/04; C07D 311/04 20060101 C07D311/04; A61K 31/336 20060101
A61K031/336; A61K 31/357 20060101 A61K031/357; A61K 31/343 20060101
A61K031/343; A61K 31/352 20060101 A61K031/352; A61P 35/00 20060101
A61P035/00 |
Claims
1. A compound having the structure (I): ##STR00483## and
pharmaceutically acceptable derivatives thereof; wherein each
occurrence of E or D is independently absent, CR.sub.A,
CR.sub.AR.sub.B, C.dbd.O, O, S, NR.sub.A, or N, wherein each
occurrence of R.sub.A and R.sub.B is independently hydrogen, a
protecting group, or an aliphatic, alicyclic, heteroaliphatic
heteroalicyclic, aryl or heteroaryl moiety; D and E are linked by a
single or double bond as valency permits; z is 0, 1, 2, 3, 4, 5 or
6; R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are each independently
hydrogen, halogen, --CN, --OR.sub.C, --SR.sub.C, --NR.sub.CR.sub.D,
--(C.dbd.O)R.sub.C or an aliphatic, alicyclic, heteroaliphatic,
heteroalicyclic, aryl or heteroaryl moiety, wherein each occurrence
of R.sub.C and R.sub.D is independently hydrogen, a protecting
group, or an aliphatic, alicyclic, heteroaliphatic,
heteroalicyclic, aryl or heteroaryl moiety, or R.sub.C and R.sub.D,
taken together, form a heteroalicyclic or heteroaryl moiety; or
wherein any two adjacent groups R.sub.1, R.sub.2, R.sub.3 and
R.sub.4, taken together, form an alicyclic or heteroalicyclic
moiety, or an aryl or heteroaryl moiety; R.sub.5 and R.sub.6 are
each independently an aliphatic, alicyclic, heteroaliphatic,
heteroalicyclic, aryl or heteroaryl moiety; and Q is an
epoxycarbonyl moiety having the structure: ##STR00484## wherein
R.sup.Q1 is hydrogen an aliphatic, alicyclic heteroaliphatic,
heteroalicyclic aryl or heteroaryl moiety, an oxygen protecting
group or a prodrug moiety.
2-62. (canceled)
63. A pharmaceutical composition comprising a compound of claim 1;
and a pharmaceutically acceptable carrier or diluent, and
optionally further comprising an additional therapeutic agent.
64-67. (canceled)
68. A method for treating cancer comprising: administering to a
subject in need thereof a therapeutically effective amount of a
compound of claim 1; and optionally further administering an
additional therapeutic agent.
69-70. (canceled)
71. The compound of claim 1, wherein the compound has the
structure: ##STR00485##
72. The compound of claim 71, wherein the compound has the
structure: ##STR00486##
73. The compound of claim 1, wherein the compound has the
structure: ##STR00487## wherein R.sub.5a is hydrogen, an aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, aryl or heteroaryl
moiety, an oxygen protecting group or a prodrug moiety.
74. The compound of claim 1, wherein the compound has the
structure: ##STR00488## wherein AR is an aryl or heteroaryl
moiety.
75. The compound of claim 1, wherein the compound has the
structure: ##STR00489## wherein R.sub.5a and R.sub.5b are each
independently hydrogen, a nitrogen protecting group, an aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, aryl or heteroaryl
moiety, or a prodrug, or R.sub.5a and R.sub.5b, taken together,
form a heteroalicyclic or heteroaryl moiety.
76. The compound of claim 1, wherein the compound has the
structure: ##STR00490##
77. The compound of claim 1, wherein the compound has the
structure: ##STR00491##
78. The compound of claim 1, wherein the compound has the
structure: ##STR00492## wherein R.sub.5a is hydrogen, an aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, aryl or heteroaryl
moiety, an oxygen protecting group or a prodrug moiety.
79. The compound of claim 1, wherein the compound has the
structure: ##STR00493## wherein AR is an aryl or heteroaryl
moiety.
80. The compound of claim 1, wherein the compound has the
structure: ##STR00494## wherein R.sub.5a and R.sub.5b are each
independently hydrogen, a nitrogen protecting group, an aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, aryl or heteroaryl
moiety, or a prodrug, or R.sub.5, and R.sub.5b, taken together,
form a heteroalicyclic or heteroaryl moiety.
81. The compound of claim 1, wherein the compound has the
structure: ##STR00495##
82. The compound of claim 1, wherein the compound has the
structure: ##STR00496## wherein R.sub.5a is hydrogen, an aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, aryl or heteroaryl
moiety, an oxygen protecting group or a prodrug moiety.
83. The compound of clam 1, wherein the compound has the structure:
##STR00497## wherein AR is an aryl or heteroaryl moiety.
84. The compound of claim 1, wherein the compound has the
structure: ##STR00498## wherein R.sub.5a and R.sub.5b are each
independently hydrogen, a nitrogen protecting group, an aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, aryl or heteroaryl
moiety, or a prodrug, or R.sub.5a and R.sub.5b, taken together,
form a heteroalicyclic or heteroaryl moiety.
85. The compound of claim 1, wherein R.sub.1, R.sub.2, R.sub.3 and
R.sub.4 are each independently hydrogen, halogen, protected or
unprotected hydroxyl, protected or unprotected thiol, protected or
unprotected amino, alkyl, alkoxy, thioalkyl, mono- or
di-substituted alkylamino, or wherein any two adjacent groups
R.sub.1, R.sub.2, R.sub.3 or R.sub.4, taken together are a
cycloalkyl, heterocycloalkyl, aryl or heteroaryl moiety, whereby
each of the alkyl moieties is independently substituted or
unsubstituted, linear or branched, cyclic or acyclic, and each of
the aryl and heteroaryl moieties is independently substituted or
unsubstituted.
86. The compound of claim 1, wherein R.sub.5 is alkyl, cycloalkyl,
alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, C.sub.1-6OR.sub.5a,
C.sub.1-6NR.sub.5aR.sub.5b, aryl or heteroaryl; wherein R.sub.5a
and R.sub.5b are each independently hydrogen, alkyl, alkenyl,
alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl,
--C(NH.sub.2).dbd.N(NO.sub.2), --C(.dbd.O)OR.sub.5s,
--C(.dbd.O)R.sub.5, or a protecting group; wherein R.sub.5c is
hydrogen, alkyl, alkenyl, alkynyl, aryl or heteroaryl.
87. The compound of claim 1, wherein R.sub.5 is alkyl, cycloalkyl,
--CH.sub.2OR.sub.5a, --CH.sub.2NR.sub.5aR.sub.5b, --CH.sub.2aryl or
--CH.sub.2heteroaryl; wherein R.sub.5, and R.sub.5b are each
independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl,
cycloalkenyl, aryl, heteroaryl, --C(NH.sub.2).dbd.N(NO.sub.2),
--C(.dbd.O)OR.sub.5c, --C(.dbd.O)R.sub.5c or a protecting group;
wherein R.sub.5c is hydrogen, alkyl, alkenyl, alkynyl, aryl or
heteroaryl.
88. The compound of claim 1, wherein R.sub.5 is alkyl, cycloalkyl,
CH.sub.2OR.sub.5a, CH.sub.2NR.sub.5aR.sub.5b or substituted or
unsubstituted --CH.sub.2Ph; wherein R.sub.5, and R.sub.5b are each
independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl,
cycloalkenyl, aryl, heteroaryl, --C(NH.sub.2).dbd.N(NO.sub.2),
--C(.dbd.O)OR.sub.5c, --C(.dbd.O)R.sub.5, or a protecting group;
wherein R.sub.5c is hydrogen, alkyl, alkenyl, alkynyl, aryl or
heteroaryl.
89. The compound of claim 1, wherein R.sub.5 is --CH.sub.2OH or
benzyl.
90. The compound of claim 1, wherein R.sub.6 is alkyl, cycloalkyl,
alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or
heteroaryl.
91. The compound of claim 1, wherein R.sub.6 is lower alkyl or
aryl.
92. The compound of claim 1, wherein R.sub.6 is
--CH.sub.2CH(CH.sub.3).sub.2.
93. The compound of claim 1 or 71, wherein Q has the structure:
##STR00499##
94. The compound of claim 1 or 71, wherein Q has the structure:
##STR00500##
95. The compound of claim 1, wherein the compound is selected from
the group consisting of the following compounds: ##STR00501##
##STR00502## ##STR00503## ##STR00504## ##STR00505## ##STR00506##
##STR00507## ##STR00508## ##STR00509## ##STR00510##
##STR00511##
96. The compound of claim 1, wherein the compound is
##STR00512##
97. The pharmaceutical of claim 63 wherein the compound is present
in an amount effective to exert an antiproliferative and/or
anticancer effect.
98. The pharmaceutical of claim 63 wherein the compound and the
additional therapeutic agent are present in an amount effective to
exert an antiproliferative and/or anticancer effect.
99. The pharmaceutical of claim 63 wherein the compound is present
in an amount effective to exert an anti-inflammatory effect.
100. The pharmaceutical of claim 63 wherein the compound and the
additional therapeutic agent are present in an amount effective to
exert an anti-inflammatory effect.
101. The method of claim 68, wherein the method is used to treat
prostate, breast, colon, bladder, cervical, skin, testicular,
kidney, ovarian, stomach, brain, liver, pancreatic or esophageal
cancer or lymphoma, leukemia, or multiple myeloma.
102. The method of claim 68, wherein the cancer is a solid
tumor.
103. A compound having the structure: ##STR00513## and
pharmaceutically acceptable derivatives thereof.
Description
PRIORITY CLAIM
[0001] The present application claims priority to U.S. Patent
Application No. 60/346,711, filed Jan. 8, 2002, and U.S. Patent
Application No. 60/373,011, filed Apr. 16, 2002, the entire
contents of each of these applications are incorporated herein by
reference.
BACKGROUND OF THE INVENTION
[0002] Epoxomicin and Eponemycin, depicted below, are natural
products isolated from unidentified actinomycete strain No. Q996-17
and from streptomyces thiogroscopicus No. P247-271, respectively
(see, Hanada et al. J. Antibiotics 1992, 45, 1746-1752; Sugawara et
al. J. Antibiotics 1990, 43, 8-18) that exhibit anti-tumor, and, in
the case of epoxomicin, anti-inflammatory activity. Interestingly,
both of these compounds have also been shown to bind covalently and
irreversibly to the 20S proteasome (see, Sin et al. Bioorg. Med.
Chem. Lett. 1999, 9, 2283-2288; Meng et al. Proc. Natl. Acad. Sci.
USA 1999, 96, 10403-10408; Meng et al. Cancer Res. 1999, 59,
2798-2801).
##STR00001##
[0003] The ability of these natural products and other compounds to
act as proteasome inhibitors has attracted significant interest
because of the wide range of cellular substrates and processes
controlled or affected by the ubiquitin-proteasome pathway. For
example, the oscillation of cyclins (cell cycle proteins required
for the orderly progression through the cell cycle) has been found
to be due to the regulated degradation mediated by the
ubiquitin-proteasome pathway, and inhibition of this pathway is
believed to result in the blockage of cell cycle progression.
Additionally, the transcription factor NF-.kappa.B is another
regulatory protein involved in a variety of cellular processes,
including immune and inflammatory responses, apoptosis, and
cellular proliferation, whose mode of action is controlled by the
ubiquitin-proteasome pathway. Furthermore, it has also been shown
that the ubiquitin-proteasome pathway is involved in retrovirus
assembly and thus may be a useful target for the development of
anti-HIV drugs. For a general discussion of the
ubiquitin-proteasome pathway and proteasome inhibitors see, Myung
et al. "The Ubiquitin-Proteasome Pathway and Proteasome Inhibitors"
Medicinal Research Reviews 2001, 21, 245-273.
[0004] As a result of the ability of the epoxyketones epoxomicin
and eponemycin to inhibit the proteasome, there has been interest
in developing the therapeutic potential of this class of compounds.
Examples of other linear peptide epoxyketones that have been
isolated recently on the basis of proteasome inhibition screening
from microbial metabolites include TMC-86A, TMC-86B, TC 1084,
TMC-89A and TMC-89B (see, Koguchi et al. J. Antibiotics, 2000, 53,
63-65; Koguchi et al. J. Antibiotics 2000, 53, 967-972).
Additionally, certain synthetic epoxyketones have been prepared and
investigated (Elofsson et al. Chem. Biol. 1999, 6, 811-822).
Although there has been significant interest in the development of
epoxyketones as proteasome inhibitors useful as potential
therapeutics, there remains a need to prepare and investigate the
biological activity of a wider range of analogues of this class of
compounds. Clearly, it would be desirable to develop analogues that
are safe and efficacious for the treatment of cancer, immune or
inflammatory disorders, or HIV, to name a few. Additionally, it
would be desirable to develop analogues that specifically target
the ubiquitin-proteasome pathway.
SUMMARY OF THE INVENTION
[0005] As discussed above, there remains a need for the development
of novel therapeutic agents and agents useful for treating
disorders mediated by proteasomes, The present invention provides
novel compounds of general formula (I),
##STR00002##
[0006] and pharmaceutical compositions thereof, as described
generally and in subclasses herein, which compounds are useful as
inhibitors of proteasomes, and thus are useful, for example, for
the treatment of various disorders involving proteasome activity,
including, for example, cancer, immune or inflammatory disorders,
or HIV.
BRIEF DESCRIPTION OF THE DRAWING
[0007] FIG. 1 is a graphical representation depicting comparative
human breast carcinoma cell growth inhibition of Paclitaxel and
exemplary inventive compounds.
DESCRIPTION OF CERTAIN PREFERRED EMBODIMENTS OF THE INVENTION
[0008] As discussed above, the demonstrated antitumor and
anti-inflammatory activity of the natural products epoxomicin and
eponemycin, as well as their ability to inhibit the 20S proteasome,
has led to increased interest in the synthesis and biological
investigation of these compounds and epoxyketones generally. In
recognition of the need to further develop the therapeutic
potential of this class of compounds, the present invention
provides novel epoxomicin and eponemycin analogs. In certain
embodiments, the compounds of the present invention can be used for
the treatment of cancer and inflammatory disorders. More generally,
in certain other embodiments, the compounds of the invention act as
proteasome inhibitors.
[0009] 1) General Description of Compounds of the Invention
[0010] The compounds of the invention include compounds of the
general formula (I) as further defined below:
##STR00003##
[0011] and pharmaceutically acceptable derivatives thereof;
[0012] wherein each occurrence of A, J, C, D or G is independently
absent, CR.sub.A, CR.sub.AR.sub.B, C.dbd.O, O, S, NR.sub.A, or N,
wherein each occurrence of R.sub.A and R.sub.B is independently
hydrogen, a protecting group, or an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety;
[0013] A and J, J and D, D and E, and D and G are each
independently linked by a single or double bond as valency
permits;
[0014] w, x, y and z are each independently 0, 1, 2, 3, 4, 5 or 6,
but the sum of x, y and z is 0-6;
[0015] R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are each independently
hydrogen, halogen, --CN, --OR.sub.C, --SR.sub.C, --NR.sub.CR.sub.D,
--(C.dbd.O)R.sub.C or an aliphatic, alicyclic, heteroaliphatic,
heteroalicyclic, aryl or heteroaryl moiety, wherein each occurrence
of R.sub.C and R.sub.D is independently hydrogen, a protecting
group, or an aliphatic, alicyclic, heteroaliphatic,
heteroalicyclic, aryl or heteroaryl moiety, or R.sub.C and R.sub.D,
taken together, form a heteroalicyclic or heteroaryl moiety; or
wherein any two adjacent groups R.sub.1, R.sub.2, R.sub.3 and
R.sub.4, taken together, form an alicyclic or heteroalicyclic
moiety, or an aryl or heteroaryl moiety;
[0016] R.sub.5 and R.sub.6 are each independently an aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, aryl or heteroaryl
moiety; and
[0017] Q is an O-containing heteroaliphatic or heteroalicyclic
moiety.
[0018] In certain embodiments, the present invention defines
certain classes of compounds which are of special interest. For
example, one class of compounds of special interest includes those
compounds of formula (I) wherein the compound has the
stereochemistry as shown in Formula (I.sup.A):
##STR00004##
[0019] Another class of compounds of special interest includes
those compounds wherein the compound has the stereochemistry as
shown in Formula (I.sup.B):
##STR00005##
[0020] Another class of compounds of special interest includes
those compounds wherein the compound has the stereochemistry as
shown in Formula (I.sup.C):
##STR00006##
[0021] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (I) above, Q is
an epoxycarbonyl moiety and the compound has the formula (II):
##STR00007##
[0022] wherein R.sup.Q1 is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0023] Another class of compounds of special interest includes
those compounds wherein the compound of formula (II) above has the
stereochemistry shown in formula (II.sup.A):
##STR00008##
[0024] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (II) above,
R.sub.5 is CH.sub.2OR.sub.5, and the compound has the formula
(II.sup.B):
##STR00009##
(II.sup.B)
[0025] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0026] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (II) above,
R.sub.5 is aryl or heteroaryl and the compound has the formula
(II.sup.C):
##STR00010##
[0027] wherein AR is an aryl or heteroaryl moiety.
[0028] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (II) above,
R.sub.5 is CH.sub.2NR.sub.5aR.sub.5b and the compound has the
formula (II.sup.D):
##STR00011##
[0029] wherein R.sub.5a, and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0030] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (II) above,
R.sub.6 is --CH.sub.2CH(CH.sub.3).sub.2 and the compound has the
formula (II.sup.E):
##STR00012##
[0031] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (II.sup.E)
above, R.sub.5 is CH.sub.2OR.sub.5a and the compound has the
formula (II.sup.F):
##STR00013##
[0032] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0033] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula II.sup.E)
above, R.sub.5 is aryl or heteroaryl and the compound has the
formula (II.sup.G):
##STR00014##
[0034] wherein AR is an aryl or heteroaryl moiety.
[0035] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (II.sup.E)
above, R.sub.5 is --CH.sub.2NR.sub.5R.sub.5b and the compound has
the formula (II.sup.H):
##STR00015##
[0036] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0037] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (I) above, Q is
a boron-containing moiety and the compound has the formula
(III):
##STR00016##
[0038] wherein R.sup.Q1 and R.sup.Q2 are each independently
hydrogen, an aliphatic, alicyclic, heteroaliphatic,
heteroalicyclic, aryl or heteroaryl moiety, or an oxygen protecting
group, or R.sup.Q1 and R.sup.Q2, taken together, form a
heteroalicyclic moiety.
[0039] Another class of compounds of special interest includes
those compounds wherein the compound of formula (III) above has the
stereochemistry shown in formula (III.sup.A)
##STR00017##
[0040] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (III) above,
R.sub.5 is CH.sub.2OR.sub.5a and the compound has the formula
(III.sup.B):
##STR00018##
[0041] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0042] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (III) above,
R.sub.5 is aryl or heteroaryl and the compound has the formula
(III.sup.C):
##STR00019##
[0043] wherein AR is an aryl or heteroaryl moiety.
[0044] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (III) above,
R.sub.5 is --CH.sub.2NR.sub.5aR.sub.5b and the compound has the
formula (III.sup.D):
##STR00020##
[0045] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety,
[0046] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (III) above,
R.sub.6 is --CH.sub.2CH(CH.sub.3).sub.2 and the compound has the
formula (III.sup.E):
##STR00021##
[0047] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (III.sup.E)
above, R.sub.5 is CH.sub.2OR.sub.5a and the compound has the
formula (III.sup.F):
##STR00022##
[0048] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0049] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (III.sup.E)
above, R.sub.5 is aryl or heteroaryl and the compound has the
formula (III.sup.G):
##STR00023##
[0050] wherein AR is an aryl or heteroaryl moiety.
[0051] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (III.sup.E)
above, R.sub.5 is CH.sub.2NR.sub.5aR.sub.5b and the compound has
the formula (III.sup.H):
##STR00024##
[0052] wherein R.sub.5a and R.sup.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0053] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (I) above, A,
J, D, and E are each CH.sub.2 and the compound has the structure of
formula IV):
##STR00025##
[0054] Another class of compounds of special interest includes
those compounds of formula (IV) above, wherein the compound has the
stereochemistry as shown in Formula (IV.sup.A):
##STR00026##
[0055] Another class of compounds of special interest includes
those compounds of formula (IV) above, wherein the compound has the
stereochemistry as shown in Formula (IV.sup.B):
##STR00027##
[0056] Another class of compounds of special interest includes
those compounds of formula (IV) above, wherein the compound has the
stereochemistry as shown in Formula (IV.sup.C):
##STR00028##
[0057] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (IV) above, Q
is an epoxycarbonyl moiety and the compound has the formula
(V):
##STR00029##
[0058] wherein R.sup.Q1 is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0059] Another class of compounds of special interest includes
those compounds wherein the compound of formula (V) above has the
stereochemistry shown in formula (V.sup.A):
##STR00030##
[0060] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (V) above,
R.sub.5 is CH.sub.2OR.sub.5a and the compound has the formula
(V.sup.B):
##STR00031##
wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0061] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (V) above,
R.sub.5 is aryl or heteroaryl and the compound has the formula
(V.sup.C):
##STR00032##
[0062] wherein AR is an aryl or heteroaryl moiety.
[0063] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (V) above,
R.sub.5 is CH.sub.2NR.sub.5aR.sub.5b and the compound has the
formula (V.sup.D):
##STR00033##
[0064] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0065] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (V) above,
R.sub.6 is --CH.sub.2CH(CH.sub.3).sub.2 and the compound has the
formula (V.sup.E):
##STR00034##
[0066] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (V.sup.E)
above, R.sub.5 is CH.sub.2OR.sub.5a and the compound has the
formula (V.sup.F):
##STR00035##
[0067] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0068] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (V.sup.E)
above, R.sub.5 is aryl or heteroaryl and the compound has the
formula (V.sup.G):
##STR00036##
[0069] wherein AR is an aryl or heteroaryl moiety.
[0070] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (V.sup.E)
above, R.sub.5 is CH.sub.2NR.sub.5aR.sub.5b and the compound has
the formula (V.sup.H):
##STR00037##
[0071] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0072] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (IV) above, Q
is a boron-containing moiety and the compound has the formula
(VI):
##STR00038##
[0073] wherein R.sup.Q1 and R.sup.Q2 are each independently
hydrogen, an aliphatic, alicyclic, heteroaliphatic,
heteroalicyclic, aryl or heteroaryl moiety, or an oxygen protecting
group, or R.sup.Q1 and R.sup.Q2, taken together, form a
heteroalicyclic moiety.
[0074] Another class of compounds of special interest includes
those compounds wherein the compound of formula (VI) above has the
stereochemistry shown in formula (VI.sup.A):
##STR00039##
[0075] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VI) above,
R.sub.5 is CH.sub.2OR.sub.5a and the compound has the formula
(VI.sup.B):
##STR00040##
[0076] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0077] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VI) above,
R.sub.5 is aryl or heteroaryl and the compound has the formula
(VI.sup.C):
##STR00041##
[0078] wherein AR is an aryl or heteroaryl moiety.
[0079] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VI) above,
R.sub.5 is CH.sub.2NR.sub.5aR.sub.5b and the compound has the
formula (VI.sup.D):
##STR00042##
[0080] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0081] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VI) above,
R.sub.6 is --CH.sub.2CH(CH.sub.3).sub.2 and the compound has the
formula (VI.sup.E):
##STR00043##
[0082] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VI.sup.E)
above, R.sub.5 is --CH.sub.2OR.sub.5a and the compound has the
formula (VI.sup.F):
##STR00044##
[0083] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0084] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VI.sup.E)
above, R.sub.5 is aryl or heteroaryl and the compound has the
formula (VI.sup.G):
##STR00045##
[0085] wherein AR is an aryl or heteroaryl moiety.
[0086] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VI.sup.E)
above, R.sub.5 is --H.sub.2NR.sub.5aR.sub.5b and the compound has
the formula (VI.sup.H):
##STR00046##
[0087] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0088] Another class of compounds of special interest consists of
compounds wherein, in the compound of formula (I) above, w, x and y
are each 0 the compound has the formula (VII):
##STR00047##
[0089] Another class of compounds of special interest consists of
those compounds of formula (VII) wherein the compound has the
stereochemistry shown in formula (VII.sup.A):
##STR00048##
[0090] Another class of compounds of special interest includes
those compounds of formula (VII) wherein the compound has the
stereochemistry shown in Formula (VII.sup.B):
##STR00049##
[0091] Another class of compounds of special interest includes
those compounds of formula (VII) wherein the compound has the
stereochemistry shown in Formula (VII.sup.C):
##STR00050##
[0092] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VII) above, Q
is an epoxycarbonyl moiety and the compound has the formula
(VIII):
##STR00051##
[0093] wherein R.sup.Q1 is hydrogen, alkyl, aryl, heteroaryl, an
oxygen protecting group or a prodrug moiety.
[0094] Another class of compounds of special interest includes
those compounds wherein the compound of formula (VIII) above has
the stereochemistry shown in formula (VIII.sup.A):
##STR00052##
[0095] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VIII) above,
R.sub.5 is CH.sub.2OR.sub.5a and the compound has the formula
(VIII.sup.B):
##STR00053##
[0096] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0097] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VIII) above,
R.sub.5 is aryl or heteroaryl and the compound has the formula
(VIII.sup.C):
##STR00054##
[0098] wherein AR is an aryl or heteroaryl moiety.
[0099] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VIII) above,
R.sub.5 is CH.sub.2NR.sub.5aR.sub.5b and the compound has the
formula (VIII.sup.D):
##STR00055##
[0100] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0101] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VIII.sup.E)
above, R.sub.5 is --CH.sub.2CH(CH.sub.3).sub.2 and the compound has
the formula (VIII.sup.E):
##STR00056##
[0102] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VIII.sup.E)
above, R.sub.5 is --CH.sub.2OR.sub.5a and the compound has the
formula (VIII.sup.F):
##STR00057##
[0103] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl Or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0104] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VIII.sup.E)
above, R.sub.5 is aryl or heteroaryl and the compound has the
formula (VIII.sup.G):
##STR00058##
[0105] wherein AR is an aryl or heteroaryl moiety.
[0106] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VIII.sup.E)
above, R.sub.5 is --CH.sub.2NR.sub.5aR.sub.5b and the compound has
the formula (VIII.sup.H):
##STR00059##
[0107] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0108] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (VII) above, Q
is a boron-containing moiety and the compound has the formula
(IX):
##STR00060##
[0109] wherein R.sup.Q1 and R.sup.Q2 are each independently
hydrogen, an aliphatic, alicyclic, heteroaliphatic,
heteroalicyclic, aryl or heteroaryl moiety, or an oxygen protecting
group, or R.sup.Q1 and R.sup.Q2, taken together, form a
heteroalicyclic moiety.
[0110] Another class of compounds of special interest includes
those compounds wherein the compound of formula (IX) above has the
stereochemistry shown in formula (IX.sup.A):
##STR00061##
[0111] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (IX) above,
R.sub.5 is CH.sub.2OR.sub.5a and the compound has the formula
(IX.sup.B):
##STR00062##
[0112] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0113] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (IX) above,
R.sub.5 is aryl or heteroaryl and the compound has the formula
(IX.sup.C):
##STR00063##
[0114] wherein AR is an aryl or heteroaryl moiety.
[0115] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (IX) above,
R.sub.5 is --CH.sub.2NR.sub.5aR.sub.5b and the compound has the
formula (IX.sup.D):
##STR00064##
[0116] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0117] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (IX) above, P,
is --CH.sub.2CH(CH.sub.13).sub.2 and the compound has the formula
(IX.sup.E):
##STR00065##
[0118] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (IX.sup.E)
above, R.sub.5 is CH.sub.2OR.sub.5a and the compound has the
formula (IX.sup.F):
##STR00066##
[0119] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0120] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (IX.sup.E)
above, R.sub.5 is aryl or heteroaryl and the compound has the
formula (IX.sup.G):
##STR00067##
[0121] wherein AR is an aryl or heteroaryl moiety.
[0122] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (IX.sup.E)
above, R.sub.5 is --CH.sub.2NR.sub.5aR.sub.5b and the compound has
the formula (IX.sup.H):
##STR00068##
[0123] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0124] Another class of compounds of special interest consists of
compounds wherein, in the compound of formula (I) above, x, y, z
and w are 0 and D is absent and the compound has the formula
(X):
##STR00069##
[0125] Another class of compounds of special interest consists of
those compounds of formula (X) wherein the compound has the
stereochemistry shown in formula (X.sup.A)
##STR00070##
[0126] Another class of compounds of special interest includes
those compounds of formula (X) wherein the compound has the
stereochemistry shown in Formula (X.sup.B):
##STR00071##
[0127] Another class of compounds of special interest includes
those compounds of formula (X) wherein the compound has the
stereochemistry shown in Formula (X.sup.C):
##STR00072##
[0128] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (X) above, Q is
an epoxycarbonyl moiety and the compound has the formula (XI):
##STR00073##
[0129] wherein R.sup.Q1 is hydrogen, alkyl, aryl, heteroaryl, an
oxygen protecting group or a prodrug moiety.
[0130] Another class of compounds of special interest includes
those compounds wherein the compound of formula (XI) above has the
stereochemistry shown in formula (XI.sup.A):
##STR00074##
[0131] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XI) above,
R.sub.5 is --CH.sub.2OR.sub.5a and the compound has the formula
(XI.sup.B):
##STR00075##
[0132] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0133] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XI) above,
R.sub.5 is aryl or heteroaryl and the compound has the formula
(XI.sup.C):
##STR00076##
[0134] wherein AR is an aryl or heteroaryl moiety.
[0135] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XI) above,
R.sub.5 is --CH.sub.2NR.sub.5aR.sub.5b and the compound has the
formula (XI.sup.D)
##STR00077##
[0136] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0137] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XI) above,
R.sub.6 is --CH.sub.2CH(CH.sub.3).sub.2 and the compound has the
formula (XI.sup.E):
##STR00078##
[0138] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XI.sup.E)
above, R.sub.5 is --CH.sub.2OR.sub.5a and the compound has the
formula (XI.sup.E):
##STR00079##
wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0139] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XI.sup.E)
above, R.sub.5 is aryl or heteroaryl and the compound has the
formula (XI.sup.G):
##STR00080##
[0140] wherein AR is an aryl or heteroaryl moiety.
[0141] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XI.sup.E)
above, R.sub.5a is CH.sub.2N.sub.5aR.sub.5b and the compound has
the formula (XI.sup.H):
##STR00081##
[0142] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0143] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (X) above, Q is
a boron-containing moiety and the compound has the formula
(XII):
##STR00082##
[0144] wherein R.sup.Q1 and P.sup.Q2 are each independently
hydrogen, an aliphatic, alicyclic, heteroaliphatic,
heteroalicyclic, aryl or heteroaryl moiety, or an oxygen protecting
group, or R.sup.Q1 and R.sup.Q2, taken together, form a
heteroalicyclic moiety.
[0145] Another class of compounds of special interest includes
those compounds wherein the compound of formula (XII) above has the
stereochemistry shown in formula (XII.sup.A):
##STR00083##
(XII.sup.A)
[0146] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XII) above,
R.sub.5 is CH.sub.2OR.sub.5a and the compound has the formula
(XII.sup.B):
##STR00084##
[0147] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0148] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XII) above,
R.sub.5 is aryl or hetetoaryl and the compound has the Formula
(XII.sup.C):
##STR00085##
[0149] wherein AR is an aryl or heteroaryl moiety.
[0150] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XII) above,
R.sub.5 is --CH.sub.2NR.sub.5aR.sub.5b and the compound has the
formula (XII.sup.D):
##STR00086##
[0151] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0152] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XII) above,
R.sub.6 is --CH.sub.2CH(CH.sub.3).sub.2 and the compound has the
formula (XII.sup.E):
##STR00087##
[0153] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XII.sup.E)
above, R.sub.5 is CH.sub.2OR.sub.5a, and the compound has the
formula (XII.sup.F):
##STR00088##
[0154] wherein R.sub.5a is hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, an
oxygen protecting group or a prodrug moiety.
[0155] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XII.sup.E)
above, R.sub.5 is aryl or heteroaryl and the compound has the
formula (XII.sup.G):
##STR00089##
[0156] wherein AR is an aryl or heteroaryl moiety.
[0157] Another class of compounds of special interest includes
those compounds wherein, in the compound of formula (XII.sup.E)
above, R.sub.5 is CH.sub.2NR.sub.5aR.sub.5b and the compound has
the formula (XII.sup.H):
##STR00090##
[0158] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety.
[0159] A number of important subclasses of each of the foregoing
classes deserve separate mention; these subclasses include
subclasses of the foregoing classes in which:
[0160] i) is CH.sub.2 and w is 0, 1, or 2;
[0161] ii) x, y and z are each 1 and A, J, D and E are each
--CH.sub.2--;
[0162] iii) x, y and z are each 1 and A-J-D-E together represent
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--;
[0163] iv) x is 0 and A is absent, y and z are each 1, and J-D-E
together represent --CH.sub.2--CH.sub.2--CH.sub.2--;
[0164] v) x is 0 and A is absent, z is 0 and E is absent, and J-D
together represents --CH.sub.2--CH.sub.2--;
[0165] vi) x, y and z are each 1 and A-J-D-E together represent
N.dbd.CH--CH.dbd.N--; [0166] vii) x, y and z are each 1 and A-J-D-E
together represent --CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2-- and G
is CH.sub.2 and w is 0, 1 or 2;
[0167] viii) x, y and z are each 0 and D is absent;
[0168] ix) R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are each
independently hydrogen, halogen, protected or unprotected hydroxyl,
protected or unprotected thiol, protected or unprotected amino,
alkyl, alkoxy, thioalkyl, mono- or di-substituted alkylamino, or
wherein any two adjacent groups R.sub.1, R.sub.2, R.sub.3 or
R.sub.4, taken together are a cycloalkyl, heterocycloalkyl, aryl or
heteroaryl moiety, whereby each of the alkyl moieties is
independently substituted or unsubstituted, linear or branched,
cyclic or acyclic, and each of the aryl and heteroaryl moieties is
independently substituted or unsubstituted;
[0169] x) R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are each
independently hydrogen or lower alkoxy;
[0170] xi) R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are each
independently hydrogen or methoxy;
[0171] xii) R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are each
independently methoxy;
[0172] xiii) R.sub.1 is hydrogen and each of R.sub.2, R.sub.3 and
R.sub.4 are independently lower alkoxy;
[0173] xiv) R.sub.1 is hydrogen and each of R.sub.2, R.sub.3 and
R.sub.4 are methoxy;
[0174] xv) R.sub.5 is alkyl, cycloalkyl, alkenyl, cycloalkenyl,
alkynyl, cycloalkynyl, C.sub.1-6OR.sub.5a,
C.sub.1-6NR.sub.5aR.sub.5b, aryl or heteroaryl; wherein R.sub.5a
and R.sub.5b are each independently hydrogen, alkyl, alkenyl,
alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl,
--C(NH.sub.2).dbd.N(NO.sub.2), --C(.dbd.O)OR.sub.5c,
--C(.dbd.O)R.sub.5c or a protecting group; wherein R.sub.5c is
hydrogen) alkyl, alkenyl, alkynyl, aryl or heteroaryl;
[0175] xvi) R.sub.5 is alkyl, cycloalkyl, --Cl.sub.2OR.sub.5a,
--CH.sub.2NR.sub.5aR.sub.5b, --CH.sub.2aryl or
--CH.sub.2heteroaryl; wherein R.sub.5a and R.sub.5b are each
independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl,
cycloalkenyl, aryl, heteroaryl, --C(NH.sub.2).dbd.N(NO.sub.2),
--C(.dbd.O)OR.sub.5c, --C(.dbd.O)R.sub.5c or a protecting group;
wherein R.sub.5c is hydrogen, alkyl, alkenyl, alkynyl, aryl or
heteroaryl;
[0176] xvii) R.sub.5 is alkyl, cycloalkyl, CH.sub.2OR.sub.5a,
CH.sub.2NR.sub.5aR.sub.5b or substituted or unsubstituted
--CH.sub.2Ph; wherein R.sub.5a and R.sub.5b are each independently
hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl,
heteroaryl, --C(NH.sub.2).dbd.N(NO.sub.2), --C(.dbd.O)OR.sub.5c,
--C(.dbd.O)R.sub.5c, or a protecting group; wherein R.sub.5c is
hydrogen, alkyl, alkenyl, alkynyl, aryl or heteroaryl;
[0177] xviii) R.sub.5 is --CH.sub.2OH or benzyl;
[0178] xix) R.sub.6 is alkyl, cycloalkyl, alkenyl, cycloalkenyl,
alkynyl, cycloalkynyl, aryl or heteroaryl;
[0179] xx) R.sub.6 is lower alkyl or aryl;
[0180] xxi) R.sub.6 is --CH.sub.2CH(CH.sub.3).sub.2;
[0181] xxii) Q is an epoxycarbonyl moiety;
[0182] xxiii) Q is an epoxycarbonyl moiety having the
structure:
##STR00091##
[0183] wherein R.sup.Q1 is hydrogen, alkyl, aryl, heteroaryl, an
oxygen protecting group or a prodrug moiety;
[0184] xxiv) Q is an epoxycarbonyl moiety having the structure:
##STR00092##
[0185] wherein R.sup.Q1 is hydrogen, alkyl, aryl, heteroaryl, an
oxygen protecting group or a prodrug moiety;
[0186] xxv) Q is an epoxycarbonyl moiety having the structure:
##STR00093##
[0187] xxvi) Q is an epoxycarbonyl moiety having the structure:
##STR00094##
[0188] xxvii) Q is a Boron-containing moiety;
[0189] xxviii) Q is --B(OH).sub.2;
[0190] xxix) Q is a Boron-containing moiety having the
structure:
##STR00095##
[0191] wherein R.sup.Q3 is lower alkyl and p is an integer from
0-4; and
[0192] xxxi) Q is a Boron-containing moiety having the
structure:
##STR00096##
[0193] As the reader will appreciate, compounds of particular
interest include, among others, those which share the attributes of
one or more of the foregoing subclasses. Some of those subclasses
are illustrated by the following sorts of compounds:
[0194] I) Compounds of the Formula (and Pharmaceutically Acceptable
Derivatives Thereof):
##STR00097##
[0195] wherein w is 0, 1 or 2; and R.sub.1, R.sub.2, R.sub.3 and
R.sub.4 are each independently hydrogen, OR.sub.c, halogen, or
NR.sub.CR.sub.D, wherein each occurrence of R.sub.C and R.sub.D is
independently hydrogen or lower alkyl.
[0196] In certain embodiments, each of R.sub.1, R.sub.2, R.sub.3
and R.sub.4 is a substituent other than hydrogen.
[0197] In certain other embodiments, one of R.sub.1, R.sub.2,
R.sub.3 or R.sub.4 is hydrogen, and each of the other groups is
substituted with a substituent other than hydrogen.
[0198] In still other embodiments, two of R.sub.1, R.sub.2,
R.sub.3, or R.sub.4 are hydrogen, and the other two are substituted
with a substituent other than hydrogen.
[0199] In yet other embodiments, three of R.sub.1, R.sub.2, R.sub.3
or R.sub.4 are hydrogen, and the remaining group is substituted
with a substituent other than hydrogen.
[0200] In certain other embodiments, w is 0 or 1. In still other
embodiments, one or more of R.sub.1-R.sub.4 is OR.sub.C, where
R.sub.C is lower alkyl.
[0201] II) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00098##
[0202] wherein AR is an aryl or heteroaryl moiety; w is 0, 1 or 2;
and R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are each independently
hydrogen, OR.sub.C, halogen, or NR.sub.CR.sub.D, wherein each
occurrence of R.sub.C and R.sub.D is independently hydrogen or
lower alkyl.
[0203] In certain embodiments, AR is substituted or unsubstituted
phenyl.
[0204] In certain embodiments, each of R.sub.1, R.sub.2, R.sub.3
and R.sub.4 is a substituent other than hydrogen.
[0205] In certain other embodiments, one of R.sub.1, R.sub.2,
R.sub.3 or R.sub.4 is hydrogen, and each of the other groups is
substituted with a substituent other than hydrogen.
[0206] In still other embodiments, two of R.sub.1, R.sub.2,
R.sub.3, or R.sub.4 are hydrogen, and the other two are substituted
with a substituent other than hydrogen.
[0207] In yet other embodiments, three of R.sub.1, R.sub.2, R.sub.3
or R.sub.4 are hydrogen, and the remaining group is substituted
with a substituent other than hydrogen.
[0208] In certain other embodiments, w is 0 or 1. In still other
embodiments, one or more of R.sub.1-R.sub.4 is OR.sub.C, where
R.sub.C is lower alkyl.
[0209] III) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00099##
[0210] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety; w is 0, 1 or 2; and R.sub.1,
R.sub.2, R.sub.3 and R.sub.4 are each independently hydrogen,
OR.sub.C, halogen, or NR.sub.CR.sub.D, wherein each occurrence of
R.sub.C and R.sub.D is independently hydrogen or lower alkyl.
[0211] In certain embodiments, R.sub.5a and R.sub.5b are each
independently hydrogen, --C(NH.sub.2).dbd.N(NO.sub.2),
--C(.dbd.O)OR.sub.5s, --C(--O)R.sub.5c, wherein R.sub.5c is alkyl,
alkenyl, aryl or heteroaryl.
[0212] In certain embodiments, each of R.sub.1, R.sub.2, R.sub.3
and R.sub.4 is a substituent other than hydrogen.
[0213] In certain other embodiments, one of R.sub.1, R.sub.2,
R.sub.3 or R.sub.4 is hydrogen, and each of the other groups is
substituted with a substituent other than hydrogen.
[0214] In still other embodiments, two of R.sub.1, R.sub.2,
R.sub.3, or R.sub.4 are hydrogen, and the other two are substituted
with a substituent other than hydrogen.
[0215] In yet other embodiments, three of R.sub.1, R.sub.2, R.sub.3
or R.sub.4 are hydrogen, and the remaining group is substituted
with a substituent other than hydrogen.
[0216] In certain other embodiments, w is 0 or 1. In still other
embodiments, one or more of R.sub.1-R.sub.4 is OR.sub.C, where
R.sub.C is lower alkyl.
[0217] IV) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00100##
[0218] wherein R.sup.Q1 and R.sup.Q2 are each independently
hydrogen, an aliphatic, alicyclic, heteroaliphatic,
heteroalicyclic, aryl or heteroaryl moiety, or an oxygen protecting
group, or R.sup.Q1 and R.sup.Q2, taken together, form a
heteroalicyclic moiety; w is 0, 1 or 2; and R.sub.1, R.sub.2,
R.sub.3 and R.sub.4 are each independently hydrogen, OR.sub.C,
halogen, or NR.sub.CR.sub.D, wherein each occurrence of R.sub.C and
R.sub.D is independently hydrogen or lower alkyl.
[0219] In certain embodiments, R.sup.Q1 and R.sup.Q2 are each
independently hydrogen, lower alkyl, or, taken together, form a 5-
to 6-membered alicyclic moiety.
[0220] In certain embodiments, each of R.sub.1, R.sub.2, R.sub.3
and R.sub.4 is a substituent other than hydrogen.
[0221] In certain other embodiments, one of R.sub.1, R.sub.2,
R.sub.3 or R.sub.4 is hydrogen, and each of the other groups is
substituted with a substituent other than hydrogen.
[0222] In still other embodiments, two of R.sub.1, R.sub.2,
R.sub.3, or R.sub.4 are hydrogen, and the other two are substituted
with a substituent other than hydrogen.
[0223] In yet other embodiments, three of R.sub.1, R.sub.2, R.sub.3
or R.sub.4 are hydrogen, and the remaining group is substituted
with a substituent other than hydrogen.
[0224] In still other embodiments, the compound has the
structure:
##STR00101##
[0225] In yet other embodiments, the compound has the
structure:
##STR00102##
[0226] wherein R.sup.Q3 is lower alkyl and p is an integer from
0-4.
[0227] In certain other embodiments, R.sup.Q3 is methyl and p is
4.
[0228] In certain other embodiments, w is 0 or 1. In still other
embodiments, one or more of R.sub.1-R.sub.4 is OR.sub.C, where
R.sub.C is lower alkyl.
[0229] V) Compounds of the Formula (and Pharmaceutically Acceptable
Derivatives Thereon:
##STR00103##
[0230] wherein AR is an aryl or heteroaryl moiety; R.sup.Q1 and
R.sup.Q2 are each independently hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or an
oxygen protecting group, or R.sup.Q1 and R.sup.Q2, taken together,
form a heteroalicyclic moiety; w is 0, 1 or 2; and R.sub.1,
R.sub.2, R.sub.3 and R.sub.4 are each independently hydrogen,
OR.sub.C, halogen, or NR.sub.CR.sub.D, wherein each occurrence of
R.sub.C and R.sub.D is independently hydrogen or lower alkyl.
[0231] In certain embodiments, AR is substituted or unsubstituted
phenyl.
[0232] In certain embodiments, R.sup.Q1 and R.sup.Q2 are each
independently hydrogen, lower alkyl, or, taken together, form a 5-
to 6-membered alicyclic moiety.
[0233] In certain embodiments, each of R.sub.1, R.sub.2, R.sub.3
and R.sub.4 is a substituent other than hydrogen.
[0234] In certain other embodiments, one of R.sub.1, R.sub.2,
R.sub.3 or R.sub.4 is hydrogen, and each of the other groups is
substituted with a substituent other than hydrogen.
[0235] In still other embodiments, two of R.sub.1, R.sub.2,
R.sub.3, or R.sub.4 are hydrogen, and the other two are substituted
with a substituent other than hydrogen.
[0236] In yet other embodiments, three of R.sub.1, R.sub.2, R.sub.3
or R.sub.4 are hydrogen, and the remaining group is substituted
with a substituent other than hydrogen.
[0237] In still other embodiments, the compound has the
structure:
##STR00104##
[0238] In yet other embodiments, the compound has the
structure:
##STR00105##
[0239] wherein R.sup.Q3 is lower alkyl and p is an integer from
0-4.
[0240] In certain other embodiments, R.sup.Q3 is methyl and p is
4.
[0241] In certain other embodiments, w is 0 or 1. In still other
embodiments, one or more of R.sub.1-R.sub.4 is OR.sub.C, where
R.sub.C is lower alkyl.
[0242] In certain other embodiments, w is 0 or 1. In still other
embodiments, one or more of R.sub.1-R.sub.4 is OR.sub.C, where
R.sub.C is lower alkyl.
[0243] VI) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00106##
[0244] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety; R.sup.Q1 and R.sup.Q2 are
each independently hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or an
oxygen protecting group, or R.sup.Q1 and R.sup.Q2, taken together,
form a heteroalicyclic moiety; w is 0, 1 or 2; and R.sub.1,
R.sub.2, R.sub.3 and R.sub.4 are each independently hydrogen,
OR.sub.C, halogen, or NR.sub.CR.sub.D, wherein each occurrence of
R.sub.C and R.sub.D is independently hydrogen or lower alkyl.
[0245] In certain embodiments, R.sub.5a and R.sub.5b are each
independently hydrogen, --C(NH.sub.2).dbd.N(NO.sub.2),
--C(.dbd.O)OR.sub.5c, --C(.dbd.O)R.sub.5c, wherein R.sub.5c is
alkyl, alkenyl, aryl or heteroaryl.
[0246] In certain embodiments, R.sup.Q1 and R.sup.Q2 are each
independently hydrogen, lower alkyl, or, taken together, form a 5-
to 6-membered alicyclic moiety.
[0247] In certain embodiments, each of R.sub.1, R.sub.2, R.sub.3
and R.sub.4 is a substituent other than hydrogen.
[0248] In certain other embodiments, one of R.sub.1, R.sub.2,
R.sub.3 or R.sub.4 is hydrogen, and each of the other groups is
substituted with a substituent other than hydrogen.
[0249] In still other embodiments, two of R.sub.1, R.sub.2,
R.sub.3, or R.sub.4 are hydrogen, and the other two are substituted
with a substituent other than hydrogen.
[0250] In yet other embodiments, three of R.sub.1, R.sub.2, R.sub.3
or R.sub.4 are hydrogen, and the remaining group is substituted
with a substituent other than hydrogen.
[0251] In still other embodiments, the compound has the
structure:
##STR00107##
[0252] In yet other embodiments, the compound has the
structure:
##STR00108##
[0253] wherein R.sup.Q3 is lower alkyl and p is an integer from
0-4.
[0254] In certain other embodiments, R.sup.Q3 is methyl and p is
4.
[0255] In certain other embodiments, w is 0 or 1. In still other
embodiments, one or more of R.sub.1-R.sub.4 is OR.sub.C, where
R.sub.C is lower alkyl.
[0256] VII) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00109##
[0257] wherein w is 0, 1 or 2, each occurrence of R.sub.C is
independently lower alkyl, and q is 0, 1, 2, 3 or 4.
[0258] In certain embodiments, w is 0 or 1.
[0259] In certain other embodiments, q is 1. In still other
embodiments, q is 2.
[0260] In yet other embodiments, q is 3. In still other
embodiments, q is 3.
[0261] In yet other embodiments, each occurrence of R.sub.C is
methyl,
[0262] VIII) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof).
##STR00110##
[0263] wherein AR is an aryl or heteroaryl moiety; w is 0, 1 or 2,
each occurrence of R.sub.C is independently lower alkyl, and q is
0, 1, 2, 3 or 4.
[0264] In certain embodiments, AR is substituted or unsubstituted
phenyl.
[0265] In certain embodiments, w is 0 or 1.
[0266] In certain other embodiments, q is 1. In still other
embodiments, q is 2.
[0267] In yet other embodiments, q is 3. In still other
embodiments, q is 3.
[0268] In yet other embodiments, each occurrence of R.sub.C is
methyl.
[0269] IX) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof)
##STR00111##
[0270] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety; w is 0, t or 2, each
occurrence of R.sub.C is independently lower alkyl, and q is 0, 1,
2, 3 or 4.
[0271] In certain embodiments, R.sub.5a and R.sub.5b are each
independently hydrogen, --C(--NH.sub.2)--N(NO.sub.2),
--C(.dbd.O)OR.sub.5c, --C(--O)R.sub.5c, wherein R.sub.5c is alkyl,
alkenyl, aryl or heteroaryl.
[0272] In certain embodiments, w is 0 or 1.
[0273] In certain other embodiments, q is 1. Instill other
embodiments, q is 2. In yet other embodiments, q is 3. In still
other embodiments, q is 3.
[0274] In yet other embodiments, each occurrence of R.sub.C is
methyl.
[0275] X) Compounds of the Formula (and Pharmaceutically Acceptable
Derivatives Thereof)
##STR00112##
[0276] wherein R.sup.Q1 and R.sup.Q2 are each independently
hydrogen, an aliphatic, alicyclic, heteroaliphatic,
heteroalicyclic, aryl or heteroaryl moiety, or an oxygen protecting
group, or R.sup.Q1 and R.sup.Q2, taken together, form a
heteroalicyclic moiety; w is 0, 1 or 2, each occurrence of R.sub.C
is independently lower alkyl, and q is 0, 1, 2, 3 or 4.
[0277] In certain embodiments, R.sup.Q1 and R.sup.Q2 are each
independently hydrogen, lower allyl, or, taken together, form a 5-
to 6-membered alicyclic moiety.
[0278] In still other embodiments, the compound has the
structure:
##STR00113##
[0279] In yet other embodiments, the compound has the
structure:
##STR00114##
[0280] wherein R.sup.Q3 is lower alkyl and p is an integer from
0-4.
[0281] In certain other embodiments, R.sup.Q3 is methyl and p is
4.
[0282] In certain embodiments, w is 0 or 1.
[0283] In certain other embodiments, q is 1. In still other
embodiments, q is 2.
[0284] In yet other embodiments, q is 3. In still other
embodiments, q is 3.
[0285] In yet other embodiments, each occurrence of R.sub.C is
methyl.
[0286] XI) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof:
##STR00115##
[0287] wherein AR is an aryl or heteroaryl moiety; R.sup.Q1 and
R.sup.Q2 are each independently hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or an
oxygen protecting group, or R.sup.Q1 and R.sup.Q2, taken together,
form a heteroalicyclic moiety; w is 0, 1 or 2, each occurrence of
R.sub.C is independently lower alkyl, and q is 0, 1, 2, 3 or 4.
[0288] In certain embodiments, AR is substituted or unsubstituted
phenyl.
[0289] In certain embodiments, R.sup.Q1 and R.sup.Q2 are each
independently hydrogen, lower alkyl, or, taken together, form a 5-
to 6-membered alicyclic moiety.
[0290] In still other embodiments, the compound has the
structure:
##STR00116##
[0291] In yet other embodiments, the compound has the
structure:
##STR00117##
[0292] wherein R.sup.Q3 is lower alkyl and p is an integer from
0-4.
[0293] In certain other embodiments, R.sup.Q3 is methyl and p is
4.
[0294] In certain embodiments, w is 0 or 1.
[0295] In certain other embodiments, q is 1. In still other
embodiments, q is 2. In yet other embodiments, q is 3. In still
other embodiments, q is 3.
[0296] In yet other embodiments, each occurrence of R.sub.C is
methyl.
[0297] XII) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof
##STR00118##
[0298] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b, taken together, form a
heteroalicyclic or heteroaryl moiety; R.sup.Q1 and R.sup.Q2 are
each independently hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or an
oxygen protecting group, or R.sup.Q1 and R.sup.2, taken together,
form a heteroalicyclic moiety; w is 0, 1 or 2, each occurrence of
R.sub.C is independently lower alkyl, and q is 0, 1, 2, 3 or 4.
[0299] In certain embodiments, R.sub.5a and R.sub.5b are each
independently hydrogen, --C(NH.sub.2).dbd.N(NO.sub.2),
--C(.dbd.O)OR.sub.5c, --C(.dbd.O)R.sub.5c, wherein R.sub.5c is
alkyl, alkenyl, aryl or heteroaryl.
[0300] In certain embodiments, R.sup.Q1 and R.sup.Q2 are each
independently hydrogen, lower alkyl, or, taken together, form a 5-
to 6-membered alicyclic moiety.
[0301] In still other embodiments, the compound has the
structure:
##STR00119##
[0302] In yet other embodiments, the compound has the
structure:
##STR00120##
[0303] wherein R.sup.Q3 is lower alkyl and p is an integer from
0-4.
[0304] In certain other embodiments, R.sup.Q3 is methyl and p is
4.
[0305] In certain embodiments, w is 0 or 1.
[0306] In certain other embodiments, q is 1. In still other
embodiments, q is 2. In yet other embodiments, q is 3. In still
other embodiments, q is 3.
In yet other embodiments, each occurrence of R.sub.C is methyl.
[0307] XIII) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof).
##STR00121##
[0308] wherein w is 0, 1 or 2; and q is 0, 1, 2, 3 or 4.
[0309] In certain embodiments, q is 1. In certain other
embodiments, q is 2. In still other embodiments, q is 3. In yet
other embodiments q is 4.
[0310] XIV) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00122##
[0311] wherein AR is an aryl or heteroaryl moiety; w is 0, 1 or 2;
and q is 0, 1, 2, 3 or 4.
[0312] In certain embodiments, AR is substituted or unsubstituted
phenyl.
[0313] In certain embodiments, q is 1. In certain other
embodiments, q is 2.
[0314] In still other embodiments, q is 3. In yet other embodiments
q is 4.
[0315] XV) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00123##
[0316] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
prodrug, or R.sub.5a and R.sub.5b taken together, form a
heteroalicyclic or heteroaryl moiety; w is 0, 1 or 2; and q is 0,
1, 2, 3 or 4.
[0317] In certain embodiments, R.sub.5a and R.sub.5b are each
independently hydrogen, --C(NH.sub.2).dbd.N(NO.sub.2),
--C(.dbd.O)OR.sub.5c, --C(.dbd.O)R.sub.5c, wherein R.sub.5c is
alkyl, alkenyl, aryl or heteroaryl.
[0318] In certain embodiments, q is 1. In certain other
embodiments, q is 2. In still other embodiments, q is 3. In yet
other embodiments q is 4.
[0319] XVI) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00124##
[0320] wherein R.sup.Q1 and R.sup.Q2 are each independently
hydrogen, an aliphatic, alicyclic, heteroaliphatic,
heteroalicyclic, aryl or heteroaryl moiety, or an oxygen protecting
group, or R.sup.Q1 and R.sup.Q2, taken together, form a
heteroalicyclic moiety; w is 0, 1 or 2; and q is 0, 1, 2, 3 or
4.
[0321] In certain embodiments, R.sup.Q1 and R.sup.Q2 are each
independently hydrogen, lower alkyl, or, taken together, form a 5-
to 6-membered alicyclic moiety.
[0322] In still other embodiments, the compound has the
structure:
##STR00125##
[0323] In yet other embodiments, the compound has the
structure:
##STR00126##
wherein R.sup.Q3 is lower alkyl and p is an integer from 0-4.
[0324] In certain other embodiments, R.sup.Q3 is methyl and p is
4.
[0325] In certain embodiments, q is 1. In certain other
embodiments, q is 2. In still other embodiments, q is 3. In yet
other embodiments q is 4.
[0326] XXV) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00127##
[0327] wherein AR is an aryl or heteroaryl moiety; R.sup.Q1 and
R.sup.Q2 are each independently hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or an
oxygen protecting group, or R.sup.Q1 and R.sup.Q2, taken together,
form a heteroalicyclic moiety; w is 0, 1 or 2; and q is 0, 1, 2, 3
or 4.
[0328] In certain embodiments, AR is substituted or unsubstituted
phenyl.
[0329] In certain embodiments, R.sup.Q1 and R.sup.Q2 are each
independently hydrogen, lower alkyl, or, taken together, form a 5-
to 6-membered alicyclic moiety.
[0330] In still other embodiments, the compound has the
structure:
##STR00128##
[0331] In yet other embodiments, the compound has the
structure:
##STR00129##
[0332] wherein R.sup.Q3 is lower alkyl and p is an integer from
0-4.
[0333] In certain other embodiments, R.sup.Q3 is methyl and p is
4.
[0334] In certain embodiments, q is 1. In certain other
embodiments, q is 2. In still other embodiments, q is 3. ID yet
other embodiments q is 4.
[0335] XV) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00130##
[0336] wherein R.sub.5a and R.sub.5b are each independently
hydrogen, a nitrogen protecting group, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or a
pro drug, or R.sub.5a and R.sub.5b taken tog ether, form a
heteroalicyclic or heteroaryl moiety; R.sup.Q1 and R.sup.Q2 are
each independently hydrogen, an aliphatic, alicyclic,
heteroaliphatic, heteroalicyclic, aryl or heteroaryl moiety, or an
oxygen protecting group, or R.sup.Q1 and R.sup.Q2 taken together,
form a hetero alicyclic moiety; w is 0, 1 or 2; and q is 0, 2, 3 or
4
[0337] In certain embodiments, R.sub.5a and R.sub.5b are each
independently hydrogen, --C(NH.sub.2).dbd.N(NO.sub.2),
--C(.dbd.O)OR.sub.5c, --C(.dbd.O)R.sub.5c, wherein R.sub.5c is
alkyl, alkenyl, aryl or heteroaryl.
[0338] In certain embodiments, R.sup.Q1 and R.sup.Q2 are each
independently hydrogen, lower alkyl, or, taken together, form a 5-
to 6-membered alicyclic moiety.
[0339] In still other embodiments, the compound has the
structure:
##STR00131##
[0340] In yet other embodiments, the compound has the
structure:
##STR00132##
[0341] wherein R.sup.Q3 is lower alkyl and p is an integer from
0-4.
[0342] In certain other embodiments, R.sup.Q3 is methyl and p is
4.
[0343] In certain embodiments, q is 1. In certain other
embodiments, q is 2. In still other embodiments, q is 3. In yet
other embodiments q is 4.
[0344] It will also be appreciated that for each of the subgroups
I-XV described above, a variety of other subclasses are of special
interest, including, but not limited to those classes described
above i)-xxxi) and classes, subclasses and species of compounds
described above and in the examples herein.
[0345] Some of the foregoing compounds can comprise one or more
asymmetric centers, and thus can exist in various isomeric forms,
e.g., stereoisomers and/or diastereomers. Thus, inventive compounds
and pharmaceutical compositions thereof may be in the form of an
individual enantiomer, diastereomer or geometric isomer, or may be
in the form of a mixture of stereoisomers. In certain embodiments,
the compounds of the invention are enantiopure compounds. In
certain other embodiments, mixtures of stereoisomers or
diastereomers are provided.
[0346] Furthermore, certain compounds, as described herein may have
one or more double bonds that can exist as either the Z or E
isomer, unless otherwise indicated. The invention additionally
encompasses the compounds as individual isomers substantially free
of other isomers and alternatively, as mixtures of various isomers,
e.g., racemic mixtures of stereoisomers. In addition to the
above-mentioned compounds per se, this invention also encompasses
pharmaceutically acceptable derivatives of these compounds and
compositions comprising one or more compounds of the invention and
one or more pharmaceutically acceptable excipients or
additives.
[0347] Compounds of the invention may be prepared by
crystallization of compound of formula (I) under different
conditions and may exist as one or a combination of polymorphs of
compound of general formula (I) forming part of this invention. For
example, different polymorphs may be identified and/or prepared
using different solvents, or different mixtures of solvents for
recrystallization; by performing crystallizations at different
temperatures; or by using various modes of cooling, ranging from
very fast to very slow cooling during crystallizations. Polymorphs
may also be obtained by heating or melting the compound followed by
gradual or fast cooling. The presence of polymorphs may be
determined by solid probe NMR spectroscopy, IR spectroscopy,
differential scanning calorimetry, powder X-ray diffractogram
and/or other techniques. Thus, the present invention encompasses
inventive compounds, their derivatives, their tautomeric forms,
their stereoisomers, their polymorphs, their pharmaceutically
acceptable salts their pharmaceutically acceptable solvates and
pharmaceutically acceptable compositions containing them.
[0348] 2) Compounds and Definitions
[0349] As discussed above, this invention provides novel compounds
with a range of biological properties. Compounds of this invention
have biological activities relevant for the treatment of cancer
and/or inflammatory disorders, and/or disorders caused by
activation of certain regulatory subunits of the proteasome.
[0350] Compounds of this invention include those specifically set
forth above and described herein, and are illustrated in part by
the various classes, subgenera and species disclosed elsewhere
herein.
[0351] Additionally, the present invention provides
pharmaceutically acceptable derivatives of the inventive compounds,
and methods of treating a subject using these compounds,
pharmaceutical compositions thereof, or either of these in
combination with one or more additional therapeutic agents. The
phrase, "pharmaceutically acceptable derivative", as used herein,
denotes any pharmaceutically acceptable salt, ester, or salt of
such ester, of such compound, or any other adduct or derivative
which, upon administration to a patient, is capable of providing
(directly or indirectly) a compound as otherwise described herein,
or a metabolite or residue thereof. Pharmaceutically acceptable
derivatives thus include among others pro-drugs. A pro-drug is a
derivative of a compound, usually with significantly reduced
pharmacological activity, which contains an additional moiety that
is susceptible to removal in vivo yielding the parent molecule as
the pharmacologically active species. An example of a pro-drug is
an ester which is cleaved in vivo to yield a compound of interest.
Pro-drugs of a variety of compounds, and materials and methods for
derivatizing the parent compounds to create the pro-drugs, are
known and may be adapted to the present invention. Certain
exemplary pharmaceutical compositions and pharmaceutically
acceptable derivatives will be discussed in more detail herein
below.
[0352] Numerous suitable prodrug moieties, and information
concerning their selection, synthesis and use are well known in the
art. Examples of prodrug moieties of interest include, among
others, prodrug moieties that can be attached to primary or
secondary anine-containing functionalities. Examples of such
prodrug moieties include the following:
##STR00133##
For the synthesis of the prodrug groups, see Borchardt, R. T. et.
al., J. Org. Chem. 1997, 43, 3641-3652.
[0353] R.sup.1=all natural,
[0354] unnatural amino acids
##STR00134##
For the synthesis of the prodrug groups, see Zhou, X-X. et. al.,
PCT WO 99/51613.
[0355] R.sup.1.dbd.C1-C4 alkyl, cycloalkyl, oxyalkyl,
[0356] aminoalkyl, etc.
[0357] R.sup.2=all natural, unnatural amino acids
##STR00135##
For the synthesis of the prodrug groups, see Ezra, A. et. al., J.
Med. Chem. 2000, 43, 3641-3652.
[0358] R.sup.1, R.sup.2=all natural, unnatural amino acids
[0359] Other examples of prodrug moieties of interest include
prodrug moieties that can be attached to hydroxyl-containing
functionalities. Such prodrug moieties a well-known in the art, and
will be readily identified by a person skilled in the relevant art.
The present invention encompasses any prodrug form of the compounds
described herein.
[0360] Certain compounds of the present invention, and definitions
of specific functional groups are also described in more detail
below. For purposes of this invention, the chemical elements are
identified in accordance with the Periodic Table of the Elements,
CAS version, Handbook of Chemistry and Physics, 75 Ed., inside
cover, and specific functional groups are generally defined as
described therein. Additionally, general principles of organic
chemistry, as well as specific functional moieties and reactivity,
are described in "Organic Chemistry", Thomas Sorrell, University
Science Books, Sausalito: 1999, the entire contents of which are
incorporated herein by reference. Furthermore, it will be
appreciated by one of ordinary skill in the art that the synthetic
methods, as described herein, utilize a variety of protecting
groups. By the term "protecting group", has used herein, it is
meant that a particular functional moiety, e.g., O, S, or N, is
temporarily blocked so that a reaction can be carried out
selectively at another reactive site in a multifunctional compound.
In preferred embodiments, a protecting group reacts selectively in
good yield to give a protected substrate that is stable to the
projected reactions; the protecting group must be selectively
removed in good yield by readily available, preferably nontoxic
reagents that do not attack the other functional groups; the
protecting group forms an easily separable derivative (more
preferably without the generation of new stereogenic centers); and
the protecting group has a minimum of additional functionality to
avoid further sites of reaction. As detailed herein, oxygen,
sulfur, nitrogen and carbon protecting groups may be utilized. For
example, in certain embodiments, as detailed herein, certain
exemplary oxygen protecting groups are utilized. These oxygen
protecting groups include, but are not limited to methyl ethers,
substituted methyl ethers (e.g., MOM (methoxymethyl ether), MTM
(methylthiomethyl ether), BOM (benzyloxymethyl ether), PMBM
(p-methoxybenzyloxymethyl ether), to name a few), substituted ethyl
ethers, substituted benzyl ethers, silyl ethers (e.g., TMS
(trimethylsilyl ether), TES (triethylsilylether), TIPS
(triisopropylsilyl ether), TBDMS (t-butyldimethylsilyl ether),
tribenzyl silyl ether, TBDPS (t-butyldiphenyl silyl ether), to name
a few), esters (e.g., formate, acetate, benzoate (Bz),
trifluoroacetate, dichloroacetate, to name a few), carbonates,
cyclic acetals and ketals. In certain other exemplary embodiments,
nitrogen protecting groups are utilized. These nitrogen protecting
groups include, but are not limited to, carbamates (including
methyl, ethyl and substituted ethyl carbamates (e.g., Troc), to
name a few) amides, cyclic imide derivatives, N-Alkyl and N-Aryl
amines, imine derivatives, and enamine derivatives, to name a few.
Certain other exemplary protecting groups are detailed herein,
however, it will be appreciated that the present invention is not
intended to be limited to these protecting groups; rather, a
variety of additional equivalent protecting groups can be readily
identified using the above criteria and utilized in the present
invention. Additionally, a variety of protecting groups are
described in "Protective Groups in Organic Synthesis" Third Ed.
Greene, T. W. and Wuts, P. G., Eds., John Wiley & Sons, New
York: 1999, the entire contents of which are hereby incorporated by
reference.
[0361] It will be appreciated that the compounds, as described
herein, may be substituted with any number of substituents or
functional moieties. In general, the term "substituted" whether
preceded by the term "optionally" or not, and substituents
contained in formulas of this invention, refer to the replacement
of hydrogen radicals in a given structure with the radical of a
specified substituent. When more than one position in any given
structure may be substituted with more than one substituent
selected from a specified group, the substituent may be either the
same or different at every position. As used herein, the term
"substituted" is contemplated to include all permissible
substituents of organic compounds. In a broad aspect, the
permissible substituents include acyclic and cyclic, branched and
unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic
substituents of organic compounds. For purposes of this invention,
heteroatoms such as nitrogen may have hydrogen substituents and/or
any permissible substituents of organic compounds described herein
which satisfy the valencies of the heteroatoms. Furthermore, this
invention is not intended to be limited in any manner by the
permissible substituents of organic compounds. Combinations of
substituents and variables envisioned by this invention are
preferably those that result in the formation of stable compounds
useful in the treatment, for example of inflammatory disorders,
cancer, and other disorders, as described generally above. The term
"stable", as used herein, preferably refers to compounds which
possess stability sufficient to allow manufacture and which
maintain the integrity of the compound for a sufficient period of
time to be detected and preferably for a sufficient period of time
to be useful for the purposes detailed herein.
[0362] The term "aliphatic", as used herein, includes both
saturated and unsaturated, straight chain (i.e., unbranched) or
branched aliphatic hydrocarbons, which are optionally substituted
with one or more functional groups. As will be appreciated by one
of ordinary skill in the art, "aliphatic" is intended herein to
include, but is not limited to, alkyl, alkenyl, alkynyl moieties.
Thus, as used herein, the term "alkyl" includes straight and
branched alkyl groups. An analogous convention applies to other
generic terms such as "alkenyl", "alkynyl" and the like.
Furthermore, as used herein, the terms "alkyl", "alkenyl",
"alkynyl" and the like encompass both substituted and unsubstituted
groups. In certain embodiments, as used herein, "lower alkyl" is
used to indicate those alkyl groups (cyclic, acyclic, substituted,
unsubstituted, branched or unbranched) having 1-6 carbon atoms.
[0363] In certain embodiments, the alkyl, alkenyl and alkynyl
groups employed in the invention contain 1-20 aliphatic carbon
atoms. In certain other embodiments, the alkyl, alkenyl, and
alkynyl groups employed in the invention contain 1-10 aliphatic
carbon atoms. In yet other embodiments, the alkyl, alkenyl, and
alkynyl groups employed in the invention contain 1-8 aliphatic
carbon atoms. In still other embodiments, the alkyl, alkenyl, and
alkyl groups employed in the invention contain 1-6 aliphatic carbon
atoms. In yet other embodiments, the alkyl, alkenyl, and alkynyl
groups employed in the invention contain 1-4 carbon atoms.
Illustrative aliphatic groups thus include, but are not limited to,
for example, methyl, ethyl, n-propyl, isopropyl, allyl, n-butyl,
sec-butyl, isobutyl, tert-butyl, n-pentyl, sec-pentyl, isopentyl,
tert-pentyl, n-hexyl, sec-hexyl, moieties and the like, which
again, may bear one or more substituents. Alkenyl groups include,
but are not limited to, for example, ethenyl, propenyl, butenyl,
1-methyl-2-buten-1-yl, and the like. Representative alkynyl groups
include, but are not limited to, ethynyl, 2-propynyl (propargyl),
1-propynyl and the like.
[0364] The term "alicyclic", as used herein, refers to compounds
which combine the properties of aliphatic and cyclic compounds and
include but are not limited to cyclic, or polycyclic aliphatic
hydrocarbons and bridged cycloalkyl compounds, which are optionally
substituted with one or more functional groups. As will be
appreciated by one of ordinary skill in the art, "alicyclic" is
intended herein to include, but is not limited to, cycloalkyl,
cycloalkenyl, and cycloalkynyl moieties, which are optionally
substituted with one or more functional groups. Illustrative
alicyclic groups thus include, but are not limited to, for example,
cyclopropyl, --CH.sub.2-cyclopropyl, cyclobutyl,
--CH.sub.2-cyclobutyl, cyclopentyl, --CH.sub.2-cyclopentyl-n,
cyclohexyl, --CH.sub.2-cyclohexyl, cyclohexenylethyl,
cyclohexanylethyl, norborbyl moieties and the like, which again,
may bear one or more substituents.
[0365] The term "alkoxy" (or "alkyloxy"), or "thioalkyl" as used
herein refers to an alkyl group, as previously defined, attached to
the parent molecular moiety through an oxygen atom or through a
sulfur atom. In certain embodiments, the alkyl group contains 1-20
aliphatic carbon atoms. In certain other embodiments, the alkyl
group contains 1-10 aliphatic carbon atoms. In yet other
embodiments, the alkyl, alkenyl, and alkynyl groups employed in the
invention contain 1-8 aliphatic carbon atoms. In still other
embodiments, the alkyl group contains 1-6 aliphatic carbon atoms.
In yet other embodiments, the alkyl group contains 1-4 aliphatic
carbon atoms. Examples of alkoxy, include but are not limited to,
methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, tert-butoxy,
neopentoxy and n-hexoxy. Examples of thioalkyl include, but are not
limited to, methylthio, ethylthio, propylthio, isopropylthio,
n-butylthio, and the like.
[0366] The term "alkylamino" refers to a group having the structure
--NHR' wherein R' is alkyl, as defined herein. The term
"dialkylamino" refers to a group having the structure
--N(R').sub.2, wherein R' is alkyl, as defined herein. The term
"aminoalkyl" refers to a group having the structure NH.sub.2R',
wherein R' is alkyl, as defined herein. In certain embodiments, the
alkyl group contains 1-20 aliphatic carbon atoms. In certain other
embodiments, the alkyl group contains 1-10 aliphatic carbon atoms.
In yet other embodiments, the alkyl, alkenyl, and alkynyl groups
employed in the invention contain 1-8 aliphatic carbon atoms. In
still other embodiments, the alkyl group contains 1-6 aliphatic
carbon atoms. In yet other embodiments, the alkyl group contains
1-4 aliphatic carbon atoms. Examples of alkylamino include, but are
not limited to, methylamino, ethylamino, iso-propylamino and the
like.
[0367] Some examples of substituents of the above-described
aliphatic (and other) moieties of compounds of the invention
include, but are not limited to aliphatic; alicyclic;
heteroaliphatic; heteroalicyclic; aryl; heteroaryl; alkylaryl;
alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy;
alkylthio; arylthio; heteroalkylthio; heteroarylthio; F; Cl; Br; I;
--OH; --NO.sub.2; --CN; --CF.sub.3; --CH.sub.2CF.sub.3;
--CHCl.sub.2; --CH.sub.2OH; --CH.sub.2CH.sub.2OH;
--CH.sub.2NH.sub.2; --CH.sub.2SO.sub.2CH.sub.3; --C(O)R.sub.x;
--CO.sub.2(R.sub.x); --CON(R.sub.x).sub.2; --OC(O)R.sub.x;
--OCO.sub.2R.sub.x; --OCON(R.sub.x).sub.2; --N(R.sub.x).sub.2;
--S(O).sub.2R.sub.x; --NR.sub.x(CO)R.sub.x wherein each occurrence
of R.sub.x independently includes, but is not limited to,
aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl,
heteroaryl, alkylaryl, or alkylheteroaryl, wherein any of the
aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, alkylaryl,
or alkylheterol substituents described above and herein may be
substituted or unsubstituted, branched or unbranched, cyclic or
acyclic, and wherein any of the aryl or heteroaryl substituents
described above and herein may be substituted or unsubstituted.
Additional examples of generally applicable substituents are
illustrated by the specific embodiments shown in the Examples that
are described herein.
[0368] In general, the terms "aryl" and "heteroaryl", as used
herein, refer to stable mono- or polycyclic, heterocyclic,
polycyclic, and polyheterocyclic unsaturated moieties having
preferably 3-14 carbon atoms, each of which may be substituted or
unsubstituted. It will also be appreciated that aryl and heteroaryl
moieties, as defined herein may be attached via an aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, alkyl or heteroalkyl
moiety and thus also include -aliphatic)aryl,
-(heteroaliphatic)aryl, -(aliphatic)heteroaryl,
-(heteroaliphatic)heteroaryl, -alkyl)aryl, -(heteroalkyl)aryl,
-(heteroalkyl)aryl, and (heteroalkyl)heteroaryl moieties. Thus, as
used herein, the phrases "aryl or heteroaryl" and "aryl,
heteroaryl, [aliphatic)aryl, -(heteroaliphatic)aryl,
-(aliphatic)heteroaryl, -(heteroaliphatic)heteroaryl, -alkyl)aryl,
-(heteroalkyl)aryl, -(heteroalkyl)aryl, and heteroalkyl)heteroaryl"
are interchangeable. Substituents include, but are not limited to,
any of the previously mentioned substitutents, i.e., the
substituents recited for aliphatic moieties, or for other moieties
as disclosed herein, resulting in the formation of a stable
compound. In certain embodiments of the present invention, "aryl"
refers to a mono- or bicyclic carbocyclic ring system having one or
two aromatic rings including, but not limited to, phenyl, naphthyl,
tetrahydronaphthyl, indanyl, indenyl and the like. In certain
embodiments of the present invention, the term "heteroaryl", as
used herein, refers to a cyclic aromatic radical having from five
to ten ring atoms of which one ring atom is selected from S, O and
N; zero, one or two ring atoms are additional heteroatoms
independently selected from S, O and N; and the remaining ring
atoms are carbon, the radical being joined to the rest of the
molecule via any of the ring atoms, such as, for example, pyridyl,
pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl,
oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl,
furanyl, quinolinyl, isoquinolinyl, and the like.
[0369] It will be appreciated that aryl and heteroaryl groups
(including bicyclic aryl groups) can be unsubstituted or
substituted, wherein substitution includes replacement of one or
more of the hydrogen atoms thereon independently with any one or
more of the following moieties including, but not limited to:
aliphatic; alicyclic; heteroaliphatic; heteroalicyclic; aryl;
heteroaryl; alkylaryl; alkylheteroaryl; alkoxy; aryloxy;
heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio;
heteroarylthio; F; Cl; Br; I; --OH; --NO.sub.2; --CN; --CF.sub.3;
--CH.sub.2CF.sub.3; --CHCl.sub.2; --CH.sub.2OH;
--CH.sub.2CH.sub.2OH; --CH.sub.2NH.sub.2;
--CH.sub.2SO.sub.2CH.sub.3; --C(O)R.sub.x; --CO.sub.2(R.sub.x);
--CON(R.sub.x).sub.2; --OC(O)R.sub.x; --OCO.sub.2R.sub.x;
--OCON(R.sub.x).sub.2; --N(R.sub.x).sub.2; --S(O).sub.2R.sub.x;
--NR.sub.x(CO)R.sub.x wherein each occurrence of R.sub.x
independently includes, but is not limited to, aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl,
alkylaryl, or alkylheteroaryl, wherein any of the aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, alkylaryl, or
alkylheteroaryl substituents described above and herein may be
substituted or unsubstituted, branched or unbranched, cyclic or
acyclic, and wherein any of the aryl or heteroaryl substituents
described above and herein may be substituted or unsubstituted.
Additional examples of generally applicable substituents are
illustrated by the specific embodiments shown in the Examples that
are described herein.
[0370] The term "cycloalkyl", as used herein, refers specifically
to groups having three to seven, preferably three to ten carbon
atoms. Suitable cycloalkyls include, but are not limited to
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and
the like, which, as in the case of other aliphatic, heteroaliphatic
or heterocyclic moieties, may optionally be substituted with
substituents including, but not limited to aliphatic; alicyclic;
heteroaliphatic; heteroalicyclic; aryl; heteroaryl; alkylaryl;
alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy;
alkylthio; arylthio; heteroalkylthio; heteroarylthio; F; Cl; Br; I;
--OH; --NO.sub.2; --CN; --CF.sub.3; --CH.sub.2CF.sub.3;
--CHCl.sub.2; --CH.sub.2OH; --CH.sub.2CH.sub.2OH;
--CH.sub.2NH.sub.2; --CH.sub.2SO.sub.2CH.sub.3; --C(O)R.sub.x;
--CO.sub.2(R--); --CON(R.sub.x).sub.2; --OC(O)R.sub.x;
--OCO.sub.2R.sub.x; --OCON(R.sub.x).sub.2; --N(R.sub.x).sub.2;
--S(O).sub.2R.sub.x; --NR.sub.x(CO)R.sub.x wherein each occurrence
of R.sub.x independently includes, but is not limited to,
aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl,
heteroaryl, alkylaryl, or alkylheteroaryl, wherein any of the
aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, alkylaryl,
or alkylheteroaryl substituents described above and herein may be
substituted or unsubstituted, branched or unbranched, cyclic or
acyclic, and wherein any of the aryl or heteroaryl substituents
described above and herein may be substituted or unsubstituted.
Additional examples of generally applicable substituents are
illustrated by the specific embodiments shown in the Examples that
are described herein.
[0371] The term "heteroaliphatic", as used herein, refers to
aliphatic moieties in which one or more carbon atoms in the main
chain have substituted with an heteroatom. Thus, a heteroaliphatic
group refers to an aliphatic chain which contains one or more
oxygen sulfur, nitrogen, phosphorus or silicon atoms, e.g. in place
of carbon atoms. Heteroaliphatic moieties may be branched or linear
unbranched. In certain embodiments, heteroaliphatic moieties are
substituted by independent replacement of one or more of the
hydrogen atoms thereon with one or more moieties including, but not
limited to aliphatic; alicyclic; heteroaliphatic; heteroalicyclic;
aryl; heteroaryl; alkylaryl; alkylheteroaryl; alkoxy; aryloxy;
heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio;
heteroarylthio; F; Cl; Br; I; --OH; --NO.sub.2; --CN; --CF.sub.3;
--CH.sub.2CF.sub.3; --CHCl.sub.2; --CH.sub.2OH;
--CH.sub.2CH.sub.2OH; --CH.sub.2NH.sub.2;
--CH.sub.2SO.sub.2CH.sub.3; --C(O)R.sub.x; --CO.sub.2(R.sub.x);
--CON(R.sub.x).sub.2; --OC(O)R.sub.x; --OCO.sub.2R.sub.x;
--OCON(R.sub.x).sub.2; --N(R.sub.x).sub.2; --S(O).sub.2R.sub.x;
--NR.sub.x(CO)R.sub.x wherein each occurrence of R.sub.x
independently includes, but is not limited to, aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl,
alkylaryl, or alkylheteroaryl, wherein any of the aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, alkylaryl, or
alkylheteroaryl substituents described above and herein may be
substituted or unsubstituted, branched or unbranched, cyclic or
acyclic, and wherein any of the aryl or heteroaryl substituents
described above and herein may be substituted or unsubstituted.
Additional examples of generally applicable substituents are
illustrated by the specific embodiments shown in the Examples that
are described herein.
[0372] The term "heteroalicyclic", as used herein, refers to
compounds which combine the properties of heteroaliphatic and
cyclic compounds and include but are not limited to saturated and
unsaturated mono- or polycyclic heterocycles such as morpholino,
pyrrolidinyl, furanyl, thiofuranyl, pyrrolyl etc., which are
optionally substituted with one or more functional groups.
[0373] The terms "halo" and "halogen" as used herein refer to an
atom selected from fluorine, chlorine, bromine and iodine.
[0374] The term "haloalkyl" denotes an alkyl group, as defined
above, having one, two, or three halogen atoms attached thereto and
is exemplified by such groups as chloromethyl, bromoethyl,
trifluoromethyl, and the like.
[0375] The term "heterocycloalkyl" or "heterocycle", as used
herein, refers to a non-aromatic 5-, 6- or 7-membered ring or a
polycyclic group, including, but not limited to a bi- or tri-cyclic
group comprising fused six-membered rings having between one and
three heteroatoms independently selected from oxygen, sulfur and
nitrogen, wherein (i) each 5-membered ring has 0 to 1 double bonds
and each 6-membered ring has 0 to 2 double bonds, (ii) the nitrogen
and sulfur heteroatoms may be optionally be oxidized, (iii) the
nitrogen heteroatom may optionally be quaternized, and (iv) any of
the above heterocyclic rings may be fused to a benzene ring.
Representative heterocycles include, but are not limited to,
pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl,
imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl,
isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, and
tetrahydrofuryl. In certain embodiments, a "substituted
heterocycloalkyl or heterocycle" group is utilized and as used
herein, refers to a heterocycloalkyl or heterocycle group, as
defined above, substituted by the independent replacement of one or
more of the hydrogen atoms thereon with but are not limited to
aliphatic; alicyclic; heteroaliphatic; heteroalicyclic; aryl;
heteroaryl; alkylaryl; alkylheteroaryl; alkoxy; aryloxy;
heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio;
heteroarylthio; F; Cl; Br; I; --OH; --NO.sub.2; --CN; --CF.sub.3;
--CH.sub.2CF.sub.3; --CHCl.sub.2; --CH.sub.2OH;
--CH.sub.2CH.sub.2OH; --CH.sub.2NH.sub.2;
--CH.sub.2SO.sub.2CH.sub.3; --C(O)R.sub.x; --CO.sub.2(R.sub.x);
--CON(R.sub.x).sub.2; --OC(O)R.sub.x; --OCO.sub.2R.sub.x;
--OCON(R).sub.2; --N(R.sub.x).sub.2; --S(O).sub.2R.sub.x;
--NR.sub.x(CO)R.sub.x wherein each occurrence of R.sub.x
independently includes, but is not limited to, aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl,
alkylaryl, or alkylheteroaryl, wherein any of the aliphatic,
alicyclic, heteroaliphatic, heteroalicyclic, alkylaryl, or
alkylheteroaryl substituents described above and herein may be
substituted or unsubstituted, branched or unbranched, cyclic or
acyclic, and wherein any of the aryl or heteroaryl substitutents
described above and herein may be substituted or unsubstituted.
Additional examples or generally applicable substituents are
illustrated by the specific embodiments shown in the Examples which
are described herein.
[0376] 3) Research Uses, Formulation and Administration
[0377] According to the present invention, the inventive compounds
may be assayed in any of the available assays known in the art for
identifying compounds having a pre-determined biological activity.
For example, the assay may be cellular or non-cellular, in vivo or
in vitro, high- or low-throughput format, etc. In certain exemplary
embodiments, the inventive compounds are tested in assays to
identify those compounds having proteasome inhibitory activity,
antiproliferative/anticancer activity and/or anti-inflammatory
effect.
[0378] Thus, in one aspect, compounds of this invention which are
of particular interest include those which: [0379] exhibit activity
generally as proteasome inhibitors; [0380] exhibit an
antiproliferative and/or anticancer effect on suitable cell lines
maintained in vitro, or in animal studies using a scientifically
acceptable model; [0381] exhibit an anti-inflammatory effect on
suitable cell lines maintained in vitro, or in animal studies using
a scientifically acceptable model; [0382] exhibit in vivo efficacy
vs. human cancer xenografts; and [0383] exhibit a favorable
therapeutic profile (e.g., safety, efficacy, and stability).
[0384] As detailed in the exemplification herein in assays to
determine the ability of exemplary compounds to effect growth
inhibition, certain of the compounds exhibit IC.sub.50s (growth
inhibition vs. adherent HT-29, OVCAR-3, or MDA-MB-435) less than 1
.mu.M. In certain other embodiments, compounds exhibit IC.sub.50s
(growth inhibition vs. adherent HT-29, OVCAR-3, or MDA-MB-435) less
than 100 nM. In still other embodiments, compounds exhibit
IC.sub.50s (growth inhibition vs. adherent HT-29, OVCAR-3, or
MDA-MB-435) less than 10 nM. As also described in the
exemplification herein, inventive compounds are also useful as
proteasome inhibitors. In certain embodiments, compounds exhibit
IC.sub.50s (human leukocyte lysates, HT-29 colon carcinoma, and 20S
proteasomes) less than 1 .mu.M. In certain other embodiments,
compounds exhibit IC.sub.50s (human leukocyte lysates, HT-29 colon
carcinoma, and 20S proteasomes) less then 100 nM. In still other
embodiments, compounds exhibit IC.sub.50s (human leukocyte lysates,
HT-29 colon carcinoma, and 20S proteasomes) less than 10 nM.
[0385] In yet other embodiments, for compounds as described
generally above where A-B-D-E together represent
CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2, w is 0, and R.sub.1,
R.sub.2, R.sub.3 and R.sub.4 are each independently hydrogen,
OR.sub.C, halogen, or NR.sub.CR.sub.D, wherein each occurrence of
R.sub.C and R.sub.D is independently hydrogen or lower alkyl, or
wherein R.sub.C and R.sub.D taken together, or two occurrences of
R.sub.C or R.sub.D, taken together are a cycloaliphatic, or
heterocycloaliphatic moiety, IC.sub.50s (20S inhibition) in the
range of 0.50 to 5.0 nM are observed and IC.sub.50s (growth
inhibition) in the range of 1.0 to 11.0 nM are observed.
[0386] As discussed above, certain of the compounds as described
herein exhibit activity generally as proteasome inhibitors. More
specifically, compounds of the invention demonstrate
anti-inflammatory and/or antitumor activity and thus the invention
further provides a method for treating an inflammatory disorder or
a proliferative disorder, such as cancer. In certain other
embodiments, the compounds of the invention are useful for treating
cancer, where the cancer is a solid tumor.
[0387] The method involves the administration of a therapeutically
effective amount of the compound or a pharmaceutically acceptable
derivative thereof to a subject (including, but not limited to a
human or animal) in need of it. In certain embodiments, the
inventive compounds as useful for the treatment of cancer
(including, but not limited to, prostate cancer, breast cancer,
lung cancer, colon cancer, lymphoma, bladder cancer, cervical
cancer, uterine cancer, melanoma and/or skin cancer, kidney cancer,
testicular cancer, ovarian cancer, stomach cancer, leukemia, brain
cancer, multiple myeloma, liver cancer, pancreatic cancer or
esophageal cancer). In other embodiments, the inventive compounds
are useful for the treatment of inflammatory disorders, and/or
disorders caused by activation of certain regulatory subunits of
the proteasome (e.g., diseases caused by the activation of the
NF.kappa.B pathway, or certain genetic diseases involving the
misfolding of proteins, to name a few). These disorders include,
but are not limited to inflammation, autoimmune diseases (e.g.,
rheumatoid arthritis, lupus erythematosus, multiple sclerosis),
respiratory distress syndrome, neurological disease (e.g.,
Alzheimer's Disease), ischemia, cachexia, cystic fibrosis,
neoplasm, and HIV infection.
[0388] Pharmaceutical Compositions
[0389] As discussed above this invention provides novel compounds
that have biological properties useful for the treatment of cancer
and/or inflammatory disorders, and, in certain embodiments, more
generally are useful as proteaseome inhibitors. In certain
embodiments, the inventive compounds as useful for the treatment of
cancer (including, but not limited to, prostate cancer, breast
cancer, lung cancer, colon cancer, lymphoma, bladder cancer,
cervical cancer, uterine cancer, melanoma and/or skin cancer,
kidney cancer, testicular cancer, ovarian cancer, stomach cancer,
leukemia, brain cancer, multiple myeloma, liver cancer, pancreatic
cancer or esophageal cancer). In other embodiments, the inventive
compounds are useful for the treatment of inflammatory disorders,
and/or disorders caused by activation of the regulatory subunits of
the proteasome. These disorders include, but are not limited to
inflammation, autoimmune diseases (e.g., rheumatoid arthritis,
lupus erythematosus, multiple sclerosis), respiratory distress
syndrome, neurological disease (e.g., Alzheimer's Disease),
ischemia, cachexia, cystic fibrosis, neoplasm, and HIV
infection.
[0390] Accordingly, in another aspect of the present invention,
pharmaceutical compositions are provided, which comprise any one of
the compounds described herein (or a prodrug, pharmaceutically
acceptable salt or other pharmaceutically acceptable derivative
thereof, and optionally comprise a pharmaceutically acceptable
carrier. In certain embodiments, these compositions optionally
further comprise one or more additional therapeutic agents.
Alternatively, a compound of this invention may be administered to
a patient in need thereof in combination with the administration of
one or more other therapeutic agents. For example, additional
therapeutic agents for conjoint administration or inclusion in a
pharmaceutical composition with a compound of this invention may be
an anti-inflammatory agent (e.g., an agent for the treatment of
rheumatoid arthritis or psoriasis) or cytotoxic agent or anticancer
agent approved for the treatment of cancer, as discussed in more
detail herein, or it may be any one of a number of agents
undergoing approval in the Food and Drug Administration that
ultimately obtain approval for the treatment of cancer or an
inflammatory disorder. It will also be appreciated that certain of
the compounds of present invention can exist in free form for
treatment, or where appropriate, as a pharmaceutically acceptable
derivative thereof. According to the present invention, a
pharmaceutically acceptable derivative includes, but is not limited
to, pharmaceutically acceptable salts, esters, salts of such
esters, or a prodrug or other adduct or derivative of a compound of
this invention which upon administration to a patient in need is
capable of providing, directly or indirectly, a compound as
otherwise described herein, or a metabolite or residue thereof.
[0391] As used herein, the term "pharmaceutically acceptable salt"
refers to those salts which are, within the scope of sound medical
judgment, suitable for use in contact with the tissues of humans
and lower animals without undue toxicity, irritation, allergic
response and the like, and are commensurate with a reasonable
benefit/risk ratio. Pharmaceutically acceptable salts of amines,
carboxylic acids, and other types of compounds, are well known in
the art. For example, S. M. Berge, et al. describe pharmaceutically
acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19
(1977), incorporated herein by reference. The salts can be prepared
in situ during the final isolation and purification of the
compounds of the invention, or separately by reacting a free base
or free acid function with a suitable reagent, as described
generally below. For example, a free base function can be reacted
with a suitable acid. Furthermore, where the compounds of the
invention carry an acidic moiety, suitable pharmaceutically
acceptable salts thereof may, include metal salts such as alkali
metal salts, e.g. sodium or potassium salts; and alkaline earth
metal salts, e.g. calcium or magnesium salts. Examples of
pharmaceutically acceptable, nontoxic acid addition salts are salts
of an amino group formed with inorganic acids such as hydrochloric
acid, hydrobromic acid, phosphoric acid, sulfuric acid and
perchloric acid or with organic acids such as acetic acid, oxalic
acid, maleic acid, tartaric acid, citric acid, succinic acid or
malonic acid or by using other methods used in the art such as ion
exchange. Other pharmaceutically acceptable salts include adipate,
alginate, ascorbate, aspartate, benzenesulfonate, benzoate,
bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,
cyclopentanepropionate, digluconate, dodecylsulfate,
ethanesulfonate, formate, fumarate, glucoheptonate,
glycerophosphate, gluconate, hernisulfate, heptanoate, hexanoate,
hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate,
laurate, lauryl sulfate, malate, maleate, malonate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,
oleate, oxalate, palmitate, pamoate, pectinate, persulfate,
3-phenylpropionate, phosphate, picrate, pivalate, propionate,
stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, valerate salts, and the like.
Representative alkali or alkaline earth metal salts include sodium,
lithium, potassium, calcium, magnesium, and the like. Further
pharmaceutically acceptable salts include, when appropriate,
nontoxic ammonium, quaternary ammonium, and amine cations formed
using counterions such as halide, hydroxide, carboxylate, sulfate,
phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.
[0392] Additionally, as used herein, the term "pharmaceutically
acceptable ester" refers to esters that hydrolyze in vivo and
include those that break down readily in the human body to leave
the parent compound or a salt thereof. Suitable ester groups
include, for example, those derived from pharmaceutically
acceptable aliphatic carboxylic acids, particularly alkanoic,
alkenoic, cycloalkanoic and alkanedioic acids, in which each alkyl
or alkenyl moiety advantageously has not more than 6 carbon atoms.
Examples of particular esters include formates, acetates,
propionates, butyrates, acrylates and ethylsuccinates.
[0393] Furthermore, the term "pharmaceutically acceptable prodrugs"
as used herein refers to those prodrugs of the compounds of the
present invention which are, within the scope of sound medical
judgment, suitable for use in contact with the issues of humans and
lower animals with undue toxicity, irritation, allergic response,
and the like, commensurate with a reasonable benefit/risk ratio,
and effective for their intended use, as well as the zwitterionic
forms, where possible, of the compounds of the invention. The term
"prodrug" refers' to compounds that are rapidly transformed in vivo
to yield the parent compound of the above formula, for example by
hydrolysis in blood. A thorough discussion is provided in T.
Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14
of the A.C.S. Symposium Series, and in Edward B. Roche, ed.,
Bioreversible Carriers in Drug Design, American Pharmaceutical
Association and Pergamon Press, 1987, both of which are
incorporated herein by reference.
[0394] As described above, the pharmaceutical compositions of the
present invention additionally comprise a pharmaceutically
acceptable carrier, which, as used herein, includes any and all
solvents, diluents, or other liquid vehicle, dispersion or
suspension aids, surface active agents, isotonic agents, thickening
or emulsifying agents, preservatives, solid binders, lubricants and
the like, as suited to the particular dosage form desired.
Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W.
Martin (Mack Publishing Co., Easton, Pa., 1980) discloses various
carriers used in formulating pharmaceutical compositions and known
techniques for the preparation thereof. Except insofar as any
conventional carrier medium is incompatible with the compounds of
the invention, such as by producing any undesirable biological
effect or otherwise interacting in a deleterious manner with any
other component(s) of the pharmaceutical composition, its use is
contemplated to be within the scope of this invention. Some
examples of materials which can serve as pharmaceutically
acceptable carriers include, but are not limited to, sugars such as
lactose, glucose and sucrose; starches such as corn starch and
potato starch; cellulose and its derivatives such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
powdered tragacanth; malt; gelatine; talc; excipients such as cocoa
butter and suppository waxes; oils such as peanut oil, cottonseed
oil; safflower oil, sesame oil; olive oil; corn oil and soybean
oil; glycols; such as propylene glycol; esters such as ethyl oleate
and ethyl laurate; agar; buffering agents such as magnesium
hydroxide and aluminum hydroxide; alginic acid; pyrogenfree water;
isotonic saline; Ringer's solution; ethyl alcohol, and phosphate
buffer solutions, as well as other non-toxic compatible lubricants
such as sodium lauryl sulfate and magnesium stearate, as well as
coloring agents, releasing agents, coating agents, sweetening,
flavoring and perfuming agents, preservatives and antioxidants can
also be present in the composition, according to the Judgment of
the formulator.
[0395] Uses and Formulations of Compounds of the Invention
[0396] As described in more detail herein, in general, the present
invention provides compounds useful for the treatment of cancer and
inflammatory disorders. Without wishing to be bound by any
particular theory, more generally, the compounds of the invention
have also been shown to act as proteaseome inhibitors and thus may
be useful more generally for a variety of disorders that are
affected by processes regulated by the proteasome (e.g., cell
cycle, activation of NF.kappa.B, to name a few).
[0397] As discussed above, compounds of the invention exhibit
antiproliferative and antitumor activity. As such, compounds of the
invention are particularly useful for the treatment of cancer, and
in certain embodiments for the treatment of solid tumors.
Additionally, the inventive compounds are useful as proteasome
inhibitors and can thus be used for the treatment of a variety of
disorders, as discussed herein, that are affected by the
proteasome.
[0398] Thus, as described above, in another aspect of the
invention, a method for the treatment of cancer is provided
comprising administering a therapeutically effective amount of a
compound of formula (I), as described herein, to a subject in need
thereof. It will be appreciated that the compounds and
compositions, according to the method of the present invention, may
be administered using any amount and any route of administration
effective for the treatment of cancer. Thus, the expression
"effective amount" as used herein, refers to a sufficient amount of
agent to kill or inhibit the growth of tumor cells. In other
embodiments, compounds may be useful for the treatment of
inflammatory disorders, or other disorders affected by proteasome
inhibition and thus "effective amount" refers to a sufficient
amount of agent to treat or ameliorate the symptoms of the
inflammatory disorder, or alternatively, refers to a sufficient
amount to effect proteasome inhibition. The exact amount required
will vary from subject to subject, depending on the species, age,
and general condition of the subject, the severity of the
infection, the particular therapeutic agent, its mode of
administration, and the like. The compounds of the invention are
preferably formulated in dosage unit form for ease of
administration and uniformity of dosage. The expression "dosage
unit form" as used herein refers to a physically discrete unit of
therapeutic agent appropriate for the patient to be treated. It
will be understood, however, that the total daily usage of the
compounds and compositions of the present invention will be decided
by the attending physician within the scope of sound medical
judgment. The specific therapeutically effective dose level for any
particular patient or organism will depend upon a variety of
factors including the disorder being treated and the severity of
the disorder; the activity of the specific compound employed; the
specific composition employed; the age, body weight, general
health, sex and diet of the patient; the time of administration,
route of administration, and rate of excretion of the specific
compound employed; the duration of the treatment; drugs used in
combination or coincidental with the specific compound employed;
and like factors well known in the medical arts.
[0399] Furthermore, after formulation with an appropriate
pharmaceutically acceptable carrier in a desired dosage, the
pharmaceutical compositions of this invention can be administered
to humans and other animals orally, rectally, parenterally,
intracistemally, intravaginally, intraperitoneally, topically (as
by powders, ointments, or drops), bucally, as an oral or nasal
spray, as an aerosol, or the like, depending on the severity of the
infection being treated. In certain embodiments, the compounds of
the invention may be administered at dosage levels of about 0.001
mg/kg to about 50 mg/kg, from about 0.01 mg/kg to about 25 mg/kg,
or from about 0.1 mg/kg to about 10 mg/kg of subject body weight
per day, one or more times a day, to obtain the desired therapeutic
effect. It will also be appreciated that dosages smaller than 0.001
mg/kg or greater than 50 mg/kg (for example 50-100 mg/kg) can be
administered to a subject. In certain embodiments, compounds are
administered orally or parenterally.
[0400] Liquid dosage forms for oral administration include, but are
not limited to, pharmaceutically acceptable emulsions,
microemulsions, solutions, suspensions, syrups and elixirs. In
addition to the active compounds, the liquid dosage forms may
contain inert diluents commonly used in the art such as, for
example, water or other solvents, solubilizing agents and
emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in
particular, cottonseed, groundnut, corn, germ, olive, castor, and
sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene
glycols and fatty acid esters of sorbitan, and mixtures thereof.
Besides inert diluents, the oral compositions can also include
adjuvants such as wetting agents, emulsifying and suspending
agents, sweetening, flavoring, and perfuming agents.
[0401] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions may be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution, suspension or emulsion in a nontoxic
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution, U.S.P.
and isotonic sodium chloride solution. In addition, sterile, fixed
oils are conventionally employed as a solvent or suspending medium.
For this purpose any bland fixed oil can be employed including
synthetic mono- or diglycerides. In addition, fatty acids such as
oleic acid are used in the preparation of injectables.
[0402] The injectable formulations can be sterilized, for example,
by filtration through a bacterial-retaining filter, or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium prior to use.
[0403] In order to prolong the effect of a drug, it is often
desirable to slow the absorption of the drug from subcutaneous or
intramuscular injection. This may be accomplished by the use of a
liquid suspension or crystalline or amorphous material with poor
water solubility. The rate of absorption of the drug then depends
upon its rate of dissolution that, in turn, may depend upon crystal
size and crystalline form. Alternatively, delayed absorption of a
parenterally administered drug form is accomplished by dissolving
or suspending the drug in an oil vehicle. Injectable depot forms
are made by forming microencapsule matrices of the drug in
biodegradable polymers such as polylactide-polyglycolide. Depending
upon the ratio of drug to polymer and the nature of the particular
polymer employed, the rate of drug release can be controlled.
Examples of other biodegradable polymers include (poly(orthoesters)
and poly(anhydrides). Depot injectable formulations are also
prepared by entrapping the drug in liposomes or microemulsions
which are compatible with body tissues.
[0404] Compositions for rectal or vaginal administration are
preferably suppositories which can be prepared by mixing the
compounds of this invention with suitable non-irritating excipients
or carriers such as cocoa butter, polyethylene glycol or a
suppository wax which are solid at ambient temperature but liquid
at body temperature and therefore melt in the rectum or vaginal
cavity and release the active compound.
[0405] Solid dosage forms for oral administration include capsules,
tablets, pills, powders, and granules. In such solid dosage forms,
the active compound is mixed with at least one inert,
pharmaceutically acceptable excipient or carrier such as sodium
citrate or dicalcium phosphate and/or a) fillers or extenders such
as starches, lactose, sucrose, glucose, mannitol, and silicic acid,
b) binders such as, for example, carboxymethylcellulose, alginates,
gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants
such as glycerol, d) disintegrating agents such as agar-agar,
calcium carbonate, potato or tapioca starch, alginic acid, certain
silicates, and sodium carbonate, e) solution retarding agents such
as paraffin, f) absorption accelerators such as quaternary ammonium
compounds, g) wetting agents such as, for example, cetyl alcohol
and glycerol monostearate, h) absorbents such as kaolin and
bentonite clay, and i) lubricants such as talc, calcium stearate,
magnesium stearate, solid polyethylene glycols, sodium lauryl
sulfate, and mixtures thereof. In the case of capsules, tablets and
pills, the dosage form may also comprise buffering agents.
[0406] Solid compositions of a similar type may also be employed as
fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugar as well as high molecular
weight polyethylene glycols and the like. The solid dosage forms of
tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings and other
coatings well known in the pharmaceutical formulating art. They may
optionally contain opacifying agents and can also be of a
composition that they release the active ingredient(s) only, or
preferentially, in a certain part of the intestinal tract,
optionally, in a delayed manner. Examples of embedding compositions
that can be used include polymeric substances and waxes. Solid
compositions of a similar type may also be employed as fillers in
soft and hard-filled gelatin capsules using such excipients as
lactose or milk sugar as well as high molecular weight polyethylene
glycols and the like.
[0407] The active compounds can also be in micro-encapsulated form
with one or more excipients as noted above. The solid dosage forms
of tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings, release
controlling coatings and other coatings well known in the
pharmaceutical formulating art. In such solid dosage forms the
active compound may be admixed with at least one inert diluent such
as sucrose, lactose and starch. Such dosage forms may also
comprise, as in normal practice, additional substances other than
inert diluents, e.g., tableting lubricants and other tableting aids
such as magnesium stearate and microcrystalline cellulose. In the
case of capsules, tablets and pills, the dosage forms may also
comprise buffering agents. They may optionally contain opacifying
agents and can also be of a composition that they release the
active ingredient(s) only, or preferentially, in a certain part of
the intestinal tract, optionally, in a delayed manner. Examples of
embedding compositions which can be used include polymeric
substances and waxes.
[0408] Dosage forms for topical or transdermal administration of a
compound of this invention include ointments, pastes, creams,
lotions, gels, powders, solutions, sprays, aerosols, inhalants or
patches. The active component is admixed under sterile conditions
with a pharmaceutically acceptable carrier and any needed
preservatives or buffers as may be required. Ophthalmic
formulation, ear drops, and eye drops are also contemplated as
being within the scope of this invention. Additionally, the present
invention contemplates the use of transdermal patches, which have
the added advantage of providing controlled delivery of a compound
to the body. Such dosage forms are made by dissolving or dispensing
the compound in the proper medium. Absorption enhancers can also be
used to increase the flux of the compound across the skin. The rate
can be controlled by either providing a rate controlling membrane
or by dispersing the compound in a polymer matrix or gel.
[0409] It will also be appreciated that the compounds and
pharmaceutical compositions of the present invention can be
formulated and employed in combination therapies, that is, the
compounds and pharmaceutical compositions can be formulated with or
administered concurrently with, prior to, or subsequent to, one or
more other desired therapeutics or medical procedures. The
particular combination of therapies (therapeutics or procedures) to
employ in a combination regimen will take into account
compatibility of the desired therapeutics and/or procedures and the
desired therapeutic effect to be achieved. For example, other
therapies or anticancer agents that may be used in combination with
the inventive anticancer agents of the present invention include
surgery, radiotherapy (in but a few examples, .gamma.-radiation,
neutron beam radiotherapy, electron beam radiotherapy, proton
therapy, brachytherapy, and systemic radioactive isotopes, to name
a few), endocrine therapy, biologic response modifiers
(interferons, interleukins, and tumor necrosis factor (TNF) to name
a few), hyperthermia and cryotherapy, agents to attenuate any
adverse effects (e.g., antiemetics), and other approved
chemotherapeutic drugs, including, but not limited to, alkylating
drugs (mechlorethamine, chlorambucil, Cyclophosphamide, Melphalan,
Ifosfamide), antimetabolites (Methotrexate), purine antagonists and
pyrimidine antagonists (6-Mercaptopurine, 5-Fluorouracil,
Cytarabile, Gemcitabine), spindle poisons (Vinblastine,
Vincristine, Vinorelbine, Paclitaxel), podophyllotoxins (Etoposide,
Irinotecan, Topotecan), antibiotics (Doxorubicin, Bleomycin,
Mitomycin), nitrosoureas (Carmustine, Lomustine), inorganic ions
(Cisplatin, Carboplatin), enzymes (Asparaginase), and hormones
(Tamoxifen, Leuprolide, Flutamide, and Megestrol), to name a few.
For a more comprehensive discussion of updated cancer therapies
see, http://www.nci.nih.gov/, a list of the FDA approved oncology
drugs at http://www.fda.gov/cder/cancer/druglistframe.htm, and The
Merck Manual, Seventeenth Ed. 1999, the entire contents of which
are hereby incorporated by reference. It will also be appreciated
that the therapies employed may achieve a desired effect for the
same disorder (for example, an inventive compound may be
administered concurrently with another anti-inflammatory agent or
anticancer agent), or they may achieve different effects (e.g.,
control of any adverse effects).
[0410] In certain embodiments, the pharmaceutical compositions of
the present invention further comprise one or more additional
therapeutically active ingredients (e.g., chemotherapeutic and/or
palliative). For purposes of the invention, the term "Palliative"
refers to treatment that is focused on the relief of symptoms of a
disease and/or side effects of a therapeutic regimen, but is not
curative. For example, palliative treatment encompasses
painkillers, antinausea medications and anti-sickness drugs. In
addition, chemotherapy, radiotherapy and surgery can all be used
palliatively (that is, to reduce symptoms without going for cure;
e.g., for shrinking tumors and reducing pressure, bleeding, pain
and other symptoms of cancer).
Treatment Kits
[0411] In other embodiments, the present invention relates to a kit
for conveniently and effectively carrying out the methods in
accordance with the present invention. In general, the
pharmaceutical pack or kit comprises one or more containers filled
with one or more of the ingredients of the pharmaceutical
compositions of the invention. Such kits are especially suited for
the delivery of solid oral forms such as tablets or capsules. Such
a kit preferably includes a number of unit dosages, and may also
include a card having the dosages oriented in the order of their
intended use. If desired, a memory aid can be provided, for example
in the form of numbers, letters, or other markings or with a
calendar insert, designating the days in the treatment schedule in
which the dosages can be administered. Alternatively, placebo
dosages, or calcium dietary supplements, either in a form similar
to or distinct from the dosages of the pharmaceutical compositions,
can be included to provide a kit in which a dosage is taken every
day. Optionally associated with such container(s) can be a notice
in the form prescribed by a governmental agency regulating the
manufacture, use or sale of pharmaceutical products, which notice
reflects approval by the agency of manufacture, use or sale for
human administration.
Equivalents
[0412] The representative examples that follow are intended to help
illustrate the invention, and are not intended to, nor should they
be construed to, limit the scope of the invention. Indeed, various
modifications of the invention and many further embodiments
thereof, in addition to those shown and described herein, will
become apparent to those skilled in the art from the full contents
of this document, including the examples which follow and the
references to the scientific and patent literature cited herein. It
should further be appreciated that the contents of those cited
references are incorporated herein by reference to help illustrate
the state of the art.
[0413] The following examples contain important additional
information, exemplification and guidance that can be adapted to
the practice of this invention in its various embodiments and the
equivalents thereof.
Exemplification
[0414] The practitioner has a well-established literature of
peptide chemistry to draw upon, in combination with the information
contained herein, for guidance on synthetic strategies, protecting
groups, and other materials and methods useful for the synthesis of
the compounds of this invention.
[0415] The various references cited herein provide helpful
background information on preparing compounds similar to the
inventive compounds described herein or relevant intermediates, as
well as information on formulation, uses, and administration of
such compounds which may be of interest.
[0416] Moreover, the practitioner is directed to the specific
guidance and examples provided in this document relating to various
exemplary compounds and intermediates thereof.
[0417] The compounds of this invention and their preparation can be
understood further by the examples that illustrate some of the
processes by which these compounds are prepared or used. It will be
appreciated, however, that these examples do not limit the
invention. Variations of the invention, now known or further
developed, are considered to fall within the scope of the present
invention as described herein and as hereinafter claimed.
[0418] According to the present invention, any available techniques
can be used to make or prepare the inventive compounds or
compositions including them. For example, a variety of solution
phase synthetic methods such as those discussed in detail below may
be used. Alternatively or additionally, the inventive compounds may
be prepared using any of a variety combinatorial techniques,
parallel synthesis and/or solid phase synthetic methods known in
the art.
[0419] It will be appreciated as described below, that a variety of
inventive compounds can be synthesized according to the methods
described herein. The starting materials and reagents used in
preparing these compounds are either available from commercial
suppliers such as Aldrich Chemical Company (Milwaukee, Wis.),
Bachem (Torrance, Calif.), Sigma (St. Louis, Mo.), or are prepared
by methods well known to a person of ordinary skill in the art
following procedures described in such references as Fieser and
Fieser 1991, "Reagents for Organic Synthesis", vols 1-17, John
Wiley and Sons, New York, N.Y., 1991; Rodd 1989 "Chemistry of
Carbon Compounds", vols. 1-5 and supps, Elsevier Science
Publishers, 1989; "Organic Reactions", vols 1-40, John Wiley and
Sons, New York, N.Y., 1991; March 2001, "Advanced Organic
Chemistry", 5th ed. John Wiley and Sons, New York, N.Y.; and Larock
1990, "Comprehensive Organic Transformations: A Guide to Functional
Group Preparations", 2.sup.nd ed. VCH Publishers. These schemes are
merely illustrative of some methods by which the compounds of this
invention can be synthesized, and various modifications to these
schemes can be made and will be suggested to a person of ordinary
skill in the art having regard to this disclosure.
[0420] The starting materials, intermediates, and compounds of this
invention may be isolated and purified using conventional
techniques, including filtration, distillation, crystallization,
chromatography, and the like. They may be characterized using
conventional methods, including physical constants and spectral
data.
[0421] 1) Experimental Procedures:
[0422] As described above, the present invention provides novel
epoxomicin and eponemycin analogs having formula (I) as described
above and in certain classes and subclasses herein. The synthesis
of certain exemplary compounds is described in detail below. It
will be appreciated that the methods as described herein can be
applied to each of the compounds as disclosed herein and
equivalents thereof. Additionally, certain regents and starting
materials are well known to those skilled in the art. Although the
following examples describe certain exemplary compounds, it will be
appreciated that the use of alternate starting materials will
readily yield other analogues encompassed by the invention.
[0423] The Examples that follow describe exemplary synthetic
methodologies for the preparation of inventive compounds wherein Q
is an epoxycarbonyl moiety. In addition, the skilled practitioner
has a well-established literature of boron and peptide chemistry to
draw upon, in combination with the information contained in the
many examples which follow, for guidance on synthetic strategies,
protecting groups, and other materials and methods useful for the
synthesis of those inventive compounds wherein Q is a
boron-containing moiety. References of particular interest include
U.S. Pat. No. 6,297,217 and references cited therein, each of which
is incorporated herein by reference in its entirety. Thus, a person
of ordinary skill in the art may draw upon the cited literature and
the synthetic guidance provided herein (e.g., for P2 and P3
fragments) to prepare the inventive boron-containing compounds. The
practitioner, armed with knowledge in the relevant art and the
teaching and guidance provided herein and the cited references will
know how to select available relevant chemical transformations,
combined with protection and deprotection schemes, as desired or
required, to prepare compounds of the invention wherein Q is a
boron-containing moiety.
[0424] In certain embodiments, as depicted generally below in
Schemes 1-5, the three components of the compounds, P1, P2 and P3
can each be varied to generate a variety of analogues. In certain
embodiments, P1 and P2 are constant, as depicted in Scheme 6, and
P3 can be varied as desired as depicted generally below and in the
exemplification herein.
##STR00136## ##STR00137## ##STR00138##
##STR00139## ##STR00140## ##STR00141##
##STR00142##
##STR00143## ##STR00144## ##STR00145##
##STR00146##
[0425] As discussed above, in certain embodiments of the invention,
P1 and P2 are held constant while P3 is varied. One exemplary
embodiment is depicted below. Certain other embodiments are
described generally in the experimentals below.
##STR00147##
[0426] As depicted above in Scheme 6, in general, the components Pt
and P2 are prepared and coupled to yield the epoxyalcohol. Scheme 6
also depicts one exemplary embodiment where P3 represents a
bicyclic carboxylic acid which is then coupled via amide coupling
to the P1-P2 component. Subsequent oxidation of the alcohol and
deprotection yields the subclass of compounds depicted above. It
will be appreciated, however, that a variety of P3 components can
be utilized to generate inventive compounds, many of which are
depicted herein. Furthermore, although the preparation of the P1-P2
component as an epoxyalcohol is depicted above, it will be
appreciated that this component can also be prepared as the
epoxyketone, as detailed in the exemplification herein.
[0427] A) General Reaction Procedures:
[0428] Unless mentioned specifically, reaction mixtures were
stirred using a magnetically driven stirrer bar. An inert
atmosphere refers to either dry argon or dry nitrogen. Reactions
were monitored either by thin layer chromatography, or by proton
nuclear magnetic resonance, of a suitably worked up sample of the
reaction mixture.
[0429] B) General Work Up Procedures,
[0430] Unless mentioned specifically, reaction mixtures were cooled
to room temperature or below then quenched, when necessary, with
either water or a saturated aqueous solution of ammonium chloride.
Desired products were extracted by partitioning between water and a
suitable water-immiscible solvent (e.g. ethyl acetate,
dichloromethane, diethyl ether). The desired product containing
extracts were washed appropriately with water followed by a
saturated solution of brine. On occasions where the product
containing extract was deemed to contain residual oxidants, the
extract was washed with a 10% solution of sodium sulphite in
saturated aqueous sodium bicarbonate solution, prior to the
aforementioned washing procedure. On occasions where the product
containing extract was deemed to contain residual acids, the
extract was washed with saturated aqueous sodium bicarbonate
solution, prior to the aforementioned washing procedure (except in
those cases where the desired product itself had acidic character).
On occasions where the product containing extract was deemed to
contain residual bases, the extract was washed with 10% aqueous
citric acid solution, prior to the aforementioned washing procedure
(except in those cases where the desired product itself had basic
character). Post washing, the desired product containing extracts
were dried over anhydrous magnesium sulphate, then filtered. The
crude products were then isolated by removal of solvent(s) by
rotary evaporation under reduced pressure, at an appropriate
temperature (generally less than 45.degree. C.).
[0431] C) General Purification Procedures:
[0432] Unless mentioned specifically, chromatographic purification
refers to flash column chromatography on silica, using a single
solvent or mixed solvent as eluent. Suitably purified desired
product containing elutes were combined and concentrated under
reduced pressure at an appropriate temperature (generally less than
45.degree. C.) to constant mass. Final compounds were dissolved in
50% aqueous acetonitrile, filtered and transferred to vials, then
freeze-dried under high vacuum before submission for biological
testing.
[0433] D) Experimentals for Certain Exemplary Compounds:
[0434] All commercial solvent was used without any purification.
Flush column chromatography was performed on Merck silica gel 60
(230-400 mesh) using hexane-ethyl acetate solvent.
[0435] According to the general procedure below mentioned, and
referring to the listing of compounds following these procedures,
compounds A are synthesized with the modification of P3 part,
compounds J are synthesized with the modification of P2, compounds
C are synthesized with the modification of P1, and compounds D are
synthesized with the modification of P1, P2 and P3.
##STR00148##
[0436] SOCl.sub.2 (130 ml) was added to methanol (500 ml) at
-10.degree. C., after 30 minutes L-Cbz-leucine (133 g) was added
and stirred for overnight at RT. Methanol was evaporated and
partitioned between ethyl acetate and NaHCO.sub.3 aqueous solution.
The organics were washed with brine, dried and evaporated.
[0437] To the above crude material in dichloromethane (1000 ml) was
added DIBAL (500 ml) at -78.degree. C., 30 minutes after the
addition, was added NH4Cl (50 ml) at -78.degree. C. and raised up
to O.degree. C., partitioned between ether and potassium sodium
tartrate, dried, evaporated to give 100 g of aldehyde.
##STR00149##
[0438] To the suspension of CrCl.sub.2 (20.4 g) and NiCl.sub.2
(0.054 g,) in DMF was added aldehyde (22.8 g) and bromide (25.3 g)
at 0.degree. C. After the addition was completed the reaction
mixture was stirred for overnight at RT, extracted with ethyl
acetate, dried and chromatographed to give 25.0 g of alcohol.
##STR00150##
[0439] Allyl alcohol (16.5 g) was dissolved into 150 ml of toluene.
VO(acac).sub.2 (0.42 g,) was added followed by tBuOOH (15.6 ml of a
5-6M solution). The resulting solution was stirred for 3 hours at
room temperature, extracted with ethyl acetate, dried and
chromatographed to give 15.4 g of epoxide.
##STR00151##
[0440] Epoxide (10 g) was dissolved in 100 ml of ethyl acetate,
followed by the addition of catalytic 5% Pd--C, and the mixture was
stirred for 4 hours at RT in hydrogen atmosphere.
[0441] The reaction mixture was then filtered through Celite and
the organics concentrated to obtain 6.9 g of amine. The crude
material was used without further purification.
##STR00152##
[0442] The solution of amine (16.43 g), carboxylic acid (25 g), DCC
(22.9 g) and N-hydroxy-succinimide (12.6 g) in dichloromethane (400
ml) was stirred at -20.degree. C. for overnight and the reaction
mixture was loaded on chromatography to give 24.8 g of desired
compound.
##STR00153##
[0443] To the solution of starting material (24.7 g) in DMF (130
ml) was added piperazine (29 g) at .degree. C. and stirred for 1
hour. The reaction mixture was partitioned between water and ethyl
acetate, dried and chromatographed to give 16.7 g of desired
amine.
##STR00154##
[0444] A solution of unsaturated ester (17.5 g) in 30 ml of
methanol were added to the
(R,R)-(-)-1,2-bis-[(o-methoxyphenyl)(phenyl)phosphino]ethane(1,5-cyclooct-
adiene)rhodium(I) tetrafluoroborate (100 mg) and triethylamine (4.3
g). The reaction mixture was stirred at room temperature for 24
hours in the hydrogen atmosphere. After removing solvent, the
residue was partitioned between 0.1 M NaOH and dichloromethane. The
aqueous layer was acidified and extracted with ethyl acetate, dried
to give 10 g of pure product.
##STR00155##
[0445] Starting acid (5 g) was dissolved in trifluoromethane
sulfonic acid (20 ml) at 0.degree. C., stirred for 2 hours,
partitioned between ethyl acetate and sodium bicarbonate solution.
Organic layer was dried to give 4.5 g of ketone.
##STR00156##
[0446] To the ketone (4.5 g) in trifluoro acetic acid (20 ml) was
added triethyl silane (15 ml) at 0.degree. C. After stirring for 4
hours at RT, the reaction mixture was partitioned between ethyl
acetate and sodium bicarbonate solution, the organic layer was
dried, evaporated to give 4 g of eater. This ester was hydrolyzed
with lithium hydroxide in THF: MEeOH: H2O (3:1:1) at RT for 2 hours
to give 3.5 g of acid.
##STR00157##
[0447] The solution of carboxylic acid (266 mg), amine (502 mg) and
HBTU (760 mg) in dichloromethane (10 ml) was stirred for overnight
at RT. Then the mixture was partitioned between ethyl acetate and
water, the organics was dried and chromatographed to give 525 mg of
desired amide.
##STR00158##
[0448] To the solution of alcohol (525 mg) in dichloromethane (20
ml) was added tetrapropylammonium perruthenate (10 mg) and 4-methyl
morpholine oxide (408 mg). After stirred for 5 hours at RT, the
mixture was chromatographed to give ketone (500 mg).
##STR00159##
[0449] To the solution of ketone (500 mg) in THF (30 ml) was added
hydrogen fluoride-pyridine (4 ml) at RT. After 2 hours, the mixture
was partitioned between ethyl acetate and sodium bicarbonate
solution, the organics was dried, chromatographed to give alcohol
(312 mg).
[0450] I) Alternative Procedure to Prepare Oxidized P1-P2
Component:
##STR00160##
Synthesis of (3)
[0451] To the 25 g (0.0566 mol) of (1), 12.66 g (0.11 mol) of
N-hydroxysuccinimide and 22.8 g (0.11 mol) of DCC were dissolved in
300 ml of dichloromethane at -78.degree. C. After sting for 1 hour
16.43 g (0.0541 mol) of (2) in 20 ml of dichloromethane was added
and stirred at -20.degree. C. for overnight. The reaction mixture
was evaporated and the crude material was purified on silica get
chromatography to afford 24.8 g (63% yield) of (3) as pale yellow
foam.
[0452] Synthesis of (4)
[0453] 24.8 g (0.034 mol) of (3) was dissolved in 136 ml of
anhydrous DMF at 0.degree. C. To this solution was added 29 g
(0.344 mol) of piperazine and stirred at 0.degree. C. for 2 hours
and then the reaction mixture was poured into ethylacetate. The
organic layer was washed successively water and brine and dried
over MgSO.sub.4. The crude material laws purified on silica gel
chromatography to afford 16.72, (97%) of (4) as colorless oil.
[0454] F) Synthesis of ER-804191:
##STR00161## ##STR00162## ##STR00163##
[0455] Step 1
[0456] Compound 1.
[0457] 20 g of methyl 4-hydroxybenzoate (131 mmol) was dissolved in
dry DMF (200 mL). Solid potassium carbonate (36.3 g, 263 mmol) then
3-chloro-2-methylpropene (23.8 g, 263 mmol) were added. The
reaction mixture was stirred for two hours at room temperature. The
reaction mixture was diluted in water and extracted several times
with MTBE. The combined organic layers were washed with NaOH 1N and
with brine then dried over magnesium sulfate. The crude material
(26 g) was used directly in the next step.
[0458] Step 2
[0459] Compound 2
[0460] Compound 1 (15 g, 73 mmol) was dissolved into 120 mL of
aniline. The mixture was stirred for four hours at 196C. The
aniline was removed by distillation under vacuum. The residue was
poured into 1N HCl, and extracted ethyl acetate. The combined
organic layers were washed twice with water and dried over
MgSO.sub.4. The organics were then concentrated and purified by
flash column chromatography using hexane/ethyl acetate 90/10.
[0461] Step 3
[0462] Compound 3
[0463] Compound 2 (22 g, 106 mmol) was dissolved in 125 mL of dry
CH.sub.2Cl.sub.2, followed by the addition of dry pyridine (11.23
ml, 139 mmol). The mixture was cooled at 0 C and triflic anhydride
(21.6 ml 28.16 mmol) was added. After five minutes the reaction
mixture was warmed to room temperature and stirred for 1 hour. The
reaction mixture was concentrated under vacuum and suspended in
ethyl ether. The suspension was filtered and salts washed with
ethyl ether. The solution was concentrated under vacuum to give
34.91 g of compound 3.
[0464] Step 4
[0465] Compound 4
[0466] Compound 3 (34.37 g, 102 mmol), Pd(OAc).sub.2 (6.9 g, 30.06
mmol) and 5% palladium on charcoal (6.9 g) were suspended in 680 mL
of MeOH. The reaction mixture was cooled to 0 C and
NH.sub.4CO.sub.2H (20.5 g, 325 mmol) was added. After 15 minutes
the reaction mixture was warmed to room temperature and stirred for
3 hours. The reaction mixture was then filtered through celite and
the organics concentrated to obtain an oil. The crude material was
diluted with ethyl ether, and the salts removed by filtration.
Purification on silicagel (Hexane/Ethyl Acetate: 98/2) afforded
19.6 g of compound 4.
[0467] Step 5
[0468] Compound 5
[0469] Compound 4 (3 g, 15.6 mmol) was dissolved in ethanol (32 ml)
and a solution of potassium hydroxide (4.4 g, 78.1 mmol) in water
(16 ml) was added. The reaction mixture was stirred overnight at
room temperature. The reaction mixture was poured into 1N HCL and
extracted with MTBE. The combined organic layers were washed once
with 1N sodium hydroxide and brine. The crude material (compound 4,
28 g) was used in the next step without further purification.
[0470] Step 6
[0471] Compound 6
[0472] L-Serine Benzylester HCl (14.14 g, 61 mmol) was dissolved
into 170 mL of CH.sub.2Cl.sub.2 followed by the addition of HBTU
(22.36 g, 72.14 mmol), Et.sub.3N (29 mL, 166.47 mmol), and compound
4 (2.8 g, 11.93 mmol). The reaction mixture was stirred at room
temperature for 18 hours and quenched by pouring into 1N HCl. The
mixture was extracted four times with CH.sub.2Cl.sub.2. The
combined organic layers were dried over MgSO.sub.4 and then
concentrated. Purification of the crude material by flash column
chromatography using Hexane/Ethyl acetate: 70/30 afforded 18.9 g of
compound 6.
[0473] Step 7
[0474] Compound 7.
[0475] Compound 6 (18.33 g, 51.57 mmol) was dissolved in 50 mL of
DMF and TBSCl (8.16 g, 54.14 mmol) and imidazole (8.78 mg, 128-93
mmol) were added and stirred overnight at room temperature. The
reaction mixture was poured into water and extracted three times
with MTBE. The combined organic layers were washed with 1N HCl,
water and with a saturated solution of sodium bicarbonate. The
crude material was purified by flash column chromatography with
Hexane/Ethyl Acetate: 80/20 to give 23.8 g of compound 7.
[0476] Step 8.
[0477] Compound 8.
[0478] Compound 7 (2.0 g, 4.3 mmol) was dissolved in 20 mL of MeOH,
followed by the addition of Pd/C 10%/wt (20 mg). The solution was
then flushed with hydrogen and stirred for 2 hours under hydrogen
at room temperature. The reaction mixture was then filtered through
celite and the organics concentrated to obtain 1.60 g of compound
8. The crude material is used without further purification.
[0479] Step 9.
[0480] Compound 9.
[0481] N-benzyloxycarbonyl leucine methyl ester (79 g, 285 mmol)
was dissolved in 3000 ml of dry toluene. The solution was cooled at
-78 C and a 1M solution of DLBAI-H (598 ml, 598 mmol) was added
dropwise. After 1 hour at -78 C, 108 ml of a saturated aqueous
solution of NH.sub.4Cl were added at -78 C, The reaction mixture
was warmed to 0 C and poured into 3000 ml of Ethyl Ether. The
resulting slurry was vigorously stirred for 1 hour, and then the
aluminum salts were filtered out and washed several times with
ethyl ether. The combined organic layers were concentrated to give
75 g of crude compound 9. The crude material is used in the next
step without further purification.
[0482] Step 10.
[0483] Compound 10.
[0484] Compound 9 (285 mmol) and
3-tert-buthyldimethylsilyloxy-2-bromopropen (71.3 g, 64 mmol) were
then dissolved in dry DMF (1600 mL) under nitrogen. The mixture was
cooled at 0 C and CrCl.sub.2 (64 g) and NiCl.sub.2 (0.64 g) were
added. The reaction was stirred overnight. The reaction mixture was
poured into 4000 ml of water and extracted five times with MTBE.
Combined organic layers were washed three times with water and once
with a saturated solution of sodium bicarbonate then dried over
MgSO.sub.4 and concentrated. The crude green oil was then purified
by column chromatography using Hexane/MTBE/i-PrOH (90/10/1) to
afford 16.5 g of compound 10.
[0485] Step 11.
[0486] Compound 11.
[0487] Compound 10 (16.5 g mmol) was dissolved into 156 mL of
toluene. VO(acac).sub.2 (0.42 g, 0.1.9 mmol) was added followed by
tBuOOH (15.6 mL of a 5-6M solution). The resulting deep red
solution was stirred for 3 hours at room temperature. The reaction
was quenched with sat. sodium sulfite, washed with MTBE, and dried
with MgSO.sub.4. Crude was purified by flash column chromatography
using hexanes:EtOAc (80/20) to give 14 g of compound 11.
[0488] Step 12.
[0489] Compound 12.
[0490] Compound 11 (1 g, 2.3 mmol) was dissolved in 25 mL of ethyl
acetate, followed by the addition of Pd/C.sub.5%/wt (160 mg). The
solution was then flushed with hydrogen and stirred for 2 hours
under hydrogen at room temperature. The reaction mixture was then
filtered through celite and the organics concentrated to obtain
0.82 g of compound 12. The crude material was used without further
purification.
[0491] Step 13
[0492] Compound 13
[0493] Compound 8 (1.02 g, 2.7 mmol) was dissolved into 10 mL of
dry THF, DEDBT (1.02 g, 3.4 mmol), diisopropylethylamine (0.48 mL,
3.4 mmol) were added at room temperature. The reaction mixture was
stirred 15 minutes and a solution of compound 12 (0.82 g, 2.2 mmol)
in 10 ml of dry THF was added. The reaction mixture was stirred at
room temperature overnight. The reaction was concentrated under
vacuum and the crude material purified by flash column
chromatography using Hexane/MTBE/EtOH (80/19/1) to give 0.69 g of
compound 13.
[0494] Step 14.
[0495] Compound 14.
[0496] DMSO (0.084 mL, 1.1 mmol) was dissolved in 5 mL of dry
CH.sub.2Cl.sub.2 and the solution was cooled to -78.degree. C. A 2M
solution of (COCl).sub.2 in methylene chloride (0.4 mL, 0.68 mmol)
was then added dropwise, followed by the addition of compound 13
(0.34 , 0.52 mmol) in 5 mL of CH.sub.2Cl.sub.2. The reaction
mixture was stirred at -78 C for 45 minutes and triethylamine (0.36
ml, 2.3 mmol) was added dropwise and then reaction allowed to warn
to 0 C. The reaction was quenched with a saturated aqueous solution
of ammonium chloride and extracted with MTBE. The combined organic
layers were washed with 1N HCl, water and with a saturated aqueous
solution of sodium bicarbonate. The crude material was purified by
flash column chromatography using Hexane:EtOAc, (80/20) to afford
0.2 g of compound 14.
[0497] Step 15.
[0498] Compound 15.
[0499] Compound 14 (0.25 g, 0.38 mmol) was dissolved in THF then 3
ml of HF:pyridine complex were added at room temperature. After 105
minutes the reaction mixture was poured into a saturated solution
of sodium bicarbonate. The resulting solution was extracted several
times with MTBE. Combined organic layers were washed once with 1N
HCl, twice with water and once with a saturated solution of sodium
bicarbonate. The crude material was purified by flash column
chromatography using CH.sub.2Cl.sub.2/EtOH (95/5) to give 0.156 g
of compound 15 (ER-804191).
[0500] 2) Exemplary Compounds:
TABLE-US-00001 ER-# Structure ER-804191 ##STR00164## A ER-803894
##STR00165## A ER-804146 ##STR00166## A ER-804384 ##STR00167## A
ER-804385 ##STR00168## A ER-804445 ##STR00169## B ER-804486
##STR00170## B ER-805319 ##STR00171## A ER-805320 ##STR00172## A
ER-805321 ##STR00173## A ER-805322 ##STR00174## A ER-805323
##STR00175## A ER-805516 ##STR00176## A ER-805555 ##STR00177## A
ER-805556 ##STR00178## A ER-805557 ##STR00179## A ER-805558
##STR00180## A ER-805591 ##STR00181## B ER-805592 ##STR00182## B
ER-805617 ##STR00183## A ER-805642 ##STR00184## A ER-805643
##STR00185## A ER-805644 ##STR00186## A ER-805646 ##STR00187## A
ER-805647 ##STR00188## A ER-805648 ##STR00189## A ER-805704
##STR00190## A ER-805705 ##STR00191## A ER-805706 ##STR00192## A
ER-805707 ##STR00193## A ER-805708 ##STR00194## A ER-805714
##STR00195## A ER-805716 ##STR00196## C ER-805741 ##STR00197## A
ER-805742 ##STR00198## A ER-805743 ##STR00199## A ER-805744
##STR00200## A ER-805746 ##STR00201## A ER-805747 ##STR00202## A
ER-805748 ##STR00203## A ER-805749 ##STR00204## A ER-805750
##STR00205## A ER-805751 ##STR00206## A ER-805752 ##STR00207## A
ER-805753 ##STR00208## A ER-805960 ##STR00209## A ER-805961
##STR00210## C ER-805978 ##STR00211## C ER.about.805979
##STR00212## A ER-805981 ##STR00213## A ER-805982 ##STR00214## A
ER-806017 ##STR00215## B ER-806018 ##STR00216## B ER-806082
##STR00217## A ER-806121 ##STR00218## C ER-806156 ##STR00219## C
ER-806334 ##STR00220## A ER-806335 ##STR00221## A ER-806337
##STR00222## A ER-806338 ##STR00223## A ER-806339 ##STR00224## A
ER-806408 ##STR00225## A ER-806561 ##STR00226## B ER-806562
##STR00227## B ER-806564 ##STR00228## D ER-806565 ##STR00229## D
ER-806566 ##STR00230## D ER-806567 ##STR00231## D ER-806581
##STR00232## A ER-806612 ##STR00233## A ER-806613 ##STR00234## A
ER-806614 ##STR00235## A ER-806615 ##STR00236## A ER-805616
##STR00237## A ER-806622 ##STR00238## A ER-806627 ##STR00239## A
ER-806631 ##STR00240## A ER-806642 ##STR00241## B ER-806649
##STR00242## B ER-806650 ##STR00243## A ER-806664 ##STR00244## A
ER-806665 ##STR00245## A ER-806696 ##STR00246## B ER-806697
##STR00247## B ER-806698 ##STR00248## B ER-806699 ##STR00249## A
ER-806700 ##STR00250## A ER-806701 ##STR00251## A ER-806709
##STR00252## A ER-806750 ##STR00253## A ER-806751 ##STR00254## A
ER-806771 ##STR00255## A ER-806779 ##STR00256## A ER-806780
##STR00257## A ER-806783 ##STR00258## A ER-806784 ##STR00259## A
ER-806785 ##STR00260## A ER-806786 ##STR00261## A ER-806787
##STR00262## A ER-806788 ##STR00263## A ER-806789 ##STR00264## A
ER-806805 ##STR00265## A ER-806807 ##STR00266## A ER-806808
##STR00267## A ER-806811 ##STR00268## A ER-806812 ##STR00269## A
ER-806813 ##STR00270## A ER-806814 ##STR00271## A ER-806815
##STR00272## A ER-806816 ##STR00273## A ER-806817 ##STR00274## A
ER-806845 ##STR00275## A ER-806846 ##STR00276## A ER-806857
##STR00277## A ER-806859 ##STR00278## A ER-806872 ##STR00279## B
ER-806873 ##STR00280## B ER-806883 ##STR00281## B ER-806884
##STR00282## B ER-806910 ##STR00283## A ER-806911 ##STR00284## A
ER-806913 ##STR00285## A ER-806914 ##STR00286## A ER-806915
##STR00287## A
ER-806916 ##STR00288## A ER-806917 ##STR00289## A ER-806918
##STR00290## A ER-806919 ##STR00291## A ER-806988 ##STR00292## A
ER-806989 ##STR00293## A ER-806990 ##STR00294## A ER-806991
##STR00295## A ER-806994 ##STR00296## A ER-806995 ##STR00297## A
ER-806996 ##STR00298## A ER-806997 ##STR00299## D ER-806998
##STR00300## D ER-807005 ##STR00301## A ER-806706 ##STR00302## A
ER-807007 ##STR00303## A ER-807009 ##STR00304## A ER-807042
##STR00305## A ER-807043 ##STR00306## A ER-807048 ##STR00307## A
ER-807049 ##STR00308## A ER-807050 ##STR00309## A ER-807051
##STR00310## A ER-807052 ##STR00311## B ER-807053 ##STR00312## B
ER-807054 ##STR00313## B ER-807055 ##STR00314## B ER-807056
##STR00315## B ER-807057 ##STR00316## B ER-807058 ##STR00317## B
ER-807059 ##STR00318## B ER-807060 ##STR00319## B ER-807061
##STR00320## B ER-807062 ##STR00321## B ER-807063 ##STR00322## A
ER-807064 ##STR00323## A ER-807065 ##STR00324## B ER-807066
##STR00325## B ER-807067 ##STR00326## B ER-807068 ##STR00327## B
ER-807069 ##STR00328## B ER-807070 ##STR00329## B ER-807071
##STR00330## B ER-807072 ##STR00331## A ER-807073 ##STR00332## B
ER-807074 ##STR00333## B ER-807075 ##STR00334## B ER-807076
##STR00335## B ER-807105 ##STR00336## A ER-807106 ##STR00337## A
ER-807107 ##STR00338## A ER-807109 ##STR00339## B ER-807110
##STR00340## B ER-807111 ##STR00341## B ER-807112 ##STR00342## B
ER-807117 ##STR00343## A ER-807118 ##STR00344## A ER-807119
##STR00345## A ER-807120 ##STR00346## A ER-807121 ##STR00347## A
ER-807122 ##STR00348## A ER-807124 ##STR00349## A ER-807125
##STR00350## A ER-807126 ##STR00351## A ER-807130 ##STR00352## A
ER-807131 ##STR00353## A ER-807136 ##STR00354## A ER-807137
##STR00355## A ER-807149 ##STR00356## A ER-807150 ##STR00357## A
ER-807151 ##STR00358## A ER-807152 ##STR00359## A ER-807153
##STR00360## A ER-807154 ##STR00361## A ER-807155 ##STR00362## A
ER-807156 ##STR00363## A ER-807159 ##STR00364## A ER-807162
##STR00365## A ER-807163 ##STR00366## A ER-807165 ##STR00367## A
ER-807167 ##STR00368## A ER-807181 ##STR00369## A ER-807189
##STR00370## A ER-807190 ##STR00371## A ER-807191 ##STR00372## A
ER-807207 ##STR00373## A ER-807208 ##STR00374## A ER-807220
##STR00375## A ER-807221 ##STR00376## A ER-807223 ##STR00377## A
ER-807224 ##STR00378## A ER-807241 ##STR00379## A ER-807268
##STR00380## A ER-807325 ##STR00381## A ER-807359 ##STR00382## A
ER-807395 ##STR00383## A ER-807396 ##STR00384## A ER-807415
##STR00385## A ER-807416 ##STR00386## A ER-807455 ##STR00387## A
ER-807456 ##STR00388## A ER-807446 ##STR00389## A ##STR00390##
##STR00391## ##STR00392## ##STR00393## ##STR00394## ER-807359
##STR00395## ER-807395 ##STR00396## ER-807396 ##STR00397##
ER-807415 ##STR00398## ER-807416 ##STR00399## ER-807455
##STR00400## ER-807456 ##STR00401## ER-807446 ##STR00402##
##STR00403## ##STR00404## ##STR00405## ##STR00406## ##STR00407##
##STR00408## ##STR00409## ##STR00410## ##STR00411##
##STR00412##
##STR00413## ##STR00414## ##STR00415## ##STR00416## ##STR00417##
##STR00418## ##STR00419## ##STR00420## ##STR00421## ##STR00422##
##STR00423## ##STR00424## ##STR00425## ##STR00426## ##STR00427##
##STR00428## ##STR00429## ##STR00430## ##STR00431## ##STR00432##
##STR00433## ##STR00434## ##STR00435## ##STR00436## ##STR00437##
##STR00438## ##STR00439## ##STR00440## ##STR00441## ##STR00442##
##STR00443## ##STR00444## ##STR00445## ##STR00446## ##STR00447##
##STR00448## ##STR00449## ##STR00450## ##STR00451## ##STR00452##
##STR00453## ##STR00454## ##STR00455## ##STR00456## ##STR00457##
##STR00458## ##STR00459## ##STR00460## ##STR00461## ##STR00462##
##STR00463## ##STR00464## ##STR00465## ##STR00466## ##STR00467##
##STR00468## ##STR00469## ##STR00470## ##STR00471## ##STR00472##
##STR00473## ##STR00474## ##STR00475## ##STR00476## ##STR00477##
##STR00478## ##STR00479## ##STR00480## ##STR00481##
##STR00482##
[0501] 3) Biological Assays.
[0502] A) Growth Inhibition of Cultured Cancer Cell Lines
[0503] HT-29 human colon adenocarcinoma cells were grown in McCoy's
5A modified medium (GIBCO) supplemented with 10% fetal bovine serum
(FBS) and antibiotics. Cells were seeded at 7.5.times.10.sup.3
cells/well into 96-well plates, in 100 .mu.l medium. After 3-4
hours at 37.degree. C., 2.times.-concentrated compounds were added,
and cells were incubated Pt 37.degree. C. One plate was harvested
after a 3-day incubation; the other after 4 days. At harvest, media
were aspirated, and 100 .mu.l methylene blue (5 mg/ml in 50%
ethanol) added to each well. Incubation proceeded at room
temperature for 30 minutes, after which the methylene blue is
removed, and the wells were washed by immersion in water four
times. After plates were air-dried, 100 .mu.l of sarcosine (10
mg/ml in PBS) was added to each well, and plates were agitated for
1-2 hr at room temperature. Absorbance was then measured on a
Titertek Multiscan plate-reader at 600 and 405 nm. Corrected data
were graphed and compound IC.sub.50 values calculated.
[0504] This procedure has also been extended to NIH:OVCAR-3 human
ovarian adenocarcinoma cells, obtained from ATCC, and grown in
RPMI-1640 supplemented with 20% FBS, 10 mM HEPES, 1 mM sodium
pyruvate, and 10 .mu.g/ml bovine insulin and antibiotics. The assay
was modified by starting with 2.25.times.10.sup.4 cells/well, since
these cells grow less rapidly than the HT-29s.
[0505] MDA-MB-435 human breast carcinoma cells were also tested in
an analogous assay. Cells were grown in DMEM medium with HEPES,
supplemented with 10% FBS and antibiotics, and plated at
7.5.times.10.sup.3 cells/well.
[0506] B) Cytotoxicity of Quiescent IMR-90 Human Fibroblasts:
[0507] As an indicator of undesirable cytotoxicity towards
non-cancerous cells, compounds were tested on quiescent IMR-90
normal human fibroblasts. These cells were deprived of serum to
halt replication, then treated with test compound for 24 hours.
ATP, a marker of cell viability, was then measured in a
luciferase-based luminescence assay.
[0508] Methods. IMR-90 cells were maintained in MEM medium
supplemented with non-essential amino acids, 1 mM sodium pyruvate,
glutamine, and 10% FBS, and antibiotics. At the start of the assay,
96-well plates were seeded with 8.times.10.sup.3 cells/well in 200
.mu.l medium, and incubated at 37.degree. C. Four days later, cells
were washed and medium was replaced with a low serum (0.1% FBS)
version of the same medium. Three days later test compound was
added, and incubation continues for another 24 hours. Finally, ATP
concentration was measured following lysis of cells and addition of
luminescent substrate (ATPLite kit, Packard).
[0509] C) Inhibition of Proteasome Enzymatic Activity
[0510] Assays have been established to measure peptide hydrolyzing
enzymatic activities of mammalian proteasomes. These assays were
originally adapted from published work (Dubiel et al., J. Biol.
Chem. 1992; 267:22369; Gardner et al., Biochem. J. 2000; 346: 447;
Hough et al., J. Biol. Chem. 1987; 262:8303; Ma et al., J. Biol.
Chem. 1992; 267:10515; Reidinger et al., J. Biol. Chem. 1997; 227:
24899; Riveft et al., Met. Enzmmol. 1994; 244:331; Stein et al.,
Biochemistry. 1996; 35: 3899).
[0511] Methods: Purified 20S proteasomes. Chymotrypsin-like
proteasome activity employs commercially available 20S proteasomes
purified from rabbit skeletal muscle (Calbiochem). The assay relies
on the proteasome's intrinsic ability to cleave a conjugated
fluorogenic peptide substrate (Suc-Leu-Leu-Val-Tyr-AMC;
Calbiochem), releasing the fluorescent product AMC
(amidomethylcoumarin). Inhibitor dilutions were first prepared at
4.times. concentrations in pre-warmed buffer (50 mM Hepes/KOH, 0.5
mM EDTA, pH 7.5; just prior to use, SDS was added to a final
concentration of 0.035%, and DTT was added to 5 mM), and then 20S
proteasomes were added, also at 4.times. final concentration, in a
total volume of 50 .mu.l per well. Inhibitor, buffer, and 20S
proteasomes were pre-incubated together for 15 minutes at
37.degree. C. A 4.times. solution of substrate was also made with
pre-warned buffer, and 25 .mu.l/well of this substrate was added
after the 15 minute incubation. Final reaction volume was 100
.mu.l. The final reaction concentration of proteasomes was 0.5 nM;
substrate was 50 .mu.M. The plate was then placed in the
fluorimeter, which was pre-equilibrated to 37.degree. C. After 5
minutes, readings were collected every 5 minutes for an hour, using
the excitation wavelength of 355 nm and emission wavelength of 460
nm, and kinetic curves were constructed.
[0512] Methods: proteasomes in human leukocyte lysates. The
peptidyl-glutamyl peptide hydrolyzing (PGPH) activity of
proteasomes was also measured. The method reported by Adams et al.
(Cancer Res. 1999; 59:2615) was utilized which relies upon simple
cell lysates from human buffy coat leukocytes purified from whole
blood. Briefly, washed cell pellets were aliquoted, then lysed with
5 mM EDTA, pH 8.0, frozen on ice, thawed, and centrifuged at
10,000.times.g. Resulting supernatants were stored at -80.degree..
Titration experiments suggested that the lysate prepared from
7.times.10.sup.4 leukocytes gave good activity. Substrate was again
a conjugated fluorogenic peptide substrate (Z-Leu-Leu-Glu-AMC;
Calbiochem), which also releases the fluorescent product AMC
(amidomethylcoumarin) upon hydrolysis.
[0513] Methods: proteasomes in intact HT-29 colon carcinoma cells.
This assay tests for inhibition of proteasome enzymatic activities
in whole HT-29 cells, and uses a cell-permeable peptide substrate
coupled to a fluorophore. Specifically, HT-29 cells were plated and
grown to confluence overnight in a black 96-well plate with clear
bottom. The following morning, media were replaced with 50 mM
TRIS-HCl (pH 7.9) with or without proteasome inhibitors. Cells were
treated with the inhibitors for 15 min at 37.degree. C. The
lipophilic fluorogenic substrate, MeOSuc-Phe-Leu-Phe-AFC (Enzyme
Systems Products, Livermore Calif.), was added to a final
concentration of 100 .mu.M. Substrate hydrolysis was followed in a
fluorimeter for 1 hr, detecting the product
7-amino-4-trifluoromethyl coumarin via its emission at 505 nm after
excitation at 400 nm.
[0514] D) In Vivo Efficacy
[0515] To assess compound efficacy in vivo, MDA-MB-435, a human
breast carcinoma, was grown subcutaneously in nude mice. In this
model, compound was typically administered intravenously (i.p.), on
a Mon-Weds-Fri schedule, for four weeks, beginning 10-13 days after
implantation of 1.times.10.sup.6 cells. Mice were weighed once
weekly, and their tumors were measured by electronic calipers
(BioMedic), with volumes calculated according to the formula:
V=0.5.times.1.33.pi.r.sup.3, which assumes that xenografts grow as
semi-spheres. Exemplary data for ER-805751 and ER-805616 in this
model is shown in FIG. 1.
Sequence CWU 1
1
114PRTArtificialSynthetic fluorogenic peptide substrate 1Leu Leu
Val Tyr1
* * * * *
References