U.S. patent application number 15/064198 was filed with the patent office on 2016-11-03 for halofuginone analogs for inhibition of trna synthetases and uses thereof.
This patent application is currently assigned to President and Fellows of Harvard College. The applicant listed for this patent is Children's Medical Center Corporation, President and Fellows of Harvard College. Invention is credited to Tracy Keller, Jinbo Lee, Ralph Mazitschek, Anjana Rao, Mark S. Sundrud, Malcolm Whitman.
Application Number | 20160317498 15/064198 |
Document ID | / |
Family ID | 41669525 |
Filed Date | 2016-11-03 |
United States Patent
Application |
20160317498 |
Kind Code |
A1 |
Keller; Tracy ; et
al. |
November 3, 2016 |
HALOFUGINONE ANALOGS FOR INHIBITION OF TRNA SYNTHETASES AND USES
THEREOF
Abstract
The present invention provides novel analogs and derivatives of
halofuginone. The invention also provides pharmaceutical and
cosmetic compositions thereof and methods for using halofuginone
analogs in treating chronic inflammatory diseases, autoimmune
diseases, dry eye syndrome, fibrosis, scar formation, angiogenesis,
viral infections, ischemic damage, transplant and implant
rejection, neurodegenerative diseases, and cosmetic
applications.
Inventors: |
Keller; Tracy; (Jamaica
Plain, MA) ; Mazitschek; Ralph; (Belmont, MA)
; Whitman; Malcolm; (Jamaica Plain, MA) ; Lee;
Jinbo; (Andover, MA) ; Sundrud; Mark S.;
(Jupiter, FL) ; Rao; Anjana; (La Jolla,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
President and Fellows of Harvard College
Children's Medical Center Corporation |
Cambridge
Boston |
MA
MA |
US
US |
|
|
Assignee: |
President and Fellows of Harvard
College
Cambridge
MA
Children's Medical Center Corporation
Boston
MA
|
Family ID: |
41669525 |
Appl. No.: |
15/064198 |
Filed: |
March 8, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13058486 |
May 27, 2011 |
9284297 |
|
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PCT/US2009/004581 |
Aug 11, 2009 |
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15064198 |
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61153866 |
Feb 19, 2009 |
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61188740 |
Aug 11, 2008 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 29/02 20180101;
C07D 405/14 20130101; A61P 25/00 20180101; A61P 27/02 20180101;
A61K 31/4184 20130101; A61P 29/00 20180101; A61K 31/685 20130101;
C07D 403/06 20130101; A61P 3/10 20180101; A61P 37/06 20180101; A61K
31/695 20130101; A61P 43/00 20180101; C07D 491/04 20130101; C07D
401/06 20130101; A61P 9/00 20180101; C07D 401/14 20130101; A61P
33/02 20180101; C07F 9/65583 20130101; A61P 31/00 20180101; A61P
35/00 20180101; A61P 11/06 20180101; C07D 495/04 20130101; A61P
31/12 20180101 |
International
Class: |
A61K 31/4184 20060101
A61K031/4184; A61K 31/685 20060101 A61K031/685; A61K 31/695
20060101 A61K031/695 |
Goverment Interests
GOVERNMENT SUPPORT
[0002] This invention was made with U.S. Government support under
HD029468 and CA078048 awarded by National Institutes of Health. The
U.S. Government has certain rights in the invention.
Claims
1-172. (canceled)
173. A method of inhibiting glutamyl-prolyl tRNA synthetase
comprising contacting glutamyl-prolyl tRNA synthetase with a
compound of Formula (I): ##STR00219## or a pharmaceutically
acceptable salt thereof, or a pharmaceutical composition thereof,
wherein: j is an integer between 0 and 10, inclusive; p is an
integer between 0 and 6, inclusive; q is an integer between 0 and
6, inclusive; m is 1 or 2; v is an integer between 1 and 3,
inclusive; X is N or CR.sub.X, wherein R.sub.X is hydrogen halogen;
substituted or unsubstituted aliphatic substituted or unsubstituted
heteroaliphatic; substituted or unsubstituted aryl; substituted or
unsubstituted heteroaryl; --OR.sub.F; -SR.sub.F;
--N(R.sub.F).sub.2; and --C(R.sub.F).sub.3; wherein each occurrence
of R.sub.F is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; each occurrence of Y is
independently S, O, N, NR.sub.Y, or CR.sub.Y, wherein each
occurrence of R.sub.Y is independently hydrogen; halogen; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
aliphatic; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched heteroaliphatic; substituted or
unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; each occurrence of T and G is
independently --S--, --O--, --NR.sub.E--, or C(R.sub.E).sub.2--,
wherein each occurrence of R.sub.E is independently hydrogen;
halogen; substituted or unsubstituted aliphatic substituted or
unsubstituted heteroaliphatic; substituted or unsubstituted aryl;
substituted or unsubstituted heteroaryl; --OR.sub.G; --SR.sub.G;
--N(R.sub.G).sub.2; and --C(R.sub.G).sub.3; wherein each occurrence
of R.sub.G, is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.1 is hydrogen; a
protecting group; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl;
--C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3 wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; each occurrence of
R.sub.2 is independently hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.B; --C(.dbd.O)R.sub.B;
--CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2; --CN; --SCN;
--SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B; --NO.sub.2;
--N(R.sub.B).sub.2; --NHC(O)R.sub.B; or --C(R.sub.B).sub.3; wherein
each occurrence of R.sub.B is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.3 is hydrogen
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.C;
--C(.dbd.O)R.sub.C; --CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C); --CN;
--SCN; --SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2;
--N(R.sub.C).sub.2; --NHC(O)R.sub.C; or --C(R.sub.C).sub.3; wherein
each occurrence of R.sub.C is independently a hydrogen a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.4 and R.sub.5 are
independently hydrogen halogen; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched aliphatic cyclic or acyclic,
substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl: substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.D; --C(.dbd.O)R.sub.D;
--CO.sub.2R.sub.D; --C(.dbd.O)N(R.sub.D).sub.2; --CN; --SCN;
--SR.sub.D; --SOR.sub.D; --SO.sub.2R.sub.D; --NO.sub.2;
--N(R.sub.D).sub.2; --NHC(O)R.sub.D; or --C(R.sub.D).sub.3; wherein
each occurrence of RR is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; or R.sub.4 and R.sub.5
may optionally be taken together to form .dbd.O, .dbd.S,
.dbd.NR.sub.D, .dbd.N--OR.sub.D, .dbd.N--NHR.sub.D,
.dbd.N--N(R.sub.D).sub.2, .dbd.C(R.sub.D).sub.2; or R.sub.4 and
R.sub.5 may optionally be taken together with the intervening atom
to form a saturated or unsaturated, substituted or unsubstituted
cyclic or heterocyclic structure; and R.sub.6 is hydrogen halogen;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.K;
--C(.dbd.O)R.sub.K; --CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2;
--CN; --SCN; --SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K;
--NO.sub.2; --N(R.sub.K).sub.2; --NHC(O)R.sub.K; or
--C(R.sub.K).sub.3; wherein each occurrence of R.sub.K is
independently a hydrogen a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety, a heteroaryl moiety, alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety.
174. The method of claim 173, wherein the glutamyl-prolyl tRNA
synthetase is contacted in vitro.
175. The method of claim 174, wherein the glutamyl-prolyl tRNA
synthetase is purified.
176. The method of claim 173, wherein the glutamyl-prolyl tRNA
synthetase in contacted in a cell.
177. A method of treating a Th17-mediated condition comprising
administering to a subject a compound of Formula (I): ##STR00220##
or a pharmaceutically acceptable salt thereof, or a pharmaceutical
composition thereof, wherein: j is an integer between 0 and 10,
inclusive; p is an integer between 0 and 6, inclusive; q is an
integer between 0 and 6, inclusive; m is 1 or 2; v is an integer
between 1 and 3, inclusive; X is N or CR.sub.X, wherein R.sub.X is
hydrogen, halogen; substituted or unsubstituted aliphatic;
substituted or unsubstituted heteroaliphatic; substituted or
unsubstituted aryl; substituted or unsubstituted heteroaryl;
--OR.sub.F; --SR.sub.F; --N(R.sub.F).sub.2; and --C(R.sub.F).sub.3;
wherein each occurrence of R.sub.F is independently a hydrogen; a
halogen; a protecting group; an aliphatic moiety; a heteroaliphatic
moiety; an acyl moiety; an aryl moiety; a heteroaryl moiety;
alkoxy; aryloxy; alkylthioxy; arylthioxy; amino; alkylamino;
dialkylamino; heteroaryloxy; or heteroarylthioxy moiety; each
occurrence of Y is independently S, O, N, NR.sub.Y, or CR.sub.Y,
wherein each occurrence of R.sub.Y is independently hydrogen;
halogen, cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic, cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.C is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; each occurrence of T and G is
independently --S--, --O--, --NR.sub.E--, or C(R.sub.E).sub.2--,
wherein each occurrence of R.sub.E is independently hydrogen;
halogen, substituted or unsubstituted aliphatic; substituted or
unsubstituted heteroaliphatic; substituted or unsubstituted aryl;
substituted or unsubstituted heteroaryl; --OR; --SR.sub.G;
--N(R.sub.G).sub.2; and --C(R.sub.G).sub.3; wherein each occurrence
of R.sub.G is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.1 is hydrogen a
protecting group; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl;
--C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3 wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; each occurrence of
R.sub.2 is independently hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.B; --C(.dbd.O)R.sub.B;
--CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2; --CN; --SCN;
--SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B; --NO.sub.2;
--N(R.sub.B).sub.2; --NHC(O)R.sub.B; or --C(RB).sub.2; wherein each
occurrence of R.sub.B is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.3 is hydrogen
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.C;
--C(.dbd.O)R.sub.C; --CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2;
--CN; --SCN; --SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C;
--NO.sub.2; --N(R.sub.C).sub.2; --NHC(O)R.sub.C; or
--C(R.sub.C).sub.3; wherein each occurrence of R.sub.C is
independently a hydrogen a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; R.sub.4 and R.sub.5 are independently
hydrogen halogen; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.D;
--C(.dbd.O)R.sub.D; --CO.sub.2R.sub.D; --C(.dbd.O)N(R.sub.D).sub.2;
--CN; --SCN; --SR.sub.D; --SOR.sub.D; --SO.sub.2R.sub.D;
--NO.sub.2; --N(R.sub.D).sub.2; --NHC(O)R.sub.D; or
--C(R.sub.D).sub.3; wherein each occurrence of R.sub.D is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety, an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; or R.sub.4 and R.sub.5 may optionally be
taken together to form .dbd.O, .dbd.S, .dbd.NR.sub.D,
.dbd.N--OR.sub.D, .dbd.N--NHR.sub.D, .dbd.N--N(R.sub.D).sub.2,
.dbd.C(R.sub.D).sub.2; or R.sub.4 and R.sub.5 may optionally be
taken together with the intervening atom to form a saturated or
unsaturated, substituted or unsubstituted cyclic or heterocyclic
structure; and R.sub.6 is hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
178. The method of claim 177, wherein the Th17-mediated condition
is selected from the group consisting of autoimmune diseases, dry
eye syndrome, fibrosis, scar formation, angiogenesis, ischemic
damage, inflammatory diseases, and neurodegenerative diseases.
179. The method of claim 177, wherein the Th17-mediated condition
is selected from cellulite or stretch marks.
180. The method of claim 177 further comprising administering a
second agent that inhibits expression or activity of a
proinflammatory cytokine.
181. The method of claim 180, wherein the proinflammatory cytokine
is selected from one or more of TNF.alpha., IFN.gamma., GM-CSF,
MIP-2, IL-12, IL-1.alpha., IL-I .beta., and IL-23.
182-183. (canceled)
184. The method of claim 180, wherein the proinflammatory cytokine
is IL-6 or IL-21.
185-192. (canceled)
193. A method of treating a glutamyl-prolyl tRNA
synthetase-mediated condition comprising administering to a subject
a compound of Formula (I): ##STR00221## or a pharmaceutically
acceptable salt thereof, or a pharmaceutical composition thereof,
wherein: j is an integer between 0 and 10, inclusive; p is an
integer between 0 and 6, inclusive; q is an integer between 0 and
6, inclusive; m is 1 or 2; v is an integer between 1 and 3,
inclusive; X is N or CR.sub.X, wherein R.sub.X is hydrogen,
halogen; substituted or unsubstituted aliphatic; substituted or
unsubstituted heteroaliphatic; substituted or unsubstituted aryl;
substituted or unsubstituted heteroaryl; --OR.sub.F; --SR.sub.F;
--N(R.sub.F).sub.2; and --C(R.sub.F).sub.3; wherein each occurrence
of R.sub.F is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; each occurrence of Y is
independently S, O, N, NR.sub.Y, or CR.sub.Y, wherein each
occurrence of R is independently hydrogen halogen; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
aliphatic cyclic or acyclic, substituted or unsubstituted, branched
or unbranched heteroaliphatic; substituted or unsubstituted,
branched or unbranched acyl; substituted or unsubstituted, branched
or unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.G; --C(.dbd.O)R.sub.G;
--CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2; --CN; --SCN;
--SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G; --NO.sub.2;
--N(R.sub.G).sub.2; --NHC(O)R.sub.G; or --C(R.sub.G).sub.3; wherein
each occurrence of R.sub.G is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; each occurrence of T and
G is independently --S--, --O--, --NR.sub.E--, or
C(R.sub.E).sub.2--, wherein each occurrence of R.sub.E is
independently hydrogen; halogen; substituted or unsubstituted
aliphatic substituted or unsubstituted heteroaliphatic; substituted
or unsubstituted aryl; substituted or unsubstituted heteroaryl;
--OR.sub.G; --SR.sub.G; --N(R.sub.G).sub.2; and --C(R.sub.G).sub.3;
wherein each occurrence of R.sub.G is independently a hydrogen; a
halogen; a protecting group; an aliphatic moiety; a heteroaliphatic
moiety; an acyl moiety; an aryl moiety; a heteroaryl moiety;
alkoxy; aryloxy; alkylthioxy; arylthioxy; amino; alkylamino;
dialkylamino; heteroaryloxy; or heteroarylthioxy moiety; R.sup.1 is
hydrogen a protecting group; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched aliphatic cyclic or acyclic,
substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; each occurrence of
R.sub.2 is independently hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.B; --C(.dbd.O)R.sub.B;
--CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2; --CN; --SCN;
--SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B; --NO.sub.2;
--N(R.sub.B).sub.2; --NHC(O)R.sub.B; or --C(R.sub.B).sub.3; wherein
each occurrence of R.sub.B is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.3 is hydrogen
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.C;
--C(.dbd.O)R.sub.C; --CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.B).sub.2;
--CN; --SCN; --SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C;
--NO.sub.2; --N(R.sub.C).sub.2; --NHC(O)R.sub.C; or
--C(R.sub.C).sub.3; wherein each occurrence of RP is independently
a hydrogen a halogen; a protecting group; an aliphatic moiety; a
heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; alkoxy; aryloxy; alkylthioxy; arylthioxy; amino;
alkylamino; dialkylamino; heteroaryloxy; or heteroarylthioxy
moiety; R.sub.4 and R.sub.5 are independently hydrogen halogen;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.D;
--C(.dbd.O)R.sub.D; --CO.sub.2R.sub.D; --C(.dbd.O)N(R.sub.D).sub.2;
--CN; --SCN; --SR.sub.D; --SOR.sub.D; --SO.sub.2R.sub.D;
--NO.sub.2; --N(R.sub.D).sub.2; --NHC(O)R.sub.D; or
--C(R.sub.D).sub.3; wherein each occurrence of R.sub.D is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; or R.sub.4 and R.sub.5 may optionally be
taken together to form .dbd.O, --S, .dbd.NR.sub.D,
.dbd.N--OR.sub.D, .dbd.N--NHR.sub.D, .dbd.N--N(R.sub.D).sub.2,
--C(R.sub.D).sub.2; or R.sub.4 and R.sub.5 may optionally be taken
together with the intervening atom to form a saturated or
unsaturated, substituted or unsubstituted cyclic or heterocyclic
structure; and R.sub.6 is hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic,
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN; --SR;
--SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2; --N(R.sub.K).sub.2;
--NHC(O)R.sub.K; or --C(R.sub.K).sub.3, wherein each occurrence of
R.sub.K is independently a hydrogen, a halogen; a protecting group;
an aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an
aryl moiety, a heteroaryl moiety, alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety.
194. The method of claim 193, wherein the glutamyl-prolyl tRNA
synthetase-mediated condition involves collagen synthesis.
195. A method of treating fibrosis comprising administering to a
subject a compound of Formula (I): ##STR00222## or a
pharmaceutically acceptable salt thereof, or a pharmaceutical
composition thereof, wherein: j is an integer between 0 and 10,
inclusive; p is an integer between 0 and 6, inclusive; q is an
integer between 0 and 6, inclusive; m is 1 or 2; v is an integer
between 1 and 3, inclusive; X is N or CR.sub.X, wherein R.sub.X is
hydrogen halogen; substituted or unsubstituted aliphatic
substituted or unsubstituted heteroaliphatic; substituted or
unsubstituted aryl; substituted or unsubstituted heteroaryl;
--OR.sub.F; --SR.sub.F; --N(R.sub.F).sub.2; and --C(R.sub.F).sub.3;
wherein each occurrence of R.sub.F is independently a hydrogen; a
halogen; a protecting group; an aliphatic moiety; a heteroaliphatic
moiety; an acyl moiety; an aryl moiety; a heteroaryl moiety;
alkoxy; aryloxy; alkylthioxy; arylthioxy; amino; alkylamino;
dialkylamino; heteroaryloxy; or heteroarylthioxy moiety; each
occurrence of Y is independently S, O, N, NR.sub.Y, or CR.sub.Y,
wherein each occurrence of R.sub.Y is independently hydrogen
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; each occurrence of T and G is
independently --S--, --O--, --NR.sub.E--, or C(R.sub.E).sub.2--,
wherein each occurrence of R.sub.E is independently hydrogen;
halogen; substituted or unsubstituted aliphatic substituted or
unsubstituted heteroaliphatic; substituted or unsubstituted aryl;
substituted or unsubstituted heteroaryl; --OR.sub.G; --SR.sub.G;
--N(R.sub.G).sub.2; and --C(R.sub.G).sub.3; wherein each occurrence
of R.sub.G is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.1 is hydrogen a
protecting group; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl;
--C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; each occurrence of R, is
independently hydrogen halogen; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched aliphatic cyclic or acyclic,
substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.B; --C(.dbd.O)R.sub.B;
--CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2; --CN; --SCN;
--SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B; --NO.sub.2;
--N(R.sub.B).sub.2; --NHC(O)R.sub.B; or --C(R.sub.B).sub.3; wherein
each occurrence of R.sub.B is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.3 is hydrogen
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.C;
--C(.dbd.O)R.sub.C; --CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2;
--CN; --SCN; --SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C;
--NO.sub.2; --N(R.sub.C).sub.2; --NHC(O)R.sub.C; or
--C(R.sub.C).sub.3; wherein each occurrence of R.sub.C is
independently a hydrogen a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; R.sub.4 and R.sub.5 are independently
hydrogen halogen; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.D;
--C(.dbd.O)R.sub.D; --CO.sub.2R.sub.D; --C(.dbd.O)N(R.sub.D).sub.2;
--CN; --SCN; --SR.sub.D; --SOR.sub.D; --SO.sub.2R.sub.D;
--NO.sub.2; --N(R.sub.D).sub.2; --NHC(O)R.sub.D; or
--C(R.sub.D).sub.3; wherein each occurrence of R.sub.D is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; or R.sub.4 and R.sub.5 may optionally be
taken together to form .dbd.O, .dbd.S, .dbd.NR.sub.D,
.dbd.N--OR.sub.D, .dbd.N--NHR.sub.D, .dbd.N--N(R.sub.n).sub.2,
.dbd.C(R.sub.D).sub.2; or R.sub.4 and R.sub.5 may optionally be
taken together with the intervening atom to form a saturated or
unsaturated, substituted or unsubstituted cyclic or heterocyclic
structure; and R.sub.6 is hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
196. A method of protecting a donor organ for transplantation,
comprising contacting said organ with a compound of Formula (I):
##STR00223## or a pharmaceutically acceptable salt thereof, or a
pharmaceutical composition thereof, wherein: j is an integer
between 0 and 10, inclusive; p is an integer between 0 and 6,
inclusive; q is an integer between 0 and 6, inclusive; m is 1 or 2;
v is an integer between 1 and 3, inclusive; X is N or CR.sub.X,
wherein R.sub.X is hydrogen, halogen; substituted or unsubstituted
aliphatic; substituted or unsubstituted heteroaliphatic;
substituted or unsubstituted aryl; substituted or unsubstituted
heteroaryl; --OR.sub.F; --SR.sub.F; --N(R.sub.F).sub.2; and
--C(R.sub.F).sub.3; wherein each occurrence of R.sub.F is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; each occurrence of Y is independently S,
O, N, NR.sub.Y, or CR.sub.Y, wherein each occurrence of R.sub.Y is
independently hydrogen; halogen, cyclic or acyclic, substituted or
unsubstituted, branched or unbranched aliphatic, cyclic or acyclic,
substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.G; --C(.dbd.O)R.sub.G;
--CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2; --CN; --SCN;
--SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G; --NO.sub.2;
--N(R.sub.G).sub.2; --NHC(O)R.sub.G; or --C(R.sub.G).sub.3; wherein
each occurrence of R.sub.G is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; each occurrence of T and
G is independently --S--, --O--, --NR.sub.E--, or
C(R.sub.E).sub.2--, wherein each occurrence of R.sub.E is
independently hydrogen; halogen, substituted or unsubstituted
aliphatic; substituted or unsubstituted heteroaliphatic;
substituted or unsubstituted aryl; substituted or unsubstituted
heteroaryl; --OR.sub.G; --SR.sub.G; --N(R.sub.G).sub.2; and
--C(R.sub.G).sub.3, wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; R.sub.1 is hydrogen; a protecting group;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched aliphatic, cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl;
--C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3 wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; each occurrence of
R.sub.2 is independently hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.B; --C(.dbd.O)R.sub.B;
--CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2; --CN; --SCN;
--SR.sub.B; --SOR.sub.B; --SOR.sub.2R.sub.B; --NO.sub.2;
--N(R.sub.B).sub.2; --NHC(O)R.sub.B; or --C(R.sub.B).sub.3; wherein
each occurrence of R.sub.B is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.3 is hydrogen
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.C;
--C(.dbd.O)R.sub.C; --CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2;
--CN; --SCN; --SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C;
--NO.sub.2; --N(R.sub.C).sub.2; --NHC(O)R.sub.C; or
--C(R.sub.C).sub.3; wherein each occurrence of R.sub.C is
independently a hydrogen a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; R.sub.4 and R.sub.5 are independently
hydrogen halogen; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.D;
--C(.dbd.O)R.sub.D; --CO.sub.2R.sub.D; --C(.dbd.O)N(R.sub.D).sub.2;
--CN; --SCN; --SR.sub.D; --SOR.sub.D; --SO.sub.2R.sub.D;
--NO.sub.2; --N(R.sub.D).sub.2; --NHC(O)R.sub.D; or
--C(R.sub.D).sub.3); wherein each occurrence of R.sub.D is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; or R.sub.4 and R.sub.5 may optionally be
taken together to form .dbd.O, .dbd.S, .dbd.NR.sub.D,
.dbd.N--OR.sub.D, .dbd.N--NHR.sub.D, .dbd.N--N(R.sub.D).sub.2,
.dbd.C(R.sub.D).sub.2; or R.sub.4 and R.sub.5 may optionally be
taken together with the intervening atom to form a saturated or
unsaturated, substituted or unsubstituted cyclic or heterocyclic
structure; and R.sub.6 is hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R_); --CN; --SCN; --SR.sub.K;
--SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2; --N(R.sub.K).sub.2;
--NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein each occurrence of
R.sub.K is independently a hydrogen a halogen; a protecting group;
an aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an
aryl moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety.
197. A method of treating an AAR-mediated condition comprising
administering to a subject a compound of Formula (I): ##STR00224##
or a pharmaceutically acceptable salt thereof, or a pharmaceutical
composition thereof, wherein: j is an integer between 0 and 10,
inclusive; p is an integer between 0 and 6, inclusive; q is an
integer between 0 and 6, inclusive; m is 1 or 2; v is an integer
between 1 and 3, inclusive; X is N or CR.sub.X, wherein R.sub.X is
hydrogen halogen; substituted or unsubstituted aliphatic
substituted or unsubstituted heteroaliphatic; substituted or
unsubstituted aryl; substituted or unsubstituted heteroaryl;
--OR.sub.F; --SR.sub.F; --N(R.sub.F).sub.2; and --C(R.sub.F).sub.3;
wherein each occurrence of R.sub.F is independently a hydrogen; a
halogen; a protecting group; an aliphatic moiety; a heteroaliphatic
moiety; an acyl moiety; an aryl moiety; a heteroaryl moiety;
alkoxy; aryloxy; alkylthioxy; arylthioxy; amino; alkylamino;
dialkylamino; heteroaryloxy; or heteroarylthioxy moiety; each
occurrence of Y is independently S, O, N, NR.sub.Y, or CR.sub.Y,
wherein each occurrence of R.sub.Y is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; each occurrence of T and G is
independently --S--, --O--, --NR.sub.E--, or C(R.sub.E).sub.2--,
wherein each occurrence of R.sub.E is independently hydrogen;
halogen; substituted or unsubstituted aliphatic substituted or
unsubstituted heteroaliphatic; substituted or unsubstituted aryl;
substituted or unsubstituted heteroaryl; --OR.sub.G; --SR.sub.G;
--N(R.sub.G).sub.2; and --C(R.sub.G).sub.3; wherein each occurrence
of R.sub.G is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.1 is hydrogen; a
protecting group; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl;
--C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3 wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; each occurrence of
R.sub.2 is independently hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.B; --C(.dbd.O)R.sub.B;
--CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2; --CN; --SCN;
--SR.sub.B; --SOR.sub.B; --SOR.sub.2R.sub.B; --NO.sub.2;
--N(R.sub.B).sub.2; --NHC(O)R.sub.B; or --C(R.sub.B).sub.3; wherein
each occurrence of R.sub.B is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.3 is hydrogen
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.C;
--C(.dbd.O)R.sub.C; --CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2;
--CN; --SCN; --SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C;
--NO.sub.2; --N(R.sub.C).sub.2; --NHC(O)R.sub.C; or
--C(R.sub.C).sub.3; wherein each occurrence of R.sub.C is
independently a hydrogen a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; R.sub.4 and R.sub.5 are independently
hydrogen halogen; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.D;
--C(.dbd.O)R.sub.D; --CO.sub.2R.sub.D; --C(.dbd.O)N(R.sub.D).sub.2;
--CN; --SCN; --SR.sub.D; --SOR.sub.D; --SO.sub.2R.sub.D;
--NO.sub.2; --N(R.sub.D).sub.2; --NHC(O)R.sub.D; or
--C(R.sub.D).sub.3); wherein each occurrence of R.sub.D is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; or R.sub.4 and R.sub.5 may optionally be
taken together to form .dbd.O, .dbd.S, .dbd.NR.sub.D,
.dbd.N--OR.sub.D, .dbd.N--NHR.sub.D, .dbd.N--N(R.sub.D).sub.2,
--C(R.sub.D).sub.2; or R.sub.4 and R.sub.5 may optionally be taken
together with the intervening atom to form a saturated or
unsaturated, substituted or unsubstituted cyclic or heterocyclic
structure; and R.sub.6 is hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
198. The method of claim 197, wherein the AAR-mediated condition is
selected from the group consisting of fibrosis, scar formation,
autoimmune disease, dry eye syndrome, graft versus host disease,
angiogenesis, cancer, macular degeneration, choroidal
neovascularization, asthma, chronic inflammation, cardiovascular
disease, diabetes, viral infection, protozol infection, fungal
infection, cellulite, and stretch marks.
199. The method of claim 198, wherein the autoimmune disease is
selected from multiple sclerosis, rheumatoid arthritis, lupus,
psoriasis, scleroderma, or dry eye syndrome.
200. (canceled)
201. A method of treating cellulite comprising administering to a
subject a compound of Formula (I): ##STR00225## or a
pharmaceutically acceptable salt thereof, or a pharmaceutical
composition thereof, wherein: j is an integer between 0 and 10,
inclusive; p is an integer between 0 and 6, inclusive; q is an
integer between 0 and 6, inclusive; m is 1 or 2; v is an integer
between 1 and 3, inclusive; X is N or CR.sub.X, wherein R.sub.X is
hydrogen halogen; substituted or unsubstituted aliphatic
substituted or unsubstituted heteroaliphatic; substituted or
unsubstituted aryl; substituted or unsubstituted heteroaryl;
--OR.sub.F; --SR.sub.F; --N(R.sub.F).sub.2; and --C(R.sub.F).sub.3;
wherein each occurrence of R.sub.F is independently a hydrogen; a
halogen; a protecting group; an aliphatic moiety; a heteroaliphatic
moiety; an acyl moiety; an aryl moiety; a heteroaryl moiety;
alkoxy; aryloxy; alkylthioxy; arylthioxy; amino; alkylamino;
dialkylamino; heteroaryloxy; or heteroarylthioxy moiety; each
occurrence of Y is independently S, O, N, NR.sub.Y, or CR.sub.Y,
wherein each occurrence of R.sub.Y is independently hydrogen
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; each occurrence of T and G is
independently --S--, --O--, --NR.sub.E--, or C(R.sub.E).sub.2--,
wherein each occurrence of R.sub.E is independently hydrogen;
halogen; substituted or unsubstituted aliphatic substituted or
unsubstituted heteroaliphatic; substituted or unsubstituted aryl;
substituted or unsubstituted heteroaryl; --OR.sub.G; --SR.sub.G;
--N(R.sub.G).sub.2; and --C(R.sub.G).sub.3; wherein each occurrence
of R.sub.G is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sup.1 is hydrogen a
protecting group; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl;
--C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; each occurrence of
R.sub.2 is independently hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.B; --C(.dbd.O)R.sub.B;
--CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2; --CN; --SCN;
--SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B; --NO.sub.2;
--N(R.sub.B).sub.2; --NHC(O)R.sub.B; or --C(R.sub.B).sub.3; wherein
each occurrence of R.sub.B is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.3 is hydrogen
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.C;
--C(.dbd.O)R.sub.C; --CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2;
--CN; --SCN; --SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C;
--NO.sub.2; --N(R.sub.C).sub.2; --NHC(O)R.sub.C; or
--C(R.sub.C).sub.3; wherein each occurrence of R.sub.C is
independently a hydrogen a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; R.sub.4 and R.sub.5 are independently
hydrogen halogen; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.D;
--C(.dbd.O)R.sub.D; --CO.sub.2R.sub.D; --C(.dbd.O)N(R.sub.D).sub.2;
--CN; --SCN; --SR.sub.D; --SOR.sub.D; --SO.sub.2R.sub.D;
--NO.sub.2; --N(R.sub.D).sub.2; --NHC(O)R.sub.D; or
--C(R.sub.D).sub.3); wherein each occurrence of R.sub.D is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; or R.sub.4 and R.sub.5 may optionally be
taken together to form .dbd.O, .dbd.S, .dbd.NR.sub.D,
.dbd.N--OR.sub.D, .dbd.N--NHR.sub.D, .dbd.N--N(R.sub.D).sub.2,
.dbd.C(R.sub.D).sub.2; or R.sub.4 and R.sub.5 may optionally be
taken together with the intervening atom to form a saturated or
unsaturated, substituted or unsubstituted cyclic or heterocyclic
structure; and R.sub.6 is hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
202. A method of treating a protozoal disease comprising
administering to a subject a compound of Formula (I): ##STR00226##
or a pharmaceutically acceptable salt thereof, or a pharmaceutical
composition thereof, wherein: j is an integer between 0 and 10,
inclusive; p is an integer between 0 and 6, inclusive; q is an
integer between 0 and 6, inclusive; m is 1 or 2; v is an integer
between 1 and 3, inclusive; X is N or CR.sub.X, wherein R.sub.X is
hydrogen halogen; substituted or unsubstituted aliphatic
substituted or unsubstituted heteroaliphatic; substituted or
unsubstituted aryl; substituted or unsubstituted heteroaryl;
--OR.sub.F; --SR.sub.F; --N(R.sub.F).sub.2; and --C(R.sub.F).sub.3;
wherein each occurrence of R.sub.F is independently a hydrogen; a
halogen; a protecting group; an aliphatic moiety; a heteroaliphatic
moiety; an acyl moiety; an aryl moiety; a heteroaryl moiety;
alkoxy; aryloxy; alkylthioxy; arylthioxy; amino; alkylamino;
dialkylamino; heteroaryloxy; or heteroarylthioxy moiety; each
occurrence of Y is independently S, O, N, NR.sub.Y, or CR.sub.Y,
wherein each occurrence of R.sub.Y is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3, wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; each occurrence of T and G is
independently --S--, --O--, --NR.sub.E--, or C(R.sub.E).sub.2--,
wherein each occurrence of R.sub.E is independently hydrogen;
halogen; substituted or unsubstituted aliphatic substituted or
unsubstituted heteroaliphatic; substituted or unsubstituted aryl;
substituted or unsubstituted heteroaryl; --OR.sub.G; --SR.sub.G;
--N(R.sub.G).sub.2; and --C(R.sub.G).sub.3; wherein each occurrence
of R.sub.G is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.1 is hydrogen; a
protecting group; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl;
--C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A; --C(.dbd.O)N(R.sub.A); or
--C(R.sub.A).sub.3 wherein each occurrence of R.sub.A is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; each occurrence of R.sub.2 is
independently hydrogen halogen; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched aliphatic; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.B; --C(.dbd.O)R.sub.B;
--CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2; --CN; --SCN;
--SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B; --NO.sub.2;
--N(R.sub.B).sub.2; --NHC(O)R.sub.B; or --C(R.sub.B).sub.3; wherein
each occurrence of R.sub.B is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.3 is hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.C;
--C(.dbd.O)R.sub.C; --CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2;
--CN; --SCN; --SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C;
--NO.sub.2; --N(R.sub.C).sub.2; --NHC(O)R.sub.C; or
--C(R.sub.C).sub.3; wherein each occurrence of R.sub.C is
independently a hydrogen a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; R.sub.4 and R.sub.5 are independently
hydrogen halogen; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.D;
--C(.dbd.O)R.sub.D; --CO.sub.2R.sub.D; --C(.dbd.O)N(R.sub.D).sub.2;
--CN; --SCN; --SR.sub.D; --SOR.sub.D; --SO.sub.2R.sub.D;
--NO.sub.2; --N(R.sub.D).sub.2; --NHC(O)R.sub.D; or
--C(R.sub.D).sub.3; wherein each occurrence of R.sub.D is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; or R.sub.4 and R.sub.5 may optionally be
taken together to form .dbd.O, .dbd.S, .dbd.NR.sub.D,
.dbd.N--OR.sub.D, .dbd.N--NHR.sub.D, .dbd.N--N(R.sub.D).sub.2,
--C(R.sub.D).sub.2; or R.sub.4 and R.sub.5 may optionally be taken
together with the intervening atom to form a saturated or
unsaturated, substituted or unsubstituted cyclic or heterocyclic
structure; and R.sub.6 is hydrogen halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
203. The method of claim 178, wherein the disease is a
neurodegenerative disease.
204. The method of claim 203, wherein the disease is multiple
sclerosis.
Description
RELATED APPLICATIONS
[0001] The present application is a divisional of and claims
priority under 35 U.S.C. .sctn.120 to U.S. application, U.S. Ser.
No. 13/058,486, filed May 27, 2011, which is a national stage
filing under 35 U.S.C. .sctn.371 of international PCT application,
PCT/US2009/004581, filed Aug. 11, 2009, which claims priority under
35 U.S.C. .sctn.119(e) to U.S. provisional patent applications,
U.S. Ser. No. 61/188,740, filed Aug. 11, 2008, and U.S. Ser. No.
61/153,866, filed Feb. 19, 2009, each of which is incorporated
herein by reference.
BACKGROUND OF THE INVENTION
[0003] Halofuginone (1) is a halogenated derivative of febrifugine
(3), a natural product extracted from the roots of the hydrangea
Dichroa febrifuga. Dichroa febrifuga is one of the "fifty
fundamental herbs" of traditional Chinese medicine, originally used
as an anti-malarial remedy (Jiang et al., Antimicrob. Agents
Chemother. (2005) 49:1169-1176). Halofuginone, otherwise known as
7-bromo-6-chloro-3-[3-(3-hydroxy-2-piperidinyl)-2-oxopropyl]-4(3H)-quinaz-
olinone, and halofuginone derivatives were first described in U.S.
Pat. No. 2,694,711, incorporated herein by reference. Febrifugine
has been shown to be the active ingredient in Dichroa febrifuga
extracts; halofuginone was originally synthesized in search of less
toxic anti-malarial derivatives of febrifugine. In addition to its
anti-malarial properties, however, halofuginone has striking
anti-fibrotic properities in vivo (Pines, et al., Biol. Blood
Marrow Transplant (2003) 9: 417-425; U.S. Pat. No. 6,028,075,
incorporated herein by reference). Halofuginone shows some toxicity
in humans, such as nausea, vomiting, and fatigue, and possibly
bleeding complications (de Jonge et al., Eur. J. Cancer (2006) 42:
1768-1774).
##STR00001##
[0004] Since halofuginone has shown promising biological
activities, there remains a need for identifying further related
compounds with useful biological activities, especially those that
may be less toxic than halofuginone or febrifugine.
SUMMARY OF THE INVENTION
[0005] The present invention stems from the recognition that
analogs of the quinazolinone alkaloid halofuginone are inhibitors
of metazoan glutamyl-prolyl tRNA synthetase (EPRS), particularly
mammalian EPRS (FIGS. 21 and 22), and/or non-metazoan prolyl tRNA
synthetase, particularly protozoan tRNA synthetase. Inhibitors of
EPRS such as halofuginone and halofuginone analogs can inhibit the
pro-fibrotic behavior of fibroblasts (FIG. 1 and Table 1) and
therefore may be useful in treating disorders associated with
fibrosis. In addition, halofuginone and analogs of halofuginone are
useful in the modification of effector T-cell differentiation (FIG.
2 and Table 2) and therefore may be useful in treating diseases
such as inflammatory diseases and autoimmune diseases. Furthermore,
halofuginone and halofuginone analogs can inhibit angiogenesis and
may be useful in treating tumor neovascularization and ocular
diseases involving choroidal neovascularization and retinal edema.
Halofuginone and halofuginone analogs may also be useful as
antiprotozoal agents given their ability to inhibit protozoal
prolyl tRNA synthetase. The present invention provides novel
classes of quinazolinones, quinolinones, and heteroaryl derivatives
of halofuginone and analogs thereof. The inventive compounds may
have more desirable properties than halofuginone. For example, the
inventive compounds may be less toxic than halofuginone or
febrifugine and/or the inventive compounds may be more potent than
halofuginone or febrifugine.
[0006] In one aspect, the inventive compounds are generally of the
formula:
##STR00002##
wherein:
[0007] j is an integer between 0 and 10, inclusive;
[0008] p is an integer between 0 and 6, inclusive;
[0009] q is an integer between 0 and 6, inclusive;
[0010] m is 1 or 2;
[0011] v is an integer between 1 and 3, inclusive;
[0012] X is N or CR.sub.X, wherein R.sub.X is hydrogen; halogen;
substituted or unsubstituted aliphatic; substituted or
unsubstituted heteroaliphatic; substituted or unsubstituted aryl;
substituted or unsubstituted heteroaryl; --OR.sub.F; --SR.sub.F;
--N(R.sub.F).sub.2; and --C(R.sub.F).sub.3; wherein each occurrence
of R.sub.F is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0013] each occurrence of Y is independently S, O, N, NR.sub.Y, or
CR.sub.Y, wherein each occurrence of R.sub.Y is independently
hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0014] each occurrence of T and G is independently --S--, --O--,
--NR.sub.E--, or C(R.sub.E).sub.2--, wherein R.sub.E is selected
from the group consisting of hydrogen, halogen, substituted or
unsubstituted aliphatic, substituted or unsubstituted
heteroaliphatic, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl, --OR.sub.G, --SR.sub.G,
--N(R.sub.G).sub.2, and --C(R.sub.G).sub.3; wherein each occurrence
of R.sub.G is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0015] R.sub.1 is hydrogen; a protecting group; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0016] each occurrence of R.sub.2 is independently selected from
the group consisting of hydrogen; halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.B; --C(.dbd.O)R.sub.B;
--CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2; --CN; --SCN;
--SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B; --NO.sub.2;
--N(R.sub.B).sub.2; --NHC(O)R.sub.B; or --C(R.sub.B).sub.3; wherein
each occurrence of R.sub.B is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0017] R.sub.3 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.C; --C(.dbd.O)R.sub.C;
--CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2; --CN; --SCN;
--SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2;
--N(R.sub.C).sub.2; --NHC(O)R.sub.C; or --C(R.sub.C).sub.3; wherein
each occurrence of R.sub.C is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0018] R.sub.4 and R.sub.5 are independently hydrogen; halogen;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.D;
--C(.dbd.O)R.sub.D; --CO.sub.2R.sub.D; --C(.dbd.O)N(R.sub.D).sub.2;
--CN; --SCN; --SR.sub.D; --SOR.sub.D; --SO.sub.2R.sub.D;
--NO.sub.2; --N(R.sub.D).sub.2; --NHC(O)R.sub.D; or
--C(R.sub.D).sub.3; wherein each occurrence of R.sub.D is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; or
[0019] R.sub.4 and R.sub.5 may optionally be taken together to form
.dbd.O, .dbd.S, .dbd.NR.sub.D, .dbd.N--OR.sub.D, .dbd.N--NHR.sub.D,
.dbd.N--N(R.sub.D).sub.2, or --C(R.sub.D).sub.2; or
[0020] R.sub.4 and R.sub.5 may optionally be taken together with
the intervening atom to form a saturated or unsaturated,
substituted or unsubstituted cyclic or heterocyclic structure; and
R.sub.6 is independently hydrogen; halogen; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety, or a salt thereof. In
certain embodiments, the inventive compound is of the
stereochemistry shown in formula:
##STR00003##
In other embodiments, the inventive compound is of the
stereochemistry shown in formula:
##STR00004##
Without wishing to be bound by any particular theory, these
compounds are thought to act by binding in the active site of the
tRNA synthetase, thereby inhibiting the incorporation of proline
into tRNA. The invention also provides methods of preparing the
inventive compounds. The inventive compounds may be prepared via a
total synthesis from commercially available starting materials or
may be prepared via a semi-synthetic process starting from a
compound such as halofuginone or febrifugine.
[0021] In another aspect, the present invention provides methods of
treatment comprising administering an inventive compound to a
subject. Without wishing to be bound by a particular theory, the
compounds of the invention are thought to act by inhibiting
glutamyl-prolyl tRNA synthetase (EPRS) or prolyl tRNA synthetase.
See FIGS. 21 and 22. The compounds of the invention or
pharmaceutical compositions thereof may be used to treat any
disease including autoimmune diseases, such as multiple sclerosis,
rheumatoid arthritis, lupus, psoriasis, scleroderma, or dry eye
syndrome, inflammatory diseases, cardiovascular diseases,
neurodegenerative diseases, protein aggregation disorders, and
disorders involving angiogenesis, such as cancer, restenosis,
macular degeneration, and choroidal neovascularization. The
compounds of the invention may also be used to promote wound
healing and/or prevent scarring and may be useful cosmetically,
such as for the treatment of cellulite or stretch marks. Therefore,
the inventive compounds may be used in cosmetic as well as
pharmaceutical treatments. The compounds of the invention may be
used to treat or prevent disease in humans and other animals
including domesticated animals. In certain embodiments, the
compounds of the invention may be used to inhibit pro-fibrotic
behavior in fibroblasts or inhibit the differentiation of Th17
cells. Therefore, the inventive compounds may be useful in
preventing fibrosis. The inventive compounds may also be used as
probes of biological pathways. The inventive compounds may also be
used in studying the differentiation of T cells.
[0022] In some embodiments of the method, a second agent which
inhibits the expression or activity of a proinflammatory cytokine
is administered to the subject. In some embodiments, the
proinflammatory cytokine is selected from one or more of
TNF.alpha., IFN.gamma., GM-CSF, MIP-2, IL-12, IL-1.alpha.,
IL-1.beta., and IL-23. In some embodiments of the method, a second
agent which is an agent that inhibits expression or activity of
IL-6 or IL-21 is administered to the subject. In some embodiments,
a second agent which is an agent that inhibits TNF.alpha. is
administered to the subject. In some embodiments, the agent that
inhibits TNF.alpha. is an anti-TNF.alpha. antibody. In some
embodiments, the agent that inhibits TNF.alpha. is a soluble TNF
receptor. In other embodiments of the method, a second agent which
is an immunomodulatory agent (e.g., steroids, non-steroidal
anti-inflammatory agent, rapamycin, FK506, cyclosporine, HDAC
inhibitors) is administered to the subject.
[0023] In another aspect, the present invention provides
pharmaceutical and cosmetic compositions comprising the inventive
compounds. The compositions may comprise an inventive compound in a
therapeutically effective amount to suppress Th17 differentiation
and/or treat or prevent autoimmune diseases, inflammatory diseases,
cardiovascular diseases, neurodegenerative diseases, protein
aggregation disorders, fibrosis, cellulite, stretch marks, or
disorders involving angiogenesis, such as cancer, restenosis,
macular degeneration, and choroidal neovascularization. The
pharmaceutical compositions may optionally include a
pharmaceutically acceptable excipient. The cosmetic compositions
may optionally include a cosmetically acceptable excipient. In some
embodiments, the pharmaceutical composition further comprises a
second agent that inhibits expression or activity of a
proinflammatory cytokine. In some embodiments, the proinflammatory
cytokine is selected from one or more of IL-6, I-21, TNF.alpha.,
IFN.gamma., GM-CSF, MIP-2, IL-12, IL-1.alpha., IL-.beta., and
IL-23. Any mode of administration including oral, parenteral, and
topical administration of the inventive compound or a
pharmaceutical composition thereof may be used.
[0024] References, scientific articles, patent applications, and
patents cited in this application are incorporated herein by
reference.
DEFINITIONS
[0025] Definitions of specific functional groups and chemical terms
are 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.sup.th 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; Smith and March March's Advanced Organic Chemistry, 5.sup.th
Edition, John Wiley & Sons, Inc., New York, 2001; Larock,
Comprehensive Organic Transformations, VCH Publishers, Inc., New
York, 1989; Carruthers, Some Modern Methods of Organic Synthesis,
3.sup.rd Edition, Cambridge University Press, Cambridge, 1987.
[0026] The compounds of the present invention may exist in
particular geometric or stereoisomeric forms. The present invention
contemplates all such compounds, including cis- and trans-isomers,
R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the
racemic mixtures thereof, and other mixtures thereof, as falling
within the scope of the invention.
[0027] Where an isomer/enantiomer is preferred, it may, in some
embodiments, be provided substantially free of the corresponding
enantiomer, and may also be referred to as "optically enriched."
"Optically enriched," as used herein, means that the compound is
made up of a significantly greater proportion of one enantiomer. In
certain embodiments the compound of the present invention is made
up of at least about 90% by weight of a preferred enantiomer. In
other embodiments the compound is made up of at least about 95%,
98%, or 99% by weight of a preferred enantiomer. Preferred
enantiomers may be isolated from racemic mixtures by any method
known to those skilled in the art, including chiral high pressure
liquid chromatography (HPLC) and the formation and crystallization
of chiral salts or prepared by asymmetric syntheses. See, for
example, Jacques et al., Enantiomers, Racemates and Resolutions
(Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron
33:2725 (1977); Eliel, Stereochemistry of Carbon Compounds
(McGraw-Hill, N Y, 1962); Wilen, Tables of Resolving Agents and
Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ. of Notre Dame
Press, Notre Dame, Ind. 1972).
[0028] It will be appreciated that the compounds of the present
invention, 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 substitution with all
permissible substituents of organic compounds, any of the
substituents described herein (for example, aliphatic, alkyl,
alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl,
acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro,
hydroxyl, thiol, halo, etc.), and any combination thereof (for
example, aliphaticamino, heteroaliphaticamino, alkylamino,
heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl,
aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy,
aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy,
alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy,
acyloxy, and the like) that results in the formation of a stable
moiety. The present invention contemplates any and all such
combinations in order to arrive at a stable substituent/moiety.
Additional examples of generally applicable substitutents are
illustrated by the specific embodiments shown in the Examples,
which are described herein. For purposes of this invention,
heteroatoms such as nitrogen may have hydrogen substituents and/or
any suitable substituent as described herein which satisfy the
valencies of the heteroatoms and results in the formation of a
stable moiety.
[0029] As used herein, substituent names which end in the suffix
"-ene" refer to a biradical derived from the removal of two
hydrogen atoms from the substitutent. Thus, for example, acyl is
acylene; alkyl is alkylene; alkeneyl is alkenylene; alkynyl is
alkynylene; heteroalkyl is heteroalkylene, heteroalkenyl is
heteroalkenylene, heteroalkynyl is heteroalkynylene, aryl is
arylene, and heteroaryl is heteroarylene.
[0030] The term "acyl," as used herein, refers to a group having
the general formula --C(.dbd.O)R.sup.X1, --C(.dbd.O)OR.sup.X1,
--C(.dbd.O)--C(.dbd.O)R.sup.X1, --C(.dbd.O)SR.sup.X1,
--C(.dbd.O)N(R.sup.X1).sub.2, --C(.dbd.S)R.sup.X1,
--C(.dbd.S)N(R.sup.X1).sub.2, and --C(.dbd.S)S(R.sup.X1),
--C(.dbd.NR.sup.X1)R.sup.X1, --C(.dbd.NR.sup.X1)OR.sup.X1,
--C(.dbd.NR.sup.X1)SR.sup.X1, and
--C(.dbd.NR.sup.X1)N(R.sup.X1).sub.2, wherein R.sup.X1 is hydrogen;
halogen; substituted or unsubstituted hydroxyl; substituted or
unsubstituted thiol; substituted or unsubstituted amino;
substituted or unsubstituted acyl, cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched alkyl; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched alkenyl; substituted or
unsubstituted alkynyl; substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, aliphaticoxy,
heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy,
heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy,
heteroalkylthioxy, arylthioxy, heteroarylthioxy, mono- or
di-aliphaticamino, mono- or di-heteroaliphaticamino, mono- or
di-alkylamino, mono- or di-heteroalkylamino, mono- or di-arylamino,
or mono- or di-heteroarylamino; or two R.sup.X1 groups taken
together form a 5- to 6-membered heterocyclic ring. Exemplary acyl
groups include aldehydes (--CHO), carboxylic acids (--CO.sub.2H),
ketones, acyl halides, esters, amides, imines, carbonates,
carbamates, and ureas. Acyl substituents include, but are not
limited to, any of the substituents described herein, that result
in the formation of a stable moiety (e.g., aliphatic, alkyl,
alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl,
acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro,
hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino,
alkylamino, heteroalkylamino, arylamino, heteroarylamino,
alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy,
heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy,
heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy,
heteroarylthioxy, acyloxy, and the like, each of which may or may
not be further substituted).
[0031] The term "acyloxy" refers to a "substituted hydroxyl" of the
formula (--OR.sup.i), wherein R.sup.i is an optionally substituted
acyl group, as defined herein, and the oxygen moiety is directly
attached to the parent molecule.
[0032] The term "aliphatic," as used herein, includes both
saturated and unsaturated, nonaromatic, straight chain (i.e.,
unbranched), branched, acyclic, and cyclic (i.e., carbocyclic)
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, cycloalkyl, cycloalkenyl, and
cycloalkynyl moieties. Thus, as used herein, the term "alkyl"
includes straight, branched and cyclic 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, "aliphatic" is used to indicate those aliphatic groups
(cyclic, acyclic, substituted, unsubstituted, branched or
unbranched) having 1-20 carbon atoms. Aliphatic group substituents
include, but are not limited to, any of the substituents described
herein, that result in the formation of a stable moiety (e.g.,
aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic,
aryl, heteroaryl, acyl, oxo, imino, thiooxo, cyano, isocyano,
amino, azido, nitro, hydroxyl, thiol, halo, aliphaticamino,
heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino,
heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy,
heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy,
heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy,
heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the
like, each of which may or may not be further substituted).
[0033] The term "alkyl," as used herein, refers to saturated,
straight- or branched-chain hydrocarbon radicals derived from a
hydrocarbon moiety containing between one and twenty carbon atoms
by removal of a single hydrogen atom. In some embodiments, the
alkyl group employed in the invention contains 1-20 carbon atoms.
In another embodiment, the alkyl group employed contains 1-15
carbon atoms. In another embodiment, the alkyl group employed
contains 1-10 carbon atoms. In another embodiment, the alkyl group
employed contains 1-8 carbon atoms. In another embodiment, the
alkyl group employed contains 1-5 carbon atoms. Examples of alkyl
radicals include, but are not limited to, methyl, ethyl, n-propyl,
isopropyl, n-butyl, iso-butyl, sec-butyl, sec-pentyl, iso-pentyl,
tert-butyl, n-pentyl, neopentyl, n-hexyl, sec-hexyl, n-heptyl,
n-octyl, n-decyl, n-undecyl, dodecyl, and the like, which may bear
one or more sustitutents. Alkyl group substituents include, but are
not limited to, any of the substituents described herein, that
result in the formation of a stable moiety (e.g., aliphatic, alkyl,
alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl,
acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro,
hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino,
alkylamino, heteroalkylamino, arylamino, heteroarylamino,
alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy,
heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy,
heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy,
heteroarylthioxy, acyloxy, and the like, each of which may or may
not be further substituted).
[0034] The term "alkenyl," as used herein, denotes a monovalent
group derived from a straight- or branched-chain hydrocarbon moiety
having at least one carbon-carbon double bond by the removal of a
single hydrogen atom. In certain embodiments, the alkenyl group
employed in the invention contains 2-20 carbon atoms. In some
embodiments, the alkenyl group employed in the invention contains
2-15 carbon atoms. In another embodiment, the alkenyl group
employed contains 2-10 carbon atoms. In still other embodiments,
the alkenyl group contains 2-8 carbon atoms. In yet other
embodiments, the alkenyl group contains 2-5 carbons. Alkenyl groups
include, for example, ethenyl, propenyl, butenyl,
1-methyl-2-buten-1-yl, and the like, which may bear one or more
substituents. Alkenyl group substituents include, but are not
limited to, any of the substituents described herein, that result
in the formation of a stable moiety (e.g., aliphatic, alkyl,
alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl,
acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro,
hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino,
alkylamino, heteroalkylamino, arylamino, heteroarylamino,
alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy,
heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy,
heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy,
heteroarylthioxy, acyloxy, and the like, each of which may or may
not be further substituted).
[0035] The term "alkynyl," as used herein, refers to a monovalent
group derived from a straight- or branched-chain hydrocarbon having
at least one carbon-carbon triple bond by the removal of a single
hydrogen atom. In certain embodiments, the alkynyl group employed
in the invention contains 2-20 carbon atoms. In some embodiments,
the alkynyl group employed in the invention contains 2-15 carbon
atoms. In another embodiment, the alkynyl group employed contains
2-10 carbon atoms. In still other embodiments, the alkynyl group
contains 2-8 carbon atoms. In still other embodiments, the alkynyl
group contains 2-5 carbon atoms. Representative alkynyl groups
include, but are not limited to, ethynyl, 2-propynyl (propargyl),
1-propynyl, and the like, which may bear one or more substituents.
Alkynyl group substituents include, but are not limited to, any of
the substituents described herein, that result in the formation of
a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl,
heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino,
thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol,
halo, aliphaticamino, heteroaliphaticamino, alkylamino,
heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl,
aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy,
aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy,
alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy,
acyloxy, and the like, each of which may or may not be further
substituted).
[0036] The term "amino," as used herein, refers to a group of the
formula (--NH.sub.2). A "substituted amino" refers either to a
mono-substituted amine (--NHR.sup.h) of a disubstitued amine
(--NR.sup.h.sub.2), wherein the R.sup.h substituent is any
substitutent as described herein that results in the formation of a
stable moiety (e.g., a suitable amino protecting group; aliphatic,
alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl,
heteroaryl, acyl, amino, nitro, hydroxyl, thiol, halo,
aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino,
arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy,
heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy,
heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy,
heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the
like, each of which may or may not be further substituted). In
certain embodiments, the R.sup.h substituents of the di-substituted
amino group(--NR.sup.h.sub.2) form a 5- to 6-membered heterocyclic
ring.
[0037] The term "alkoxy" refers to a "substituted hydroxyl" of the
formula (--OR.sup.i), wherein R.sup.i is an optionally substituted
alkyl group, as defined herein, and the oxygen moiety is directly
attached to the parent molecule.
[0038] The term "alkylthioxy" refers to a "substituted thiol" of
the formula (--SR.sup.r), wherein R.sup.r is an optionally
substituted alkyl group, as defined herein, and the sulfur moiety
is directly attached to the parent molecule.
[0039] The term "alkylamino" refers to a "substituted amino" of the
formula (--NR.sup.h.sub.2), wherein R.sup.h is, independently, a
hydrogen or an optionally subsituted alkyl group, as defined
herein, and the nitrogen moiety is directly attached to the parent
molecule.
[0040] The term "aryl," as used herein, refer to stable aromatic
mono- or polycyclic ring system having 3-20 ring atoms, of which
all the ring atoms are carbon, and which may be substituted or
unsubstituted. In certain embodiments of the present invention,
"aryl" refers to a mono, bi, or tricyclic C.sub.4-C.sub.20 aromatic
ring system having one, two, or three aromatic rings which include,
but not limited to, phenyl, biphenyl, naphthyl, and the like, which
may bear one or more substituents. Aryl substituents include, but
are not limited to, any of the substituents described herein, that
result in the formation of a stable moiety (e.g., aliphatic, alkyl,
alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl,
acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro,
hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino,
alkylamino, heteroalkylamino, arylamino, heteroarylamino,
alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy,
heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy,
heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy,
heteroarylthioxy, acyloxy, and the like, each of which may or may
not be further substituted).
[0041] The term "arylalkyl," as used herein, refers to an aryl
substituted alkyl group, wherein the terms "aryl" and "alkyl" are
defined herein, and wherein the aryl group is attached to the alkyl
group, which in turn is attached to the parent molecule. An
exemplary arylalkyl group includes benzyl.
[0042] The term "aryloxy" refers to a "substituted hydroxyl" of the
formula (--OR.sup.i), wherein R.sup.i is an optionally substituted
aryl group, as defined herein, and the oxygen moiety is directly
attached to the parent molecule.
[0043] The term "arylamino," refers to a "substituted amino" of the
formula (--NR.sup.h.sub.2), wherein R.sup.h is, independently, a
hydrogen or an optionally substituted aryl group, as defined
herein, and the nitrogen moiety is directly attached to the parent
molecule.
[0044] The term "arylthioxy" refers to a "substituted thiol" of the
formula (--SR.sup.r), wherein R.sup.r is an optionally substituted
aryl group, as defined herein, and the sulfur moiety is directly
attached to the parent molecule.
[0045] The term "azido," as used herein, refers to a group of the
formula (--N.sub.3).
[0046] The term "cyano," as used herein, refers to a group of the
formula (--CN).
[0047] The terms "halo" and "halogen" as used herein refer to an
atom selected from fluorine (fluoro, --F), chlorine (chloro, --Cl),
bromine (bromo, --Br), and iodine (iodo, --I).
[0048] The term "heteroaliphatic," as used herein, refers to an
aliphatic moiety, as defined herein, which includes both saturated
and unsaturated, nonaromatic, straight chain (i.e., unbranched),
branched, acyclic, cyclic (i.e., heterocyclic), or polycyclic
hydrocarbons, which are optionally substituted with one or more
functional groups, and that contain one or more oxygen, sulfur,
nitrogen, phosphorus, or silicon atoms, e.g., in place of carbon
atoms. In certain embodiments, heteroaliphatic moieties are
substituted by independent replacement of one or more of the
hydrogen atoms thereon with one or more substituents. As will be
appreciated by one of ordinary skill in the art, "heteroaliphatic"
is intended herein to include, but is not limited to, heteroalkyl,
heteroalkenyl, heteroalkynyl, heterocycloalkyl, heterocycloalkenyl,
and heterocycloalkynyl moieties. Thus, the term "heteroaliphatic"
includes the terms "heteroalkyl," "heteroalkenyl", "heteroalkynyl",
and the like. Furthermore, as used herein, the terms "heteroalkyl",
"heteroalkenyl", "heteroalkynyl", and the like encompass both
substituted and unsubstituted groups. In certain embodiments, as
used herein, "heteroaliphatic" is used to indicate those
heteroaliphatic groups (cyclic, acyclic, substituted,
unsubstituted, branched or unbranched) having 1-20 carbon atoms.
Heteroaliphatic group substituents include, but are not limited to,
any of the substituents described herein, that result in the
formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl,
alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl,
sulfinyl, sulfonyl, oxo, imino, thiooxo, cyano, isocyano, amino,
azido, nitro, hydroxyl, thiol, halo, aliphaticamino,
heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino,
heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy,
heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy,
heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy,
heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the
like, each of which may or may not be further substituted).
[0049] The term "heteroalkyl," as used herein, refers to an alkyl
moiety, as defined herein, which contain one or more oxygen,
sulfur, nitrogen, phosphorus, or silicon atoms, e.g., in place of
carbon atoms.
[0050] The term "heteroalkenyl," as used herein, refers to an
alkenyl moiety, as defined herein, which contain one or more
oxygen, sulfur, nitrogen, phosphorus, or silicon atoms, e.g., in
place of carbon atoms.
[0051] The term "heteroalkynyl," as used herein, refers to an
alkynyl moiety, as defined herein, which contain one or more
oxygen, sulfur, nitrogen, phosphorus, or silicon atoms, e.g., in
place of carbon atoms.
[0052] The term "heteroalkylamino" refers to a "substituted amino"
of the formula (--NR.sup.h.sub.2), wherein R.sup.h is,
independently, a hydrogen or an optionally substituted heteroalkyl
group, as defined herein, and the nitrogen moiety is directly
attached to the parent molecule.
[0053] The term "heteroalkyloxy" refers to a "substituted hydroxyl"
of the formula (--OR.sup.i), wherein R.sup.i is an optionally
substituted heteroalkyl group, as defined herein, and the oxygen
moiety is directly attached to the parent molecule.
[0054] The term "heteroalkylthioxy" refers to a "substituted thiol"
of the formula (--SR.sup.r), wherein R.sup.r is an optionally
substituted heteroalkyl group, as defined herein, and the sulfur
moiety is directly attached to the parent molecule.
[0055] The term "heterocyclic," "heterocycles," or "heterocyclyl,"
as used herein, refers to a cyclic heteroaliphatic group. A
heterocyclic group refers to a non-aromatic, partially unsaturated
or fully saturated, 3- to 12-membered ring system, which includes
single rings of 3 to 8 atoms in size, and bi- and tri-cyclic ring
systems which may include aromatic five- or six-membered aryl or
heteroaryl groups fused to a non-aromatic ring. These heterocyclic
rings include those having from one to three heteroatoms
independently selected from oxygen, sulfur, and nitrogen, in which
the nitrogen and sulfur heteroatoms may optionally be oxidized and
the nitrogen heteroatom may optionally be quaternized. In certain
embodiments, the term heterocylic refers to a non-aromatic 5-, 6-,
or 7-membered ring or polycyclic group wherein at least one ring
atom is a heteroatom selected from O, S, and N (wherein the
nitrogen and sulfur heteroatoms may be optionally oxidized), and
the remaining ring atoms are carbon, the radical being joined to
the rest of the molecule via any of the ring atoms. Heterocycyl
groups include, but are not limited to, a bi- or tri-cyclic group,
comprising fused five, six, or seven-membered rings having between
one and three heteroatoms independently selected from the oxygen,
sulfur, and nitrogen, wherein (i) each 5-membered ring has 0 to 2
double bonds, each 6-membered ring has 0 to 2 double bonds, and
each 7-membered ring has 0 to 3 double bonds, (ii) the nitrogen and
sulfur heteroatoms may be optionally oxidized, (iii) the nitrogen
heteroatom may optionally be quaternized, and (iv) any of the above
heterocyclic rings may be fused to an aryl or heteroaryl ring.
Exemplary heterocycles include azacyclopropanyl, azacyclobutanyl,
1,3-diazatidinyl, piperidinyl, piperazinyl, azocanyl, thiaranyl,
thietanyl, tetrahydrothiophenyl, dithiolanyl, thiacyclohexanyl,
oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropuranyl, dioxanyl,
oxathiolanyl, morpholinyl, thioxanyl, tetrahydronaphthyl, and the
like, which may bear one or more substituents. Substituents
include, but are not limited to, any of the substituents described
herein, that result in the formation of a stable moiety (e.g.,
aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic,
aryl, heteroaryl, acyl, sulfinyl, sulfonyl, oxo, imino, thiooxo,
cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo,
aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino,
arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy,
heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy,
heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy,
heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the
like, each of which may or may not be further substituted).
[0056] The term "heteroaryl," as used herein, refer to stable
aromatic mono- or polycyclic ring system having 3-20 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.
Exemplary heteroaryls include, but are not limited to pyrrolyl,
pyrazolyl, imidazolyl, pyridinyl, pyrimidinyl, pyrazinyl,
pyridazinyl, triazinyl, tetrazinyl, pyyrolizinyl, indolyl,
quinolinyl, isoquinolinyl, benzoimidazolyl, indazolyl, quinolinyl,
isoquinolinyl, quinolizinyl, cinnolinyl, quinazolynyl,
phthalazinyl, naphthridinyl, quinoxalinyl, thiophenyl,
thianaphthenyl, furanyl, benzofuranyl, benzothiazolyl, thiazolynyl,
isothiazolyl, thiadiazolynyl, oxazolyl, isoxazolyl, oxadiaziolyl,
oxadiaziolyl, and the like, which may bear one or more
substituents. Heteroaryl substituents include, but are not limited
to, any of the substituents described herein, that result in the
formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl,
alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl,
sulfinyl, sulfonyl, oxo, imino, thiooxo, cyano, isocyano, amino,
azido, nitro, hydroxyl, thiol, halo, aliphaticamino,
heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino,
heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy,
heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy,
heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy,
heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the
like, each of which may or may not be further substituted).
[0057] The term "heteroarylene," as used herein, refers to a
biradical derived from an heteroaryl group, as defined herein, by
removal of two hydrogen atoms. Heteroarylene groups may be
substituted or unsubstituted. Additionally, heteroarylene groups
may be incorporated as a linker group into an alkylene, alkenylene,
alkynylene, heteroalkylene, heteroalkenylene, or heteroalkynylene
group, as defined herein. Heteroarylene group substituents include,
but are not limited to, any of the substituents described herein,
that result in the formation of a stable moiety (e.g., aliphatic,
alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl,
heteroaryl, acyl, oxo, imino, thiooxo, cyano, isocyano, amino,
azido, nitro, hydroxyl, thiol, halo, aliphaticamino,
heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino,
heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy,
heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy,
heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy,
heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the
like, each of which may or may not be further substituted).
[0058] The term "heteroarylamino" refers to a "substituted amino"
of the (--NR.sup.h.sub.2), wherein R.sup.h is, independently, a
hydrogen or an optionally substituted heteroaryl group, as defined
herein, and the nitrogen moiety is directly attached to the parent
molecule.
[0059] The term "heteroaryloxy" refers to a "substituted hydroxyl"
of the formula (--OR.sup.i), wherein R.sup.i is an optionally
substituted heteroaryl group, as defined herein, and the oxygen
moiety is directly attached to the parent molecule.
[0060] The term "heteroarylthioxy" refers to a "substituted thiol"
of the formula (--SR.sup.r), wherein R.sup.r is an optionally
substituted heteroaryl group, as defined herein, and the sulfur
moiety is directly attached to the parent molecule.
[0061] The term "hydroxy," or "hydroxyl," as used herein, refers to
a group of the formula (--OH). A "substituted hydroxyl" refers to a
group of the formula (--OR.sup.i), wherein R.sup.i can be any
substitutent which results in a stable moiety (e.g., a suitable
hydroxyl protecting group; aliphatic, alkyl, alkenyl, alkynyl,
heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, nitro,
alkylaryl, arylalkyl, and the like, each of which may or may not be
further substituted).
[0062] The term "imino," as used herein, refers to a group of the
formula (.dbd.NR.sup.r), wherein R.sup.r corresponds to hydrogen or
any substitutent as described herein, that results in the formation
of a stable moiety (for example, a suitable amino protecting group;
aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic,
aryl, heteroaryl, acyl, amino, hydroxyl, alkylaryl, arylalkyl, and
the like, each of which may or may not be further substituted). In
certain embodiments, imino refers to .dbd.NH wherein R.sup.r is
hydrogen.
[0063] The term "isocyano," as used herein, refers to a group of
the formula (--NC).
[0064] The term "nitro," as used herein, refers to a group of the
formula (--NO.sub.2).
[0065] The term "oxo," as used herein, refers to a group of the
formula (.dbd.O).
[0066] The term "stable moiety," as used herein, preferably refers
to a moiety which possess stability sufficient to allow
manufacture, and which maintains its integrity for a sufficient
period of time to be useful for the purposes detailed herein.
[0067] A "suitable amino-protecting group," as used herein, is well
known in the art and include those described in detail in Greene's
Protective Groups in Organic Synthesis, P. G. M. Wuts and T. W.
Greene, 4.sup.th edition, Wiley-Interscience, 2006, the entirety of
which is incorporated herein by reference. Suitable
amino-protecting groups include methyl carbamate, ethyl carbamante,
9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl
carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate,
2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl
carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc),
2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl
carbamate (Teoc), 2-phenylethyl carbamate (hZ),
1-(1-adamantyl)-1-methylethyl carbamate (Adpoc),
1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl
carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate
(TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc),
1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2'-
and 4'-pyridyl)ethyl carbamate (Pyoc),
2-(N,N-dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate
(BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl
carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl
carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl
carbamate, N-hydroxypiperidinyl carbamate, alkyldithio carbamate,
benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz),
p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl
carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl
carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl
carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl
carbamate, 2-(p-toluenesulfonyl)ethyl carbamate,
[2-(1,3-dithianyl)]methyl carbamate (Dmoc), 4-methylthiophenyl
carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc),
2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl
carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate,
m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl
carbamate, 5-benzisoxazolylmethyl carbamate,
2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc),
m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate,
o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate,
phenyl(o-nitrophenyl)methyl carbamate, phenothiazinyl-(10)-carbonyl
derivative, N'-p-toluenesulfonylaminocarbonyl derivative,
N'-phenylaminothiocarbonyl derivative, t-amyl carbamate, S-benzyl
thiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate,
cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl
carbamate, p-decyloxybenzyl carbamate, 2,2-dimethoxycarbonylvinyl
carbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate,
1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate,
1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate,
2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl
carbamate, isobutyl carbamate, isonicotinyl carbamate,
p-(p'-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl
carbamate, 1-methylcyclohexyl carbamate,
1-methyl-1-cyclopropylmethyl carbamate,
1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate,
1-methyl-1-(p-phenylazophenyl)ethyl carbamate,
1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl
carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate,
2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl
carbamate, 2,4,6-trimethylbenzyl carbamate, formamide, acetamide,
chloroacetamide, trichloroacetamide, trifluoroacetamide,
phenylacetamide, 3-phenylpropanamide, picolinamide,
3-pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide,
p-phenylbenzamide, o-nitophenylacetamide, o-nitrophenoxyacetamide,
acetoacetamide, (N'-dithiobenzyloxycarbonylamino)acetamide,
3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide,
2-methyl-2-(o-nitrophenoxy)propanamide,
2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide,
3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine
derivative, o-nitrobenzamide, o-(benzoyloxymethyl)benzamide,
4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide
(Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole,
N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE),
5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one,
5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexan-2-one,
1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine,
N-[2-(trimethylsilyl)ethoxy]methylamine (SEM),
N-3-acetoxypropylamine,
N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary
ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine,
N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr),
N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr),
N-9-phenylfluorenylamine (PhF),
N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino
(Fcm), N-2-picolylamino N'-oxide, N-1,1-dimethylthiomethyleneamine,
N-benzylideneamine, N-p-methoxybenzylideneamine,
N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine,
N--(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine,
N-p-nitrobenzylideneamine, N-salicylideneamine,
N-5-chlorosalicylideneamine,
N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine,
N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine,
N-borane derivative, N-diphenylborinic acid derivative,
N-[phenyl(pentacarbonylchromium- or tungsten)carbonyl]amine,
N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosoamine,
amine N-oxide, diphenylphosphinamide (Dpp),
dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt),
dialkyl phosphoramidates, dibenzyl phosphoramidate, diphenyl
phosphoramidate, benzenesulfenamide, o-nitrobenzenesulfenamide
(Nps), 2,4-dinitrobenzenesulfenamide,
pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide,
triphenylmethylsulfenamide, 3-nitropyridinesulfenamide (Npys),
p-toluenesulfonamide (Ts), benzenesulfonamide,
2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr),
2,4,6-trimethoxybenzenesulfonamide (Mtb),
2,6-dimethyl-4-methoxybenzenesulfonamide (Pme),
2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte),
4-methoxybenzenesulfonamide (Mbs),
2,4,6-trimethylbenzenesulfonamide (Mts),
2,6-dimethoxy-4-methylbenzenesulfonamide (iMds),
2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc),
methanesulfonamide (Ms), .beta.-trimethylsilylethanesulfonamide
(SES), 9-anthracenesulfonamide,
4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),
benzylsulfonamide, trifluoromethylsulfonamide, and
phenacylsulfonamide.
[0068] A "suitable carboxylic acid protecting group," or "protected
carboxylic acid," as used herein, are well known in the art and
include those described in detail in Greene (1999). Examples of
suitably protected carboxylic acids further include, but are not
limited to, silyl-, alkyl-, alkenyl-, aryl-, and
arylalkyl-protected carboxylic acids. Examples of suitable silyl
groups include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl,
t-butyldiphenylsilyl, triisopropylsilyl, and the like. Examples of
suitable alkyl groups include methyl, benzyl, p-methoxybenzyl,
3,4-dimethoxybenzyl, trityl, t-butyl, tetrahydropyran-2-yl.
Examples of suitable alkenyl groups include allyl. Examples of
suitable aryl groups include optionally substituted phenyl,
biphenyl, or naphthyl. Examples of suitable arylalkyl groups
include optionally substituted benzyl (e.g., p-methoxybenzyl (MPM),
3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl,
2,6-dichlorobenzyl, p-cyanobenzyl), and 2- and 4-picolyl.
[0069] A "suitable hydroxyl protecting group" as used herein, is
well known in the art and includes those described in detail in
Greene (1999). Suitable hydroxyl protecting groups include methyl,
methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl,
(phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM),
p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM),
guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM),
siloxymethyl, 2-methoxyethoxymethyl (MEM),
2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl,
2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP),
3-bromotetrahydropyranyl, tetrahydrothiopyranyl,
1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP),
4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl
S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl
(CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl,
2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl,
1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl,
1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,
2,2,2-trichloroethyl, 2-trimethylsilylethyl,
2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl,
p-methoxyphenyl, 2,4-dinitrophenyl, benzyl, p-methoxybenzyl,
3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl,
2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl,
4-picolyl, 3-methyl-2-picolyl N-oxido, diphenylmethyl,
p,p'-dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl,
.alpha.-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl,
di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl,
4-(4'-bromophenacyloxyphenyl)diphenylmethyl,
4,4',4''-tris(4,5-dichlorophthalimidophenyl)methyl,
4,4',4'-tris(levulinoyloxyphenyl)methyl,
4,4',4'-tris(benzoyloxyphenyl)methyl,
3-(imidazol-1-yl)bis(4',4'-dimethoxyphenyl)methyl,
1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl,
9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,
1,3-benzodithiolan-2-yl, benzisothiazolyl S,S-dioxido,
trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl
(TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl
(DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS),
t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl,
triphenylsilyl, diphenylmethylsilyl (DPMS),
t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate,
acetate, chloroacetate, dichloroacetate, trichloroacetate,
trifluoroacetate, methoxyacetate, triphenylmethoxyacetate,
phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate,
4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate
(levulinoyldithioacetal), pivaloate, adamantoate, crotonate,
4-methoxycrotonate, benzoate, p-phenylbenzoate,
2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate,
9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl
2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl
carbonate (TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec),
2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutyl
carbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkyl
p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl
p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate,
alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl
S-benzyl thiocarbonate, 4-ethoxy-1-napththyl carbonate, methyl
dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate,
4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate,
2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl,
4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate,
2,6-dichloro-4-methylphenoxyacetate,
2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,
2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,
isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate,
o-(methoxycarbonyl)benzoate, .alpha.-naphthoate, nitrate, alkyl
N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate,
borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate,
sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate
(Ts). For protecting 1,2- or 1,3-diols, the protecting groups
include methylene acetal, ethylidene acetal, 1-t-butylethylidene
ketal, 1-phenylethylidene ketal, (4-methoxyphenyl)ethylidene
acetal, 2,2,2-trichloroethylidene acetal, acetonide,
cyclopentylidene ketal, cyclohexylidene ketal, cycloheptylidene
ketal, benzylidene acetal, p-methoxybenzylidene acetal,
2,4-dimethoxybenzylidene ketal, 3,4-dimethoxybenzylidene acetal,
2-nitrobenzylidene acetal, methoxymethylene acetal, ethoxymethylene
acetal, dimethoxymethylene ortho ester, 1-methoxyethylidene ortho
ester, 1-ethoxyethylidine ortho ester, 1,2-dimethoxyethylidene
ortho ester, a-methoxybenzylidene ortho ester,
1-(N,N-dimethylamino)ethylidene derivative,
.alpha.-(N,N'-dimethylamino)benzylidene derivative,
2-oxacyclopentylidene ortho ester, di-t-butylsilylene group (DTBS),
1,3-(1,1,3,3-tetraisopropyldisiloxanylidene) derivative (TIPDS),
tetra-t-butoxydisiloxane-1,3-diylidene derivative (TBDS), cyclic
carbonates, cyclic boronates, ethyl boronate, and phenyl
boronate.
[0070] A "suitable thiol protecting group," as used herein, is well
known in the art and include those described in detail in
Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M.
Wuts, 3.sup.rd edition, John Wiley & Sons, 1999, the entirety
of which is incorporated herein by reference. Examples of suitably
protected thiol groups further include, but are not limited to,
thioesters, carbonates, sulfonates allyl thioethers, thioethers,
silyl thioethers, alkyl thioethers, arylalkyl thioethers, and
alkyloxyalkyl thioethers. Examples of suitable ester groups include
formates, acetates, proprionates, pentanoates, crotonates, and
benzoates. Specific examples of suitable ester groups include
formate, benzoyl formate, chloroacetate, trifluoroacetate,
methoxyacetate, triphenylmethoxyacetate, p-chlorophenoxyacetate,
3-phenylpropionate, 4-oxopentanoate,
4,4-(ethylenedithio)pentanoate, pivaloate (trimethylacetate),
crotonate, 4-methoxy-crotonate, benzoate, p-benylbenzoate,
2,4,6-trimethylbenzoate. Examples of suitable carbonates include
9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl,
2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl, allyl, and
p-nitrobenzyl carbonate. Examples of suitable silyl groups include
trimethylsilyl, triethylsilyl, t-butyldimethylsilyl,
t-butyldiphenylsilyl, triisopropylsilyl ether, and other
trialkylsilyl ethers. Examples of suitable alkyl groups include
methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl,
t-butyl, and allyl ether, or derivatives thereof. Examples of
suitable arylalkyl groups include benzyl, p-methoxybenzyl (MPM),
3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl,
2,6-dichlorobenzyl, p-cyanobenzyl, 2- and 4-picolyl ethers.
[0071] The term "thio," or "thiol," as used herein, refers to a
group of the formula (--SH). A "substituted thiol" refers to a
group of the formula (--SR.sup.r), wherein R.sup.r can be any
substituent that results in the formation of a stable moiety (e.g.,
a suitable thiol protecting group; aliphatic, alkyl, alkenyl,
alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl,
sulfinyl, sulfonyl, cyano, nitro, alkylaryl, arylalkyl, and the
like, each of which may or may not be further substituted).
[0072] 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 are well
known in the art. For example, Berge et al., describe
pharmaceutically acceptable salts in detail in J. Pharmaceutical
Sciences, 1977, 66, 1-19, incorporated herein by reference.
Pharmaceutically acceptable salts of the compounds of this
invention include those derived from suitable inorganic and organic
acids and bases. 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, hemisulfate, 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.
Salts derived from appropriate bases include alkali metal, alkaline
earth metal, ammonium and N.sup.+(C.sub.1-4alkyl).sub.4 salts.
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.
[0073] The term "subject," as used herein, refers to any animal. In
certain embodiments, the subject is a mammal. In certain
embodiments, the term "subject", as used herein, refers to a human
(e.g., a man, a woman, or a child).
[0074] The terms "administer," "administering," or
"administration," as used herein refers to implanting, absorbing,
ingesting, injecting, or inhaling the inventive compound.
[0075] As used herein, the terms "treatment," "treat," and
"treating" refer to reversing, alleviating, delaying the onset of,
or inhibiting the progress of a disease or disorder, or one or more
symptoms thereof, described herein. In some embodiments, treatment
may be administered after one or more symptoms have developed. In
other embodiments, treatment may be administered in the absence of
symptoms. For example, treatment may be administered to a
susceptible individual prior to the onset of symptoms (e.g., in
light of a history of symptoms and/or in light of genetic or other
susceptibility factors). Treatment may also be continued after
symptoms have resolved, for example to delay or prevent
recurrence.
[0076] The terms "effective amount" and "therapeutically effective
amount," as used herein, refer to the amount or concentration of an
inventive compound, that, when administered to a subject, is
effective to at least partially treat a condition from which the
subject is suffering (e.g., a neurodegenerative disease).
BRIEF DESCRIPTION OF THE DRAWINGS
[0077] FIG. 1 depicts western blot analysis showing that
halofuginone (HF) stimulates the amino acid response (AAR) in a
fibroblastic cell line.
[0078] FIG. 2 depicts western blot analysis showing that
phosphorylation of elF2alpha by halofuginone is GCN2-dependent.
[0079] FIG. 3 is a graph showing that proline rescues translational
inhibition by halofuginone.
[0080] FIG. 4A depicts western blot analysis showing that
halofuginone induced elF2alpha phosphorylation.
[0081] FIG. 4B is a graph showing that halofuginone-inhibited Th17
differentiation is blocked by added proline.
[0082] FIGS. 5A-5G. Selective inhibition of Th17 cell development
by halofuginone. FIG. 5A, left, is a graph showing dose response
analyses performed on activated carboxyfluorescein succinimidyl
ester (CFSE)-labeled CD4.sup.+ CD25.sup.- T cells in the presence
of DMSO, 40 nM compound 9 (MAZ1310), or increasing concentrations
of halofuginone (HF) (1.25-40 nM). CFSE dilution and cell surface
CD25 expression were determined 48 hours after activation.
Intracellular cytokine production was determined on day 4 or 5
following restimulation with phorbol myristate acetate (PMA) and
ionomycin. CFSE dilution and percentages of cells expressing CD25,
IFN.gamma..sup.+ IL4.sup.- (Th1 cells), IL-4.sup.+ IFN.gamma..sup.-
(Th2 cells) or IL-17.sup.+ IFN.gamma..sup.- (Th17 cells) cells are
displayed and the values are normalized to T cells treated with 40
nM MAZ1310.+-.SD. FIG. 5A, right, is a graph showing dose-response
analyses of HF effects on CD8.sup.+ T cell or B cell function.
Cells were activated in the presence of DMSO, 40 nM compound 9
(MAZ1310), or increasing concentrations of HF (1.25-40 nM). CFSE
dilution, cell surface CD25 expression, and intracellular cytokine
production were determined as above 2-5 days after activation. CFSE
dilution and percentages of CD8.sup.+ T cells expressing CD25,
IFN.gamma..sup.+ granzyme B.sup.+ (cytotoxic T lymphocytes) or
IL-6.sup.+ B cells are displayed and the values are normalized to
cells treated with 40 nM MAZ1310.+-.SD.
##STR00005##
[0083] FIG. 5B is a table showing IC.sub.50 values calculated for
the effects of halofuginone on CD4.sup.+ CD25.sup.- T cell
functions as indicated.
[0084] FIG. 5C is a graph showing data for experiments in which a
racemic mixture of halofuginone (HF) or HPLC-purified D- or
L-enantiomers of halofuginone (HF-D, or HF-L) were added to
CD4.sup.+ CD25.sup.- T cells activated in the presence of TGF.beta.
plus IL-6 and the percent of Th17 cells (IL-17.sup.+
IFN.gamma..sup.-) was determined by intracellular cytokine staining
on day 4. Values are normalized to cells treated with 40 nM
compound 9 (MAZ1310).+-.SD.
[0085] FIG. 5D is a graph showing the percent of Th17 cells
(IL-17.sup.+ IFN.gamma..sup.-) determined by intracellular staining
4 days after activation as above and values are presented as mean
percent of Th17 cells.+-.SD. CD4.sup.+ CD25.sup.- T cells were
activated in the indicated cytokine conditions, and 10 nM
halofuginone was added at the indicated times following activation.
Asterisks indicate statistical significance (p<0.005) relative
to T cells treated with 10 nM compound 9 (MAZ1310) at the time of
activation.
[0086] FIG. 5E is a set of FACS analyses of CFSE-labeled T cells
activated in the indicated cytokine conditions in the presence of
DMSO, 5 nM halofuginone (HF), 10 nM HF, 10 nM compound 9 (MAZ1310),
or 10 .mu.M SB-431542. Foxp3 intracellular staining was performed 3
days after T cell activation and intracellular cytokine staining
was performed on day 4. Cells with equivalent CFSE fluorescence are
gated on as indicated and intracellular Foxp3 or cytokine
expression is shown within each gated population.
[0087] FIG. 5F is a set of FACS analyses of purified primary human
memory T cells (CD4.sup.+ CD45RO.sup.+) activated in co-culture
with CD14.sup.+ monocytes and treated with DMSO, 100 nM HF or 100
nM MAZ1310. T cells were expanded for 6 days and intracellular
cytokine expression was determined following restimuation with PMA
plus ionomycin.
[0088] FIG. 5G is a graph depicting the percent of IL-17.sup.-
(shaded bars) or IFN.gamma..sup.- (white bars) expressing T cells
upon treatment with the indicated additives. The data were
normalized to T cells treated with MAZ1310 and are displayed as
mean values.+-.SD. Asterisk indicates statistical significance
(p<0.05). All data represent at least three similar
experiments.
[0089] FIGS. 6A-6E. HF inhibits Th17 differentiation through
effects on STAT3 phosphorylation. FIG. 6A is a set of
representative histograms displaying the kinetics of STAT3
phosphorylation in developing Th17 cells treated with or without
halofuginone. Resting naive T cells (thick black line), T cells
activated in the presence of TGF.beta. plus IL-6 (TGF.beta./IL-6)
treated with 10 nM MAZ1310 (thin black line), 5 nM HF (medium black
line), or 10 nM HF (thick black line). T cells were fixed at the
indicated times and intracellular phospho-STAT3 staining was
performed.
[0090] FIG. 6B depicts western blot analysis of CD4.sup.+
CD25.sup.- T cells treated with 10 nM HF or 10 nM MAZ1310 and
activated in the presence or absence of TGF.beta. plus IL-6. Whole
cell lysates were generated at the indicated times following
activation.
[0091] FIG. 6C is a set of FACS analyses of CD4.sup.+ CD25.sup.- T
cells from YFPfl/fl or STAT3C-GFPfl/fl mice treated with
recombinant TAT-Cre which were activated in the presence or absence
of TGF.beta. plus IL-6 and treated with DMSO, 5 nM HF, 10 nM HF, or
10 nM MAZ1310 as indicated. T cells were restimulated after 4 days
and intracellular cytokine staining was performed. T cells
expressing YFP or GFP are gated on as shown.
[0092] FIG. 6D is a bar graph displaying the percent of Th17 cells
(IL-17.sup.+ IFN.gamma..sup.-) within YFP.sup.- cells (black bars),
YFP.sup.+ cells (etched bars), STAT3CGFP.sup.- cells (white bars)
or STAT3C-GFP.sup.+ (etched bars). The data are normalized to
DMSO-treated cultures and are presented as mean values.+-.SD on
duplicate samples. Asterisks indicate statistical differences
between STAT3C-GFP.sup.+ cells and YFP.sup.+ cells (p<0.05).
[0093] FIG. 6E is a set of FACS analyses of CD4.sup.+ CD25.sup.- T
cells activated in medium or TGF.beta. plus IL-6, treated with
DMSO, 10 nM HF, 10 nM MAZ1310, or 10 nM HF plus 10 .mu.M SB-431542.
Foxp3 expression was determined on day 3 by intracellular staining.
All experiments were performed at least three times with similar
results.
[0094] FIGS. 7A-7F. Halofuginone activates the amino acid
starvation response pathway in T cells. FIG. 7A shows dot plot
analyses of microarray data from CD4.sup.+ CD25.sup.- T cells
treated with 10 nM HF or 10 nM MAZ1310 and activated in Th17
polarizing cytokine conditions for 3 or 6 hours. White dots
indicate transcripts increased at least 2-fold by HF treatment at
both 3 and 6 hours. Hallmark amino acid starvation response genes
are identified by text and arrowheads.
[0095] FIG. 7B is a graph showing chi-squared analysis of
microarray data from FIG. 4A, which shows the expression
distribution of genes previously found to be regulated by ATF4 in
tunicamycin-treated mouse embryonic fibroblasts (white dots--see
the table in FIG. 14).
[0096] FIG. 7C is a graph depicting results of quantitative
real-time PCR performed on cDNA generated from unstimulated naive T
cells or those activated for 4 hours in the presence of 10 nM
MAZ1310 or 10 nM HF. Asns, Gpt2 or eIF4Ebp1 mRNA expression was
normalized to Hprt levels and data are presented as mean
values.+-.SD in duplicate samples.
[0097] FIG. 7D depicts western blot analysis of purified CD4.sup.+
CD25.sup.- T cells either unstimulated, or TCR-activated without
exogenous cytokines in the presence of DMSO, 40 nM MAZ1310 or
titrating concentrations of HF (1.25-40 nM). Whole cell lysates
were prepared 4 hours-post TCR activation and immunoblotting was
performed with the indicated antibodies. ATF4 protein is indicated
by arrowhead. NS--non-specific band.
[0098] FIG. 7E depicts western blot analysis of purified CD4.sup.+
CD25.sup.- T cells activated through the TCR for the indicated
times without exogenous cytokines in the presence of either 10 nM
MAZ1310 or 10 nM HF as indicated. Whole cell lysates were prepared
during the timecourse and immunoblotting was performed.
[0099] FIG. 7F depicts western blot analysis of CD4.sup.+
CD25.sup.- T cells either left unstimulated or were TCR-activated
in the absence or presence of the indicated polarizing cytokine
conditions and 10 nM MAZ1310 or 10 nM HF as indicated. Whole cell
lysates were generated 4 hours after activation and immunoblotting
was performed. Microarray data were generated from three
independent experiments and all other data are representative of at
least two similar experiments.
[0100] FIGS. 8A-8F. Amino acid deprivation inhibits Th17
differentiation. FIG. 8A depicts western blot and Xbp1 splicing
analysis of CD4.sup.+ CD25.sup.- T cells left unstimulated, or
activated through the TCR for 4 hours in complete medium
(complete--200 .mu.M Cys/100 .mu.M Met), medium lacking Cys/Met
(-Cys/Met) or complete medium containing 1 .mu.g/ml tunicamycin, 10
nM HF, or 10 nM MAZ1310. Western blotting was performed on whole
cell extracts with the indicated antibodies. Xbp-1 splicing assay
was performed on cDNA synthesized from T cell cultures.
[0101] FIG. 8B is a graph depicting dose-response analyses of the
effects of limiting Cys/Met concentrations on T cell activation and
differentiation. Activated CD4.sup.+ CD25.sup.- T cells were
cultured in the absence or presence of polarizing cytokines to
induce Th1, Th2, iTreg or Th17 differentiation. Titrating
concentrations of Cys/Met are indicated. CD25 and Foxp3 expression
was determined 3 days post activation, cytokine production
determined by intracellular staining on day 4 or 5. Percentages of
cells expressing CD25, Foxp3, IFN.gamma..sup.+ IL4.sup.- (Th1
cells), IL-4.sup.+ IFN.gamma..sup.- (Th2 cells), or IL-17.sup.+
IFN.gamma..sup.- (Th17 cells) are displayed, and the values are
normalized to T cells cultured in complete medium (200 .mu.M
Cys/100 .mu.M Met).
[0102] FIG. 8C is a set of representative histograms show the
kinetics of STAT3 phosphorylation in CD4.sup.+ CD25.sup.- T cells
activated in the presence of TGF.beta. plus IL-6. Resting naive T
cells (thick black line), T cells cultured in complete medium (200
.mu.M Cys/100 .mu.M Met--thick black line), low Cys/Met
concentrations (10 .mu.M Cys/5 .mu.M Met--medium black line) or
complete medium with 10 nM HF (thin black line). T cells were fixed
at the indicated times and intracellular phospho-STAT3 staining was
performed.
[0103] FIG. 8D is a graph depicting quantification of the
intracellular phospho-STAT3 data. Data are presented as the percent
of phospho-STAT3.sup.+ T cells in each condition multiplied by mean
fluorescence intensity (MFI). Mean values from duplicate samples
are displayed.+-.SD.
[0104] FIG. 8E is a set of FACS analyses of activated T cells
cultured in the indicated cytokine condition in complete medium
(complete--200 .mu.M Cys/100 .mu.M Met/4 mM Leucine), medium
containing 0.1.times. cysteine and methionine (Cys/Met), medium
containing 0.1.times. leucine (Leu) or complete medium plus 0.2 mM
L-tryptophanol. Cells were expanded for 4 days and restimulated
with PMA and ionomycin for intracellular cytokine staining.
[0105] FIG. 8F is a graph depicting analyses of CD4.sup.+
CD25.sup.- T cells cultured in the presence of titrating
concentrations of tunicamycin as indicated. These cells were
analyzed for CD25 upregulation or differentiation into Th1, Th2,
iTreg, or Th17 cells. All experiments were performed 3 times with
similar results.
[0106] FIGS. 9A-9C. Effects of halofuginone on T cell activation
and effector function. FIG. 9A is a set of FACS analyses of CFSE
labeled CD4.sup.+ CD25.sup.- T cells activated in the absence or
presence of polarizing cytokines. DMSO, 5 nM HF, or 5 nM MAZ1310
was added to the cells at the time of T cell activation.
Intracellular Foxp3 staining was performed on expanded cells 3 days
after activation. Cytokine expression was determined by
intracellular staining after PMA and ionomycin restimulation on day
4-5.
[0107] FIG. 9B is a set of FACS and graphical analyses of CFSE
labeled CD4.sup.+ CD25.sup.- T cells treated with DMSO, 5 nM HF, or
5 nM MAZ1310 activated in the absence of polarizing cytokines. CFSE
dilution and CD25 cell surface expression was determined on day 2
by FACS analyses. T cells were activated as above without exogenous
cytokines and supernatants were harvested at the indicated
time-points following activation. Cytokine secretion was determined
using a cytometric bead array (CBA) on duplicate samples. Cytokine
concentrations were determined by comparison to standard curves and
data are presented as the mean cytokine concentrations.+-.SD.
[0108] FIG. 9C is a set of graphs depicting HF effects on IL-17 and
IL-17f mRNA expression in Th17 cells. CD4.sup.+ CD25.sup.- T cells
were differentiated under Th17 cytokine conditions in the presence
of DMSO, 10 nM HF, or 10 nM MAZ1310 for 4 days as above. Cells were
harvested and restimulated with PMA and ionomycin as above, and
cDNA was generated for Sybrgreen real-time PCR analysis. Data
indicate fold changes in mRNA expression normalized to Hprt and are
presented as mean expression.+-.SD. Asterisks indicate statistical
significance for Il17 mRNA (p<0.001) and Il7f mRNA (p<0.05)
for HF-treated T cells relative to those treated with MAZ1310. All
data are representative of at least three independent
experiments.
[0109] FIGS. 10A-10D. Halofuginone does not regulate TGF.beta.
signaling in T and B cells. FIG. 10A is a set of FACS analyses of
CD4.sup.+ CD25.sup.- T cells activated in Th or Th2 polarizing
conditions, either in the presence or absence of TGF.beta.. DMSO,
10 nM HF, 10 nM MAZ1310, or 10 .mu.M SB-431542 added as indicated
at the time of activation. Intracellular cytokine staining was
performed on expanded T cells on day 5.
[0110] FIG. 10B is a set of FACS analyses of CD8.sup.+ T cells
activated in the presence or absence of TGF.beta. and cultured with
DMSO, 10 nM HF, 10 nM MAZ1310, or 10 .mu.M SB-431542. Expanded
cells were restimulated on day 5 and intracellular staining was
performed.
[0111] FIG. 10C is a set of FACS analyses of CFSE-labeled B cells
activated by LPS stimulation in the presence or absence of
TGF.beta. plus DMSO, 10 nM HF, 10 nM MAZ1310, or 10 .mu.M
SB-431542. Intracellular IL-6 production in B cells restimulated
with PMA plus ionomycin, or cell-surface IgA expression was
determined 4 days after activation.
[0112] FIG. 10D depicts western blot analyses of purified CD4.sup.+
CD25.sup.- T cells treated with DMSO, 40 nM MAZ1310, titrating
concentrations of HF (2.5-40 nM) or 10 .mu.M SB-431542 for 30
minutes in complete medium supplemented with 0.1% fetal calf serum.
T cells were then activated in the presence or absence of 3 ng/ml
TGF.beta.. Whole cell extracts were prepared after 1 hour of
stimulation and western blot analyses were performed using the
indicated antibodies. These data are representative of three
similar experiments.
[0113] FIGS. 11A-11C. Halofuginone inhibits ROR.gamma.t function,
not expression. FIG. 11A is a graph depicting analyses of CD4.sup.+
CD25.sup.- T cells treated with DMSO (if no indication), 10 nM HF,
or 10 nM MAZ1310 as indicated and activated in the presence of
cytokines as noted. T cells were harvested at the indicated times
following activation, RNA was isolated and quantitative real-time
PCR was performed on cDNA. ROR.gamma.t expression was normalized to
Gapdh levels and the data are presented as fold changes relative to
unstimulated T cells.
[0114] FIG. 11B is a set of FACS analyses of CD4.sup.+ CD25.sup.- T
cells activated in the presence or absence of TGF.beta. plus IL-6,
which were transduced with empty (MIG) or ROR.gamma.t-expressing
(MIG.ROR.gamma.t) retroviruses 12 hours-post activation. Infected T
cells were expanded and restimulated on day 4 for intracellular
staining. MIG and MIG.ROR.gamma.t-transduced cells were gated based
on GFP fluorescence.
[0115] FIG. 11C is a graph depicting the percent of Th17 cells
(IL-17.sup.+ IFN.gamma..sup.-) in cultures of MIG-transduced
(shaded bars) or MIG.ROR.gamma.t-transduced (white bars) T cells as
determined by intracellular cytokine staining were normalized to
DMSO-treated cultures. The data are presented as mean values.+-.SD
on duplicate samples. These data are representative of three
similar experiments.
[0116] FIGS. 12A-12B. Halofuginone-enforced Foxp3 expression is not
necessary or sufficient for the inhibition of Th17 differentiation.
FIG. 12A is a set of FACS analyses of CD4.sup.+ CD25.sup.- T cells
activated in the presence or absence of TGF.beta. plus IL-6 which
were transduced with empty (pRV) or FOXP3-expressing (pRV.FOXP3)
retroviruses 12 hours after activation. Intracellular FOXP3 and
cytokine expression was determined 3 days after infection (4 days
after activation). IFN.gamma. and IL-17 expression in pRV- and
pRV.FOXP3-transduced cells was determined by FACS analyses after
gating on GFP.sup.+ cells.
[0117] FIG. 12B is a set of FACS analyses of FACS sorted naive
CD4.sup.+ T cells from wild-type (WT) or Foxp3-deficient (Foxp3 KO)
male mice, treated with DMSO, 10 nM HF, or 10 nM MAZ1310 as
indicated and activated in the absence or presence of TGF.beta.
plus IL-6. T cells were expanded and were restimulated on day 4 for
intracellular cytokine staining. These results are representative
of cells purified from two pairs of WT and Foxp3 KO mice.
[0118] FIG. 13. Halofuginone induces a stress response in
fibroblasts. SV-MES mesangial cells were stimulated for 2 hours
with DMSO, 20 nM MAZ1310, or 20 nM HF. Whole cell lysates were
analyzed for expression of phosphorylated or total eIF2.alpha. by
western blotting. These data represent at least two similar
experiments.
[0119] FIGS. 14A-14D. Amino acid deprivation mimics the effects of
halofuginone on T cell differentiation. FIG. 14A depicts western
blot analyses of CD4.sup.+ CD25.sup.- T cells activated through the
TCR for the indicated times without polarizing cytokines in the
presence or absence of cysteine and methionine (Cys/Met). Whole
cell lysates were prepared and immunoblotting was performed.
[0120] FIG. 14B is a graph depicting results of quantitative
real-time PCR performed on cDNA generated from naive T cells,
either left unstimulated or activated through the TCR for 4 hours
without exogenous cytokines in the presence or absence of Cys/Met
as indicated. Asns, Gpt2 or eIF4Ebp1 mRNA expression was normalized
to Hprt levels, and data are presented as mean expression
values.+-.SD in duplicate samples.
[0121] FIG. 14C is a set of FACS analyses of CD4.sup.+ CD25.sup.- T
cells cultured in complete medium (200 .mu.M Cys/100 .mu.M Met),
medium containing limiting concentrations of Cys/Met (0.1.times.-20
.mu.M Cys/10 .mu.M Met), or complete medium plus 31.25 ng/ml
tunicamycin. Cells were activated through the TCR in the absence or
presence of polarizing cytokines to induce Th1, Th2, iTreg, or Th17
differentiation. Foxp3 intracellular staining was performed on day
3-post activation, and intracellular cytokine expression was
determined on cells restimulated with PMA plus ionomycin on day
4-5.
[0122] FIG. 14D is a set of FACS analyses of CD4.sup.+ CD25.sup.- T
cells labeled with CFSE, cultured in medium containing the
indicated concentrations of Cys/Met and activated in the absence or
presence of TGF.beta. plus IL-6. Cells were expanded until day 4
when CFSE dilution and intracellular cytokine production was
determined on restimulated cells. Cells with equivalent CFSE
fluorescence are gated on as indicated, and intracellular cytokine
expression is shown within each gated population.
[0123] FIG. 15. Genes induced by halofuginone treatment in T cells.
Gene symbols and names of transcripts increased at least 2-fold by
HF treatment at both 3 and 6 hours. Mean fold increases.+-.SD from
triplicate samples of HF-versus MAZ1310-treated T cells is shown at
3 and 6 hours.
[0124] FIGS. 16A-16C. Probe IDs of known stress response genes.
Affymetrix probe IDs and gene names previously identified as ATF4
responsive during tunicamycin-induced ER stress in mouse embryonic
fibroblasts.
[0125] FIG. 17 is a set of FACS analyses showing that depletion of
amino acids or tRNA synthetase inhibition with L-tryptophanol
inhibits Th17 differentiation. T cells were cultured in the
presence of medium containing 0.1.times., 0.2.times., and 1.times.
cysteine and methionine (Cys/Met), medium containing 0.1.times.,
0.2.times., and 1.times. leucine (Leu), or complete medium plus 0.1
mM, 0.2 mM, 0.4 mM, or 0.8 mM tryptophanol. Cells were activated
and cultured under Th17 differentiating conditions, restimulated,
and assayed for IL-17 and IFN.gamma. expression.
[0126] FIGS. 18A-18F. Inhibition of IL-17-associated autoimmune
inflammation in vivo. FIG. 18A is a set of FACS analyses of
CNS-infiltrating mononuclear cells which were isolated from myelin
oligodendrocyte glycoprotein (MOG)-immunized mice during active
disease (day 19-clinical score=2) and stimulated ex vivo with PMA
and ionomycin. Expression of IFN.gamma. (left panel) and IL-17
(right panel) was determined in CD4.sup.+ TCR.beta..sup.+ T
cells.
[0127] FIG. 18B is a graph depicting the effect of systemic HF
administration on adjuvant-driven experimental autoimmune
encephalomyelitis (EAE). Control mice were immunized with an
emulsion of PBS in Complete Freund's Adjuvant (CFA) and treated
with 2 mg HF daily (no MOG+HF (n=10)). Other mice were immunized
with MOG.sub.33-55 in CFA and treated daily with either DMSO
(MOG+DMSO (n=12)), or 2 mg HF (MOG+HF (n=14)). Disease was
monitored daily.
[0128] FIG. 18C is a set of FACS analyses of leukocytes isolated
from CNS tissue of mice with active EAE following transfer of
PLP-specific T cells. Cells were stimulated ex vivo with PMA and
ionomycin and expression of IFN.gamma. (left panel) or IL-17 (right
panel) was determined in PLP-reactive (TCRV.beta.6 gated) CD4.sup.+
T cells by intracellular staining.
[0129] FIG. 18D is a graph depicting the effect of HF in a passive
EAE model. Following the transfer of PLP-specific T cells,
recipient mice were treated daily either with 2 mg HF (n=6) or
vehicle control (n=5) and disease was monitored daily. Data are
shown as mean EAE scores.
[0130] FIG. 18E is a set of FACS analyses of cells from lymph node
or CNS of HF treated animals or control animals in an
adjuvant-driven EAE model. For FIG. 18E, left panels, paraaortic
lymph nodes were harvested from MOG-immunized mice treated with
DMSO or HF after 6 days. Cells were cultured in the absence
(resting-top panels) or presence (P+I-bottom panels) of PMA and
ionomycin and stained for intracellular cytokine expression. For
FIG. 18E, right panels, mononuclear cells were isolated from CNS
tissue of DMSO-treated (clinical score=2) or HF-treated (clinical
score=0) mice 17 days after immunization with MOG. Intracellular
staining was performed on cells following PMA and ionomycin
stimulation as above. Cytokine production is shown in
TCR.beta..sup.+ CD4.sup.+ gated cells and the percentages of
IL-17-expressing cells are indicated.
[0131] FIG. 18F, left panel, depicts western blot analysis of
protein from cells of wild-type mice injected i.p. with vehicle
(DMSO) or 2.5 mg HF. Spleens were harvested 6 hours post injection,
red blood cells were removed by NH.sub.4Cl lysis buffer and
immunoblotting for phosphorylated or total eIF2.alpha. was
performed on whole cell extracts. FIG. 18F, right panel is a graph
depicting levels of AAR-associated gene expression (Asns, Gpt2,
eIF4Ebp1) analyzed by quantitative real-time PCR using cDNA from
splenocytes of mice treated with DMSO or HF as above. Expression of
AAR-associated transcripts were normalized to Hprt levels and data
are presented as mean relative expression from duplicate
samples.+-.SD. All data represent 2-3 similar experiments.
[0132] FIGS. 19A-19B. Regulation of T cell differentiation by
halofuginone during adjuvant-driven EAE. FIG. 19A is a set of FACS
analyses of T cells from paraaortic lymph nodes (day 6). FIG. 19B,
left panel, is a set of FACS analyses of T cells from CNS tissue
(day 18). T cells analyzed in FIGS. 19A and 19B were from control-
or HF-treated mice analyzed for CD44 and CD62L expression following
induction of EAE. CD44 and CD62L expression in shown on cells gated
for CD4 and TCR.beta. expression as shown. FIG. 19B, right panel,
is a graph depicting cell numbers of CNS infiltrates in
DMSO-treated mice (clinical score=2) or HF-treated mice (clinical
score=0), which were determined during active EAE disease (day 18).
Total mononuclear cells, CD4.sup.+ TCR.beta..sup.+ T cells, Th1
cells (IFN.gamma..sup.+) or Th17 cells (IL-17.sup.+) present within
CNS preparations were quantified following FACS analyses and are
displayed as mean numbers.+-.SD. Asterisks indicate statistical
significance. These data are representative of at least 2
independent experiments analyzing at least 3 mice per group.
[0133] FIG. 20. Novel derivatives of HF selectively inhibit Th17
differentiation. Two novel active derivatives of HF (MAZ 1320 and
MAZ1686) selectively inhibit differentiation of Th17 cells without
inhibiting Th differentiation.
[0134] FIG. 21. HF inhibits the incorporation of .sup.14C proline
but not .sup.35S methionine into tRNA. This figure establishes the
specificity of HF as an inhibitor of prolyl-tRNA synthetase (EPRS)
but not a different tRNA synthetase (methionyl tRNA synthetase) in
a crude in vitro translation system.
[0135] FIG. 22. HF inhibits purified EPRS. These data establish
that HF directly inhibits purified mammalian EPRS.
[0136] FIG. 23. A tRNA synthetase inhibitor structurally unrelated
to HF selectively inhibits Th17 differentiation. Borrelidin, a
threonyl tRNA synthetase inhibitor structurally unrelated to HF was
tested for its ability to alter T-cell differentiation. Borrelidin
inhibits Th17 differentiation without affecting Th1, Th2, or Treg
differentiation or cell number, a selectivity identical to that of
HF. tRNA synthetase inhibition therefore provides a general
approach to the selective inhibition of Th17 differentiation
without generalized immunosuppression.
[0137] FIG. 24. HF Has Therapeutic Benefit Even After Symptom
Onset. HF was injected into mice immunized to produce experimental
autoimmune encephalomyelitis (EAE) as described in FIG. 18, with
the exception that HF was not introduced into the animals until day
10 following immunization. These data demonstrate that HF controls
autoimmune inflammation even after inflammatory pathogenesis is
evident, providing a more accurate representation of autoimmune
disease in humans.
[0138] FIG. 25. Transient treatment with HF prevents the
development of EAE symptoms. HF was injected into mice immunized to
produce EAE as described in FIG. 18, with the exception that HF
injection was terminated at day 10 following immunization. These
data demonstrate that HF exerts a protective effect that extends
well beyond the time of treatment, consistent with its proposed
role in preventing the differentiation of pro-inflammatory Th17
cells.
DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION
[0139] The present invention provides novel analogs of
halofuginone. In some embodiments, the inventive compounds include
a quinazolinone moiety connected via a linker to a piperidine,
pyrrolidine, or other heterocycle as shown herein. The compounds of
the present invention are useful in the treatment of disorders
associated with glutamyl-prolyl tRNA synthetase (EPRS) inhibition,
Th17 differentiation, and amino acid starvation response (AAR)
induction, such as chronic inflammation, fibrosis, autoimmune
diseases, scarring, angiogenesis, transplant, implant, or device
rejection, ischemic damage, viral infections, and neurodegenerative
disorders. The compounds may also be used in treating protozoal
infections by inhibiting the prolyl tRNA synthetase of protozoa.
The present invention also provides pharmaceutical compositions and
methods of using the inventive compounds for the treatment of
various diseases (e.g., neurodegenerative diseases), as well as
methods for synthesizing halofuginone analogs.
Compounds
[0140] Compounds of the present invention include quinazolinones,
quinolinones, and analogs and derivatives of halofuginone.
Particularly useful compounds of the present invention include
those with biological activity. The inventive compounds have been
found to have a variety of biological activities. In some
embodiments, the compounds of the invention inhibit tRNA
synthetase. In particular, the compounds of the invention inhibit
glutamyl-prolyl tRNA synthetase (EPRS) (e.g., mammalian EPRS, human
EPRS). In certain embodiments, the compounds of the invention
inhibit non-metazoan prolyl tRNA synthetase (e.g., protozoal prolyl
tRNA synthease). In certain embodiments, the compounds of the
invention suppress the differentiation of a subset of effector
T-cells (i.e., Th17 cells). In certain embodiments, the compounds
of the invention suppress IL-17 production. In certain embodiments,
the compounds of the invention activate the amino acid starvation
response (AAR). In some embodiments, the compounds of the
inventions are useful in the treatment of a disease associated with
IL-17 production, such as arthritis, inflammatory bowel disease,
psoriasis, multiple sclerosis, lupus, asthma, dry eye syndrome, and
other autoimmune and/or inflammatory diseases. In certain other
embodiments, the compounds of the invention suppress pro-fibrotic
gene expression; therefore, they are useful in treating or
preventing fibrosis. In some embodiments, the compounds inhibit
viral gene expression, replication, and maturation. In other
embodiments, the compounds protect organs from stress. In certain
embodiments, the compounds suppress the synthesis of toxic proteins
such as polyglutamine-containing proteins that cause
neurodegenerative diseases such as Huntington's disease. In some
embodiments, the compounds promote autophagy. In certain
embodiments, the compounds inhibit the synthesis of proline-rich
proteins such as collagen. In certain other embodiments, the
compounds inhibit angiogenesis. In certain embodiments, the
compounds are used to treat protozoal infections. In certain
embodiments, the compound have an IC.sub.50 of less than
approximately 10 .mu.M, e.g., less than approximately 1 .mu.M,
e.g., less than approximately 0.1 .mu.M, or e.g., less than
approximately 0.01 .mu.M. The inventive compounds are useful in the
treatment of a variety of diseases. Certain compounds of the
invention are useful in treating inflammatory diseases or
autoimmune diseases, such as inflammatory bowel disease, multiple
sclerosis, rheumatoid arthritis, lupus, psoriasis, scleroderma, or
dry eye syndrome. In certain embodiments, the compounds are useful
in the treatment of cardiovascular diseases, diseases involving
angiogenesis, neurodegenerative diseases, or protein aggregation
disorders. Certain compounds of the invention are also useful as
anti-scarring agents. In some embodiments, inventive compounds are
useful in treating viral infections. In other embodiments, the
compounds are useful in the treatment or prevention of restenosis.
In certain embodiments, an inventive compound is less toxic than
halofuginone, febrifuginone, or other related natural products. In
certain other embodiments, an inventive compound is more potent
than halofuginone, febrifuginone, or other related natural
products.
[0141] It will be noted that structures of some of the compounds of
the invention include asymmetric centers. It is to be understood
accordingly that the isomers arising from such asymmetry are
included within the scope of the invention, unless indicated
otherwise. Such isomers can be obtained in substantially pure form
by purification techniques and/or by stereochemically controlled
synthesis. The compounds of this invention may exist in
stereoisomeric form, and therefore can be produced as individual
stereoisomers or as mixtures thereof.
[0142] In certain embodiments, the invention provides a compound of
formula (I) or a pharmaceutically acceptable salt thereof:
##STR00006##
wherein
[0143] j is an integer between 0 and 10, inclusive;
[0144] p is an integer between 0 and 6, inclusive;
[0145] q is an integer between 0 and 6, inclusive;
[0146] m is 1 or 2;
[0147] v is an integer between 1 and 3, inclusive;
[0148] X is N or CR.sub.X, wherein R.sub.X is hydrogen; halogen;
substituted or unsubstituted aliphatic; substituted or
unsubstituted heteroaliphatic; substituted or unsubstituted aryl;
substituted or unsubstituted heteroaryl; --OR.sub.F; --SR.sub.F;
--N(R.sub.F).sub.2; and --C(R.sub.F).sub.3; wherein each occurrence
of R.sub.F is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0149] each occurrence of Y is independently S, O, N, NR.sub.Y, or
CR.sub.Y, wherein each occurrence of R.sub.Y is independently
hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0150] each occurrence of T and G is independently --S--, --O--,
--NR.sub.E--, or C(R.sub.E).sub.2--, wherein each occurrence of
R.sub.E is independently hydrogen; halogen; substituted or
unsubstituted aliphatic; substituted or unsubstituted
heteroaliphatic; substituted or unsubstituted aryl; substituted or
unsubstituted heteroaryl; --OR.sub.G; --SR.sub.G;
--N(R.sub.G).sub.2; and --C(R.sub.G).sub.3; wherein each occurrence
of R.sub.G is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0151] R.sub.1 is hydrogen; a protecting group; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0152] each occurrence of R.sub.2 is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.B;
--C(.dbd.O)R.sub.B; --CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2;
--CN; --SCN; --SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B;
--NO.sub.2; --N(R.sub.B).sub.2; --NHC(O)R.sub.B; or
--C(R.sub.B).sub.3; wherein each occurrence of R.sub.B is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0153] R.sub.3 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.C; --C(.dbd.O)R.sub.C;
--CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2; --CN; --SCN;
--SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2;
--N(R.sub.C).sub.2; --NHC(O)R.sub.C; or --C(R.sub.C).sub.3; wherein
each occurrence of R.sub.C is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0154] R.sub.4 and R.sub.5 are independently hydrogen; halogen;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.D;
--C(.dbd.O)R.sub.D; --CO.sub.2R.sub.D; --C(.dbd.O)N(R.sub.D).sub.2;
--CN; --SCN; --SR.sub.D; --SOR.sub.D; --SO.sub.2R.sub.D;
--NO.sub.2; --N(R.sub.D).sub.2; --NHC(O)R.sub.D; or
--C(R.sub.D).sub.3; wherein each occurrence of R.sub.D is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; or
[0155] R.sub.4 and R.sub.5 may optionally be taken together to form
.dbd.O, --S, .dbd.NR.sub.D, .dbd.N--OR.sub.D, .dbd.N--NHR.sub.D,
.dbd.N--N(R.sub.D).sub.2, --C(R.sub.D).sub.2; or
[0156] R.sub.4 and R.sub.5 may optionally be taken together with
the intervening atom to form a saturated or unsaturated,
substituted or unsubstituted cyclic or heterocyclic structure; and
R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched aliphatic; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
[0157] In some embodiments, when T and G are each --CH.sub.2--, p
and q are each 1, X is N, and v is 2, R.sub.4 and R.sub.5 are not
taken together to form .dbd.O or .dbd.N--NHR.sub.D.
[0158] In some embodiments, when T and G are each --CH.sub.2--, p
and q are each 1, X is N, m is 2, and v is 1, R.sub.4 and R.sub.5
are not --OH and --H, and R.sub.4 and R.sub.5 are not taken
together to form .dbd.O.
[0159] In certain embodiments, the compound is of the
stereochemistry of formula (Ia):
##STR00007##
[0160] In certain embodiments, the compound is of the
stereochemistry of formula (Ib):
##STR00008##
[0161] In certain embodiments, the compound is of one of the
formulae:
##STR00009##
[0162] In certain embodiments, the compound is of one of the
formulae:
##STR00010##
[0163] In certain embodiments, the compound is of one of the
formulae:
##STR00011##
[0164] In certain embodiments, the compound is of one of the
formulae:
##STR00012##
[0165] In certain embodiments, the compound is of one of the
formulae:
##STR00013##
[0166] In certain embodiments, the compound is of one of the
formulae:
##STR00014##
[0167] In certain embodiments, the compound is of one of the
formulae:
##STR00015##
[0168] In certain embodiments, the compound is of one of the
formulae:
##STR00016##
[0169] In certain embodiments, the compound is of one of the
formulae:
##STR00017##
[0170] In certain embodiments, R.sub.1 of formula I, Ia, or Ib is
hydrogen. In some embodiments, R.sub.1 of formula I, Ia, or Ib is
C.sub.1-6 alkyl. In certain other embodiments, R.sub.1 of formula
I, Ia, or Ib is a suitable amino protecting group, as defined
herein.
[0171] In some embodiments, R.sub.2 of formula I, Ia, or Ib is
hydrogen. In certain embodiments, all instances of R.sub.2 of
formula I, Ia, or Ib are hydrogen. In other embodiments, R.sub.2 of
formula I, Ia, or Ib is a halogen. In certain embodiments, R.sub.2
of formula I, Ia, or Ib is chloro, bromo, or iodo. In certain
embodiments, R.sub.2 of formula I, Ia, or Ib is a hydroxyl or
alkoxyl group. In some embodiments, R.sub.2 of formula I, Ia, or Ib
is a substituted or unsubstituted aliphatic group. In certain
embodiments, R.sub.2 of formula I, Ia, or Ib is a substituted or
unsubstituted aryl group. In certain embodiments, R.sub.2 of
formula I, Ia, or Ib is an amino group. In certain other
embodiments, R.sub.2 of formula I, Ia, or Ib is a cyano group. In
some embodiments, R.sub.2 of formula I, Ia, or Ib is a carboxylic
acid or ester group.
[0172] In certain embodiments, R.sub.3 of formula I, Ia, or Ib is
hydrogen. In certain other embodiments, R.sub.3 of formula I, Ia,
or Ib is hydroxyl. In certain embodiments, R.sub.3 of formulae I,
Ia, or Ib is alkoxy. In certain embodiments, R.sub.3 of formula I,
Ia, or Ib is a protected hydroxyl group. In certain embodiments,
R.sub.3 of formula I, Ia, or Ib is phosphate. In certain
embodiments, R.sub.3 of formula I, Ia, or Ib is sulfate. In certain
other embodiments, R.sub.3 of formula I, Ia, or Ib is acetate
(--OAc). In some embodiments, R.sub.3 of formula I, Ia, or Ib is a
thioxy group. In some embodiments, R.sub.3 of formula I, Ia, or Ib
is an amino group. In some embodiments, R.sub.3 of formula I, Ia,
or Ib is a protected amino group.
[0173] In certain embodiments, R.sub.4 of formula I, Ia, or Ib is
hydrogen. In certain other embodiments, R.sub.4 of formula I, Ia,
or Ib is hydroxyl. In certain embodiments, R.sub.4 of formula I,
Ia, or Ib is alkoxy. In certain embodiments, R.sub.4 of formula I,
Ia, or Ib is a protected hydroxyl group. In certain embodiments,
R.sub.4 of formula I, Ia, or Ib is a substituted or unsubstituted
aliphatic or heteroaliphatic group. In some embodiments, R.sub.4 of
formula I, Ia, or Ib is an amino group. In some embodiments,
R.sub.4 of formula I, Ia, or Ib is a protected amino group.
[0174] In certain embodiments, R.sub.5 of formula I, Ia, or Ib is
hydrogen. In certain other embodiments, R.sub.5 of formula I, Ia,
or Ib is hydroxyl. In certain embodiments, R.sub.5 of formula I,
Ia, or Ib is alkoxy. In certain embodiments, R.sub.5 of formule I,
Ia, or Ib is a protected hydroxyl group. In certain embodiments,
R.sub.5 of formula I, Ia, or Ib is a substituted or unsubstituted
aliphatic or heteroaliphatic group. In some embodiments, R.sub.5 of
formula I, Ia, or Ib is an amino group. In some embodiments,
R.sub.5 of formula I, Ia, or Ib is a protected amino group.
[0175] In certain embodiments, R.sub.4 and R.sub.5 of formula I,
Ia, or Ib are taken together to form .dbd.O. In some embodiments,
R.sub.4 and R.sub.5 of formula I, Ia, or Ib are taken together to
form .dbd.S. In other embodiments, R.sub.4 and R.sub.5 of formula
I, Ia, or Ib are taken together to form .dbd.NR.sub.D, and R.sub.D
is as described herein. In certain embodiments, R.sub.4 and R.sub.5
of formula I, Ia, or Ib are taken together to form
.dbd.N--OR.sub.D. In certain other embodiments, R.sub.4 and R.sub.5
of formula I, Ia, or Ib are taken together to form
.dbd.N--NHR.sub.D. In certain other embodiments, R.sub.4 and
R.sub.5 of formula I, Ia, or Ib are taken together to form
.dbd.N--N(R.sub.D).sub.2. In some embodiments, R.sub.4 and R.sub.5
of formula I, Ia, or Ib are taken together to form
.dbd.C(R.sub.D).sub.2. In certain embodiments, R.sub.4 and R.sub.5
of formula I, Ia, or Ib are taken together to form
.dbd.CH.sub.2.
[0176] In some embodiments, R.sub.4 and R.sub.5 of formula I, Ia,
or Ib are taken together with the intervening carbon to form a
ring. In some embodiments, the ring formed is an oxetane ring. In
certain embodiments, the ring formed is an aziridine ring. In
certain embodiments, the ring formed is an azetidine ring. In
certain embodiments, the ring formed is an epoxide ring. In certain
other embodiments, the ring formed is a cyclopropyl ring. In some
embodiments, the ring formed is a cyclic acetal. In other
embodiments, the ring formed is a 5-membered cyclic acetal. In yet
other embodiments, the ring formed is a 6-membered cyclic
acetal.
[0177] In certain embodiments, R.sub.6 of formula I, Ia, or Ib is
hydrogen. In certain other embodiments, R.sub.6 of formula I, Ia,
or Ib is aliphatic. In certain embodiments, R.sub.6 of formula I,
Ia, or Ib is alkyl.
[0178] In certain embodiments, j of formula I, Ia, or Ib is 0. In
certain embodiments, j of formula I, Ia, or Ib is 1. In certain
embodiments, j of formula I, Ia, or Ib is 2. In certain
embodiments, j of formula I, Ia, or Ib is 3, 4, 5, 6, 7, 8, 9, or
10.
[0179] In some embodiments, m of formula I, Ia, or Ib is 1. In
other embodiments, m of formula I, Ia, or Ib is 2.
[0180] In some embodiments, X of formula I, Ia, or Ib is N. In
other embodiments, X of formula I, Ia, or Ib is CR.sub.x, wherein
R.sub.x is as defined herein. In certain embodiments, X of formula
I, Ia, or Ib is CH.
[0181] In some embodiments, at least one instance of Y of formula
I, Ia, or Ib is CH. In some embodiments, Y of formula I, Ia, or Ib
is CR.sub.Y, where R.sub.Y is as defined herein. In other
embodiments, Y of formula I, Ia, or Ib is S. In certain
embodiments, Y of formula I, Ia, or Ib is N. In certain other
embodiments, Y is NR.sub.Y. In other embodiments, Y of formula I,
Ia, or Ib is O. In yet other embodiments, Y of formula I, Ia, or Ib
is S. In some embodiments, all instances of Y of formula I, Ia, or
Ib are CR.sub.Y. In other embodiments, at least one instance of Y
of formula I, Ia, or Ib is not CR.sub.Y. In yet other embodiments,
at least two instances of Y of formula I, Ia, or Ib are not
CR.sub.Y.
[0182] In some embodiments, R.sub.Y of formula I, Ia, or Ib is
hydrogen. In other embodiments, R.sub.Y of formula I, Ia, or Ib is
a halogen. In certain embodiments, R.sub.Y of formula I, Ia, or Ib
is chloro, bromo, or iodo. In certain embodiments, R.sub.Y of
formula I, Ia, or Ib is a hydroxyl or alkoxy group. In some
embodiments, R.sub.Y of formula I, Ia, or Ib is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sub.Y of
formula I, Ia, or Ib is a substituted or unsubstituted alkyl group.
In certain embodiments, R.sub.Y of formula I, Ia, or Ib is a
substituted or unsubstituted aryl group. In certain embodiments,
R.sub.Y of formula I, Ia, or Ib is a substituted or unsubstituted
alkenyl group. In certain other embodiments, R.sub.Y of formula I,
Ia, or Ib is a substituted or unsubstituted alkynyl group. In some
embodiments, R.sub.Y of formula I, Ia, or Ib is an acyl group. In
other embodiments, R.sub.Y of formula I, Ia, or Ib is an amino
group. In certain embodiments, R.sub.Y of formula I, Ia, or Ib is a
protected amino group.
[0183] In certain embodiments, Y of formula I, Ia, or Ib is
CR.sub.Y and R.sub.Y is hydrogen. In some embodiments, Y of formula
I, Ia, or Ib is CR.sub.Y and R.sub.Y is bromo. In other
embodiments, Y of formula I, Ia, or Ib is CR.sub.Y and R.sub.Y is
chloro. In certain embodiments, Y of formula I, Ia, or Ib is
CR.sub.Y and R.sub.Y is --CN. In certain embodiments, Y of formula
I, Ia, or Ib is CR.sub.Y and R.sub.Y is alkyl. In certain
embodiments, Y of formula I, Ia, or Ib is CR.sub.Y and R.sub.Y is
alkenyl. In certain embodiments, Y of formula I, Ia, or Ib is
CR.sub.Y and R.sub.Y is alkynyl. In certain embodiments, Y of
formula I, Ia, or Ib is CR.sub.Y and R.sub.Y is aryl. In certain
embodiments, Y of formula I, Ia, or Ib is CR.sub.Y and R.sub.Y is
phenyl. In certain embodiments, Y of formula I, Ia, or Ib is
CR.sub.Y and R.sub.Y is benzylic. In certain embodiments, Y of
formula I, Ia, or Ib is CR.sub.Y and R.sub.Y is heteroaryl. In
certain embodiments, Y of formula I, Ia, or Ib is CR.sub.Y and
R.sub.Y is pyridinyl. In certain embodiments, Y of formula I, Ia,
or Ib is CR.sub.Y and R.sub.Y is carbocyclic. In certain
embodiments, Y of formula I, Ia, or Ib is CR.sub.Y and R.sub.Y is
heterocyclic. In certain embodiments, Y of formula I, Ia, or Ib is
CR.sub.Y and R.sub.Y is morpholinyl. In certain embodiments, Y of
formula I, Ia, or Ib is CR.sub.Y and R.sub.Y is piperidinyl.
[0184] In certain embodiments, T of formula I, Ia, or Ib is
--C(R.sub.E).sub.2--, where R.sub.E is as defined herein. In
certain other embodiments, T of formula I, Ia, or Ib is
--CH.sub.2--. In some embodiments, T of formula I, Ia, or Ib is
--O--. In other embodiments, T of formula I, Ia, or Ib is --S--. In
certain embodiments, T of formula I, Ia, or Ib is --NR.sub.E--.
[0185] In certain embodiments, p of formula I, Ia, or Ib is 1. In
some embodiments, p of formula I, Ia, or Ib is 2. In other
embodiments, p of formula I, Ia, or Ib is 3, 4, 5, or 6.
[0186] In certain embodiments, G of formula I, Ia, or Ib is
--C(R.sub.E).sub.2--, where R.sub.E is as defined herein. In
certain other embodiments, G of formula I, Ia, or Ib is
--CH.sub.2--. In some embodiments, G of formula I, Ia, or Ib is
--O--. In other embodiments, G of formula I, Ia, or Ib is --S--. In
certain embodiments, G of formula I, Ia, or Ib is --NR.sub.E--.
[0187] In certain embodiments, q of formula I, Ia, or Ib is 1. In
some embodiments, q of formula I, Ia, or Ib is 2. In other
embodiments, q of formula I, Ia, or Ib is 3, 4, 5, or 6.
[0188] In certain embodiments, at least one of T and G of formula
I, Ia, or Ib is --CH.sub.2--. In other embodiments, both T and G of
formula I, Ia, or Ib are --CH.sub.2--. In certain other
embodiments, only one of T and G of formula I, Ia, or Ib is
--CH.sub.2--. In certain embodiments, at least one of T and G of
formula I, Ia, or Ib is --NR.sub.E--. In certain embodiments, at
least one of T and G of formula I, Ia, or Ib is --O--.
[0189] In certain embodiments, v of formula I, Ia, or Ib is 2 to
form a piperidine ring. In certain embodiments, v of formula I, Ia,
or Ib is 1 to form a pyrrolidine ring. In other embodiments, v of
formula I, Ia, or Ib is 3 to form a homopiperidine ring.
[0190] In some embodiments, compounds of formula I, Ia, or Ib are
of the following formulae:
##STR00018## ##STR00019##
[0191] In some embodiments, compounds of formula I, Ia, or Ib are
of the following formulae:
##STR00020## ##STR00021##
[0192] In some embodiments, compounds of formula I, Ia, or Ib are
of the following formulae:
##STR00022## ##STR00023## ##STR00024##
[0193] In certain embodiments, compounds of formula I, la, or Ib
are of the following formulae:
##STR00025## ##STR00026## ##STR00027##
[0194] In certain embodiments, compounds of formula I or Ia are of
the following formulae:
##STR00028## ##STR00029##
[0195] In certain embodiments, compounds of formula I or Ib are of
the following formulae:
##STR00030## ##STR00031##
[0196] In certain embodiments, the invention provides a compound of
formula (II) or a pharmaceutically acceptable salt thereof:
##STR00032##
wherein
[0197] j is an integer between 0 and 6, inclusive;
[0198] each occurrence of Y is independently S, O, N, or CR.sub.Y,
wherein each occurrence of R.sub.Y is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0199] R.sub.1 is hydrogen; a protecting group; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; provided that R.sub.1 is
not a tert-butoxycarbonyl group;
[0200] each occurrence of R.sub.2 is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.B;
--C(.dbd.O)R.sub.B; --CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2;
--CN; --SCN; --SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B;
--NO.sub.2; --N(R.sub.B).sub.2; --NHC(O)R.sub.B; or
--C(R.sub.B).sub.3; wherein each occurrence of R.sub.B is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0201] R.sub.3 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.C; --C(.dbd.O)R.sub.C;
--CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2; --CN; --SCN;
--SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2;
--N(R.sub.C).sub.2; --NHC(O)R.sub.C; or --C(R.sub.C).sub.3; wherein
each occurrence of R.sub.C is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino, alkylamino, dialkylamino,
heteroaryloxy; or heteroarylthioxy moiety; provided that R.sub.3 is
not --OCH.sub.2Ph;
[0202] R.sub.4 and R.sub.5 are independently hydrogen or --OH;
or
[0203] R.sub.4 and R.sub.5 may be taken together to form .dbd.O;
and
[0204] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
[0205] In some embodiments, R.sub.1 of formula II is not a
tert-butoxycarbonyl group.
[0206] In some embodiments, R.sub.3 of formula II is not
--OCH.sub.2Ph.
[0207] In some embodiments, the compound of formula II is not of
the formula:
##STR00033##
[0208] In certain embodiments, the compound has the stereochemistry
shown in formula (IIa):
##STR00034##
[0209] In certain embodiments, the compound has the stereochemistry
shown in formula (IIb):
##STR00035##
[0210] In certain embodiments, the compound of formula II is of the
formula:
##STR00036##
[0211] In certain embodiments, the compound of formula II is of the
formula:
##STR00037##
[0212] In certain embodiments, the compound of formula II is of the
formula:
##STR00038##
[0213] In certain embodiments, R.sub.1 of formula II, IIa, or IIb
is hydrogen. In certain other embodiments, R.sub.1 of formula II,
IIIa, or IIb is a suitable amino protecting group, as defined
herein.
[0214] In some embodiments, R.sub.2 of formula II, IIIa, or IIb is
hydrogen. In other embodiments, R.sub.2 of formula II, IIIa, or IIb
is a halogen. In certain embodiments, R.sub.2 of formula II, IIIa,
or IIb is chloro, bromo, or iodo. In certain embodiments, R.sub.2
of formula II, IIa, or IIb is a hydroxyl or alkoxyl group. In some
embodiments, R.sub.2 of formula II, IIa, or IIb is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sub.2 of
formula II, IIa, or IIb is a substituted or unsubstituted aryl
group. In certain embodiments, R.sub.2 of formula II, IIa, or IIb
is an amino group. In certain other embodiments, R.sub.2 of formula
II, IIIa, or IIb is a cyano group. In some embodiments, R.sub.2 of
formula II, IIIa, or IIb is a carboxylic acid or ester group.
[0215] In certain embodiments, R.sub.3 of formula II, IIIa, or IIb
is hydrogen. In certain other embodiments, R.sub.3 of formula II,
IIa, or IIb is hydroxyl. In certain embodiments, R.sub.3 of formula
I, IIa, or IIb is alkoxy. In certain embodiments, R.sub.3 of
formula II, IIa, or IIb is a protected hydroxyl group. In certain
embodiments, R.sub.3 of formula II, IIa, or IIb is phosphate. In
certain embodiments, R.sub.3 of formula II, IIa, or IIb is sulfate.
In certain other embodiments, R.sub.3 of formula II, IIa, or IIb is
acetate (--OAc). In some embodiments, R.sub.3 of formula II, IIa,
or IIb is a thioxy group. In some embodiments, R.sub.3 of formula
II, IIa, or IIb is an amino group. In some embodiments, R.sub.3 of
formula II, IIa, or IIb is a protected amino group.
[0216] In certain embodiments, R.sub.4 of formula II, IIa, or IIb
is hydrogen. In certain other embodiments, R.sub.4 of formula II,
IIa, or IIb is hydroxyl. In certain embodiments, R.sub.5 of formula
I, IIa, or IIb is hydrogen. In certain other embodiments, R.sub.5
of formula II, IIa, or IIb is hydroxyl. In certain embodiments,
R.sub.4 and R.sub.5 of formula II, IIa, or IIb are taken together
to form .dbd.O.
[0217] In certain embodiments, R.sub.6 of formula II, IIa, or IIb
is hydrogen. In certain other embodiments, R.sub.6 of formula II,
IIa, or IIb is aliphatic. In certain embodiments, R.sub.6 of
formula I, IIa, or IIb is alkyl.
[0218] In certain embodiments, j of formula II, IIa, or IIb is 0.
In certain embodiments, j is 1. In certain embodiments, j is 2. In
certain embodiments, j is 3, 4, 5, or 6.
[0219] In some embodiments, at least one instance of Y of formula
II, IIa, or IIb is CH. In some embodiments, Y of formula II, IIa,
or IIb is CR.sub.Y, where R.sub.Y is as defined herein. In other
embodiments, Y is S. In certain embodiments, Y is N. In certain
other embodiments, Y is NR.sub.Y. In other embodiments, Y is O. In
some embodiments, all instances of Y are CR.sub.Y. In other
embodiments, at least one instance of Y is not CR.sub.Y. In yet
other embodiments, at least two instances of Y are not
CR.sub.Y.
[0220] In some embodiments, R.sub.Y of formula II, IIa, or IIb is
hydrogen. In other embodiments, R.sub.Y of formula II, IIa, or IIb
is a halogen. In certain embodiments, R.sub.Y of formula I, IIa, or
IIb is chloro, bromo, or iodo. In certain embodiments, R.sub.Y of
formula II, IIa, or IIb is a hydroxyl or alkoxy group. In some
embodiments, R.sub.Y of formula II, IIa, or IIb is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sub.Y of
formula II, IIa, or IIb is a substituted or unsubstituted alkyl
group. In certain embodiments, R.sub.Y of formula II, IIa, or IIb
is a substituted or unsubstituted aryl group. In certain
embodiments, R.sub.Y of formula I, IIa, or Ib is a substituted or
unsubstituted alkenyl group. In certain other embodiments, R.sub.Y
of formula II, IIa, or IIb is a substituted or unsubstituted
alkynyl group. In some embodiments, R.sub.Y of formula II, IIa, or
IIb is an acyl group. In other embodiments, R.sub.Y of formula II,
IIa, or IIb is an amino group. In certain embodiments, R.sub.Y of
formula II, IIa, or IIb is a protected amino group.
[0221] In certain embodiments, Y of formula II, IIa, or IIb is
CR.sub.Y and R.sub.Y is hydrogen. In some embodiments, Y of formula
II, IIa, or IIb is CR.sub.Y and R.sub.Y is bromo. In other
embodiments, Y of formula II, IIa, or IIb is CR.sub.Y and R.sub.Y
is chloro. In certain embodiments, Y of formula II, IIa, or IIb is
CR.sub.Y and R.sub.Y is --CN. In certain embodiments, Y of formula
II, IIa, or IIb is CR.sub.Y and R.sub.Y is alkyl. In certain
embodiments, Y of formula II, IIa, or IIb is CR.sub.Y and R.sub.Y
is alkenyl. In certain embodiments, Y of formula II, IIa, or IIb is
CR.sub.Y and R.sub.Y is alkynyl. In certain embodiments, Y of
formula II, IIa, or IIb is CR.sub.Y and R.sub.Y is aryl. In certain
embodiments, Y of formula II, IIIa, or IIb is CR.sub.Y and R.sub.Y
is phenyl. In certain embodiments, Y of formula II, IIIa, or IIb is
CR.sub.Y and R.sub.Y is benzylic. In certain embodiments, Y of
formula II, IIIa, or IIb is CR.sub.Y and R.sub.Y is heteroaryl. In
certain embodiments, Y of formula II, IIIa, or IIb is CR.sub.Y and
R.sub.Y is pyridinyl. In certain embodiments, Y of formula II,
IIIa, or IIb is CR.sub.Y and R.sub.Y is carbocyclic. In certain
embodiments, Y of formula II, IIa, or IIb is CR.sub.Y and R.sub.Y
is heterocyclic. In certain embodiments, Y of formula II, IIa, or
IIb is CR.sub.Y and R.sub.Y is morpholinyl. In certain embodiments,
Y of formula II, IIa, or IIb is CR.sub.Y and R.sub.Y is
piperidinyl.
[0222] In certain embodiments, compounds of formula II, IIa, or IIb
are of the formula:
##STR00039##
[0223] In certain embodiments, compounds of formula II, IIa, or IIb
are of the formula:
##STR00040##
[0224] In certain embodiments, a compound of formula II or IIa is
of the formula:
##STR00041##
[0225] In certain embodiments, a compound of formula II or IIb is
of the formula:
##STR00042##
[0226] In certain embodiments, compounds of formula II, IIa, or IIb
are of the formula:
##STR00043##
[0227] In certain embodiments, a compound of formula II or IIa is
of the formula:
##STR00044##
[0228] In certain embodiments, a compound of formula II or IIb is
of the formula:
##STR00045##
[0229] In certain embodiments, compounds of formula I, IIa, or IIb
are of the formula:
##STR00046##
[0230] In certain embodiments, a compound of formula II or IIa is
of the formula:
##STR00047##
[0231] In certain embodiments, a compound of formula II or IIb is
of the formula:
##STR00048##
[0232] In certain embodiments, compounds of formula II, IIIa, or
IIb are of the formula:
##STR00049##
[0233] In certain embodiments, a compound of formula II or IIa is
of the formula:
##STR00050##
[0234] In certain embodiments, a compound of formula II or IIb is
of the formula:
##STR00051##
[0235] In certain embodiments, compounds of formula II, IIa, or IIb
are of the formula:
##STR00052##
[0236] In certain embodiments, a compound of formula II or IIa is
of the formula:
##STR00053##
[0237] In certain embodiments, a compound of formula II or IIb is
of the formula:
##STR00054##
[0238] In certain embodiments, compounds of formula II, IIa, or IIb
are of the formula:
##STR00055##
[0239] In certain embodiments, a compound of formula II or IIa is
of the formula:
##STR00056##
[0240] In certain embodiments, a compound of formula II or IIb is
of the formula:
##STR00057##
[0241] In certain embodiments, compounds of formula IIa, IIa, or
IIb are of the formula:
##STR00058##
[0242] In certain embodiments, a compound of formula II or IIa is
of the formula:
##STR00059##
[0243] In certain embodiments, a compound of formula II or IIb is
of the formula:
##STR00060##
[0244] In certain embodiments, compounds of formula II, IIa, or IIb
are of the formula:
##STR00061##
[0245] In certain embodiments, a compound of formula II or IIa is
of the formula:
##STR00062##
[0246] In certain embodiments, a compound of formula II or IIb is
of the formula:
##STR00063##
[0247] In certain embodiments, a compound of formula II, IIa or IIb
is of the formula:
##STR00064##
[0248] In certain embodiments, a compound of formula II or IIa is
of the formula:
##STR00065##
[0249] in certain embodiments, a compound of formula II or IIb is
of the formula:
##STR00066##
[0250] In certain embodiments, compounds of formula II, IIa, or IIb
are of the formula:
##STR00067##
[0251] In certain embodiments, a compound of formula II or IIa is
of the formula:
##STR00068##
[0252] In certain embodiments, a compound of formula II or IIb is
of the formula:
##STR00069##
[0253] In certain embodiments, the invention provides a compound of
formula (III) or a pharmaceutically acceptable salt thereof:
##STR00070##
wherein
[0254] j is an integer between 0 and 8, inclusive;
[0255] each occurrence of Y is independently S, O, N, NR.sub.Y, or
CR.sub.Y, wherein each occurrence of R.sub.Y is independently
hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0256] Z is .dbd.O or .dbd.N--NHR.sub.D, wherein R.sub.D is a
hydrogen; a halogen; a protecting group; an aliphatic moiety; a
heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; alkoxy; aryloxy; alkylthioxy; arylthioxy; amino;
alkylamino; dialkylamino; heteroaryloxy; or heteroarylthioxy
moiety;
[0257] R.sub.1 is hydrogen; a protecting group; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0258] each occurrence of R.sub.2 is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.B;
--C(.dbd.O)R.sub.B; --CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2;
--CN; --SCN; --SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B;
--NO.sub.2; --N(R.sub.B).sub.2; --NHC(O)R.sub.B; or
--C(R.sub.B).sub.3; wherein each occurrence of R.sub.B is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0259] R.sub.3 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.C; --C(.dbd.O)R.sub.C;
--CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2; --CN; --SCN;
--SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2;
--N(R.sub.C).sub.2; --NHC(O)R.sub.C; or --C(R.sub.C).sub.3; wherein
each occurrence of R.sub.C is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; and
[0260] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
[0261] In some embodiments, the compound of formula III is not of
formula
##STR00071##
wherein R.sub.2B is hydrogen, chlorine, or bromine; R.sub.C1 is
hydrogen or methyl; and R.sub.1B is hydrogen, --CO.sub.2CH.sub.3,
or --CO.sub.2CH.sub.2CH--CH.sub.2.
[0262] In certain embodiments, the compound is of the
stereochemistry of formula (IIIa):
##STR00072##
[0263] In certain embodiments, the compound is of the
stereochemistry of formula (IIIb):
##STR00073##
[0264] In certain embodiments, R.sub.1 of formula III, IIIa, or
IIIb is hydrogen. In certain other embodiments, R.sub.1 of formula
III, IIIa, or IIIb is a suitable amino protecting group, as defined
herein.
[0265] In some embodiments, R.sub.2 of formula III, IIIa, or IIIb
is hydrogen. In other embodiments, R.sub.2 of formula III, IIIa, or
IIIb is a halogen. In certain embodiments, R.sub.2 of formula III,
IIIa, or IIIb is chloro, bromo, or iodo. In certain embodiments,
R.sub.2 of formula m, IIIa, or IIIb is a hydroxyl or alkoxyl group.
In some embodiments, R.sub.2 of formula III, IIIa, or IIIb is a
substituted or unsubstituted aliphatic group. In certain
embodiments, R.sub.2 of formula III, IIIa, or IIIb is a substituted
or unsubstituted aryl group. In certain embodiments, R.sub.2 of
formula III, IIIa, or IIIb is an amino group. In certain other
embodiments, R.sub.2 of formula III, IIIa, or IIIb is a cyano
group. In some embodiments, R.sub.2 of formula III, IIIa, or IIIb
is a carboxylic acid or ester group.
[0266] In certain embodiments, R.sub.3 of formula III, IIIa, or
IIIb is hydrogen. In certain other embodiments, R.sub.3 of formula
III, IIIa, or IIIb is hydroxyl. In certain embodiments, R.sub.3 of
formula III, IIIa, or IIIb is alkoxy. In certain embodiments,
R.sub.3 of formula I, IIIa, or IIb is a protected hydroxyl group.
In certain embodiments, R.sub.3 of formula III, IIIa, or IIIb is
phosphate. In certain embodiments, R.sub.3 of formula III, IIIa, or
IIIb is sulfate. In certain other embodiments, R.sub.3 of formula
III, IIIa, or IIIb is acetate (--OAc). In some embodiments, R.sub.3
of formula III, IIIa, or IIIb is a thioxy group. In some
embodiments, R.sub.3 of formula III, IIIa, or IIIb is an amino
group. In some embodiments, R.sub.3 of formula III, IIIa, or IIIb
is a protected amino group.
[0267] In certain embodiments, R.sub.6 of formula I, IIa, or IIIb
is hydrogen. In certain other embodiments, R.sub.6 of formula III,
IIIa, or IIIb is aliphatic. In certain embodiments, R.sub.6 of
formula III, IIIa, or IIIb is alkyl.
[0268] In certain embodiments, Z of formula III, IIIa, or IIIb is
.dbd.O. In certain other embodiments, Z of formula III, IIIa, or
IIIb is .dbd.N--NHR.sub.D, wherein R.sub.D is as defined
herein.
[0269] In certain embodiments, j of formula III, IIIa, or IIIb is
0. In certain embodiments, j is 1. In certain embodiments, j is 2.
In certain embodiments, j is 3, 4, 5, 6, 7, or 8.
[0270] In some embodiments, at least one instance of Y of formula
III, IIIa, or IIIb is CH. In some embodiments, Y of formula III,
IIIa, or IIIb is CR.sub.Y, where R.sub.Y is as defined herein. In
other embodiments, Y is S. In certain embodiments, Y is N. In
certain other embodiments, Y is NR.sub.Y. In other embodiments, Y
is O. In some embodiments, all instances of Y are CR.sub.Y. In
other embodiments, at least one instance of Y is not CR.sub.Y. In
yet other embodiments, at least two instances of Y are not
CR.sub.Y.
[0271] In some embodiments, R.sub.Y of formula III, IIIa, or IIIb
is hydrogen. In other embodiments, R.sub.Y of formula III, IIIa, or
IIIb is a halogen. In certain embodiments, R.sub.Y of formula III,
IIIa, or IIIb is chloro, bromo, or iodo. In certain embodiments,
R.sub.Y of formula III, IIIa, or IIIb is a hydroxyl or alkoxyl
group. In some embodiments, R.sub.Y of formula III, IIIa, or IIIb
is a substituted or unsubstituted aliphatic group. In certain
embodiments, R.sub.Y of formula III, IIIa, or IIIb is a substituted
or unsubstituted alkyl group. In certain embodiments, R.sub.Y of
formula III, IIIa, or IIIb is a substituted or unsubstituted aryl
group. In certain embodiments, R.sub.Y of formula III, IIIa, or
IIIb is a substituted or unsubstituted alkenyl group. In certain
other embodiments, R.sub.Y of formula III, IIIa, or IIIb is a
substituted or unsubstituted alkynyl group. In some embodiments,
R.sub.Y of formula III, IIIa, or IIIb is an acyl group. In other
embodiments, R.sub.Y of formula III, IIIa, or IIIb is an amino
group. In certain embodiments, R.sub.Y of formula III, IIIa, or
IIIb is a protected amino group.
[0272] In certain embodiments, Y of formula III, IIIa, or IIIb is
CR.sub.Y and R.sub.Y is hydrogen. In some embodiments, Y of formula
III, IIIa, or IIIb is CR.sub.Y and R.sub.Y is bromo. In other
embodiments, Y of formula III, IIIa, or IIIb is CR.sub.Y and
R.sub.Y is chloro.
[0273] In some embodiments, compounds of formula III, IIIa, or IIIb
are of the following formulae:
##STR00074##
[0274] In some embodiments, compounds of formula III, IIIa, or IIIb
are of the following formulae:
##STR00075##
[0275] In certain embodiments, the invention provides a compound of
formula (IV) or a pharmaceutically acceptable salt thereof:
##STR00076##
wherein
[0276] j is an integer between 0 and 8, inclusive;
[0277] m is an integer between 1 and 2, inclusive;
[0278] each occurrence of Y is independently S, O, N, NR.sub.Y, or
CR.sub.Y, wherein each occurrence of R.sub.Y is independently
hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R; or --C(R.sub.G).sub.3;
wherein each occurrence of R.sub.G is independently a hydrogen; a
halogen; a protecting group; an aliphatic moiety; a heteroaliphatic
moiety; an acyl moiety; an aryl moiety; a heteroaryl moiety;
alkoxy; aryloxy; alkylthioxy; arylthioxy; amino; alkylamino;
dialkylamino; heteroaryloxy; or heteroarylthioxy moiety;
[0279] Z is .dbd.O or .dbd.N--NHR.sub.D, wherein R.sub.D is a
hydrogen; a halogen; a protecting group; an aliphatic moiety; a
heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; alkoxy; aryloxy; alkylthioxy; arylthioxy; amino;
alkylamino; dialkylamino; heteroaryloxy; or heteroarylthioxy
moiety;
[0280] R.sub.1 is hydrogen; a protecting group; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0281] each occurrence of R.sub.2 is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.B;
--C(.dbd.O)R.sub.B; --CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2;
--CN; --SCN; --SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B;
--NO.sub.2; --N(R.sub.B).sub.2; --NHC(O)R.sub.B; or
--C(R.sub.B).sub.3; wherein each occurrence of R.sub.B is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0282] R.sub.3 is halogen; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched aliphatic; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.C; --CO.sub.2R.sub.C;
--C(.dbd.O)N(R.sub.C).sub.2; --CN; --SCN; --SR.sub.C; --SOR.sub.C;
--SO.sub.2R.sub.C; --NO.sub.2; --N(R.sub.C).sub.2; --NHC(O)R.sub.C;
or --C(R.sub.C).sub.3; wherein each occurrence of R.sub.C is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; and
[0283] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
[0284] In certain embodiments, the compound is of the
stereochemistry of formula (IVa):
##STR00077##
[0285] In certain embodiments, the compound is of the
stereochemistry of formula (IVb):
##STR00078##
[0286] In certain embodiments, R.sub.1 of formula IV, IVa, or IVb
is hydrogen. In certain other embodiments, R.sub.1 of formula IV,
IVa, or IVb is a suitable amino protecting group, as defined
herein.
[0287] In some embodiments, R.sub.2 of formula IV, IVa, or IVb is
hydrogen. In other embodiments, R.sub.2 of formula IV, IVa, or IVb
is a halogen. In certain embodiments, R.sub.2 of formula IV, IVa,
or IVb is chloro, bromo, or iodo. In certain embodiments, R.sub.2
of formula IV, IVa, or IVb is a hydroxyl or alkoxyl group. In some
embodiments, R.sub.2 of formula IV, IVa, or IVb is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sub.2 of
formula IV, IVa, or IVb is a substituted or unsubstituted aryl
group. In certain embodiments, R.sub.2 of formula IV, IVa, or IVb
is an amino group. In certain other embodiments, R.sub.2 of formula
IV, IVa, or IVb is a cyano group. In some embodiments, R.sub.2 of
formula IV, IVa, or IVb is a carboxylic acid or ester group.
[0288] In certain embodiments, R.sub.3 of formula IV, IVa, or IVb
is halogen. In certain other embodiments, R.sub.3 of formula IV,
IVa, or IVb is hydroxymethyl. In certain embodiments, R.sub.3 of
formula IV, IVa, or IVb is alkoxymethyl. In certain embodiments,
R.sub.3 of formula IV, IVa, or IVb is an acyl group. In certain
embodiments, R.sub.3 of formula IV, IVa, or IVb is cyano. In some
embodiments, R.sub.3 of formula IV, IVa, or IVb is a thioxy group.
In some embodiments, R.sub.3 of formula IV, IVa, or IVb is an amino
group. In some embodiments, R.sub.3 of formula IV, IVa, or IVb is a
protected amino group.
[0289] In certain embodiments, R.sub.6 of formula IV, IVa, or IVb
is hydrogen. In certain other embodiments, R.sub.6 of formula IV,
IVa, or IVb is aliphatic. In certain embodiments, R.sub.6 of
formula IV, IVa, or IVb is alkyl.
[0290] In certain embodiments, Z of formula IV, IVa, or IVb is
.dbd.O. In certain other embodiments, Z of formula IV, IVa, or IVb
is .dbd.N--NHR.sub.D, where R.sub.D is as defined herein.
[0291] In certain embodiments, j of formula IV, IVa, or IVb is 0.
In certain embodiments, j is 1. In certain embodiments, j is 2, 3,
4, 5, 6, 7, or 8.
[0292] In some embodiments, m of formula IV, IVa, or IVb is 1. In
other embodiments, m of formula IV, IVa, or IVb is 2.
[0293] In some embodiments, at least one instance of Y of formula
IV, IVa, or IVb is CH. In some embodiments, Y of formula IV, IVa,
or IVb is CR.sub.Y, where R.sub.Y is as defined herein. In other
embodiments, Y is S. In certain embodiments, Y is N. In certain
other embodiments, Y is NR.sub.Y. In other embodiments, Y is O. In
some embodiments, all instances of Y are CR.sub.Y. In other
embodiments, at least one instance of Y is not CR.sub.Y. In yet
other embodiments, at least two instances of Y are not
CR.sub.Y.
[0294] In some embodiments, R.sub.Y of formula IV, IVa, or IVb is
hydrogen. In other embodiments, R.sub.Y of formula IV, IVa, or IVb
is a halogen. In certain embodiments, R.sub.Y of formula IV, IVa,
or IVb is chloro, bromo, or iodo. In certain embodiments, R.sub.Y
of formula IV, IVa, or IVb is a hydroxyl or alkoxyl group. In some
embodiments, R.sub.Y of formula IV, IVa, or IVb is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sub.Y of
formula IV, IVa, or IVb is a substituted or unsubstituted alkyl
group. In certain embodiments, R.sub.Y of formula IV, IVa, or IVb
is a substituted or unsubstituted aryl group. In certain
embodiments, R.sub.Y of formula IV, IVa, or IVb is a substituted or
unsubstituted alkenyl group. In certain other embodiments, R.sub.Y
of formula IV, IVa, or IVb is a substituted or unsubstituted
alkynyl group. In some embodiments, R.sub.Y of formula IV, IVa, or
IVb is an acyl group. In other embodiments, R.sub.Y of formula IV,
IVa, or IVb is an amino group. In certain embodiments, R.sub.Y of
formula IV, IVa, or IVb is a protected amino group.
[0295] In certain embodiments, Y of formula IV, IVa, or IVb is
CR.sub.Y and R.sub.Y is hydrogen. In some embodiments, Y of formula
IV, IVa, or IVb is CR.sub.Y and R.sub.Y is bromo. In other
embodiments, Y of formula IV, IVa, or IVb is CR.sub.Y and R.sub.Y
is chloro.
[0296] In some embodiments, compounds of formula IV, IVa, or IVb
are of the formula:
##STR00079##
[0297] In some embodiments, compounds of formula IV, IVa, or IVb
are of the formula:
##STR00080##
[0298] In some embodiments, a compound of formula IV or IVa is of
the formula:
##STR00081##
[0299] In some embodiments, a compound of formula IV or IVb is of
the formula:
##STR00082##
[0300] In some embodiments, compounds of formula IV, IVa, or IVb
are of the formula:
##STR00083##
[0301] In some embodiments, a compound of formula IV or IVa is of
the formula:
##STR00084##
[0302] In some embodiments, a compound of formula IV or IVb is of
the formula:
##STR00085##
[0303] In certain embodiments, the invention provides a compound of
formula (V) or a pharmaceutically acceptable salt thereof:
##STR00086##
wherein
[0304] j is an integer between 1 and 8, inclusive;
[0305] m is an integer between 1 and 2, inclusive;
[0306] each occurrence of Y is independently S, O, N, NR.sub.Y, or
CR.sub.Y, wherein each occurrence of R.sub.Y is independently
hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0307] Z is .dbd.O or .dbd.N--NHR.sub.D, wherein R.sub.D is a
hydrogen; a halogen; a protecting group; an aliphatic moiety; a
heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; alkoxy; aryloxy; alkylthioxy; arylthioxy; amino;
alkylamino; dialkylamino; heteroaryloxy; or heteroarylthioxy
moiety;
[0308] R.sub.1 is hydrogen; a protecting group; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0309] each occurrence of R.sub.2 is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.B;
--C(.dbd.O)R.sub.B; --CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2;
--CN; --SCN; --SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B;
--NO.sub.2; --N(R.sub.B).sub.2; --NHC(O)R.sub.B; or
--C(R.sub.B).sub.3; wherein each occurrence of R.sub.B is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0310] R.sub.3 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.C; --C(.dbd.O)R.sub.C;
--CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2; --CN; --SCN;
--SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2;
--N(R.sub.C).sub.2; --NHC(O)R.sub.C; or --C(R.sub.C).sub.3; wherein
each occurrence of R.sub.C is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; and
[0311] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
[0312] In certain embodiments, the compound is of the
stereochemistry of formula (Va):
##STR00087##
[0313] In certain embodiments, the compound is of the
stereochemistry of formula (Vb):
##STR00088##
[0314] In certain embodiments, R.sub.1 of formula V, Va, or Vb is
hydrogen. In certain other embodiments, R.sub.1 of formula V, Va,
or Vb is a suitable amino protecting group, as defined herein.
[0315] In other embodiments, R.sub.2 of formula V, Va, or Vb is a
halogen. In certain embodiments, R.sub.2 of formula V, Va, or Vb is
chloro, bromo, or iodo. In certain embodiments, R.sub.2 of formula
V, Va, or Vb is a hydroxyl or alkoxyl group. In some embodiments,
R.sub.2 of formula V, Va, or Vb is a substituted or unsubstituted
aliphatic group. In certain embodiments, R.sub.2 of formula V, Va,
or Vb is a substituted or unsubstituted aryl group. In certain
embodiments, R.sub.2 of formula V, Va, or Vb is an amino group. In
certain other embodiments, R.sub.2 of formula V, Va, or Vb is a
cyano group. In some embodiments, R.sub.2 of formula V, Va, or Vb
is a carboxylic acid or ester group.
[0316] In certain embodiments, R.sub.3 of formula V, Va, or Vb is
hydrogen. In certain other embodiments, R.sub.3 of formula V, Va,
or Vb is hydroxyl. In certain embodiments, R.sub.3 of formula V,
Va, or Vb is alkoxy. In certain embodiments, R.sub.3 of formula V,
Va, or Vb is a protected hydroxyl group. In certain embodiments,
R.sub.3 of formula V, Va, or Vb is phosphate. In certain
embodiments, R.sub.3 of formula V, Va, or Vb is sulfate. In certain
other embodiments, R.sub.3 of formula V, Va, or Vb is acetate
(--OAc). In some embodiments, R.sub.3 of formula V, Va, or Vb is a
thioxy group. In some embodiments, R.sub.3 of formula V, Va, or Vb
is an amino group. In some embodiments, R.sub.3 of formula V, Va,
or Vb is a protected amino group.
[0317] In certain embodiments, R.sub.6 of formula V, Va, or Vb is
hydrogen. In certain other embodiments, R.sub.6 of formula V, Va,
or Vb is aliphatic. In certain embodiments, R.sub.6 of formula V,
Va, or Vb is alkyl.
[0318] In certain embodiments, Z of formula V, Va, or Vb is .dbd.O.
In certain other embodiments, Z of formula V, Va, or Vb is
.dbd.N--NHR.sub.D.
[0319] In certain embodiments, j of formula V, Va, or Vb is 1. In
certain embodiments, j of formula V, Va, or Vb is 2. In certain
embodiments, j is 3, 4, 5, 6, 7, or 8.
[0320] In some embodiments, m of formula V, Va, or Vb is 1. In
other embodiments, m of formula V, Va, or Vb is 2.
[0321] In some embodiments, at least one instance of Y of formula
V, Va, or Vb is CH. In some embodiments, Y of formula V, Va, or Vb
is CR.sub.Y, where R.sub.Y is as defined herein. In other
embodiments, Y is S. In certain embodiments, Y is N. In certain
other embodiments, Y is NR.sub.Y. In other embodiments, Y is O. In
some embodiments, all instances of Y are CR.sub.Y. In other
embodiments, at least one instance of Y is not CR.sub.Y. In yet
other embodiments, at least two instances of Y are not
CR.sub.Y.
[0322] In some embodiments, R.sub.Y of formula V, Va, or Vb is
hydrogen. In other embodiments, R.sub.Y of formula V, Va, or Vb is
a halogen. In certain embodiments, R.sub.Y of formula V, Va, or Vb
is chloro, bromo, or iodo. In certain embodiments, R.sub.Y of
formula V, Va, or Vb is a hydroxyl or alkoxy group. In some
embodiments, R.sub.Y of formula V, Va, or Vb is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sub.Y of
formula V, Va, or Vb is a substituted or unsubstituted alkyl group.
In certain embodiments, R.sub.Y of formula V, Va, or Vb is a
substituted or unsubstituted aryl group. In certain embodiments,
R.sub.Y of formula V, Va, or Vb is a substituted or unsubstituted
alkenyl group. In certain other embodiments, R.sub.Y of formula V,
Va, or Vb is a substituted or unsubstituted alkynyl group. In some
embodiments, R.sub.Y of formula V, Va, or Vb is an acyl group. In
other embodiments, R.sub.Y of formula V, Va, or Vb is an amino
group. In certain embodiments, R.sub.Y of formula V, Va, or Vb is a
protected amino group.
[0323] In certain embodiments, Y of formula V, Va, or Vb is
CR.sub.Y and R.sub.Y is hydrogen. In some embodiments, Y of formula
V, Va, or Vb is CR.sub.Y and R.sub.Y is bromo. In other
embodiments, Y of formula V, Va, or Vb is CR.sub.Y and R.sub.Y is
chloro.
[0324] In some embodiments, compounds of formula V, Va, or Vb are
of the following
##STR00089##
[0325] In some embodiments, compounds of formula V, Va, or Vb are
of the following formula:
##STR00090##
[0326] In some embodiments, compounds of formula V or Va are of the
following formula:
##STR00091##
[0327] In some embodiments, compounds of formula V or Vb are of the
following formula:
##STR00092##
[0328] In certain embodiments, the invention provides a compound of
formula (VI) or a pharmaceutically acceptable salt thereof:
##STR00093##
wherein
[0329] j is an integer between 0 and 8, inclusive;
[0330] each occurrence of Y is S, O, N, or CR.sub.Y, wherein each
occurrence of R.sub.Y is independently hydrogen; halogen; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
aliphatic; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched heteroaliphatic; substituted or
unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.C;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0331] Z is .dbd.O or .dbd.N--NHR.sub.D, wherein R.sub.D is a
hydrogen; a halogen; a protecting group; an aliphatic moiety; a
heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; alkoxy; aryloxy; alkylthioxy; arylthioxy; amino;
alkylamino; dialkylamino; heteroaryloxy; or heteroarylthioxy
moiety;
[0332] R.sub.1 is hydrogen; a protecting group; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0333] each occurrence of R.sub.2 is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.B;
--C(.dbd.O)R.sub.B; --CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2;
--CN; --SCN; --SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B;
--NO.sub.2; --N(R.sub.B).sub.2; --NHC(O)R.sub.B; or
--C(R.sub.B).sub.3; wherein each occurrence of R.sub.B is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0334] R.sub.3 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.C; --C(.dbd.O)R.sub.C;
--CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2; --CN; --SCN;
--SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2;
--N(R.sub.C).sub.2; --NHC(O)R.sub.C; or --C(R.sub.C).sub.3; wherein
each occurrence of R.sub.C is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; and
[0335] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
[0336] In certain embodiments, at least one instance of Y of
formula VI is not CR.sub.Y.
[0337] In certain embodiments, the compound is of the
stereochemistry of formula (VIa):
##STR00094##
[0338] In certain embodiments, the compound is of the
stereochemistry of formula (VIb):
##STR00095##
[0339] In certain embodiments, R.sub.1 of formula VI, VIa, or VIb
is hydrogen. In certain other embodiments, R.sub.1 of formula VI,
VIa, or VIb is a suitable amino protecting group, as defined
herein.
[0340] In some embodiments, R.sub.2 of formula VI, VIa, or VIb is
hydrogen. In other embodiments, R.sub.2 of formula VI, VIa, or VIb
is a halogen. In certain embodiments, R.sub.2 of formula VI, VIa,
or VIb is chloro, bromo, or iodo. In certain embodiments, R.sub.2
of formula VI, VIa, or VIb is a hydroxyl or alkoxyl group. In some
embodiments, R.sub.2 of formula VI, VIa, or VIb is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sub.2 of
formula VI, VIa, or VIb is a substituted or unsubstituted aryl
group. In certain embodiments, R.sub.2 of formula VI, VIa, or VIb
is an amino group. In certain other embodiments, R.sub.2 of formula
VI, VIa, or VIb is a cyano group. In some embodiments, R.sub.2 of
formula VI, VIa, or VIb is a carboxylic acid or ester group.
[0341] In certain embodiments, R.sub.3 of formula VI, VIa, or VIb
is hydrogen. In certain other embodiments, R.sub.3 of formula VI,
VIa, or VIb is hydroxyl. In certain embodiments, R.sub.3 of formula
VI, VIa, or VIb is alkoxy. In certain embodiments, R.sub.3 of
formula VI, VIa, or VIb is a protected hydroxyl group. In certain
embodiments, R.sub.3 of formula VI, VIa, or VIb is phosphate. In
certain embodiments, R.sub.3 of formula VI, VIa, or VIb is sulfate.
In certain other embodiments, R.sub.3 of formula VI, VIa, or VIb is
acetate (--OAc). In some embodiments, R.sub.3 of formula VI, VIa,
or VIb is a thioxy group. In some embodiments, R.sub.3 of formula
VI, VIa, or VIb is an amino group. In some embodiments, R.sub.3 of
formula VI, VIa, or VIb is a protected amino group.
[0342] In certain embodiments, R.sub.6 of formula VI, VIa, or VIb
is hydrogen. In certain other embodiments, R.sub.6 of formula VI,
VIa, or VIb is aliphatic. In certain embodiments, R.sub.6 of
formula VI, VIa, or VIb is alkyl.
[0343] In certain embodiments, Z of formula VI, VIa, or VIb is
.dbd.O. In certain other embodiments, Z of formula VI, VIa, or VIb
is .dbd.N--NHR.sub.D, where R.sub.D is as defined herein.
[0344] In certain embodiments, j of formula VI, VIa, or VIb is
.dbd.O. In certain embodiments, j is 1. In certain embodiments, j
is 2. In certain embodiments, j is 3, 4, 5, 6, 7, or 8.
[0345] In some embodiments, at least one instance of Y of formula
VI, VIa, or VIb is CH. In some embodiments, Y of formula VI, VIa,
or VIb is CR.sub.Y, where R.sub.Y is as defined herein. In certain
embodiments, Y is N. In other embodiments, at least one instance of
Y is not CR.sub.Y. In yet other embodiments, at least two instances
of Y are not CR.sub.Y.
[0346] In some embodiments, R.sub.Y of formula VI, VIa, or VIb is
hydrogen. In other embodiments, R.sub.Y of formula VI, VIa, or VIb
is a halogen. In certain embodiments, R.sub.Y of formula VI, VIa,
or VIb is chloro, bromo, or iodo. In certain embodiments, R.sub.Y
of formula VI or VIa is a hydroxyl or alkoxyl group. In some
embodiments, R.sub.Y of formula VI, VIa, or VIb is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sub.Y of
formula VI, VIa, or VIb is a substituted or unsubstituted alkyl
group. In certain embodiments, R.sub.Y of formula VI, VIa, or VIb
is a substituted or unsubstituted aryl group. In certain
embodiments, R.sub.Y of formula VI, VIa, or VIb is a substituted or
unsubstituted alkenyl group. In certain other embodiments, R.sub.Y
of formula VI, VIa, or VIb is a substituted or unsubstituted
alkynyl group. In some embodiments, R.sub.Y of formula VI, VIa, or
VIb is an acyl group. In other embodiments, R.sub.Y of formula VI,
VIa, or VIb is an amino group. In certain embodiments, R.sub.Y of
formula VI, VIa, or VIb is a protected amino group.
[0347] In certain embodiments, Y of formula VI, VIa, or VIb is
CR.sub.Y and R.sub.Y is hydrogen. In some embodiments, Y of formula
VI, VIa, or VIb is CR.sub.Y and R.sub.Y is bromo. In other
embodiments, Y of formula VI, VIa, or VIb is CR.sub.Y and R.sub.Y
is chloro.
[0348] In some embodiments, compounds of formula VI, VIa, or VIb
are of the following formulae:
##STR00096##
[0349] In some embodiments, compounds of formula VI, VIa or VIb are
the following formulae:
##STR00097##
[0350] In certain embodiments, the invention provides a compound of
formula (VII) or a pharmaceutically acceptable salt thereof:
##STR00098##
[0351] wherein Z is .dbd.0 or .dbd.N--NHR.sub.D, wherein R.sub.D is
a hydrogen; a halogen; a protecting group; an aliphatic moiety; a
heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; alkoxy; aryloxy; alkylthioxy; arylthioxy; amino;
alkylamino; dialkylamino; heteroaryloxy; or heteroarylthioxy
moiety;
[0352] R.sub.1 is hydrogen; a protecting group; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0353] R.sub.3 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.C; --C(.dbd.O)R.sub.C;
--CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2; --CN; --SCN;
--SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2;
--N(R.sub.C).sub.2; --NHC(O)R.sub.C; or --C(R.sub.C).sub.3; wherein
each occurrence of R.sub.C is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0354] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0355] R.sup.A1, R.sup.A2, and R.sup.A3 are independently hydrogen;
halogen; substituted or unsubstituted aliphatic; substituted or
unsubstituted heteroaliphatic; substituted or unsubstituted aryl;
substituted or unsubstituted heteroaryl; --OR.sub.G; --SR.sub.G;
--N(R.sub.G).sub.2; and --C(R.sub.G).sub.3; wherein each occurrence
of R.sub.G is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; and
[0356] R.sup.A4 is halogen; substituted or unsubstituted aliphatic;
substituted or unsubstituted heteroaliphatic; substituted or
unsubstituted aryl; substituted or unsubstituted heteroaryl;
--OR.sub.G; --SR.sub.G; --N(R.sub.G).sub.2; and --C(R.sub.G).sub.3;
wherein each occurrence of R.sub.G is independently a hydrogen; a
halogen; a protecting group; an aliphatic moiety; a heteroaliphatic
moiety; an acyl moiety; an aryl moiety; a heteroaryl moiety;
alkoxy; aryloxy; alkylthioxy; arylthioxy; amino; alkylamino;
dialkylamino; heteroaryloxy; or heteroarylthioxy moiety.
[0357] In some embodiments, the compound of formula VII is not of
the formula:
##STR00099##
[0358] In certain embodiments, the compound is of the
stereochemistry of formula (VIIa):
##STR00100##
[0359] In certain embodiments, the compound is of the
stereochemistry of formula (VIIb):
##STR00101##
[0360] In certain embodiments, R.sub.1 of formula VII, VIIa, or
VIIb is hydrogen. In certain other embodiments, R.sub.1 of formula
VII, VIIa, or VIIb is a suitable amino protecting group, as defined
herein.
[0361] In some embodiments, R.sup.A1 of formula VII, VIIa, or VIIb
is hydrogen. In other embodiments, R.sup.A1 of formula VII, VIIa,
or VIIb is a halogen. In certain embodiments, R.sup.A1 of formula
VII, VIIa, or VIIb is chloro, bromo, or iodo. In certain
embodiments, R.sup.A1 of formula VII, VIIa, or VIIb is a hydroxyl
or alkoxy group. In some embodiments, R.sup.A1 of formula VII,
VIIa, or VIIb is a substituted or unsubstituted aliphatic group. In
certain embodiments, R.sup.A1 of formula VII, VIIa, or VIIb is a
substituted or unsubstituted alkyl group. In certain embodiments,
R.sup.A1 of formula VII, VIIa, or VIIb is a substituted or
unsubstituted aryl group. In certain embodiments, R.sup.A1 of
formula VII, VIIa, or VIIb is a substituted or unsubstituted
alkenyl group. In certain other embodiments, R.sup.A1 of formula
VII, VIIa, or VIIb is a substituted or unsubstituted alkynyl group.
In some embodiments, R.sup.A1 of formula VII, VIIa, or VIIb is an
acyl group. In some embodiments, R.sup.A1 of formula VII, VIIa, or
VIIb is an amino group. In other embodiments, R.sup.A1 of formula
VII, VIIa, or VIIb is a protected amino group.
[0362] In some embodiments, R.sup.A2 of formula VII, VIIa, or VIIb
is hydrogen. In other embodiments, R.sup.A2 of formula VII, VIIa,
or VIIb is a halogen. In certain embodiments, R.sup.A2 of formula
VII, VIIa, or VIIb is chloro, bromo, or iodo. In certain
embodiments, R.sup.A2 of formula VII, VIIa, or VIIb is bromo. In
certain embodiments, R.sup.A2 of formula VII, VIIa, or VIIb is a
hydroxyl or alkoxy group. In some embodiments, R.sup.A2 of formula
VII, VIIa, or VIIb is a substituted or unsubstituted aliphatic
group. In certain embodiments, R.sup.A2 of formula VII, VIIa, or
VIIb is a substituted or unsubstituted alkyl group. In certain
embodiments, R.sup.A2 of formula VII, VIIa, or VIIb is a
substituted or unsubstituted aryl group. In certain embodiments,
R.sup.A2 of formula VII, VIIa, or VIIb is a substituted or
unsubstituted alkenyl group. In certain other embodiments, R.sup.A2
of formula VII, VIIa, or VIIb is a substituted or unsubstituted
alkynyl group. In some embodiments, R.sup.A2 of formula VII, VIIa,
or VIIb is an acyl group. In some embodiments, R.sup.A2 of formula
VII, VIIa, or VIIb is an amino group. In other embodiments,
R.sup.A2 of formula VII, VIIa, or VIIb is a protected amino
group.
[0363] In some embodiments, R.sup.A3 of formula VII, VIIa, or VIIb
is hydrogen. In other embodiments, R.sup.A3 of formula VII, VIIa,
or VIIb is a halogen. In certain embodiments, R.sup.A3 of formula
VII, VIIa, or VIIb is chloro, bromo, or iodo. In certain
embodiments, R.sup.A3 of formula VII, VIIa, or VIIb is chloro. In
certain embodiments, R.sup.A3 of formula VII, VIIa, or VIIb is a
hydroxyl or alkoxy group. In some embodiments, R.sup.A3 of formula
VII, VIIa, or VIIb is a substituted or unsubstituted aliphatic
group. In certain embodiments, R.sup.A3 of formula VII, VIIa, or
VIIb is a substituted or unsubstituted alkyl group. In certain
embodiments, R.sup.A3 of formula VII, VIIa, or VIIb is a
substituted or unsubstituted aryl group. In certain embodiments,
R.sup.A3 of formula VII, VIIa, or VIIb is a substituted or
unsubstituted alkenyl group. In certain other embodiments, R.sup.A3
of formula VII, VIIa, or VIIb is a substituted or unsubstituted
alkynyl group. In some embodiments, R.sup.A3 of formula VII, VIIa,
or VIIb is an acyl group. In some embodiments, R.sup.A3 of formula
VII, VIIa, or VIIb is an amino group. In other embodiments,
R.sup.A3 of formula VII, VIIa, or VIIb is a protected amino
group.
[0364] In some embodiments, R.sup.A4 of formula VII, VIIa, or VIIb
is a halogen. In certain embodiments, R.sup.A4 of formula VII,
VIIa, or VIIb is chloro, bromo, or iodo. In certain embodiments,
R.sup.A4 of formula VII, VIIa, or VIIb is chloro. In certain
embodiments, R.sup.A4 of formula VII, VIIa, or VIIb is bromo. In
certain embodiments, R.sup.A4 of formula VII, VIIa, or VIIb is a
hydroxyl or alkoxy group. In some embodiments, R.sup.A4 of formula
VII, VIIa, or VIIb is a substituted or unsubstituted aliphatic
group. In certain embodiments, R.sup.A4 of formula VII, VIIa, or
VIIb is a substituted or unsubstituted alkyl group. In certain
embodiments, R of formula VII, VIIa, or VIIb is a substituted or
unsubstituted aryl group. In certain embodiments, R.sup.A4 of
formula VII, VIIa, or VIIb is a substituted or unsubstituted
alkenyl group. In certain other embodiments, R.sup.A4 of formula
VII, VIIa, or VIIb is a substituted or unsubstituted alkynyl group.
In some embodiments, R.sup.A4 of formula VII, VIIa, or VIIb is an
acyl group. In some embodiments, R.sup.A4 of formula VII, VIIa, or
VIIb is an amino group. In other embodiments, R.sup.A4 of formula
VII, VIIa, or VIIb is a protected amino group.
[0365] In certain embodiments, R.sub.3 of formula VII, VIIa, or
VIIb is hydrogen. In certain other embodiments, R.sub.3 of formula
VII, VIIa, or VIIb is hydroxyl. In certain embodiments, R.sub.3 of
formula VII, VIIa, or VIIb is alkoxy. In certain embodiments,
R.sub.3 of formula VII, VIIa, or VIIb is a protected hydroxyl
group. In certain embodiments, R.sub.3 of formula VII, VIIa, or
VIIb is phosphate. In certain embodiments, R.sub.3 of formula VII,
VIIa, or VIIb is sulfate. In certain other embodiments, R.sub.3 of
formula VII, VIIa, or VIIb is acetate (--OAc). In some embodiments,
R.sub.3 of formula VII, VIIa, or VIIb is a thioxy group. In some
embodiments, R.sub.3 of formula VII, VIIa, or VIIb is an amino
group. In some embodiments, R.sub.3 of formula VII, VIIa, or VIIb
is a protected amino group.
[0366] In certain embodiments, R.sub.6 of formula VII, VIIa, or
VIIb is hydrogen. In certain other embodiments, R.sub.6 of formula
VII, VIIa, or VIIb is aliphatic. In certain embodiments, R.sub.6 of
formula VII, VIIa, or VIIb is alkyl.
[0367] In certain embodiments, Z of formula VII, VIIa, or VIIb is
.dbd.O. In certain other embodiments, Z of formula VII, VIIa, or
VIIb is .dbd.N--NHR.sub.D, where R.sub.D is as defined herein.
[0368] In certain embodiments, the invention provides a compound of
formula (VIII) or a pharmaceutically acceptable salt thereof:
##STR00102##
[0369] wherein Z is .dbd.O or .dbd.N--NHR.sub.D, wherein R.sub.D is
a hydrogen; a halogen; a protecting group; an aliphatic moiety; a
heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; alkoxy; aryloxy; alkylthioxy; arylthioxy; amino;
alkylamino; dialkylamino; heteroaryloxy; or heteroarylthioxy
moiety;
[0370] R.sub.1 is hydrogen; a protecting group; an aliphatic
moiety; a heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; --SO.sub.2R.sub.A; --S(.dbd.O)R.sub.A;
--C(R.sub.A).sub.2NHC(.dbd.O)R.sub.A;
C(.dbd.O)OCH.sub.2OC(.dbd.O)R.sub.A;
C(.dbd.O)OCH.sub.2OC(.dbd.O)OR.sub.A; or
--C(R.sub.A).sub.2OC(.dbd.O)R.sub.A; wherein each occurrence of
R.sub.A is independently a hydrogen; a halogen; a protecting group;
an aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an
aryl moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0371] R.sub.C is a protecting group; an aliphatic moiety; a
heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; --C(.dbd.O)R.sub.C1; --C(.dbd.O)OR.sub.C1;
--C(.dbd.O)N(R.sub.C1).sub.2; --P(.dbd.O)(OR.sub.C1).sub.2;
--S(.dbd.O)(OR.sub.C1).sub.2; or
--C(R.sub.C1).sub.2OC(.dbd.O)R.sub.C1; wherein each occurrence of
R.sub.C1 is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; provided that R.sub.C is
not methyl, ethyl, or acetyl;
[0372] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; and
[0373] R.sup.A1, R.sup.A2, R.sup.A3, and R.sup.A4 are independently
hydrogen; halogen; substituted or unsubstituted aliphatic;
substituted or unsubstituted heteroaliphatic; substituted or
unsubstituted aryl; substituted or unsubstituted heteroaryl;
--OR.sub.G; --SR.sub.G; --N(R.sub.G).sub.2; and --C(R.sub.G).sub.3;
wherein each occurrence of R.sub.G is independently a hydrogen; a
halogen; a protecting group; an aliphatic moiety; a heteroaliphatic
moiety; an acyl moiety; an aryl moiety; a heteroaryl moiety;
alkoxy; aryloxy; alkylthioxy; arylthioxy; amino; alkylamino;
dialkylamino; heteroaryloxy; or heteroarylthioxy moiety.
[0374] In certain embodiments, the compound is of the
stereochemistry of formula (VIIIa):
##STR00103##
[0375] In certain embodiments, the compound is of the
stereochemistry of formula (VIIIb):
##STR00104##
[0376] In certain embodiments, R.sub.1 of formula VIII, VIIIa, or
VIIIb is hydrogen. In certain other embodiments, R.sub.1 of formula
VIII, VIIIa, or VIIIb is a suitable amino protecting group, as
defined herein.
[0377] In certain embodiments, R.sub.6 of formula VIII, VIIIa, or
VIIIb is hydrogen. In certain other embodiments, R.sub.6 of formula
VIII, VIIIa, or VIIIb is aliphatic. In certain embodiments, R.sub.6
of formula VIII, VIIIa, or VIIIb is alkyl.
[0378] In some embodiments, R.sup.A1 of formula VIII, VIIIa, or
VIIIb is hydrogen. In other embodiments, R.sup.A1 of formula VIII,
VIIIa, or VIIIb is a halogen. In certain embodiments, R.sup.A1 of
formula VIII, VIIIa, or VIIIb is chloro, bromo, or iodo. In certain
embodiments, R.sup.A1 of formula VIII, VIIIa, or VIIIb is a
hydroxyl or alkoxyl group. In some embodiments, R.sup.A1 of formula
VIII, VIIIa, or VIIIb is a substituted or unsubstituted aliphatic
group. In certain embodiments, R.sup.A1 of formula VIII, VIIIa, or
VIIIb is a substituted or unsubstituted alkyl group. In certain
embodiments, R.sup.A1 of formula VIII, VIIIa, or VIIIb is a
substituted or unsubstituted aryl group. In certain embodiments,
R.sup.A1 of formula VIII, VIIIa, or VIIIb is a substituted or
unsubstituted alkenyl group. In certain other embodiments, R.sup.A1
of formula VIII, VIIIa, or VIIIb is a substituted or unsubstituted
alkynyl group. In some embodiments, R.sup.A1 of formula VIII,
VIIIa, or VIIIb is an acyl group. In some embodiments, R.sup.A1 of
formula VIII, VIIIa, or VIIIb is an amino group. In other
embodiments, R.sup.A1 of formula VIII, VIIIa, or VIIIb is a
protected amino group.
[0379] In some embodiments, R.sup.A2 of formula VIII, VIIIa, or
VIIIb is hydrogen. In other embodiments, R.sup.A2 of formula VIII,
VIIIa, or VIIIb is a halogen. In certain embodiments, R.sup.A2 of
formula VIII, VIIIa, or VIIIb is chloro, bromo, or iodo. In certain
embodiments, R.sup.A2 of formula VIII, VIIIa, or VIIIb is bromo. In
certain embodiments, R.sup.A2 of formula VIII, VIIIa, or VIIIb is a
hydroxyl or alkoxyl group. In some embodiments, R.sup.A2 of formula
VIII, VIIIa, or VIIIb is a substituted or unsubstituted aliphatic
group. In certain embodiments, R.sup.A2 of formula VIII, VIIIa, or
VIIIb is a substituted or unsubstituted alkyl group. In certain
embodiments, R.sup.A2 of formula VIII, VIIa, or VIIIb is a
substituted or unsubstituted aryl group. In certain embodiments,
R.sup.A2 of formula VIII, VIIIa, or VIIIb is a substituted or
unsubstituted alkenyl group. In certain other embodiments, R.sup.A2
of formula VIII, VIIIa, or VIIIb is a substituted or unsubstituted
alkynyl group. In some embodiments, R.sup.A2 of formula VIII,
VIIIa, or VIIIb is an acyl group. In some embodiments, R.sup.A2 of
formula VIII, VIIIa, or VIIIb is an amino group. In other
embodiments, R.sup.A2 of formula VIII, Villa, or VIIIb is a
protected amino group.
[0380] In some embodiments, R.sup.A3 of formula VIII, VIIIa, or
VIIIb is hydrogen. In other embodiments, R.sup.A3 of formula VIII,
VIIIa, or VIIIb is a halogen. In certain embodiments, R.sup.A3 of
formula VIII, VIIIa, or VIIIb is chloro, bromo, or iodo. In certain
embodiments, R.sup.A3 of formula VIII, VIIIa, or VIIIb is chloro.
In certain embodiments, R.sup.A3 of formula VIII, VIIIa, or VIIIb
is a hydroxyl or alkoxyl group. In some embodiments, R.sup.A3 of
formula VIII, VIIIa, or VIIIb is a substituted or unsubstituted
aliphatic group. In certain embodiments, R.sup.A3 of formula VIII,
VIIIa, or VIIIb is a substituted or unsubstituted alkyl group. In
certain embodiments, R.sup.A3 of formula VIII, VIIa, or VIIIb is a
substituted or unsubstituted aryl group. In certain embodiments,
R.sup.A3 of formula VIII, VIIIa, or VIIIb is a substituted or
unsubstituted alkenyl group. In certain other embodiments, R.sup.A3
of formula VIII, VIIIa, or VIIIb is a substituted or unsubstituted
alkynyl group. In some embodiments, R.sup.A3 of formula VIII,
VIIIa, or VIIIb is an acyl group. In some embodiments, R.sup.A3 of
formula VIII, VIIIa, or VIIIb is an amino group. In other
embodiments, R.sup.A3 of formula VIII, VIIIa, or VIIIb is a
protected amino group.
[0381] In some embodiments, R.sup.A4 of formula VIII, VIIIa, or
VIIIb is hydrogen. In some embodiments, R.sup.A4 of formula VIII,
VIIIa, or VIIIb is a halogen. In certain embodiments, R.sup.A4 of
formula VIII, VIIIa, or VIIIb is chloro, bromo, or iodo. In certain
embodiments, R.sup.A4 of formula VIII, VIIIa, or VIIIb is chloro.
In certain embodiments, R.sup.A4 of formula VIII, VIIIa, or VIIIb
is bromo. In certain embodiments, R.sup.A4 of formula VIII, VIIIa,
or VIIIb is a hydroxyl or alkoxyl group. In some embodiments,
R.sup.A of formula VIII, VIIIa, or VIIIb is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sup.A4 of
formula VIII, VIIIa, or VIIIb is a substituted or unsubstituted
alkyl group. In certain embodiments, R.sup.A of formula VIII,
VIIIa, or VIIIb is a substituted or unsubstituted aryl group. In
certain embodiments, R.sup.A4 of formula VIII, VIIIa, or VIIIb is a
substituted or unsubstituted alkenyl group. In certain other
embodiments, R.sup.A4 of formula VIII, VIIIa, or VIIIb is a
substituted or unsubstituted alkynyl group. In some embodiments,
R.sup.A4 of formula VIII, VIIIa, or VIIIb is an acyl group. In some
embodiments, R.sup.A4 of formula VIII, VIIIa, or VIIIb is an amino
group. In other embodiments, R.sup.A4 of formula VIII, VIIIa, or
VIIIb is a protected amino group.
[0382] In certain embodiments, R.sub.C of formula VIII, VIIIa, or
VIIIb is a suitable hydroxyl protecting group, as defined herein.
In certain embodiments, R.sub.C of formula VIII, VIIIa, or VIIIb is
an acyl group. In certain embodiments, R.sub.C of formula VIII,
VIIIa, or VIIIb is an ester group. In certain embodiments, R.sub.C
of formula VIII, VIIIa, or VIIIb is an aliphatic group. In certain
embodiments, R.sub.C of formula VIII, VIIIa, or VIIIb is a
heteroaliphatic group. In certain embodiments, --OR.sub.C of
formula VIII, VIIIa, or VIIIb is a phosphate group. In certain
other embodiments, --OR.sub.C of formula VIII, VIIIa, or VIIIb is a
sulfate group.
[0383] In certain embodiments, Z of formula VIII, VIIIa, or VIIIb
is .dbd.O. In certain other embodiments, Z of formula VIII, VIIIa,
or VIIIb is .dbd.N--NHR.sub.D, wherein R.sub.D is as defined
herein.
[0384] In some embodiments, compounds of formula VIII, VIIIa, or
VIIIb are of the following formulae:
##STR00105##
[0385] In some embodiments, compounds of formula VIII, VIIIa, or
VIIIb are of the following formulae:
##STR00106##
[0386] In some embodiments, compounds of formula VIII, VIIIa, or
VIIIb are of the following formulae:
##STR00107##
[0387] In some embodiments, compounds of formula VIII or VIIIa are
of the following formulae:
##STR00108##
[0388] In some embodiments, compounds of formula VIII or VIIIb are
of the following formulae:
##STR00109##
[0389] In some embodiments, compounds of formula VIII, VIIIa, or
VIIIb are of the following formulae:
##STR00110##
[0390] In some embodiments, compounds of formula VIII or VIIIa are
of the following formulae:
##STR00111##
[0391] In some embodiments, compounds of formula VIII or VIIIb are
of the following formulae:
##STR00112##
[0392] In certain embodiments, the invention provides a compound of
formula (IX) or a pharmaceutically acceptable salt thereof:
##STR00113##
[0393] wherein Z is .dbd.O or .dbd.N--NHR.sub.D, wherein R.sub.D is
a hydrogen; a halogen; a protecting group; an aliphatic moiety; a
heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; alkoxy; aryloxy; alkylthioxy; arylthioxy; amino;
alkylamino; dialkylamino; heteroaryloxy; or heteroarylthioxy
moiety;
[0394] R.sub.1 is a protecting group; an aliphatic moiety; a
heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; --SO.sub.2R.sub.A; --S(.dbd.O)R.sub.A;
--C(R.sub.A).sub.2NHC(.dbd.O)R.sub.A;
C(.dbd.O)OCH.sub.2OC(.dbd.O)R.sub.A;
C(.dbd.O)OCH.sub.2OC(.dbd.O)OR.sub.A; or
--C(R.sub.A).sub.2OC(.dbd.O)R.sub.A; wherein each occurrence of
R.sub.A is independently a hydrogen; a halogen; a protecting group;
an aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an
aryl moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; provided that R.sub.1 is not
--CO.sub.2CH.sub.3 or --CO.sub.2CH.sub.2CH--CH.sub.2;
[0395] R.sub.C is hydrogen; a protecting group; an aliphatic
moiety; a heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; --C(.dbd.O)R.sub.C1; --C(.dbd.O)OR.sub.C1;
--C(.dbd.O)N(R.sub.C1).sub.2; --P(.dbd.O)(OR.sub.C1).sub.2;
--S(.dbd.O)(OR.sub.C1).sub.2; or
--C(R.sub.C1).sub.2OC(.dbd.O)R.sub.C1; wherein each occurrence of
R.sub.C1 is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0396] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; and
[0397] R.sup.A1, R.sup.A2, R.sup.A3, and R.sup.A4 are independently
hydrogen; halogen; substituted or unsubstituted aliphatic;
substituted or unsubstituted heteroaliphatic; substituted or
unsubstituted aryl; substituted or unsubstituted heteroaryl;
--OR.sub.G; --SR.sub.G; --N(R.sub.G).sub.2; and --C(R.sub.G).sub.3;
wherein each occurrence of R.sub.G is independently a hydrogen; a
halogen; a protecting group; an aliphatic moiety; a heteroaliphatic
moiety; an acyl moiety; an aryl moiety; a heteroaryl moiety;
alkoxy; aryloxy; alkylthioxy; arylthioxy; amino; alkylamino;
dialkylamino; heteroaryloxy; or heteroarylthioxy moiety.
[0398] In certain embodiments, the compound is of the
stereochemistry of formula (IXa):
##STR00114##
[0399] In certain embodiments, the compound is of the
stereochemistry of formula (IXb):
##STR00115##
[0400] In certain embodiments, R.sub.1 of formula IX, IXa, or IXb
is a suitable amino protecting group, as defined herein. In certain
embodiments, R.sub.1 of formula IX, IXa, or IXb is an acyl group.
In certain embodiments, R.sub.1 of formula IX, IXa, or IXb is an
ester group. In certain embodiments, R.sub.1 of formula IX, IXa, or
IXb is an aliphatic group. In certain embodiments, R.sub.1 of
formula IX, IXa, or IXb is a heteroaliphatic group.
[0401] In certain embodiments, R.sub.6 of formula IX, IXa, or IXb
is hydrogen. In certain other embodiments, R.sub.6 of formula IX,
IXa, or IXb is aliphatic. In certain embodiments, R.sub.6 of
formula IX, IXa, or IXb is alkyl.
[0402] In some embodiments, R.sup.A1 of formula IX, IXa, or IXb is
hydrogen. In other embodiments, R.sup.A1 of formula IX, IXa, or IXb
is a halogen. In certain embodiments, R.sup.A1 of formula IX, IXa,
or IXb is chloro, bromo, or iodo. In certain embodiments, R.sup.A1
of formula IX, IXa, or IXb is a hydroxyl or alkoxy group. In some
embodiments, R.sup.A1 of formula IX, IXa, or IXb is a substituted
or unsubstituted aliphatic group. In certain embodiments, R.sup.A1
of formula IX, IXa, or IXb is a substituted or unsubstituted alkyl
group. In certain embodiments, R.sup.A1 of formula IX, IXa, or IXb
is a substituted or unsubstituted aryl group. In certain
embodiments, R.sup.A1 of formula IX, IXa, or IXb is a substituted
or unsubstituted alkenyl group. In certain other embodiments,
R.sup.A1 of formula IX, IXa, or IXb is a substituted or
unsubstituted alkynyl group. In some embodiments, R.sup.A1 of
formula IX, IXa, or IXb is an acyl group. In some embodiments,
R.sup.A1 of formula IX, IXa, or IXb is an amino group. In other
embodiments, R.sup.A1 of formula IX, IXa, or IXb is a protected
amino group.
[0403] In some embodiments, R.sup.A2 of formula IX, IXa, or IXb is
hydrogen. In other embodiments, R.sup.A2 of formula IX, IXa, or IXb
is a halogen. In certain embodiments, R.sup.A2 of formula IX, IXa,
or IXb is chloro, bromo, or iodo. In certain embodiments, R.sup.A2
of formula IX, IXa, or IXb is bromo. In certain embodiments,
R.sup.A2 of formula IX, IXa, or IXb is a hydroxyl or alkoxy group.
In some embodiments, R.sup.A2 of formula IX, IXa, or IXb is a
substituted or unsubstituted aliphatic group. In certain
embodiments, R.sup.A2 of formula IX, IXa, or IXb is a substituted
or unsubstituted alkyl group. In certain embodiments, R.sup.A2 of
formula IX, IXa, or IXb is a substituted or unsubstituted aryl
group. In certain embodiments, R.sup.A2 of formula IX, IXa, or IXb
is a substituted or unsubstituted alkenyl group. In certain other
embodiments, R.sup.A2 of formula IX, IXa, or IXb is a substituted
or unsubstituted alkynyl group. In some embodiments, R.sup.A2 of
formula IX, IXa, or IXb is an acyl group. In some embodiments,
R.sup.A2 of formula IX, IXa, or IXb is an amino group. In other
embodiments, R.sup.A2 of formula IX, IXa, or IXb is a protected
amino group.
[0404] In some embodiments, R.sup.A of formula IX, IXa, or IXb is
hydrogen. In other embodiments, R.sup.A3 of formula IX, IXa, or IXb
is a halogen. In certain embodiments, R.sup.A3 of formula IX, IXa,
or IXb is chloro, bromo, or iodo. In certain embodiments, R.sup.A3
of formula IX, IXa, or IXb is chloro. In certain embodiments,
R.sup.A of formula IX, IXa, or IXb is a hydroxyl or alkoxy group.
In some embodiments, R.sup.A3 of formula IX, IXa, or IXb is a
substituted or unsubstituted aliphatic group. In certain
embodiments, R.sup.A3 of formula IX, IXa, or IXb is a substituted
or unsubstituted alkyl group. In certain embodiments, R.sup.A3 of
formula IX, IXa, or IXb is a substituted or unsubstituted aryl
group. In certain embodiments, R.sup.A3 of formula IX, IXa, or IXb
is a substituted or unsubstituted alkenyl group. In certain other
embodiments, R.sup.A3 of formula IX, IXa, or IXb is a substituted
or unsubstituted alkynyl group. In some embodiments, R.sup.A3 of
formula IX, IXa, or IXb is an acyl group. In some embodiments,
R.sup.A3 of formula IX, IXa, or IXb is an amino group. In other
embodiments, R.sup.A3 of formula IX, IXa, or IXb is a protected
amino group.
[0405] In some embodiments, R.sup.A4 of formula IX, IXa, or IXb is
hydrogen. In some embodiments, R.sup.A4 of formula IX, IXa, or IXb
is a halogen. In certain embodiments, R.sup.A4 of formula IX, IXa,
or IXb is chloro, bromo, or iodo. In certain embodiments, R.sup.A
of formula IX, IXa, or IXb is chloro. In certain embodiments,
R.sup.A4 of formula IX, IXa, or IXb is bromo. In certain
embodiments, R.sup.A4 of formula IX, IXa, or IXb is a hydroxyl or
alkoxy group. In some embodiments, R.sup.A4 of formula IX, IXa, or
IXb is a substituted or unsubstituted aliphatic group. In certain
embodiments, R.sup.A4 of formula IX, IXa, or IXb is a substituted
or unsubstituted alkyl group. In certain embodiments, R.sup.A4 of
formula IX, IXa, or IXb is a substituted or unsubstituted aryl
group. In certain embodiments, R.sup.A4 of formula IX, IXa, or IXb
is a substituted or unsubstituted alkenyl group. In certain other
embodiments, R.sup.A4 of formula IX, IXa, or IXb is a substituted
or unsubstituted alkynyl group. In some embodiments, R.sup.A4 of
formula IX, IXa, or IXb is an acyl group. In some embodiments,
R.sup.A4 of formula IX, IXa, or IXb is an amino group. In other
embodiments, R.sup.A4 of formula IX, IXa, or IXb is a protected
amino group.
[0406] In certain embodiments, R.sub.C of formula IX, IXa, or IXb
is hydrogen. In certain embodiments, R.sub.C of formula IX, IXa, or
IXb is a suitable hydroxyl protecting group, as defined herein. In
certain embodiments, R.sub.C of formula IX, IXa, or IXb is an acyl
group. In certain embodiments, R.sub.C of formula IX, IXa, or IXb
is an ester group. In certain embodiments, R.sub.C of formula IX,
IXa, or IXb is an aliphatic group. In certain embodiments, R.sub.C
of formula IX, IXa, or IXb is a heteroaliphatic group. In certain
embodiments, --OR.sub.C of formula IX, IXa, or IXb is a phosphate
group. In certain other embodiments, --OR.sub.C of formula IX, IXa,
or IXb is a sulfate group.
[0407] In certain embodiments, Z of formula IX, IXa, or IXb is
.dbd.O. In certain other embodiments, Z of formula IX, IXa, or IXb
is .dbd.N--NHR.sub.D, where R.sub.D is as defined herein.
[0408] In some embodiments, compounds of formula IX, IXa, or IXb
are of the following formulae:
##STR00116##
[0409] In some embodiments, compounds of formula IX, IXa, or IXb
are of the following formulae:
##STR00117##
[0410] In some embodiments, compounds of formula IX or IXa are of
the following formulae:
##STR00118##
[0411] In some embodiments, compounds of formula IX or IXb are of
the following formulae:
##STR00119##
[0412] In certain embodiments, the invention provides a compound of
formula (X) or a pharmaceutically acceptable salt thereof:
##STR00120##
wherein
[0413] j is an integer between 0 and 10, inclusive;
[0414] p is an integer between 0 and 6, inclusive;
[0415] q is an integer between 0 and 6, inclusive;
[0416] m is an integer between 1 and 2, inclusive;
[0417] v is an integer between 1 and 3, inclusive;
[0418] X is N or CR.sub.X, wherein R.sub.X is hydrogen; halogen;
substituted or unsubstituted aliphatic; substituted or
unsubstituted heteroaliphatic; substituted or unsubstituted aryl;
substituted or unsubstituted heteroaryl; --OR.sub.F; --SR.sub.F;
--N(R.sub.F).sub.2; and --C(R.sub.F).sub.3; wherein each occurrence
of R.sub.F is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0419] each occurrence of Y is S, O, N, NR.sub.Y, or CR.sub.Y,
wherein each occurrence of R.sub.Y is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0420] each occurrence of T and G is independently --S--, --O--,
--NR.sub.E--, or C(R.sub.E).sub.2--, wherein each occurrence of
R.sub.E is independently hydrogen; halogen; substituted or
unsubstituted aliphatic; substituted or unsubstituted
heteroaliphatic; substituted or unsubstituted aryl; substituted or
unsubstituted heteroaryl; --OR.sub.G; --SR.sub.G;
--N(R.sub.G).sub.2; and --C(R.sub.G).sub.3; wherein each occurrence
of R.sub.G is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0421] R.sub.Z is --O-- or --S--;
[0422] each occurrence of R.sub.2 is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.B;
--C(.dbd.O)R.sub.B; --CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2;
--CN; --SCN; --SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B;
--NO.sub.2; --N(R.sub.B).sub.2; --NHC(O)R.sub.B; or
--C(R.sub.B).sub.3; wherein each occurrence of R.sub.B is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0423] R.sub.3 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.C; --C(.dbd.O)R.sub.C;
--CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2; --CN; --SCN;
--SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2;
--N(R.sub.C).sub.2; --NHC(O)R.sub.C; or --C(R.sub.C).sub.3; wherein
each occurrence of R.sub.C is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0424] R.sub.4 and R.sub.5 are independently hydrogen; halogen;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.D;
--C(.dbd.O)R.sub.D; --CO.sub.2R.sub.D; --C(.dbd.O)N(R.sub.D).sub.2;
--CN; --SCN; --SR.sub.D; --SOR.sub.D; --SO.sub.2R.sub.D;
--NO.sub.2; --N(R.sub.D).sub.2; --NHC(O)R.sub.D; or
--C(R.sub.D).sub.3; wherein each occurrence of R.sub.D is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; or
[0425] R.sub.4 and R.sub.5 may optionally be taken together to form
.dbd.O, --S, .dbd.NR.sub.D, .dbd.N--OR.sub.D, .dbd.N--NHR.sub.D,
.dbd.N--N(R.sub.D).sub.2, --C(R.sub.D).sub.2; or
[0426] R.sub.4 and R.sub.5 may optionally be taken together with
the intervening atom to form a saturated or unsaturated,
substituted or unsubstituted cyclic or heterocyclic structure;
and
[0427] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
[0428] In certain embodiments, the compound is of the
stereochemistry of formula (Xa):
##STR00121##
[0429] In certain embodiments, the compound is of the
stereochemistry of formula (Xb):
##STR00122##
[0430] In certain embodiments, R.sub.z of formula X, Xa, or Xb is
--O--. In certain other embodiments, R.sub.z of formula X, Xa, or
Xb is --S--.
[0431] In some embodiments, R.sub.2 of formula X, Xa, or Xb is
hydrogen. In other embodiments, R.sub.2 of formula X, Xa, or Xb is
a halogen. In certain embodiments, R.sub.2 of formula X, Xa, or Xb
is chloro, bromo, or iodo. In certain embodiments, R.sub.2 of
formula X, Xa, or Xb is a hydroxyl or alkoxyl group. In some
embodiments, R.sub.2 of formula X, Xa, or Xb is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sub.2 of
formula X, Xa, or Xb is a substituted or unsubstituted aryl group.
In certain embodiments, R.sub.2 of formula X, Xa, or Xb is an amino
group. In certain other embodiments, R.sub.2 of formula X, Xa, or
Xb is a cyano group. In some embodiments, R.sub.2 of formula X, Xa,
or Xb is a carboxylic acid or ester group.
[0432] In certain embodiments, R.sub.3 of formula X, Xa, or Xb is
hydrogen. In certain other embodiments, R.sub.3 of formula X, Xa,
or Xb is hydroxyl. In certain embodiments, R.sub.3 of formula X,
Xa, or Xb is alkoxy. In certain embodiments, R.sub.3 of formula X,
Xa, or Xb is a protected hydroxyl group. In certain embodiments,
R.sub.3 of formula X, Xa, or Xb is phosphate. In certain
embodiments, R.sub.3 of formula X, Xa, or Xb is sulfate. In certain
other embodiments, R.sub.3 of formula X, Xa, or Xb is acetate
(--OAc). In some embodiments, R.sub.3 of formula X, Xa, or Xb is a
thioxy group. In some embodiments, R.sub.3 of formula X, Xa, or Xb
is an amino group. In some embodiments, R.sub.3 of formula X, Xa,
or Xb is a protected amino group.
[0433] In certain embodiments, R.sub.4 of formula X, Xa, or Xb is
hydrogen. In certain other embodiments, R.sub.4 of formula X, Xa,
or Xb is hydroxyl. In certain embodiments, R.sub.4 of formula X,
Xa, or Xb is alkoxy. In certain embodiments, R.sub.4 of formula X,
Xa, or Xb is a protected hydroxyl group. In certain embodiments,
R.sub.4 of formula X, Xa, or Xb is a substituted or unsubstituted
aliphatic or heteroaliphatic group. In some embodiments, R.sub.4 of
formula X, Xa, or Xb is an amino group. In some embodiments,
R.sub.4 of formula X, Xa, or Xb is a protected amino group.
[0434] In certain embodiments, R.sub.5 of formula X, Xa, or Xb is
hydrogen. In certain other embodiments, R.sub.5 of formula X, Xa,
or Xb is hydroxyl. In certain embodiments, R.sub.5 of formula X,
Xa, or Xb is alkoxy. In certain embodiments, R.sub.5 of formula X,
Xa, or Xb is a protected hydroxyl group. In certain embodiments,
R.sub.5 of formula X, Xa, or Xb is a substituted or unsubstituted
aliphatic or heteroaliphatic group. In some embodiments, R.sub.5 of
formula X, Xa, or Xb is an amino group. In some embodiments,
R.sub.5 of formula X, Xa, or Xb is a protected amino group.
[0435] In certain embodiments, R.sub.4 and R.sub.5 of formula X,
Xa, or Xb are taken together to form .dbd.O. In some embodiments,
R.sub.4 and R.sub.5 of formula X, Xa, or Xb are taken together to
form .dbd.S. In other embodiments, R.sub.4 and R.sub.5 of formula
X, Xa, or Xb are taken together to form .dbd.NR.sub.D, and R.sub.D
is as described herein. In certain embodiments, R.sub.4 and R.sub.5
of formula X, Xa, or Xb are taken together to form
.dbd.N--OR.sub.D. In certain other embodiments, R.sub.4 and R.sub.5
of formula X, Xa, or Xb are taken together to form
.dbd.N--NHR.sub.D. In certain other embodiments, R.sub.4 and
R.sub.5 of formula X, Xa, or Xb are taken together to form
.dbd.N--N(R.sub.D).sub.2. In some embodiments, R.sub.4 and R.sub.5
of formula X, Xa, or Xb are taken together to form
.dbd.C(R.sub.D).sub.2. In certain embodiments, R.sub.4 and R.sub.5
of formula X, Xa, or Xb are taken together to form
.dbd.CH.sub.2.
[0436] In some embodiments, R.sub.4 and R.sub.5 of formula X, Xa,
or Xb are taken together with the intervening carbon to form a
ring. In some embodiments, the ring formed is an oxetane ring. In
certain embodiments, the ring formed is an aziridine ring. In
certain embodiments, the ring formed is an azetidine ring. In
certain embodiments, the ring formed is an epoxide ring. In certain
other embodiments, the ring formed is a cyclopropyl ring. In some
embodiments, the ring formed is a cyclic acetal.
[0437] In certain embodiments, R.sub.6 of formula X, Xa, or Xb is
hydrogen. In certain other embodiments, R.sub.6 of formula X, Xa,
or Xb is aliphatic. In certain embodiments, R.sub.6 of formula X,
Xa, or Xb is alkyl.
[0438] In certain embodiments, j of formula X, Xa, or Xb is 0. In
certain embodiments, j is 1. In certain embodiments, j is 2, 3, 4,
5, 6, 7, 8, 9, or 10.
[0439] In some embodiments, m of formula X, Xa, or Xb is 1. In
other embodiments, m of formula X, Xa, or Xb is 2.
[0440] In some embodiments, at least one instance of Y of formula
X, Xa, or Xb is CH. In some embodiments, Y of formula X, Xa, or Xb
is CR.sub.Y, where R.sub.Y is as defined herein. In other
embodiments, Y is S. In certain embodiments, Y is N. In certain
other embodiments, Y is NR.sub.Y. In other embodiments, Y is O. In
some embodiments, all instances of Y are CR.sub.Y. In other
embodiments, at least one instance of Y is not CR.sub.Y. In yet
other embodiments, at least two instances of Y are not
CR.sub.Y.
[0441] In some embodiments, R.sub.Y of formula X, Xa, or Xb is
hydrogen. In other embodiments, R.sub.Y of formula X, Xa, or Xb is
a halogen. In certain embodiments, R.sub.Y of formula X, Xa, or Xb
is chloro, bromo, or iodo. In certain embodiments, R.sub.Y of
formula X, Xa, or Xb is a hydroxyl or alkoxy group. In some
embodiments, R.sub.Y of formula X, Xa, or Xb is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sub.Y of
formula X, Xa, or Xb is a substituted or unsubstituted alkyl group.
In certain embodiments, R.sub.Y of formula X, Xa, or Xb is a
substituted or unsubstituted aryl group. In certain embodiments,
R.sub.Y of formula X, Xa, or Xb is a substituted or unsubstituted
alkenyl group. In certain other embodiments, R.sub.Y of formula X,
Xa, or Xb is a substituted or unsubstituted alkynyl group. In some
embodiments, R.sub.Y of formula X, Xa, or Xb is an acyl group. In
other embodiments, R.sub.Y of formula X, Xa, or Xb is an amino
group. In certain embodiments, R.sub.Y of formula X, Xa, or Xb is a
protected amino group.
[0442] In certain embodiments, Y of formula X, Xa, or Xb is
CR.sub.Y and R.sub.Y is hydrogen. In some embodiments, Y of formula
X, Xa, or Xb is CR.sub.Y and R.sub.Y is bromo. In other
embodiments, Y of formula X, Xa, or Xb is CR.sub.Y and R.sub.Y is
chloro.
[0443] In certain embodiments, T of formula X, Xa, or Xb is
--C(R.sub.E).sub.2--, where R.sub.E is as defined herein. In some
embodiments, T is --O--. In other embodiments, T is --S--. In
certain embodiments, T is --NR.sub.E--. In certain embodiments, p
is 1. In other embodiments, p is 2, 3, 4, 5, or 6.
[0444] In certain embodiments, G of formula X, Xa, or Xb is
--C(R.sub.E).sub.2--, where R.sub.E is as defined herein. In some
embodiments, G is --O--. In other embodiments, G is --S--. In
certain embodiments, G is --NR.sub.E--. In certain embodiments, q
is 1. In other embodiments, q is 2, 3, 4, 5, or 6.
[0445] In some embodiments, compounds of formula X, Xa, or Xb are
of the following formulae:
##STR00123##
[0446] In some embodiments, compounds of formula X, Xa, or Xb are
of the following formulae:
##STR00124##
[0447] In some embodiments, compounds of formula X or Xa are of the
following formulae:
##STR00125##
[0448] In some embodiments, compounds of formula X or Xb are of the
following formulae:
##STR00126##
[0449] In certain embodiments, the invention provides a compound of
formula (XI) or a pharmaceutically acceptable salt thereof:
##STR00127##
wherein
[0450] represents a double or triple bond;
[0451] n is an integer between 0 and 3, inclusive;
[0452] j is an integer between 0 and 8, inclusive;
[0453] R.sub.1 is hydrogen; a protecting group; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0454] each occurrence of R.sub.Y is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0455] each occurrence of R.sub.2 is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.B;
--C(.dbd.O)R.sub.B; --CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2;
--CN; --SCN; --SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B;
--NO.sub.2; --N(R.sub.B).sub.2; --NHC(O)R.sub.B; or
--C(R.sub.B).sub.3; wherein each occurrence of R.sub.B is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0456] R.sub.3 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.C; --C(.dbd.O)R.sub.C;
--CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2; --CN; --SCN;
--SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2;
--N(R.sub.C).sub.2; --NHC(O)R.sub.C; or --C(R.sub.C).sub.3; wherein
each occurrence of R.sub.C is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
[0457] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; and
[0458] R.sub.7 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.C; --C(.dbd.O)R.sub.C;
--CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2; --CN; --SCN;
--SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2;
--N(R.sub.C).sub.2; --NHC(O)R.sub.C; --Si(R.sub.C).sub.3; or
--C(R.sub.C).sub.3; wherein each occurrence of R.sub.C is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety.
[0459] In certain embodiments, the compound is of the
stereochemistry of formula (XIa):
##STR00128##
[0460] In certain embodiments, the compound is of the
stereochemistry of formula (XIb):
##STR00129##
[0461] In certain embodiments, of formula XI, XIa, or XIb
represents a triple bond. In certain other embodiments, of formula
XI, XIa, or XIb represents a double bond.
[0462] In certain embodiments, R.sub.1 of formula XI, XIa, or XIb
is hydrogen. In certain other embodiments, R.sub.1 of formula XI,
XIa, or XIb is a suitable amino protecting group, as defined
herein.
[0463] In some embodiments, R.sub.2 of formula XI, XIa, or XIb is
hydrogen. In other embodiments, R.sub.2 of formula XI, XIa, or XIb
is a halogen. In certain embodiments, R.sub.2 of formula XI, XIa,
or XIb is chloro, bromo, or iodo. In certain embodiments, R.sub.2
of formula XI, XIa, or XIb is a hydroxyl or alkoxyl group. In some
embodiments, R.sub.2 of formula XI, XIa, or XIb is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sub.2 of
formula XI, XIa, or XIb is a substituted or unsubstituted aryl
group. In certain embodiments, R.sub.2 of formula XI, XIa, or XIb
is an amino group. In certain other embodiments, R.sub.2 of formula
XI, XIa, or XIb is a cyano group. In some embodiments, R.sub.2 of
formula XI, XIa, or XIb is a carboxylic acid or ester group.
[0464] In certain embodiments, R.sub.3 of formula XI, XIa, or XIb
is hydrogen. In certain other embodiments, R.sub.3 of formula XI,
XIa, or XIb is hydroxyl. In certain embodiments, R.sub.3 of formula
XI, XIa, or XIb is alkoxy. In certain embodiments, R.sub.3 of
formula XI, XIa, or XIb is a protected hydroxyl group. In certain
embodiments, R.sub.3 of formula XI, XIa, or XIb is phosphate. In
certain embodiments, R.sub.3 of formula XI, XIa, or XIb is sulfate.
In certain other embodiments, R.sub.3 of formula XI, XIa, or XIb is
acetate (--OAc). In some embodiments, R.sub.3 of formula XI, XIa,
or XIb is a thioxy group. In some embodiments, R.sub.3 of formula
XI, XIa, or XIb is an amino group. In some embodiments, R.sub.3 of
formula XI, XIa, or XIb is a protected amino group.
[0465] In certain embodiments, R.sub.6 of formula XI, XIa, or XIb
is hydrogen. In certain other embodiments, R.sub.6 of formula XI,
XIa, or XIb is aliphatic. In certain embodiments, R.sub.6 of
formula XI, XIa, or XIb is alkyl.
[0466] In certain embodiments, j of formula XI, XIa, or XIb is 0.
In certain embodiments, j is 1. In certain embodiments, j is 2. In
certain embodiments, j is 3, 4, 5, 6, 7, or 8.
[0467] In some embodiments, R.sub.7 of formula XI, XIa, or XIb is
hydrogen. In other embodiments, R.sub.7 of formula XI, XIa, or XIb
is a halogen. In some embodiments, R.sub.7 of formula XI, XIa, or
XIb is a silyl group. In certain embodiments, R.sub.7 of formula
XI, XIa, or XIb is a trialkylsilyl group. In some embodiments,
R.sub.7 of formula XI, XIa, or XIb is a substituted or
unsubstituted aliphatic group. In some embodiments, R.sub.7 of
formula XI, XIa, or XIb is a substituted or unsubstituted
heteroaliphatic group. In certain embodiments, R.sub.7 of formula
XI, XIa, or XIb is a substituted or unsubstituted aryl group. In
some embodiments, R.sub.7 of formula XI, XIa, or XIb is a
carboxylic acid or ester group. In other embodiments, R.sub.7 of
formula XI, XIa, or XIb is an amide group.
[0468] In some embodiments, R.sub.Y of formula XI, XIa, or XIb is
hydrogen. In other embodiments, R.sub.Y of formula XI, XIa, or XIb
is a halogen. In certain embodiments, R.sub.Y of formula XI, XIa,
or XIb is chloro, bromo, or iodo. In certain embodiments, R.sub.Y
of formula XI, XIa, or XIb is a hydroxyl or alkoxyl group. In some
embodiments, R.sub.Y of formula XI, XIa, or XIb is a substituted or
unsubstituted aliphatic group. In certain embodiments, R.sub.Y of
formula XI, XIa, or XIb is a substituted or unsubstituted alkyl
group. In certain embodiments, R.sub.Y of formula XI, XIa, or XIb
is a substituted or unsubstituted aryl group. In certain
embodiments, R.sub.Y of formula XI, XIa, or XIb is a substituted or
unsubstituted alkenyl group. In certain other embodiments, R.sub.Y
of formula XI, XIa, or XIb is a substituted or unsubstituted
alkynyl group. In some embodiments, R.sub.Y of formula XI, XIa, or
XIb is an acyl group. In other embodiments, R.sub.Y of formula XI,
XIa, or XIb is an amino group. In certain embodiments, R.sub.Y of
formula XI, XIa, or XIb is a protected amino group.
[0469] In some embodiments, n of formula XI, XIa, or XIb is 0, 1,
2, or 3. In some embodiments, n is 0. In certain embodiments, n is
1. In certain other embodiments, n is 2. In some embodiments, n is
3.
[0470] In some embodiments, compounds of formula XI, XIa, or XIb
are of the following formulae:
##STR00130##
[0471] Exemplary compounds of formula XI or XIa include:
##STR00131##
[0472] Exemplary compounds of formula XI or XIb include:
##STR00132##
[0473] In certain embodiments, the invention provides a compound of
formula (XII) or a pharmaceutically acceptable salt thereof:
##STR00133##
wherein
[0474] Ar represents a substituted or unsubstituted aryl or
heteroaryl group;
n is an integer between 0 and 3, inclusive;
[0475] j is an integer between 0 and 8, inclusive;
[0476] R.sub.1 is hydrogen; a protecting group; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety;
each occurrence of R.sub.Y is independently hydrogen; halogen;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.G).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.G).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; each occurrence of R.sub.2 is
independently hydrogen; halogen; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched aliphatic; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.B; --C(.dbd.O)R.sub.B;
--CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2; --CN; --SCN;
--SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B; --NO.sub.2;
--N(R.sub.B).sub.2; --NHC(O)R.sub.B; or --C(R.sub.B).sub.3; wherein
each occurrence of R.sub.B is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; R.sub.3 is hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.C;
--C(.dbd.O)R.sub.C; --CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2;
--CN; --SCN; --SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C;
--NO.sub.2; --N(R.sub.C).sub.2; --NHC(O)R.sub.C; or
--C(R.sub.C).sub.3; wherein each occurrence of R.sub.C is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety; and R.sub.6 is hydrogen; halogen; cyclic
or acyclic, substituted or unsubstituted, branched or unbranched
aliphatic; cyclic or acyclic, substituted or unsubstituted,
branched or unbranched heteroaliphatic; substituted or
unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.K;
--C(.dbd.O)R.sub.K; --CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2;
--CN; --SCN; --SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K;
--NO.sub.2; --N(R.sub.K).sub.2; --NHC(O)R.sub.K; or
--C(R.sub.K).sub.3; wherein each occurrence of R.sub.K is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety.
[0477] In certain embodiments, the compound is of the
stereochemistry of formula (XIa):
##STR00134##
[0478] In certain embodiments, the compound is of the
stereochemistry of formula (XIIb):
##STR00135##
[0479] In certain embodiments, R.sub.1 of formula XII, XIIa, or
XIIb is hydrogen. In certain other embodiments, R.sub.1 of formula
XII, XIIa, or XIIb is a suitable amino protecting group, as defined
herein.
[0480] In some embodiments, R.sub.2 of formula XII, XIIa, or XIIb
is hydrogen. In other embodiments, R.sub.2 of formula XII, XIIa, or
XIIb is a halogen. In certain embodiments, R.sub.2 of formula XII,
XIIa, or XIIb is chloro, bromo, or iodo. In certain embodiments,
R.sub.2 of formula XII, XIIa, or XIIb is a hydroxyl or alkoxyl
group. In some embodiments, R.sub.2 of formula XII, XIIa, or XIIb
is a substituted or unsubstituted aliphatic group. In certain
embodiments, R.sub.2 of formula XII, XIIa, or XIIb is a substituted
or unsubstituted aryl group. In certain embodiments, R.sub.2 of
formula XII, XIIa, or XIIb is an amino group. In certain other
embodiments, R.sub.2 of formula XII, XIIa, or XIIb is a cyano
group. In some embodiments, R.sub.2 of formula XII, XIIa, or XIIb
is a carboxylic acid or ester group.
[0481] In certain embodiments, R.sub.3 of formula XII, XIIa, or
XIIb is hydrogen. In certain other embodiments, R.sub.3 of formula
XII, XIIa, or XIIb is hydroxyl. In certain embodiments, R.sub.3 of
formula XII, XIIa, or XIb is alkoxy. In certain embodiments,
R.sub.3 of formula XII, XIIa, or XIIb is a protected hydroxyl
group. In certain embodiments, R.sub.3 of formula XII, XIIa, or
XIIb is phosphate. In certain embodiments, R.sub.3 of formula XII,
XIIa, or XIIb is sulfate. In certain other embodiments, R.sub.3 of
formula XII, XIIa, or XIIb is acetate (--OAc). In some embodiments,
R.sub.3 of formula XII, XIIa, or XIIb is a thioxy group. In some
embodiments, R.sub.3 of formula XII, XIIa, or XIIb is an amino
group. In some embodiments, R.sub.3 of formula XII, XIIa, or XIIb
is a protected amino group.
[0482] In certain embodiments, R.sub.6 of formula XII, XIIa, or
XIIb is hydrogen. In certain other embodiments, R.sub.6 of formula
XII, XIIa, or XIIb is aliphatic. In certain embodiments, R.sub.6 of
formula XII, XIIa, or XIIb is alkyl.
[0483] In certain embodiments, j of formula XII, XIIa, or XIIb is
0. In certain embodiments, j is 1. In certain embodiments, j is 2.
In certain embodiments, j is 3, 4, 5, 6, 7, or 8.
[0484] In some embodiments, R.sub.Y of formula XI, XIa, or XIIb is
hydrogen. In other embodiments, R.sub.Y of formula XII, XIIa, or
XIIb is a halogen. In certain embodiments, R.sub.Y of formula XII,
XIIa, or XIIb is chloro, bromo, or iodo. In certain embodiments,
R.sub.Y of formula XII, XIIa, or XIIb is a hydroxyl or alkoxyl
group. In some embodiments, R.sub.Y of formula XII, XIIa, or XIIb
is a substituted or unsubstituted aliphatic group. In certain
embodiments, R.sub.Y of formula XII, XIIa, or XIIb is a substituted
or unsubstituted alkyl group. In certain embodiments, R.sub.Y of
formula XII, XIIa, or XIIb is a substituted or unsubstituted aryl
group. In certain embodiments, R.sub.Y of formula XII, XIIa, or
XIIb is a substituted or unsubstituted alkenyl group. In certain
other embodiments, R.sub.Y of formula XII, XIIa, or XIIb is a
substituted or unsubstituted alkynyl group. In some embodiments,
R.sub.Y of formula XII, XIIa, or XIIb is an acyl group. In other
embodiments, R.sub.Y of formula XII, XIIa, or XIIb is an amino
group. In certain embodiments, R.sub.Y of formula XII, XIIa, or
XIIb is a protected amino group.
[0485] In certain embodiments, Y of formula XI, XIIa, or XIIb is
CR.sub.Y and R.sub.Y is hydrogen. In some embodiments, Y of formula
XII, XIIa, or XIIb is CR.sub.Y and R.sub.Y is bromo. In other
embodiments, Y of formula XII, XIIa, or XIIb is CR.sub.Y and
R.sub.Y is chloro.
[0486] In some embodiments, Ar of formula XII, XIIa, or XIIb is
phenyl. In other embodiments, Ar of formula XII, XIIa, or XIIb is a
nitrogen-containing heterocycle. In some embodiments, Ar of formula
XII, XIIa, or XIIb is an oxygen-containing heterocycle. In certain
embodiments, Ar of formula XII, XIIa, or XIIb is pyridyl. In some
embodiments, Ar of formula XI, XIIa, or XIIb is pyrimidinyl. In
some embodiments, Ar of formula XI, XIIa, or XIIb is triazolyl. In
certain embodiments, Ar of formula XII, XIIa, or XIIb is thiazolyl.
In some embodiments, Ar of formula XII, XIIa, or XIIb is furyl. In
other embodiments, Ar of formula XII, XIIa, or XIIb is thienyl.
[0487] In some embodiments, n of formula XII, XIa, or XIIb is 0, 1,
2, or 3. In some embodiments, n is 0. In certain embodiments, n is
1. In certain other embodiments, n is 2. In some embodiments, n is
3.
[0488] In certain embodiments, the invention provides a compound of
formula (XIII) or a pharmaceutically acceptable salt thereof:
##STR00136##
wherein
[0489] j is an integer between 0 and 6, inclusive;
[0490] each occurrence of Y is independently S, O, N, or CR.sub.Y,
wherein each occurrence of R.sub.Y is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.G;
--C(.dbd.O)R.sub.G; --CO.sub.2R.sub.G; --C(.dbd.O)N(R.sub.G).sub.2;
--CN; --SCN; --SR.sub.G; --SOR.sub.G; --SO.sub.2R.sub.G;
--NO.sub.2; --N(R.sub.C).sub.2; --NHC(O)R.sub.G; or
--C(R.sub.C).sub.3; wherein each occurrence of R.sub.G is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0491] R.sub.1 is hydrogen; a protecting group; cyclic or acyclic,
substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or
unbranched heteroaliphatic; substituted or unsubstituted, branched
or unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A).sub.2; or --C(R.sub.A).sub.3; wherein each
occurrence of R.sub.A is independently a hydrogen; a halogen; a
protecting group; an aliphatic moiety; a heteroaliphatic moiety; an
acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; provided that R.sub.1 is
not a tert-butoxycarbonyl group;
[0492] each occurrence of R.sub.2 is independently hydrogen;
halogen; cyclic or acyclic, substituted or unsubstituted, branched
or unbranched aliphatic; cyclic or acyclic, substituted or
unsubstituted, branched or unbranched heteroaliphatic; substituted
or unsubstituted, branched or unbranched acyl; substituted or
unsubstituted, branched or unbranched aryl; substituted or
unsubstituted, branched or unbranched heteroaryl; --OR.sub.B;
--C(.dbd.O)R.sub.B; --CO.sub.2R.sub.B; --C(.dbd.O)N(R.sub.B).sub.2;
--CN; --SCN; --SR.sub.B; --SOR.sub.B; --SO.sub.2R.sub.B;
--NO.sub.2; --N(R.sub.B).sub.2; --NHC(O)R.sub.B; or
--C(R.sub.B).sub.3; wherein each occurrence of R.sub.B is
independently a hydrogen; a halogen; a protecting group; an
aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an aryl
moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0493] R.sub.3 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.C; --C(.dbd.O)R.sub.C;
--CO.sub.2R.sub.C; --C(.dbd.O)N(R.sub.C).sub.2; --CN; --SCN;
--SR.sub.C; --SOR.sub.C; --SO.sub.2R.sub.C; --NO.sub.2;
--N(R.sub.C).sub.2; --NHC(O)R.sub.C; or --C(R.sub.C).sub.3; wherein
each occurrence of R.sub.C is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino, alkylamino, dialkylamino,
heteroaryloxy; or heteroarylthioxy moiety; provided that R.sub.3 is
not --OCH.sub.2Ph;
[0494] R.sub.4 and R.sub.5 are independently hydrogen or --OH;
or
[0495] R.sub.4 and R.sub.5 may be taken together to form .dbd.O;
and
[0496] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
[0497] In certain embodiments, the compound has the stereochemistry
shown in formula (XIIIa):
##STR00137##
[0498] In certain embodiments, the compound has the stereochemistry
shown in formula (XIIIb):
##STR00138##
[0499] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00139##
[0500] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00140##
[0501] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00141##
[0502] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00142##
[0503] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00143##
[0504] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00144##
[0505] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00145##
[0506] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00146##
[0507] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00147##
[0508] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00148##
[0509] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00149##
[0510] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00150##
[0511] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00151##
[0512] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00152##
[0513] In certain embodiments, the compound of formula XIII is of
the formula:
##STR00153##
[0514] In certain embodiments, R.sub.1 of formula XIII, XIIIa, or
XIIIb is hydrogen. In certain other embodiments, R.sub.1 of formula
XIII, XIIIa, or XIIIb is a suitable amino protecting group, as
defined herein.
[0515] In some embodiments, R.sub.2 of formula XIII, XIIIa, or
XIIIb is hydrogen. In other embodiments, R.sub.2 of formula XIII,
XIIIa, or XIIIb is a halogen. In certain embodiments, R.sub.2 of
formula XIII, XIIIa, or XIIIb is chloro, bromo, or iodo. In certain
embodiments, R.sub.2 of formula XIII, XIIa, or XIIIb is a hydroxyl
or alkoxyl group. In some embodiments, R.sub.2 of formula XIII,
XIIa, or XIIIb is a substituted or unsubstituted aliphatic group.
In certain embodiments, R.sub.2 of formula XIII, XIIIa, or XIIIb is
a substituted or unsubstituted aryl group. In certain embodiments,
R.sub.2 of formula XIII, XIIIa, or XIIIb is an amino group. In
certain other embodiments, R.sub.2 of formula XIII, XIIIa, or XIIIb
is a cyano group. In some embodiments, R.sub.2 of formula XIII,
XIIIa, or XIIIb is a carboxylic acid or ester group.
[0516] In certain embodiments, R.sub.3 of formula XIII, XIIIa, or
XIIIb is hydrogen. In certain other embodiments, R.sub.3 of formula
XIII, XIIIa, or XIIIb is hydroxyl. In certain embodiments, R.sub.3
of formula XIII, XIIIa, or XIIIb is alkoxy. In certain embodiments,
R.sub.3 of formula XIII, XIIIa, or XIIIb is a protected hydroxyl
group. In certain embodiments, R.sub.3 of formula XIII, XIIIa, or
XIIIb is phosphate. In certain embodiments, R.sub.3 of formula
XIII, XIIIa, or XIIIb is sulfate. In certain other embodiments,
R.sub.3 of formula XIII, XIIIa, or XIIIb is acetate (--OAc). In
some embodiments, R.sub.3 of formula XIII, XIIIa, or XIIIb is a
thioxy group. In some embodiments, R.sub.3 of formula XIII, XIIIa,
or XIIIb is an amino group. In some embodiments, R.sub.3 of formula
XIII, XIIIa, or XIIIb is a protected amino group.
[0517] In certain embodiments, R.sub.4 of formula XIII, XIIIa, or
XIIIb is hydrogen. In certain other embodiments, R.sub.4 of formula
XIII, XIIIa, or XIIIb is hydroxyl. In certain embodiments, R.sub.5
of formula XIII, XIIIa, or XIIIb is hydrogen. In certain other
embodiments, R.sub.5 of formula XIII, XIIIa, or XIIIb is hydroxyl.
In certain embodiments, R.sub.4 and R.sub.5 of formula XIII, XIIIa,
or XIIIb are taken together to form .dbd.O.
[0518] In certain embodiments, R.sub.6 of formula XIII, XIIIa, or
XIIIb is hydrogen. In certain other embodiments, R.sub.6 of formula
XIII, XIIIa, or XIIIb is aliphatic. In certain embodiments, R.sub.6
of formula XIII, XIIIa, or XIIIb is alkyl.
[0519] In certain embodiments, j of formula XIII, XIIIa, or XIIIb
is 0. In certain embodiments, j is 1. In certain embodiments, j is
2. In certain embodiments, j is 3, 4, 5, or 6.
[0520] In some embodiments, at least one instance of Y of formula
XIII, XIIIa, or XIIIb is CH. In some embodiments, Y of formula
XIII, XIIIa, or XIIIb is CR.sub.Y, where R.sub.Y is as defined
herein. In other embodiments, Y is S. In certain embodiments, Y is
N. In certain other embodiments, Y is NR.sub.Y. In other
embodiments, Y is O. In some embodiments, all instances of Y are
CR.sub.Y. In other embodiments, at least one instance of Y is not
CR.sub.Y. In yet other embodiments, at least two instances of Y are
not CR.sub.Y.
[0521] In some embodiments, R.sub.Y of formula XIII, XIIIa, or
XIIIb is hydrogen. In other embodiments, R.sub.Y of formula XIII,
XIIIa, or XIIIb is a halogen. In certain embodiments, R.sub.Y of
formula XIII, XIIIa, or XIIIb is chloro, bromo, or iodo. In certain
embodiments, R.sub.Y of formula XIII, XIIIa, or XIIIb is a hydroxyl
or alkoxy group. In some embodiments, R.sub.Y of formula XIII,
XIIIa, or XIIIb is a substituted or unsubstituted aliphatic group.
In certain embodiments, R.sub.Y of formula XIII, XIIIa, or XIIIb is
a substituted or unsubstituted alkyl group. In certain embodiments,
R.sub.Y of formula XIII, XIIIa, or XIIIb is a substituted or
unsubstituted aryl group. In certain embodiments, R.sub.Y of
formula XIII, XIIIa, or XIIIb is a substituted or unsubstituted
alkenyl group. In certain other embodiments, R.sub.Y of formula
XIII, XIIIa, or XIIIb is a substituted or unsubstituted alkynyl
group. In some embodiments, R.sub.Y of formula XIII, XIIIa, or
XIIIb is an acyl group. In other embodiments, R.sub.Y of formula
XIII, XIIIa, or XIIIb is an amino group. In certain embodiments,
R.sub.Y of formula XIII, XIIIa, or XIIIb is a protected amino
group.
[0522] In certain embodiments, Y of formula XIII, XIIIa, or XIIIb
is CR.sub.Y and R.sub.Y is hydrogen. In some embodiments, Y of
formula XIII, XIIIa, or XIIIb is CR.sub.Y and R.sub.Y is bromo. In
other embodiments, Y of formula XIII, XIIIa, or XIIIb is CR.sub.Y
and R.sub.Y is chloro. In certain embodiments, Y of formula XIII,
XIIIa, or XIIIb is CR.sub.Y and R.sub.Y is --CN. In certain
embodiments, Y of formula XIII, XIIIa, or XIIIb is CR.sub.Y and
R.sub.Y is alkyl. In certain embodiments, Y of formula XIII, XIIIa,
or XIIIb is CR.sub.Y and R.sub.Y is alkenyl. In certain
embodiments, Y of formula XIII, XIIIa, or XIIIb is CR.sub.Y and
R.sub.Y is alkynyl. In certain embodiments, Y of formula XIII,
XIIIa, or XIIIb is CR.sub.Y and R.sub.Y is aryl. In certain
embodiments, Y of formula XIII, XIIIa, or XIIIb is CR.sub.Y and
R.sub.Y is phenyl. In certain embodiments, Y of formula XIII,
XIIIa, or XIIIb is CR.sub.Y and R.sub.Y is benzylic. In certain
embodiments, Y of formula XIII, XIIIa, or XIIIb is CR.sub.Y and
R.sub.Y is heteroaryl. In certain embodiments, Y of formula XIII,
XIIIa, or XIIIb is CR.sub.Y and R.sub.Y is pyridinyl. In certain
embodiments, Y of formula XIII, XIIIa, or XIIIb is CR.sub.Y and
R.sub.Y is carbocyclic. In certain embodiments, Y of formula XIII,
XIIIa, or XIIIb is CR.sub.Y and R.sub.Y is heterocyclic. In certain
embodiments, Y of formula XIII, XIIIa, or XIIIb is CR.sub.Y and
R.sub.Y is morpholinyl. In certain embodiments, Y of formula XIII,
XIIIa, or XIIIb is CR.sub.Y and R.sub.Y is piperidinyl.
[0523] In some embodiments, compounds of formula XIII, XIIIa, or
XIIIb are of the following formulae:
##STR00154##
[0524] In some embodiments, compounds of formula XIII, XIIIa, or
XIIIb are of the following formulae:
##STR00155##
[0525] In some embodiments, compounds of formula XIII or XIIIa are
of the following formulae:
##STR00156##
[0526] In some embodiments, compounds of formula XIII or XIIIb are
of the following formulae:
##STR00157##
[0527] In some embodiments, compounds of formula XIII or XIIIa are
of the following formulae:
##STR00158##
[0528] In some embodiments, compounds of formula XIII or XIIIb are
of the following formulae:
##STR00159##
[0529] In certain embodiments, the invention provides a compound of
formula (XIV) or a pharmaceutically acceptable salt thereof:
##STR00160##
[0530] wherein Z is .dbd.O or .dbd.N--NHR.sub.D, wherein R.sub.D is
a hydrogen; a halogen; a protecting group; an aliphatic moiety; a
heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; alkoxy; aryloxy; alkylthioxy; arylthioxy; amino;
alkylamino; dialkylamino; heteroaryloxy; or heteroarylthioxy
moiety;
[0531] R.sub.1 is hydrogen; a protecting group; an aliphatic
moiety; a heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; --C(.dbd.O)R.sub.A; --C(.dbd.O)OR.sub.A;
--C(.dbd.O)N(R.sub.A2; --SO.sub.2R.sub.A; --S(.dbd.O)R.sub.A;
--C(R.sub.A).sub.2NHC(.dbd.O)R.sub.A;
C(.dbd.O)OCH.sub.2OC(.dbd.O)R.sub.A;
C(.dbd.O)OCH.sub.2OC(.dbd.O)OR.sub.A; or
--C(R.sub.A).sub.2OC(.dbd.O)R.sub.A; wherein each occurrence of
R.sub.A is independently a hydrogen; a halogen; a protecting group;
an aliphatic moiety; a heteroaliphatic moiety; an acyl moiety; an
aryl moiety; a heteroaryl moiety; alkoxy; aryloxy; alkylthioxy;
arylthioxy; amino; alkylamino; dialkylamino; heteroaryloxy; or
heteroarylthioxy moiety;
[0532] R.sub.C is a protecting group; an aliphatic moiety; a
heteroaliphatic moiety; an acyl moiety; an aryl moiety; a
heteroaryl moiety; --C(.dbd.O)R.sub.C1; --C(.dbd.O)OR.sub.C1;
--C(.dbd.O)N(R.sub.C1).sub.2; --P(.dbd.O)(OR.sub.C1).sub.2;
--S(.dbd.O)(OR.sub.C1).sub.2; or
--C(R.sub.C1).sub.2OC(.dbd.O)R.sub.C1; wherein each occurrence of
R.sub.C1 is independently a hydrogen; a halogen; a protecting
group; an aliphatic moiety; a heteroaliphatic moiety; an acyl
moiety; an aryl moiety; a heteroaryl moiety; alkoxy; aryloxy;
alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; provided that R.sub.C is
not methyl, ethyl, or acetyl;
[0533] R.sub.6 is hydrogen; halogen; cyclic or acyclic, substituted
or unsubstituted, branched or unbranched aliphatic; cyclic or
acyclic, substituted or unsubstituted, branched or unbranched
heteroaliphatic; substituted or unsubstituted, branched or
unbranched acyl; substituted or unsubstituted, branched or
unbranched aryl; substituted or unsubstituted, branched or
unbranched heteroaryl; --OR.sub.K; --C(.dbd.O)R.sub.K;
--CO.sub.2R.sub.K; --C(.dbd.O)N(R.sub.K).sub.2; --CN; --SCN;
--SR.sub.K; --SOR.sub.K; --SO.sub.2R.sub.K; --NO.sub.2;
--N(R.sub.K).sub.2; --NHC(O)R.sub.K; or --C(R.sub.K).sub.3; wherein
each occurrence of R.sub.K is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety; and
[0534] R.sup.A1, R.sup.A2, R.sup.A3, and R.sup.A4 are independently
hydrogen; halogen; substituted or unsubstituted aliphatic;
substituted or unsubstituted heteroaliphatic; substituted or
unsubstituted aryl; substituted or unsubstituted heteroaryl; --OR;
--SR.sub.G; --N(R.sub.G).sub.2; and --C(R.sub.G).sub.3; wherein
each occurrence of R.sub.G is independently a hydrogen; a halogen;
a protecting group; an aliphatic moiety; a heteroaliphatic moiety;
an acyl moiety; an aryl moiety; a heteroaryl moiety; alkoxy;
aryloxy; alkylthioxy; arylthioxy; amino; alkylamino; dialkylamino;
heteroaryloxy; or heteroarylthioxy moiety.
[0535] In certain embodiments, the compound is of the
stereochemistry of formula (XIVa):
##STR00161##
[0536] In certain embodiments, the compound is of the
stereochemistry of formula (XIVb):
##STR00162##
[0537] In certain embodiments, R.sub.1 of formula XIV, XIVa, or
XIVb is hydrogen. In certain other embodiments, R.sub.1 of formula
XIV, XIVa, or XIVb is a suitable amino protecting group, as defined
herein.
[0538] In certain embodiments, R.sub.6 of formula XIV, XIVa, or
XIVb is hydrogen. In certain other embodiments, R.sub.6 of formula
XIV, XIVa, or XIVb is aliphatic. In certain embodiments, R.sub.6 of
formula XIV, XIVa, or XIVb is alkyl.
[0539] In some embodiments, R.sup.A1 of formula XIV, XIVa, or XIVb
is hydrogen. In other embodiments, R.sup.A1 of formula XIV, XIVa,
or XIVb is a halogen. In certain embodiments, R.sup.A1 of formula
XIV, XIVa, or XIVb is chloro, bromo, or iodo. In certain
embodiments, R.sup.A1 of formula XIV, XIVa, or XIVb is a hydroxyl
or alkoxyl group. In some embodiments, R.sup.A1 of formula XIV,
XIVa, or XIVb is a substituted or unsubstituted aliphatic group. In
certain embodiments, R.sup.A1 of formula XIV, XIVa, or XIVb is a
substituted or unsubstituted alkyl group. In certain embodiments,
R.sup.A1 of formula XIV, XIVa, or XIVb is a substituted or
unsubstituted aryl group. In certain embodiments, R.sup.A1 of
formula XIV, XIVa, or XIVb is a substituted or unsubstituted
alkenyl group. In certain other embodiments, R.sup.A1 of formula
XIV, XIVa, or XIVb is a substituted or unsubstituted alkynyl group.
In some embodiments, R.sup.A1 of formula XIV, XIVa, or XIVb is an
acyl group. In some embodiments, R.sup.A1 of formula XIV, XIVa, or
XIVb is an amino group. In other embodiments, R.sup.A1 of formula
XIV, XIVa, or XIVb is a protected amino group.
[0540] In some embodiments, R.sup.A2 of formula XIV, XIVa, or XIVb
is hydrogen. In other embodiments, R.sup.A2 of formula XIV, XIVa,
or XIVb is a halogen. In certain embodiments, R.sup.A2 of formula
XIV, XIVa, or XIVb is chloro, bromo, or iodo. In certain
embodiments, R.sup.A2 of formula XIV, XIVa, or XIVb is bromo. In
certain embodiments, R.sup.A2 of formula XIV, XIVa, or XIVb is a
hydroxyl or alkoxyl group. In some embodiments, R.sup.A2 of formula
XIV, XIVa, or XIVb is a substituted or unsubstituted aliphatic
group. In certain embodiments, R.sup.A2 of formula XIV, XIVa, or
XIVb is a substituted or unsubstituted alkyl group. In certain
embodiments, R.sup.A2 of formula XIV, XIVa, or XIVb is a
substituted or unsubstituted aryl group. In certain embodiments,
R.sup.A2 of formula XIV, XIVa, or XIVb is a substituted or
unsubstituted alkenyl group. In certain other embodiments, R.sup.A2
of formula XIV, XIVa, or XIVb is a substituted or unsubstituted
alkynyl group. In some embodiments, R.sup.A2 of formula XIV, XIVa,
or XIVb is an acyl group. In some embodiments, R.sup.A2 of formula
XIV, XIVa, or XIVb is an amino group. In other embodiments,
R.sup.A2 of formula XIV, XIVa, or XIVb is a protected amino
group.
[0541] In some embodiments, R.sup.A3 of formula XIV, XIVa, or XIVb
is hydrogen. In other embodiments, R.sup.A3 of formula XIV, XIVa,
or XIVb is a halogen. In certain embodiments, R.sup.A3 of formula
XIV, XIVa, or XIVb is chloro, bromo, or iodo. In certain
embodiments, R.sup.A3 of formula XIV, XIVa, or XIVb is chloro. In
certain embodiments, R.sup.A3 of formula XIV, XIVa, or XIVb is a
hydroxyl or alkoxyl group. In some embodiments, R.sup.A3 of formula
XIV, XIVa, or XIVb is a substituted or unsubstituted aliphatic
group. In certain embodiments, R.sup.A3 of formula XIV, XIVa, or
XIVb is a substituted or unsubstituted alkyl group. In certain
embodiments, R.sup.A3 of formula XIV, XIVa, or XIVb is a
substituted or unsubstituted aryl group. In certain embodiments,
R.sup.A3 of formula XIV, XIVa, or XIVb is a substituted or
unsubstituted alkenyl group. In certain other embodiments, R.sup.A3
of formula XIV, XIVa, or XIVb is a substituted or unsubstituted
alkynyl group. In some embodiments, R.sup.A3 of formula XIV, XIVa,
or XIVb is an acyl group. In some embodiments, R.sup.A3 of formula
XIV, XIVa, or XIVb is an amino group. In other embodiments,
R.sup.A3 of formula XIV, XIVa, or XIVb is a protected amino
group.
[0542] In some embodiments, R.sup.A4 of formula XIV, XIVa, or XIVb
is hydrogen. In some embodiments, R.sup.A4 of formula XIV, XIVa, or
XIVb is a halogen. In certain embodiments, R.sup.A4 of formula XIV,
XIVa, or XIVb is chloro, bromo, or iodo. In certain embodiments,
R.sup.A4 of formula XIV, XIVa, or XIVb is chloro. In certain
embodiments, R.sup.A4 of formula XIV, XIVa, or XIVb is bromo. In
certain embodiments, R.sup.A4 of formula XIV, XIVa, or XIVb is a
hydroxyl or alkoxyl group. In some embodiments, R.sup.A4 of formula
XIV, XIVa, or XIVb is a substituted or unsubstituted aliphatic
group. In certain embodiments, R.sup.A4 of formula XIV, XIVa, or
XIVb is a substituted or unsubstituted alkyl group. In certain
embodiments, R.sup.A4 of formula XIV, XIVa, or XIVb is a
substituted or unsubstituted aryl group. In certain embodiments,
R.sup.A4 of formula XIV, XIVa, or XIVb is a substituted or
unsubstituted alkenyl group. In certain other embodiments, R.sup.A4
of formula XIV, XIVa, or XIVb is a substituted or unsubstituted
alkynyl group. In some embodiments, R.sup.A4 of formula XIV, XIVa,
or XIVb is an acyl group. In some embodiments, R.sup.A4 of formula
XIV, XIVa, or XIVb is an amino group. In other embodiments,
R.sup.A4 of formula XIV, XIVa, or XIVb is a protected amino
group.
[0543] In certain embodiments, R.sub.C of formula XIV, XIVa, or
XIVb is a suitable hydroxyl protecting group, as defined herein. In
certain embodiments, R.sub.C of formula XIV, XIVa, or XIVb is an
acyl group. In certain embodiments, R.sub.C of formula XIV, XIVa,
or XIVb is an ester group. In certain embodiments, R.sub.C of
formula XIV, XIVa, or XIVb is an aliphatic group. In certain
embodiments, R.sub.C of formula XIV, XIVa, or XIVb is a
heteroaliphatic group. In certain embodiments, --OR.sub.C of
formula XIV, XIVa, or XIVb is a phosphate group. In certain other
embodiments, --OR.sub.C of formula XIV, XIVa, or XIVb is a sulfate
group.
[0544] In certain embodiments, Z of formula XIV, XIVa, or XIVb is
.dbd.O. In certain other embodiments, Z of formula XIV, XIVa, or
XIVb is .dbd.N--NHR.sub.D, wherein R.sub.D is as defined
herein.
[0545] In some embodiments, compounds of formula XIV, XIVa, or XIVb
are of the following formulae:
##STR00163##
[0546] In some embodiments, compounds of formula XIV, XIVa, or XIVb
are of the following formulae:
##STR00164##
[0547] In some embodiments, compounds of formula XIV or XIVa are of
the following formulae:
##STR00165##
[0548] In some embodiments, compounds of formula XIV or XIVb are of
the following formulae:
##STR00166##
[0549] In some embodiments, compounds of formula XIV, XIVa, or XIVb
are of the following formulae:
##STR00167##
[0550] In some embodiments, compounds of formula XIV or XIVa are of
the following formulae:
##STR00168##
[0551] In some embodiments, compounds of formula XIV or XIVb are of
the following formulae:
##STR00169##
[0552] In some embodiments, compounds of formula XIV, XIVa, or XIVb
are of the following formulae:
##STR00170##
[0553] In some embodiments, compounds of formula XIV or XIVa are of
the following formulae:
##STR00171##
[0554] In some embodiments, compounds of formula XIV or XIVb are of
the following formulae:
##STR00172##
[0555] Exemplary compounds of the invention include:
##STR00173## ##STR00174## ##STR00175##
[0556] Exemplary compounds of the invention include:
##STR00176## ##STR00177## ##STR00178##
Synthesis of Inventive Compounds
[0557] The compounds provided by the present invention may be
prepared via any synthetic route known to one of skill in the art.
For example, the compounds may be prepared from simple,
commercially available starting materials, of the compounds may be
prepared semi-synthetically using more complex starting materials
such as halofuginone or febrifugine. The inventive compounds may be
prepared from literature procedures. U.S. Pat. No. 4,762,838; U.S.
Patent Application Publication 2008/0188498; Emmanuvel et al., "A
concise enantioselective synthesis of (+)-febrifugine" Tetrahedron:
Asymmetry 20(1):84-88, 2009; Ooi et al., "A Concise
Enantioselective Synthesis of Antimalarial Febrifugine Alkaloids"
Organic Letters 3(6):953-955, 2001; Ashoorzadeh et al., "Synthetic
evaluation of an enantiopure tetrahydropyridine N-oxide. Synthesis
of (+)-febrifugine" Tetrahedron 65(24):4671-4680, 2009; Sukemoto et
al., "Concise asymmetric synthesis of (+)-febrifugine utilizing
trans-selective intramolecular conjugate addition" Synthesis
(19):3081-3087, 2008; Kikuchi et al., "Exploration of a New Type of
Antimalarial Compounds Based on Febrifugine" Journal of Medicinal
Chemistry 49(15):4698-4706, 2006; Takaya et al., "New Type of
Febrifugine Analogues, Bearing a Quinolizidine Moiety, Show Potent
Antimalarial Activity against Plasmodium Malaria Parasite" Journal
of Medicinal Chemistry 42(16):3163-3166, 1999. The inventive
compounds may also be prepared from commercially available starting
materials using the following synthetic schemes. The following are
only meant to exemplify the routes available to a synthetic organic
chemist for preparing the inventive compounds. As would be readily
apparent to one of skill in this art, these exemplary schemes may
be modified to use different starting materials, reagents, and/or
reaction conditions.
[0558] Various groups such as cyano, alkenyl, alkynyl, aryl, and
amino may be substituted for the bromine of halofuginone or
derivatives thereof as shown in the scheme below:
##STR00179##
[0559] The starting material S1 can be prepared according to
literature procedures for closely related compounds. See, e.g.,
Kikuchi et al., J. Med. Chem. 49(15):4698-4706, 2006; Ooi et al.,
Org. Lett. 3:953-55, 2001; JP2002201192; CN1583729. The starting
material S2 can be prepared according to literature procedures for
closely related compounds. See, e.g., U.S. Patent Application
Publication US 2008/0188498.
[0560] In certain embodiments, inventive compounds are prepared
wherein the phenyl moiety of the bicyclic quinazolinone ring system
of halofuginone is replaced with a heteroaryl moiety such as
thiophenyl, furanyl, pyridinyl, or pyrimidinyl. Such compounds may
be prepared by the scheme:
##STR00180##
[0561] The starting material S3 can be prepared according to U.S.
Patent Application Publication US 2008/0188498. Corresponding
heterocycles for preparing such inventive compounds are either
commercially available or can be prepared following literature
procedures. Song, Heterocyclic Communications 13(1):33-34, 2007;
Peng et al., Journal of Combinatorial Chemistry 9(3):431-436, 2007;
Robba et al., Bulletin de la Societe Chimique de France 3-4(Pt.
2):587-91, 1975; Reigan et al., Bioorganic & Medicinal
Chemistry Letters 14(21):5247-5250, 2004; Al-Shaar et al., Journal
of the Chemical Society, Perkin Transactions 1: Organic and
Bio-Organic Chemistry 21:2789-811, 1992; Hanami et al., Tetrahedron
Letters 48(22):3801-3803, 2007; Vogel et al., Helvetica Chimica
Acta 58(3):761-71, 1975; Heim-Riether et al., Journal of Organic
Chemistry 70(18):7331-7337, 2005; Patil et al., Journal of
Heterocyclic Chemistry 31(4):781-6, 1994.
[0562] Inventive compounds with an amino group off the phenyl
moiety of halofuginone or derivatives thereof may be prepared by
the following exemplary route:
##STR00181##
[0563] Prodrugs (e.g., esters of the hydroxyproline) may be
prepared by the following exemplary scheme:
##STR00182##
[0564] In Scheme 1 below, intermediate 1A is either commercially
available or is be prepared as described in literature procedures.
Song, Heterocyclic Communications 13(1):33-34, 2007; Peng et al.,
Journal of Combinatorial Chemistry 9(3):431-436, 2007; Robba et
al., Bulletin de la Societe Chimique de France 3-4(Pt. 2):587-91,
1975; Reigan et al., Bioorganic & Medicinal Chemistry Letters
14(21):5247-5250, 2004; Al-Shaar et al., Journal of the Chemical
Society, Perkin Transactions 1: Organic and Bio-Organic Chemistry
21:2789-811, 1992. Intermediate 1B can be prepared following
literature procedures with modest variations if necessary. U.S.
Patent Application Publication 2008/0188498; Emmanuvel et al., "A
concise enantioselective synthesis of (+)-febrifugine" Tetrahedron:
Asymmetry 20(1):84-88, 2009; Ooi et al., "A Concise
Enantioselective Synthesis of Antimalarial Febrifugine Alkaloids"
Organic Letters 3(6):953-955, 2001; Ashoorzadeh et al., "Synthetic
evaluation of an enantiopure tetrahydropyridine N-oxide. Synthesis
of (+)-febrifugine" Tetrahedron 65(24):4671-4680, 2009; Sukemoto et
al., "Concise asymmetric synthesis of (+)-febrifugine utilizing
trans-selective intramolecular conjugate addition" Synthesis
(19):3081-3087, 2008; Kikuchi et al., "Exploration of a New Type of
Antimalarial Compounds Based on Febrifugine" Journal of Medicinal
Chemistry 49(15):4698-4706, 2006; Takaya et al., "New Type of
Febrifugine Analogues, Bearing a Quinolizidine Moiety, Show Potent
Antimalarial Activity against Plasmodium Malaria Parasite" Journal
of Medicinal Chemistry 42(16):3163-3166, 1999. Compound 1C can be
prepared by reacting the nucleophilic 1A with the electrophilic 1B
in the presence of a base, e.g., KH or K.sub.2CO.sub.3.
##STR00183##
[0565] When R.sub.4 and R.sub.5 are taken together to form .dbd.O,
or R.sub.4 is hydrogen and R.sub.5 is hydroxyl, compounds 2C and 2D
can be prepared following Scheme 2. Intermediate 2B is prepared
following literature procedures with modest variations if
necessary. U.S. Patent Application Publication 2008/0188498;
Kikuchi et al., "Exploration of a New Type of Antimalarial
Compounds Based on Febrifugine" Journal of Medicinal Chemistry
49(15):4698-4706, 2006. Compound 2C is prepared by reacting the
nucleophilic 1A with the electrophilic 2B in the presence of a
base, e.g., KH or K.sub.2CO.sub.3. Compound 2C is then oxidized to
provide compound 2D. In certain embodiments, the reaction
conditions comprise TPAP and NMO at room temperature in the
presence of ground molecular sieves.
##STR00184##
[0566] When R.sub.4 and R.sub.5 are taken together to form
.dbd.NR.sub.D, compound 3A is prepared according to Scheme 3.
Reaction of compound 2D with primary amine R.sub.D--NH.sub.2 in the
presence of an acid catalyst (e.g., formic acid, acetic acid,
TiCl.sub.4) yields compound 3A.
##STR00185##
[0567] When R.sub.4 and R.sub.5 are taken together to form
.dbd.N--NR.sub.D or .dbd.N--N(R.sub.D).sub.2, compound 4A or
compound 4B can be prepared according to Scheme 4. A mixture of
intermediate 2D and the corresponding hydrazine are refluxed in
methanol, ethanol, or another suitable solvent, optionally in the
presence of an acid catalyst (e.g., acetic acid) to yield compound
4A or 4B.
##STR00186##
[0568] When R.sub.4 and R.sub.5 are taken together to form
.dbd.N--OR.sub.D, compound 5A can be prepared according to Scheme
5. A mixture of intermediate 2D and the corresponding hydroxylamine
are refluxed in methanol, ethanol, or another suitable solvent,
optionally in the presence of a base (e.g., pyridine, sodium
acetate) to yield compound 5A.
##STR00187##
[0569] When R.sub.4 and R.sub.5 are taken together to form .dbd.S,
compound 6A can be prepared according to Scheme 6. Compound 6A can
be prepared by treating intermediate 2D with Lawesson's
reagent.
##STR00188##
[0570] When R.sub.4 and R.sub.5 are taken together to form
.dbd.C(R.sub.D).sub.2, compound 7A can be prepared according to
Scheme 7. Reaction of the ketone intermediate 2D with the
corresponding Wittig reagent yields compound 7A.
##STR00189##
[0571] When R.sub.4 and R.sub.5 are taken together with a
neighboring atom to form a cyclopropyl ring, compound 8A can be
prepared according to Scheme 8. Intermediate 7A can be prepared
following Scheme 7. Treatment of 7A with the corresponding carbene
reagent yields the cyclopropyl compound 8A.
##STR00190##
[0572] When R.sub.4 and R.sub.5 are taken together with a
neighboring atom to form an epoxide ring, compounds 9A can be
prepared according to Scheme 9. Reaction of intermediate 7A with
oxygen donors such as MCPBA, hydrogen peroxide, or tert-butyl
peroxide provides compound 9A as the product.
##STR00191##
[0573] When R.sub.4 and R.sub.5 are taken together with a
neighboring atom to form an aziridine ring, compound 10A can be
prepared according to Scheme 10. Compound 10A is prepared by first
reacting 7A with ICI and NaN.sub.3 in MeCN, followed by reductive
ring closure with LiAlH.sub.4 in Et.sub.2O.
##STR00192##
[0574] In Scheme 11, compounds 11A-E can be prepared starting with
the ketone intermediate 2D. Reductive amination of 2D with
corresponding amine R.sub.D--NH.sub.2 with a catalytic amount of
acetic acid and NaCNBH.sub.3 (or NaBH(OAc).sub.3) in DCE can
provide compound 11A. Subsequently, reaction of 11A with the
corresponding acid chloride yields compound 11B; reaction of 11A
with the corresponding sulfonyl chloride yields compound 11C;
reaction of 11A with the corresponding chloroformate yields
compound 11D; and reaction of 11A with the corresponding isocyanate
yields compound 11E.
##STR00193##
[0575] In Scheme 12, reduction of intermediate 7A using H.sub.2 in
the presence of palladium or platinum catalyst, yields compounds
12A.
##STR00194##
[0576] In Scheme 13, the diaziridine compound 13A can be prepared
by treating intermediate 2D with HOSO.sub.2ONH.sub.2 and NH.sub.3
in MeOH. Oxidation of 13A with Ag.sub.2O in Et.sub.2O then provides
compound 13B.
##STR00195##
[0577] In Scheme 14, treatment of intermediate 2D with the
corresponding 1,2-ethanediol in the presence of p-tosyl acid in
toluene or another suitable solvent under reflux conditions yields
compound 14A.
##STR00196##
[0578] In Scheme 15, refluxing of intermediate 2D with the
corresponding 2-aminoethanol in toluene or another suitable solvent
yields compound 15A.
##STR00197##
[0579] In Scheme 16, intermediate 1C is prepared according to
Scheme 1. Reaction of 1C with the corresponding chloroformate, acid
chloride, isocyanate, or sulfonyl chloride provides compounds 16A,
16B, 16C, and 16D, respectively.
##STR00198##
[0580] In Scheme 17, alkylation of intermediate 1C with
electrophile (R.sub.A).sub.3C--Br (the leaving group can also be I,
Cl, OMs, or OTs) provides compound 17A. Alternatively, reductive
amination of 1C with the corresponding aldehyde (or ketone)
provides compound 17B.
##STR00199##
[0581] In Scheme 18, cross-coupling of IC with aryl bromide in the
presence of a copper or palladium catalyst provides compound
18A.
##STR00200##
[0582] In Scheme 19, intermediate IC can be prepared according to
Scheme 1. Reaction of compound 1C with the corresponding acid
chloride, chloroformate, isocyanate, or phosphorochloridate yield
compounds 19A, 19B, 19C, or 19D, respectively.
##STR00201##
Inhibition of Glutamyl-Prolyl tRNA Synthetase (EPRS)
[0583] Some of the analogs of halofuginone (1) described herein act
as inhibitors of metazoan glutamyl-prolyl tRNA synthetase (EPRS) or
non-metazoan prolyl tRNA synthetase. See FIGS. 21 and 22. In
certain embodiments, the EPRS is a eukaryotic EPRS. In certain
embodiments, the EPRS is a human EPRS. In certain embodiments, the
prolyl tRNA synthetase is a protozoan prolyl tRNA synthetase. A
structural feature of these inhibitors is a piperidine or
pyrrolidine ring, or an analog thereof. Without wishing to be bound
by a particular theory, it is believed that the piperidine ring of
halofuginone (1) acts by binding in the active site of the tRNA
synthetase like the pyrrolidine ring of proline, thus preventing
the charging of the amino acid proline to the tRNA synthetase.
[0584] Inhibition of EPRS or other tRNA synthetases leads to the
accumulation of uncharged prolyl tRNAs, which in turn activates the
amino acid starvation response (AAR, FIG. 1). Activation of the AAR
in T-cells suppresses the differentiation of a subset of effector
T-cells (Th17 cells) that promote autoimmunity. AAR also suppresses
pro-fibrotic gene expression and viral gene expression,
replication, and maturation. AAR may contribute to the protection
of organs from stress (e.g., ER stress in the pancreas during the
development of diabetes).
[0585] Inhibition of EPRS suppresses the synthesis and accumulation
of proteins such as polyglutamine-containing proteins that cause
neurodegenerative diseases such as Huntington's disease. This class
of EPRS inhibitors also promotes autophagy, a process that clears
protein aggregates in diseases such as Huntington's disease,
Alzheimer's disease, Parkinson's disease, and amyotrophic lateral
sclerosis (ALS). Halofuginone and similarly active compounds are
therefore useful as promoters of autophagy.
[0586] The specific inhibition of EPRS (as opposed to other tRNA
synthetases) also inhibits the synthesis of proline-rich proteins
such as collagen, which may be useful for the inhibiton of scarring
and fibrosis due to excess collagen deposition. Inhibition of
collagen synthesis may be useful for cosmetic and therapeutic
applications. The role of collagen in fibrosis makes the inventive
compounds useful in various cosmetic and therapeutic applications
associated with the accumulation of collagen.
[0587] The synthesis of collagen and the degradation and
remodelling of the ECM are also involved in a number of
physiological and pathological conditions, including angiogenesis,
systemic sclerosis, graft-versus-host disease (GVHD), pulmonary and
hepatic fibrosis, and autoimmune diseases. These disease are many
times associated with the excessive production of connective tissue
components, particularly collagen, which results in the destruction
of normal tissue architecture and function. Therefore, the
inventive compounds may be useful in treating or preventing these
diseases associated with collagen accumulation or the degradation
and remodelling of the ECM.
Molecular Target of Halofuginone
[0588] Halofuginone activates phosphorylation of elF2alpha, a
downstream component of the amino acid starvation response. The
activation of elF2alpha leads to the stimulation of an upstream
kinase, GCN2, the activation of which is monitored by measuring its
state of phosphorylation. FIG. 1 shows that GCN2 is phosphorylated
in response to halofuginone but not an inactive derivative of
halofuginone. The ability of halofuginone to stimulate elF2alpha
phosphorylation is dependent on GCN2, as the ability to stimulate
elF2alpha is lost in cells lacking GCN2 (FIG. 2). Activation of
GCN2 is well established to be regulated by the accumulation of
uncharged tRNAs (Dong, et al., Mol. Cell. (2000) 6: 269-279). These
data indicate that halofuginone acts either to limit the
availability of amino acids for tRNA charging or to inhibit the
enzymes that charge tRNA, the amino-acyl tRNA synthetases. The data
detailed below show that halofuginone inhibits an amino-acyl tRNA
synthetase, in particular, EPRS.
[0589] To distinguish these possibilities, the effect of
halofuginone on protein translation was examined in an in vitro
system (rabbit reticulocyte lysate) in which amino acids are
already present at levels adequate to support protein synthesis. In
this system, halofuginone at 400 nM inhibits translation of
luciferase by 5-6 orders of magnitude (FIG. 3), ruling out the
possibility that halofuginone modifies amino acid synthesis or
transport and indicating that it blocks tRNA synthetase activity.
Distinct tRNA synthetases catalyze the charging of individual tRNA
species with their cognate amino acid. To determine which tRNA
synthetase is inhibited by halofuginone, several amino acid
mixtures were added to the in vitro translation system to determine
which, if any, could reverse the inhibition of translation by
halofuginone (FIG. 3). When a mixture containing serine,
phenylalanine, and proline was found to reverse the effect of
halofuginone (FIG. 3, Mix 4), the effect of each amino acid was
tested. Only proline could rescue halofuginone inhibition,
indicating that glutamyl-prolyl tRNA synthetase (EPRS), the enzyme
responsible for proline tRNA aminoacylation, is specifically
inhibited by halofuginone. To examine whether proline could
specifically rescue the effect of halofuginone in intact cells, the
ability of each amino acid to reverse the effect of halofuginone on
elF2alpha phosphorylation in T-cells or to prevent halofuginone
inhibition of Th17 differentiation was tested (FIG. 4). Only
proline had an effect in these assays, confirming that EPRS is the
target for halofuginone in intact cells. Inhibition of EPRS with
compounds of the current invention can inhibit Th17
differentiation.
In Vitro Methods
[0590] Compounds of the invention may be screened to identify
biological activity, e.g., the ability to modulate the development
and/or expansion of Th17 cells by inhibiting EPRS, e.g., IL-17
secreting cells, in a subject. An assay for screening selective
inhibitors of IL-17 expressing cell development and/or expansion,
such as IL-17 expressing effector T-cell development and/or
expansion, e.g., Th17 development and/or expansion includes
contacting a naive T-cell population with a test compound under
conditions sufficient to allow T-cell development and/or expanion,
culturing the cell population, and detecting the level of IL-17
expression and/or the number of Th17 cells in the cell population,
wherein no change or a decrease in the level of IL-17 expression in
the cell population indicates that the test compound is a selective
Th17 inhibitor and/or wherein no change or a decrease in the number
of Th17 cells in the cell population indicates that the test
compound is a selective Th17 inhibitor. Determining the level of
IL-17 expression and/or the number of Th17 cells in the cell
population can be accomplished for example by using a detection
agent that binds to IL-17 or other marker for Th17 cells, for
example, the Th17-specific transcription factor RORgammat
(ROR.gamma.t). The detection agent is, for example, an antibody.
The detection agent can be coupled with a radioisotope or enzymatic
label such that binding of the detection agent to IL-17 or other
Th17 marker can be determined by detecting the labeled compound.
For example, the detection agent can be labeled with .sup.125I,
.sup.35S, .sup.14C, or .sup.3H, either directly or indirectly, and
the radioisotope detected by direct counting of radioemission or by
scintillation counting. Alternatively, detection agents can be
enzymatically labeled with, for example, horseradish peroxidase,
alkaline phosphatase, or luciferase, and the enzymatic label
detected by determination of conversion of an appropriate substrate
to product.
Methods of Modulating Th17 Cell Differentiation and/or
Proliferation and Other Cellular Functions using Halofuginone
Analogs and Compositions Thereof
[0591] Halofuginone and analogs thereof have been found to
specifically alter the development of T-cells away from the Th17
lineage, which is associated with cell-mediated damage, persistent
inflammation, and autoimmunity.
[0592] Th17 cells secrete several cytokines that may have a role in
promoting inflammation and fibrosis, including IL-17, IL-6, IL-21,
and GM-CSF. Of these cytokines, IL-17 is a specific product of Th17
cells, and not other T-cells. Whether Th17 cells are the only
source of IL-17 during inflammatory response is not clear, but
elevated IL-17 levels are in general thought to reflect expansion
of the Th17 cell population.
[0593] Diseases that have been associated with expansion of a Th17
cell population or increased IL-17 production include, but are not
limited to, rheumatoid arthritis, multiple sclerosis, Crohn's
disease, inflammatory bowel disease, dry eye syndrome, Lyme
disease, airway inflammation, transplantation rejection, graft
versus host disease, lupus, psoriasis, scleroderma, periodontitis,
systemic sclerosis, coronary artery disease, myocarditis,
atherosclerosis, diabetes, and inflammation associated with
microbial infection (e.g., viral, protazoal, fungal, or bacterial
infection).
[0594] Halofuginone analogs can be useful for treatment of any of
these diseases by suppressing the chronic inflammatory activity of
IL-17 expressing cells, such as IL-17 expressing effector T-cells,
e.g., Th17 cells. In some instances, this may address the root
cause of the disease (e.g., self-sustaining inflammation in
rheumatoid arthritis); in other cases (e.g., diabetes,
periodontitis) it may not address the root cause but may ameloriate
the symptoms associated with the disease.
[0595] IL-17 expressing effector T-cells, e.g., Th17 cells, and
their associated cytokine IL-17 provide a broad framework for
predicting or diagnosing diseases potentially treatable by
halofuginone analogs. Specifically, pre-clinical fibrosis and/or
transplant/graft rejection could be identified and treated with a
halofuginone analog, or with a halofuginone analog in combination
with other Th17 antagonists. Additionally, diseases that are not
currently associated with Th17 cell damage and persistence of
inflammation may be identified through the measurement of Th17 cell
expansion, or of increased IL-17 levels (e.g., in serum or synovial
fluid). Alternatively, or in addition, the use of gene profiling to
characterize sets of genes activated subsequent to Th17
differentiation may allow detection of Th17-affected tissues, prior
to histological/pathologic changes in tissues.
[0596] Halofuginone analogs could be used in combination with other
agents that act to suppress Th17 development to achieve synergistic
therapeutic effects. Current examples of potential synergistic
agents would include anti-IL-21 antibodies or antigen binding
fragments thereof, retinoic acid, or anti-IL-6 antibodies or
antigen binding fragments thereof, all of which can reduce Th17
differentiation.
[0597] Halofuginone analogs could be used in combination with other
agents that act to suppress inflammation and/or immunological
reactions, such as steroids (e.g., cortisol (hydrocortisone),
dexamethasone, methylprednisolone, and/or prednisolone),
non-steroidal anti-inflammatory drugs (NSAIDs; e.g., ibuprofen,
acetominophin, aspirin, celecoxib, valdecoxib, etoricoxib,
lumiracoxib, parecoxib, rofecoxib, nimesulide, and/or naproxen), or
immunosuppressants (e.g., cyclosporine, rapamycin, and/or FK506).
In certain embodiments, halofuginone analogs are used in
combination with agents that are immunomodulatory (e.g., modulators
of the mTOR pathway; thalidomide and derivatives thereof such as
lenalidomide and actimid; biguanides such as metformin, phenformin,
buformin, and proguanil; and HDAC inhibitors such as trichostatin
A, depsipeptide, SAHA, PXD101, LAQ824, LBH589, MS275, CI994,
MGCD0103, and valproic acid. In some embodiments, an agent that
inhibits a tRNA synthetase is used in combination with an inhibitor
of a proinflammatory cytokine. Proinflammatory cytokines that can
be targeted (in addition to IL-6 and IL-21, discussed above)
include TNF.alpha., IFN.gamma., GM-CSF, MIP-2, IL-12, IL-1.alpha.,
IL-I.beta., and IL-23. Examples of such inhibitors include
antibodies that bind to the cytokine or that bind to a receptor of
the cytokine and block its activity, agents that reduce expression
of the cytokine (e.g., small interfering RNA (siRNA) or antisense
agents), soluble cytokine receptors, and small molecule inhibitors
(see, e.g., WO 2007/058990).
[0598] In some embodiments, agents that inhibit tRNA synthetases
are used in combination with an inhibitor of TNF.alpha.. In some
embodiments, an inhibitor of TNF.alpha. comprises an
anti-TNF.alpha. antibody or antigen binding fragment thereof. In
some embodiments, the anti-TNF.alpha. antibody is adalimumab
(Humira.TM.). In some embodiments, the anti-TNF.alpha. antibody is
infliximab (Remicade.TM.). In some embodiments, the anti-TNF.alpha.
antibody is CDP571. In some embodiments, an inhibitor of TNF.alpha.
comprises a TNF.alpha. receptor. For example, in some embodiments,
the TNF.alpha. inhibitor is etanercept (Enbrel.TM.), which is a
recombinant fusion protein having two soluble TNF receptors joined
by the Fc fragment of a human IgG1 molecule. In some embodiments,
an inhibitor of TNF.alpha. comprises an agent that inhibit
expression of TNF.alpha., e.g., such as nucleic acid molecules that
mediate RNA interference (RNAi) (e.g., a TNF.alpha. selective siRNA
or shRNA) or antisense oligonucleotides. For example, a TNF.alpha.
inhibitor can include, e.g., a short interfering nucleic acid
(siNA), a short interfering RNA (siRNA), a double-stranded RNA
(dsRNA), or a short hairpin RNA (shRNA)(see, e.g., U.S. Patent
Application No. 20050227935, incorporated herein by reference).
[0599] Halofuginone analogs can be evaluated in animal models. To
determine whether a particular halofuginone analog suppresses graft
rejection, allogeneic or xenogeneic grafting (e.g., skin grafting,
organ transplantion, or cell implantation) can be performed on an
animal such as a rat, mouse, rabbit, guinea pig, dog, or non-human
primate. Strains of mice such as C57B1-10, B10.BR, and B10.AKM
(Jackson Laboratory, Bar Harbor, Me.), which have the same genetic
background but are mismatched for the H-2 locus, are well suited
for assessing various organ grafts.
[0600] In another example, heart transplantation is performed,
e.g., by performing cardiac grafts by anastomosis of the donor
heart to the great vessels in the abdomen of the host as described
by Ono et al., J. Thorac. Cardiovasc. Surg. 57:225, 1969. See also
Corry et al., Transplantation 16:343, 1973. Function of the
transplanted heart can be assessed by palpation of ventricular
contractions through the abdominal wall. Rejection is defined as
the cessation of myocardial contractions. A halofuginone analog
would be considered effective in reducing organ rejection if
animals treated with the inhibitor experience a longer period of
myocardial contractions of the donor heart than do untreated
hosts.
[0601] In another example, effectiveness of a halofuginone analog
at reducing skin graft rejection is assessed in an animal model. To
perform skin grafts on a rodent, a donor animal is anesthetized and
a full thickness skin is removed from a part of the tail. The
recipient animal is also anesthetized, and a graft bed is prepared
by removing a patch of skin (e.g., 0.5.times.0.5 cm) from the
shaved flank. Donor skin is shaped to fit the graft bed,
positioned, covered with gauze, and bandaged. Grafts are inspected
daily beginning on the sixth post-operative day and are considered
rejected when more than half of the transplanted epithelium appears
to be non-viable. A halofuginone analog that causes a host to
experience a longer period of engraftment than seen in an untreated
host would be considered effective in this type of experiment.
[0602] In another example, a halofuginone analog is evaluated in a
pancreatic islet cell allograft model. DBA/2J islet cell allografts
can be transplanted into rodents, such as 6-8 week-old B6 AFI mice
rendered diabetic by a single intraperitoneal injection of
streptozotocin (225 mg/kg; Sigma Chemical Co., St. Louis, Mo.). As
a control, syngeneic islet cell grafts can be transplanted into
diabetic mice. Islet cell transplantation can be performed by
following published protocols (for example, see Emamaullee et al.,
Diabetes 56(5):1289-98, 2007). Allograft function can be followed
by serial blood glucose measurements (Accu-Check III.TM.;
Boehringer, Mannheim, Germany). A rise in blood glucose exceeding
normal levels (on each of at least 2 successive days) following a
period of primary graft function is indicative of graft rejection.
The NOD (non-obese diabetic) mouse model is another model that can
be used to evaluate ability of a halofuginone analog to treat or
prevent type I diabetes.
[0603] In another example, a tRNA synthetase inhibitor is evaluated
in a model of dry eye disease (DED). In one such model, DED is
induced in mice in a controlled environment chamber by
administering scopolamine hydrobromide into the skin four times
daily. Chamber conditions include a relative humidity<30%,
airlflow of 15 L/min, and constant temperature (21-23.degree. C.).
Induction of dry eye can be confirmed by measuring changes in
corneal integrity with corneal fluorescein staining (see, e.g.,
Chauhan et al., J. Immunol. 182:1247-1252, 2009; Barabino et al.,
Invest. Ophthamol. Visual Sci. 46:2766-2771, 2005; and Rashid et
al., Arch. Ophthamol. 126: 219-225, 2008).
[0604] Numerous autoimmune diseases have been modeled in animals,
including rheumatic diseases, such as rheumatoid arthritis and
systemic lupus erythematosus (SLE), type I diabetes, dry eye
syndrome, and autoimmune diseases of the thyroid, gut, and central
nervous system. For example, animal models of SLE include MRL mice,
BXSB mice, and NZB mice and their F1 hybrids. The general health of
the animal as well as the histological appearance of renal tissue
can be used to determine whether the administration of a
halofuginone analog can effectively suppress the immune response in
an animal model of one of these diseases.
[0605] Animal models of intestinal inflammation are described, for
example, by Elliott et al. (Elliott et al., 1998, Inflammatory
Bowel Disease and Celiac Disease. In: The Autoimmune Diseases,
Third ed., N. R. Rose and I. R. MacKay, eds. Academic Press, San
Diego, Calif.). Some mice with genetically engineered gene
deletions develop chronic bowel inflammation similar to IBD. See,
e.g., Elson et al., Gastroenterology 109:1344, 1995; Ludviksson et
al., J. Immunol. 158:104,1997; and Mombaerts et al., Cell 75:274,
1993). One of the MRL strains of mice that develops SLE,
MRL-1pr/1pr, also develops a form of arthritis that resembles
rheumatoid arthritis in humans (Theofilopoulos et al., Adv.
Immunol. 37:269, 1985).
[0606] Models of autoimmune disease in the central nervous system
(CNS), such as experimental allergic encephalomyelitis (EAE), can
also be experimentally induced, e.g., by injection of brain or
spinal cord tissue with adjuvant into the animal (see, e.g.,
Steinman and Zamvil, Ann Neurol. 60:12-21, 2006). In one EAE model,
C57B/6 mice are injected with an immunodominant peptide of myelin
basic protein in Complete Freund's Adjuvant. EAE disease correlates
such as limp tail, weak/altered gait, hind limb paralysis, forelimb
paralysis, and morbidity are monitored in animals treated with a
halofuginone analog as compared to controls.
[0607] In addition to T cell differentiation processes,
halofuginone analogs can specifically alter processes such as
fibrosis and angiogenesis. Fibrosis can be assayed in vitro by
observing the effect of a halofuginone analog on fibroblast
behavior. In one exemplary assay for use in evaluating halofuginone
analogs, primary dermal fibroblasts are cultured in a matrix of
Type I collagen, which mimics the interstitial matrix of the dermis
and hypodermis, such that fibroblasts attach to the substratum and
spread. Inhibition of fibroblast attachment and spreading in the
presence of a halofuginone analog indicates that the halofuginone
analog has anti-fibrotic properties. Biological effects of
halofuginone analogs on non-immune cell functions can also be
evaluated in vivo. In some embodiments, a halofuginone analog
reduces extracellular matrix deposition (e.g., in an animal model
of wound healing; see, e.g., Pines et al., Biol. Bone Marrow
Transplant 9:417-425, 2003). In some embodiments, a halofuginone
analog reduces extracellular matrix deposition at a concentration
lower than the concentration at which it inhibits another cellular
function, such as cell proliferation or protein synthesis.
[0608] The invention further provides methods of treating a disease
using a halofuginone analog. The inventive method involves the
administration of a therapeutically effective amount of a
halofuginone analog to a subject (including, but not limited to a
human or other animal).
[0609] Compounds and compositions described herein are useful for
the inhibition of glutamyl-prolyl tRNA synthetase (EPRS).
Inhibition of EPRS leads to the accumulation of uncharged tRNAs,
which in turn activate the amino acid starvation response (AAR).
Activation of this response suppresses 1) pro-fibrotic gene
expression; 2) the differentiation of naive T-cells into Th17 cells
that promote autoimmunity; 3) viral gene expression, replication,
and maturation; and/or 4) stress to organs (e.g., during
transplantation).
[0610] In some embodiments, halofuginone analog that inhibits EPRS
has anti-fibrotic properties in vivo. For example, an EPRS
inhibitor, halofuginone, potently reduces dermal extracellular
matrix (ECM) deposition (Pines et al., Biol. Blood Marrow
Transplant 9: 417-425, 2003). Halofuginone inhibits the
transcription of a number of components and modulators of ECM
function, including Type I collagen, fibronectin, the matrix
metallopeptidases MMP-2 and MMP-9, and the metalloprotease
inhibitor TIMP-2 (Li et al., World J. Gastroenterol. 11: 3046-3050,
2005; Pines et al., Biol. Blood Marrow Transplant 9: 417-425,
2003). The major cell types responsible for altered ECM deposition,
tissue thickening, and contracting during fibrosis are fibroblasts
and myofibroblasts. Myofibroblasts mature/differentiate from their
precursor fibroblasts in response to cytokine release, often
following tissue damage and mechanical stress, and can be
distinguished from fibroblasts in a wide range of organs and
pathological conditions (Border et al., New Eng. J. Med. 331:
1286-1292, 1994; Branton et al., Microbes Infect. 1: 1349-1365,
1999; Flanders, Int. J. Exp. Pathol. 85: 47-64, 2004). Halofuginone
has been studied extensively as a potential anti-fibrotic
therapeutic and has progressed to phase 2 clinical trials for
applications stemming from these properties.
[0611] In animal models of wound healing and fibrotic disease,
halofuginone reduces excess dermal ECM deposition when introduced
intraperitoneally, added to food, or applied locally (Pines et al.,
Biol. Blood Marrow Transplant 9: 417-425, 2003). Halofuginone is
currently in phase 2 clinical trials as a treatment for scleroderma
(Pines et al., Biol. Blood Marrow Transplant 9: 417-425, 2003),
bladder cancer (Elkin et al., Cancer Res. 59: 4111-4118, 1999), and
angiogenesis during Kaposi's sarcoma, as well as in earlier stages
of clinical investigation for a wide range of other
fibrosis-associated disorders (Nagler et al., Am. J. Respir. Crit.
Care Med. 154: 1082-1086, 1996; Nagler et al., Arterioscler.
Thromb. Vasc. Biol. 17: 194-202, 1997; Nagler et al., Eur. J.
Cancer 40: 1397-1403, 2004; Ozcelik et al., Am. J. Surg. 187:
257-260, 2004). The results presented herein indicate that the
inhibition of fibrosis may be due at least in part to the
inhibition of glutamyl-prolyl tRNA synthetase (EPRS).
[0612] In some embodiments, a halofuginone analog inhibits
pro-fibrotic activities of fibroblasts. Thus, in certain
embodiments, the present invention provides a method for treating a
fibroblast-associated disorder comprising the step of administering
to a patient in need thereof a halofuginone analog or
pharmaceutically acceptable composition thereof.
[0613] As used herein, the term "fibroblast-associated" disorders
means any disease or other deleterious condition in which
fibroblasts are known to play a role. Accordingly, another
embodiment of the present invention relates to treating or
lessening the severity of one or more diseases in which fibroblasts
are known to play a role including, but not limited to,
fibrosis.
[0614] While halofuginone at high concentrations (between 20-40 nM)
does generally inhibit CD.sup.4+ T cell, CD.sup.8+ T cell, and
B220.sup.+ B cell activation, halofuginone also specifically
inhibits the development of Th17 cells, i.e., the T helper subset
that exclusively expresses high levels of the pro-inflammatory
cytokine interleukin IL-17, at low concentrations (PCT/US08/09774,
filed Aug. 15, 2008, which claims priority to U.S. Ser. No.
60/964,936, filed Aug. 15, 2007, the entirety of each of which is
incorporated herein by reference). Th17 cells, as a function of
their IL-17 secretion, play causal roles in the pathogenesis of two
important autoimmune diseases in the mouse, experimental autoimmune
encephalomyelitis (EAE) and type II collagen-induced arthritis
(CIA). EAE and CIA are murine models of the human autoimmune
pathologies, multiple sclerosis (MS) and rheumatoid arthritis
(R.sub.A). Halofuginone has been shown to be active in these
models. Halofuginone-mediated specific inhibition of IL-17
expressing cell development, such as IL-17 expressing effector T
cell development, e.g., Th17 cell development, takes place at
remarkably low concentrations, with 50% inhibition being achieved
around 3 nM. Therefore, halofuginone treatment specifically
inhibits the development of Th17-mediated and/or IL-17 related
diseases, including autoimmune diseases, persistent inflammatory
diseases, and infectious diseases, while not leading to profound T
cell dysfunction, either in the context of delayed-type
hypersensitivity or infection. Halofuginone analogs can also be
used to inhibit the development of Th17-mediated and/or IL-17
related diseases.
[0615] Halofuginone and analogs thereof interfere with the
differentiation of naive T-cells into IL-17-expressing Th17 cells.
Thus, in certain embodiments, the present invention provides a
method for treating a Th17-mediated or IL-17-mediated disorder
comprising the step of administering to a patient in need thereof a
halofuginone analog or a pharmaceutically acceptable composition
thereof.
[0616] As used herein, the terms "Th17-mediated" disorder and
"IL-17-mediated" disorder means any disease or other deleterious
condition in which Th17 or IL-17 is known to play a role.
Accordingly, another embodiment of the present invention relates to
treating or lessening the severity of one or more diseases in which
Th17 or IL-17 is known to play a role including, but not limited
to, autoimmune diseases, inflammatory diseases, infectious
diseases, angiogenesis, and organ protection during
transplantation.
[0617] The compounds and pharmaceutical compositions of the present
invention may be used in treating or preventing diseases or
conditions including, but not limited to, asthma, arthritis,
inflammatory diseases (e.g., Crohn's disease, rheumatoid arthritis,
psoriasis, dry eye syndrome), proliferative diseases (e.g., cancer,
benign neoplasms, diabetic retinopathy), cardiovascular diseases,
and autoimmune diseases (e.g., rheumatoid arthritis, lupus,
multiple sclerosis, psoriasis, scleroderma, or dry eye syndrome).
Halofuginone analogs and pharmaceutical compositions thereof may be
administered to animals, preferably mammals (e.g., domesticated
animals, cats, dogs, mice, rats), and more preferably humans. Any
method of administration may be used to deliver the agent or
pharmaceutical composition to the animal. In certain embodiments,
the agent or pharmaceutical composition is administered orally. In
other embodiments, the agent or pharmaceutical composition is
administered parenterally.
[0618] In certain embodiments, the present invention provides
methods for treating or lessening the severity of autoimmune
diseases including, but not limited to, acute disseminated
encephalomyelitis, alopecia universalis, alopecia areata, Addison's
disease, ankylosing spondylosis, antiphospholipid antibody
syndrome, aplastic anemia, arthritis, autoimmune diseases of the
adrenal gland, autoimmune hemolytic anemia, autoimmune hepatitis,
autoimmune oophoritis and orchitis, autoimmune thrombocytopenia,
Behcet's disease, bullous pemphigoid, cardiomyopathy, celiac
sprue-dermatitis, celiac disease, chronic fatigue immune
dysfunction syndrome (CFIDS), chronic inflammatory demyelinating
polyneuropathy, Churg-Strauss syndrome, cicatrical pemphigoid,
CREST syndrome, cold agglutinin disease, Crohn's disease, discoid
lupus, dry eye syndrome, endometriosis, dysautonomia, essential
mixed cryoglobulinemia, fibromyalgia-fibromyositis,
glomerulonephritis, idiopathic pulmonary fibrosis, Goodpasture's
syndrome, Graves' disease, Guillain-Barre syndrome, Hashimoto's
thyroiditis, IgA neuropathy, inflammatory bowel disease,
interstitial cystitis, juvenile arthritis, lichen planus, Meniere's
disease, mixed connective tissue disease, type 1 or immune-mediated
diabetes mellitus, juvenile arthritis, multiple sclerosis,
myasthenia gravis, neuromyotonia, opsoclonus-myoclonus syndrome,
optic neuritis, Ord's thyroiditis, osteoarthritis, pemphigus
vulgaris, pernicious anemia, polyarteritis nodosa, polychrondritis,
polyglandular syndromes, polymyalgia rheumatica, polymyositis and
dermatomyositis, primary agammaglobulinemia, primary biliary
cirrhosis, psoriasis, psoriatic arthritis, Raynauld's phenomenon,
Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma,
Sjogren's syndrome, stiff-man syndrome, Still's disease, systemic
lupus erythematosus, takayasu arteritis, temporal arteristis/giant
cell arteritis, idiopathic thrombocytopenic purpura, ulcerative
colitis, uveitis, vasculitides such as dermatitis herpetiformis
vasculitis, vitiligo, vulvodynia, warm autoimmune hemolytic anemia,
and Wegener's granulomatosis.
[0619] In some embodiments, the present invention provides a method
for treating or lessening the severity of one or more diseases and
conditions, wherein the disease or condition is selected from
immunological conditions or diseases, which include, but are not
limited to graft versus host disease, transplantation, transfusion,
anaphylaxis, allergies (e.g., allergies to plant pollens, latex,
drugs, foods, insect poisons, animal hair, animal dander, dust
mites, or cockroach calyx), type I hypersensitivity, allergic
conjunctivitis, allergic rhinitis, and atopic dermatitis.
[0620] In some embodiments, the present invention provides a method
for treating or lessening the severity of an inflammatory disease
including, but not limited to, asthma, appendicitis, Blau syndrome,
blepharitis, bronchiolitis, bronchitis, bursitis, cervicitis,
cholangitis, cholecystitis, chronic obstructive pulmonary disease
(COPD), chronic recurrent multifocal osteomyelitis (CRMO), colitis,
conjunctivitis, cryopyrin associated periodic syndrome (CAPS),
cystitis, dacryoadenitis, dermatitis, dermatomyositis, dry eye
syndrome, encephalitis, endocarditis, endometritis, enteritis,
enterocolitis, epicondylitis, epididymitis, familial cold-induced
autoinflammatory syndrome, familial Mediterranean fever (FMF),
fasciitis, fibrositis, gastritis, gastroenteritis, hepatitis,
hidradenitis suppurativa, laryngitis, mastitis, meningitis,
mevalonate kinase deficiency (MKD), Muckle-Well syndrome, myelitis
myocarditis, myositis, nephritis, oophoritis, orchitis, osteitis,
inflammatory osteolysis, otitis, pancreatitis, parotitis,
pericarditis, peritonitis, pharyngitis, pleuritis, phlebitis,
pneumonitis, pneumonia, proctitis, prostatitis, pulmonary fibrosis,
pyelonephritis, pyoderma gangrenosum and acne syndrome (PAPA),
pyogenic sterile arthritis, rhinitis, salpingitis, sinusitis,
stomatitis, synovitis, systemic juvenile rheumatoid arthritis,
tendonitis, TNF receptor associated periodic syndrome (TRAPS),
tonsillitis, undifferentiated spondyloarthropathy, undifferentiated
arthropathy, uveitis, vaginitis, vasculitis, vulvitis, or chronic
inflammation resulting from chronic viral or bacteria infections,
psoriasis (e.g., plaque psoriasis, pustular psoriasis,
erythrodermic psoriasis, guttate psoriasis or inverse
psoriasis).
[0621] In certain embodiments, the present invention provides
methods for treating or lessening the severity of arthropathies and
osteopathological diseases including, but not limited to,
rheumatoid arthritis, osteoarthrtis, gout, polyarthritis, and
psoriatic arthritis.
[0622] In certain embodiments, the present invention provides
methods for treating or lessening the severity of
hyperproliferative diseases including, but not limited to,
psoriasis or smooth muscle cell proliferation including vascular
proliferative disorders, atherosclerosis, and restenosis. In
certain embodiments, the present invention provides methods for
treating or lessening the severity of endometriosis, uterine
fibroids, endometrial hyperplasia, and benign prostate
hyperplasia.
[0623] In certain embodiments, the present invention provides
methods for treating or lessening the severity of acute and chronic
inflammatory diseases including, but not limited to, ulcerative
colitis, inflammatory bowel disease, Crohn's disease, dry eye
syndrome, allergic rhinitis, allergic dermatitis, cystic fibrosis,
chronic obstructive bronchitis, and asthma.
[0624] In some embodiments, the present invention provides a method
for treating or lessening the severity of a cardiovascular disorder
including, but not limited to, myocardial infarction, angina
pectoris, reocclusion after angioplasty, restenosis after
angioplasty, reocclusion after aortocoronary bypass, restenosis
after aortocoronary bypass, stroke, transitory ischemia, a
peripheral arterial occlusive disorder, pulmonary embolism, deep
venous thrombosis, ischemic stroke, cardiac hypertrophy, and heart
failure.
[0625] The present invention further includes a method for the
treatment of mammals, including humans, which are suffering from
one of the above-mentioned conditions, illnesses, disorders, or
diseases. The method comprises that a pharmacologically active and
therapeutically effective amount of one or more of the halofuginone
analogs according to this invention is administered to the subject
in need of such treatment.
[0626] The invention further relates to the use of the halofuginone
analogs according to the present invention for the production of
pharmaceutical compositions which are employed for the treatment
and/or prophylaxis and/or amelioration of the diseases, disorders,
illnesses, and/or conditions as mentioned herein.
[0627] The invention further relates to the use of halofuginone
analogs according to the present invention for the production of
pharmaceutical compositions.
[0628] The invention further relates to the use of the halofuginone
analogs according to the present invention for the production of
pharmaceutical compositions for inhibiting or treating
fibrosis.
[0629] The invention further relates to the use of halofuginone
analogs according to the present invention for the production of
pharmaceutical compositions which can be used for treating,
preventing, or ameliorating of diseases responsive to inhibiting
IL-17 production, such as autoimmune or inflammatory diseases, such
as any of those diseases mentioned herein.
[0630] The exact amount required will vary from subject to subject,
depending on the species, age, and general condition of the
subject, the particular agent, its mode of administration, its mode
of activity, and the like. The compounds of the invention are
preferably formulated in dosage unit form for ease of
administration and uniformity of dosage. It will be understood,
however, that the total daily usage of the agents 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 specific agent
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 agent employed; the duration
of the treatment; drugs used in combination or coincidental with
the specific agent employed; and like factors well known in the
medical arts.
[0631] 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,
intracisternally, intravaginally, intraperitoneally, topically (as
by powders, ointments, or drops), bucally, as an oral or nasal
spray, or the like, depending on the severity of the infection
being treated. In certain embodiments, an agent of the invention
may be administered orally or parenterally at dosage levels
sufficient to deliver from about 0.001 mg/kg to about 100 mg/kg,
from about 0.01 mg/kg to about 50 mg/kg, preferably from about 0.1
mg/kg to about 40 mg/kg, preferably from about 0.5 mg/kg to about
30 mg/kg, from about 0.01 mg/kg to about 10 mg/kg, from about 0.1
mg/kg to about 10 mg/kg, and more preferably from about 1 mg/kg to
about 25 mg/kg, of subject body weight per day, one or more times a
day, to obtain the desired therapeutic effect. The desired dosage
may be delivered three times a day, two times a day, once a day,
every other day, every third day, every week, every two weeks,
every three weeks, or every four weeks. In certain embodiments, the
desired dosage may be delivered using multiple administrations
(e.g., two, three, four, five, six, seven, eight, nine, ten,
eleven, twelve, thirteen, fourteen, or more administrations). In
certain embodiments, a halofuginone analog is administered at a
dose that is below the dose at which the agent causes non-specific
effects. In certain embodiments, a halofuginone analog is
administered at a dose that does not cause generalized
immunosuppression in a subject.
[0632] Liquid dosage forms for oral and parenteral administration
include, but are not limited to, pharmaceutically acceptable
emulsions, microemulsions, solutions, suspensions, syrups, and
elixirs. In addition to the active agents, 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, oral compositions can also include
adjuvants such as wetting agents, emulsifying and suspending
agents, sweetening, flavoring, and perfuming agents. In certain
embodiments for parenteral administration, agents of the invention
are mixed with solubilizing agents such CREMOPHOR EL.RTM.
(polyethoxylated castor oil), alcohols, oils, modified oils,
glycols, polysorbates, cyclodextrins, polymers, and combinations
thereof.
[0633] 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. 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.
[0634] 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.
[0635] 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 of crystalline or amorphous material with poor
water solubility. The rate of absorption of the drug then depends
upon its rate of dissolution which, 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 poly(lactide-co-glycolide).
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.
[0636] 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 agent.
[0637] Solid dosage forms for oral administration include capsules,
tablets, pills, powders, and granules. In such solid dosage forms,
the active agent 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.
[0638] 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
which 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 polethylene
glycols and the like.
[0639] The active agents 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 agent may be admixed with at least one inert diluent such as
sucrose, lactose or starch. Such dosage forms may also comprise, as
is normal practice, additional substances other than inert
diluents, e.g., tableting lubricants and other tableting aids such
a 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.
[0640] Formulations suitable for topical administration include
liquid or semi-liquid preparations such as liniments, lotions,
gels, applicants, oil-in-water or water-in-oil emulsions such as
creams, ointments, or pastes; or solutions or suspensions such as
drops. Formulations for topical administration to the skin surface
can be prepared by dispersing the drug with a dermatologically
acceptable carrier such as a lotion, cream, ointment, or soap.
Useful carriers are capable of forming a film or layer over the
skin to localize application and inhibit removal. For topical
administration to internal tissue surfaces, the agent can be
dispersed in a liquid tissue adhesive or other substance known to
enhance adsorption to a tissue surface. For example,
hydroxypropylcellulose or fibrinogen/thrombin solutions can be used
to advantage. Alternatively, tissue-coating solutions, such as
pectin-containing formulations can be used. 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 an agent to the body.
Such dosage forms can be made by dissolving or dispensing the agent
in the proper medium. Absorption enhancers can also be used to
increase the flux of the agent across the skin. The rate can be
controlled by either providing a rate controlling membrane or by
dispersing the agent in a polymer matrix or gel.
[0641] Additionally, the carrier for a topical formulation can be
in the form of a hydroalcoholic system (e.g., quids and gels), an
anhydrous oil or silicone based system, or an emulsion system,
including, but not limited to, oil-in-water, water-in-oil,
water-in-oil-in-water, and oil-in-water-in-silicone emulsions. The
emulsions can cover a broad range of consistencies including thin
lotions (which can also be suitable for spray or aerosol delivery),
creamy lotions, light creams, heavy creams, and the like. The
emulsions can also include microemulsion systems. Other suitable
topical carriers include anhydrous solids and semisolids (such as
gels and sticks); and aqueous based mousse systems.
[0642] It will also be appreciated that the agents and
pharmaceutical compositions of the present invention can be
employed in combination therapies, that is, the agents and
pharmaceutical compositions can be 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. It will also be appreciated that the therapies employed
may achieve a desired effect for the same disorder (for example, a
halofuginone analog may be administered concurrently with another
agent), or they may achieve different effects (e.g., control of any
adverse effects).
[0643] In still another aspect, the present invention also provides
a pharmaceutical pack or kit comprising one or more containers
filled with one or more of the ingredients of the pharmaceutical
compositions of the invention, and in certain embodiments, includes
an additional approved therapeutic agent for use as a combination
therapy. 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.
Methods of Identifying Subjects in Need of Th17 Modulation
[0644] In various embodiments of the invention, suitable in vitro
or in vivo studies are performed to determine whether
administration of a specific therapeutic that modulates the
development of IL-17 expressing cells, such as IL-17 expressing
effector T-cells, e.g., Th17 cells is indicated for treatment of a
given subject or population of subjects. For example, subjects in
need of treatment using a compound that modulates IL-17 expressing
cell development, such as IL-17 expressing effector T-cell
development, e.g., Th17 cell development, are identified by
obtaining a sample of IL-17 expressing cells, such as IL-17
expressing effector T-cells, e.g., Th17 cells from a given test
subject and expanding the sample of cells. If the concentration of
any of a variety of inflammatory cytokine markers, including IL-17,
IL-17F, IL-6, IL-21, IL-2, and TNF.alpha., also increases as the
cell population expands, then the test subject is a candidate for
treatment using any of the compounds, compositions, and methods
described herein.
[0645] Subjects in need of treatment are also identified by
detecting an elevated level of IL-17 expressing cells, such as
IL-17 expressing effector T-cells, e.g., Th17 cells or a Th17
cell-associated cytokine or a cytokine that is secreted by a Th17
cell. Cytokine levels to be evaluated include IL-17, IL-17F, IL-6,
IL-21, TNF.alpha., and GM-CSF. The cytokine IL-17, as well as other
cytokines such as IL-6, IL-21, IL-2, TNF.alpha., and GM-CSF, are
typically induced during inflammation and/or infection. Thus, any
elevated level of expression of these cytokines in a subject or
biological sample as compared to the level of expression of these
cytokines in a normal subject is useful as an indicator of a
disease state or other situation where treatment with an inventive
compound is desirable. Studies have shown that the levels of IL-17
in healthy patient serum is less than 2 .mu.g/mL (i.e., below the
detection limit of the assay used), while patients with liver
injury had levels of IL-17 expression in the range of 2-18 .mu.g/mL
and patients with rheumatoid arthritis had levels greater than 100
.mu.g/mL (see Yasumi et al., Hepatol Res. (2007) 37: 248-254, and
Ziolkowska et al., J. Immunol. (2000) 164: 2832-2838, each of which
is incorporated herein by reference). Thus, detection of an
expression level of IL-17 greater than 2 pg/mL in a subject or
biological sample is useful in identifying subjects in need of
treatment.
[0646] A subject suffering from or at risk of developing a
Th17-related and/or IL-17-related disesase such as an autoimmune
disease, a persistent inflammatory disease, or an infectious
disease is identified by methods known in the art. For example,
subjects suffering from an autoimmune disease, persistent
inflammatory disease, or an infectious disease are diagnosed based
on the presence of one or more symptoms associated with a given
autoimmune, persistent inflammatory, or infectious disease. Common
symptoms include, for example, inflammation, fever, loss of
appetite, weight loss, abdominal symptoms, such as, for example,
abdominal pain, diarrhea, or constipation, joint pain or aches
(arthralgia), fatigue, rash, anemia, extreme sensitivity to cold
(Raynaud's phenomenon), muscle weakness, muscle fatigue, change in
skin or tissue tone, shortness of breath or other abnormal
breathing patterns, chest pain or constriction of the chest
muscles, abnormal heart rate (e.g., elevated or lowered), light
sensitivity, blurry or otherwise abnormal vision, and reduced organ
function.
[0647] Subjects suffering from an autoimmune disease such as, e.g.,
multiple sclerosis, rheumatoid arthritis, Crohn's disesase, are
identified using any of a variety of clinical and/or laboratory
test such as, physical examination, radiological examination and
blood, urine, and stool analysis to evaluate immune status. For
example, subjects suffering from an infectious disease such as Lyme
disease are identified based on symptoms, objective physical
findings (such as erythema migrans, facial palsy, or arthritis),
and a history of possible exposure to infected ticks. Blood test
results are generally used to confirm a diagnosis of Lyme
disease.
Determination of the Biological Effect of Th17 Modulation
[0648] In various embodiments of the invention, suitable in vitro
or in vivo studies are performed to determine the effect of a
specific therapeutic that modulates the development of IL-17
expressing cells, such as IL-17 expressing effector T-cells, e.g.,
Th17 cells, and whether its administration is indicated for
treatment of a given subject or population of subjects. For
example, the biologial effect of a selective Th17 inhibitor
therapeutic, such as a compound of the invention, is monitored by
measuring level of IL-17 production and/or the number of IL-17
expressing cells, such as IL-17 expressing effector T-cells, e.g.,
Th17 cells in a patient-derived sample. The biolgocial effect of a
therapeutic is also measured by physical and/or clinical
observation of a patient suffering from, or at risk of developing,
a Th17-related and/or I1-17-related disease such as an autoimmune
disease, persistent inflammatory disease, and/or an infectious
disease. For example, administration of a specific Th17 inhibitor
to a patient suffering from a Th17-related disease and/or an
IL-17-related disease is considered successful if one or more of
the symptoms associated with the disorder is alleviated, reduced,
inhibited, or does not progress to a further, i.e., worse,
state.
[0649] These and other aspects of the present invention will be
further appreciated upon consideration of the following Examples,
which are intended to illustrate certain particular embodiments of
the invention but are not intended to limit its scope, as defined
by the claims.
EXAMPLES
Example 1
The Amino Acid Starvation Response (AAR) is Activated by HF in
Cultured Fibroblastic Cells
[0650] SV-MES mesangial cells were stimulated for 2 hours with
halofuginone (20 nM), or an inactive derivative of halofuginone 9
(MAZ1310, 20 nM) or control buffer, lysed and analyzed by
SDS-PAGE/Western blot for total or Ser51 phosphorylated elF2alpha,
and total or Thr 898 phosphorylated GCN2. FIG. 1 shows the results
of the experiment. Duplicate cell samples are shown.
Phosphorylation of GCN2 at Thr898 is a defining characteristic of
AAR activation, therefore the activation of GCN2 phosphorylation at
this site following HF treatment indicates that HF activates the
AAR. Activated GCN2 phosphorylates eif2alpha at Ser 51, therefore
this is an expected downstream outcome of AAR activation.
Example 2
GCN-2 Dependency of Halofuginone Stimulated elF2alpha
Phosphorylation
[0651] CD4+ CD25- T cells purified from wild type of GCN2-/- mice
were activated through TCR for 4 hours in the presence of
halofuginone (10 nM) or 9 (MAZ1310, 10 nM). Results are shown in
FIG. 2. Whole cell lysates were analyzed by SDS PAGE/Western blot
and antibodies indicated. Treatment with Halofuginone, but not the
inactive derivative 9, leads to phosphorylation of Ser51 of
eif2alpha only in wild type cells and not in GCN2-/- cells,
establishing the eif2alpha phosphorylation following halofuginone
stimulation occurs through activation of the AAR/GCN2 pathway.
Example 3
Proline Rescue of Translation Inhibition by Halofuginone
[0652] Translation in vitro was performed using rabbit reticulocyte
lysates and luciferase mRNA as template. Translation was measured
as arbitrary units of luciferase activity using a luminometer based
luminescence assay. Results are shown in FIG. 3. Log scale
presentation of background-subtracted data is shown. Translations
were performed without (dark bars) or with (light bars) 400 nM
halofuginone, in the absence of amino acids (0), or with the
following additions: Mix 1: 1 mM Asn, 1 mM Arg; Mix 2: 1 mM Lys, 1
mM Ile, 1 mM Tyr; Mix 3: 1 mM His, 1 mM Met, 1 mM Leu; Mix 4: 1 mM
Ser, 1 mM Phe, 1 mM Pro, Phe: 2 mM Phe; Pro: 2 mM Pro; Ser: 2 mM
Ser. Addition of proline, either alone or in combination with
phenylaline and serine, rescues inhibition of translation by
halofuginone, establishing that proline utilization for translation
(by glutamyl prolyl tRNA synthetase) is the critical target of
halofuginone action.
Example 4
Halofuginone-Induced elF2alpha Phosphorylation is Rescued by
Proline Addition
[0653] Naive T-cells were treated to stimulate the T-cell receptor
(TCR) in the presence or absence of 10 nM halofuginone in the
presence of 1 mM added amino acid, and then assayed for elF2alpha
activity phosphorylation by SDS PAGE/Western blot. Results are
shown in FIG. 4A. Phosphorylation induced by halofuginone is
blocked by added proline. These data establish that utilization of
proline is inhibited by halofuginone, leading to activation of the
AAR.
Example 5
Rescue of Halofuginone Inhibited Th17 Differentiation by
Proline
[0654] Naive T-cells were stimulated to differentiate in the
presence or absence of 10 nM halofuginone, with 1 mM of the
indicated amino acids added to the medium, and stained for Th17
differentiation on day 4. Results are shown in FIG. 4B. Naive
murine T cells were activated in the presence or absence of
TGF.beta. plus IL-6 as indicated, expanded in for 4 days and
restimulated with PMA and ionomycin for intracellular cytokine
staining. For intracellular cytokine staining, fixed cells were
washed twice with staining buffer (PBS/1% BSA/0.1% NaN.sub.3) and
then permeabilized with perm buffer III (BD Pharmingen) on ice for
30 minutes. Cells were then washed and stored in staining buffer
prior to data acquisitionAll FACS data was acquired on a
FACSCalibur flow cytometer (BD Pharmingen) and analyzed using
FlowJo software (Treestar, Inc., Ashland, Oreg.). FACS sorting was
performed on a FACS-Diva cytometer (BD Pharmingen). Bars indicate
percentage of cells differentiation as Th17 as indicated by IL17
expression. Proline, and no other added amino acid, rescues the
inhibition of Th17 differentiation by halofuginone, confirming that
proline utilization is the critical target for halofuginone
inhibition of Th17 differentiation.
Example 6
Inhibition of Cell Spreading by Halofuginone Analogs in a
Fibroblast Model
[0655] The ability of halofuginone and analogs thereof to inhibit
the spreading of freshly plated primary dermal fibroblasts was
examined. Human primary dermal fibroblasts were replated and
halofuginone added immediately after replating; spreading of cells
was assessed by light microscopy 28 hours after treatment with
compound. "+++" vs. "++" vs. "+" in general denotes .about.4-fold
differences in potency. "-" denotes no activity at any dose tested,
up to 1000.times. the effective dose of halofuginone. These data
establish that several analogs of halofuginone act similarly to
halofuginone in this assay, and that these activities correlate
well with activities described in Tables 2 and 3 below.
TABLE-US-00001 TABLE 1 Inhibition of cell spreading by halofuginone
analogs in a fibroblast model Compound # Fibroblast inhibition 1
+++ 2 - 3 ++ 4 - 5 ++ 6 - 7 +++ 8 + 9 - 10 +++ 11 + 12 ++ 13 ++ 15
+++ 16 -
Example 7
Inhibition of Th17 Differentiation by Halofuginone Analogs
[0656] The effect of different halofuginone and analogs thereof on
differentiation of naive T-cells was assayed as described in legend
for FIG. 4 above. Differentiation of primary naive T-cells into
Th17 cells following T-cell receptor stimulation+IL-6+TGFb. "+++"
vs. "++" vs. "+" in general denotes .about.4-fold differences in
potency. "-" denotes no activity at any dose tested, up to
1000.times. the effective dose of halofuginone. These data
establish that several analogs of halofuginone act similarly to
halofuginone in this assay.
TABLE-US-00002 TABLE 2 Inhibition of Th17 differentiation by
halofuginone analogs. Inhibition of Th17 Compound # differentiation
1 +++ 2 - 3 ++ 4 - 7 +++ 9 - 10 +++ 11 + 13 ++
Example 8
Translational Inhibition by Halofuginone Analogs in Rabbit
Reticulocyte Lysates
[0657] Inhibition of translation of luciferase RNA by halofuginone
and analogs thereof was measured as described in FIG. 3 above.
Results are shown of translation of a luciferase RNA reporter in a
rabbit reticulocyte lysate. "+++" vs. "++" vs. "+" in general
denotes .about.4-fold differences in potency. "-" denotes no
activity at any dose tested, up to 1000.times. the effective dose
of halofuginone. These data establish that several analogs of
halofuginone act similarly to halofuginone in this assay, and that
these activities correlate well with activities described in Tables
1 and 2 above.
TABLE-US-00003 TABLE 3 Translational inhibition by halofuginone
analogs in rabbit reticulocyte lysates Translational Compound #
inhibition 1 +++ 2 - 3 ++ 4 - 7 +++ 8 + 9 - 12 ++ 13 ++ 14 ++ 15 +
16 -
Example 9
Inhibition of Th17 Cell Development Through Activation of an Amino
Acid Starvation Response
[0658] This example shows that the aminoacyl tRNA synthetase
inhibitor, halofuginone (HF), imparts a selective block of Th17
differentiation in both human and mouse T cells by inducing the AAR
response.
[0659] To investigate whether HF can modulate T cell
differentiation or effector function, purified murine CD4.sup.+
CD25.sup.- T cells were treated with HF or its inactive derivative
9 (MAZ1310) and stimulated in the absence or presence of polarizing
cytokines to induce Th1, Th2, iTreg, or Th17 differentiation.
Dose-response experiments revealed a remarkably selective effect of
HF on Th17 differentiation (defined here as the percentage of
IL-17.sup.+ IFN.gamma..sup.- cells following restimulation on day
4-5). HF repressed Th17 differentiation in a dose-dependent manner
with an IC.sub.50 of 3.6 nM.+-.0.4 nM (FIG. 5A, 5B). Low
concentrations of HF (1-10 nM) that strongly reduced IL-17
production (FIG. 5A, 5B, FIG. 9A) did not affect T cell
proliferation, CD25 upregulation, or production of IL-2, TNF, or
IFN.gamma. (FIG. 9B). Low-dose HF also failed to modulate Th1, Th2
or iTreg differentiation as assessed by IFN.gamma., IL-4, or Foxp3
expression, respectively (FIG. 9A). At approximately 10-fold higher
concentrations (>20 nM), HF induced a general inhibition of T
and B cell activation, proliferation, and effector function (FIG.
5A, 5B).
[0660] The selective inhibition of Th17 differentiation by low-dose
HF was stereospecific: the HPLC-purified D-enantiomer of HF
inhibited IL-17 expression more potently than a racemic mixture,
whereas the L-enantiomer was completely inactive (FIG. 5C).
Inhibition of IL-17 expression was most pronounced when HF was
added during a 12-hour window at the start of the culture period
(FIG. 5D) and HF treatment impaired expression of both IL-17 and
IL-17f mRNA (FIG. 9C). These results suggest that HF regulates
early events, possibly involved in Th17 lineage commitment, rather
than influencing the expansion of Th17 cells or preventing acute
cytokine expression upon restimulation. Inhibition by HF was not
due to perturbation of cell cycle progression or selective
survival; HF inhibited IL-17 expression in a dose-dependent manner
even when considering only cells that had completed an equivalent
number of cell divisions based on CFSE dilution (FIG. 5E). HF also
reduced IL-17 expression in cultures where IFN.gamma. and IL-4,
cytokines known to inhibit Th17 differentiation (Park et al., Nat.
Immunol. 6:1133, 2005), were blocked by addition of neutralizing
antibodies. Thus, HF-mediated inhibition of Th17 cell development
is not secondary to effects on T cell proliferation or auxiliary
cytokine production.
[0661] In light of reports that IL-17 expression may be
differentially regulated in murine versus human T cells (Manel et
al., Nat. Immunol. 9:641, 2008; Wilson et al., Nat. Immunol. 8:950,
2007; Acosta-Rodriguez et al., Nat. Immunol. 8:942, 2007), HF
modulation of IL-17 expression by human CD4.sup.+ T cells was
investigated. These experiments showed that HF treatment greatly
reduced both the percentage of human T cells expressing IL-17 and
the amount of IL-17 produced (FIG. 5F, 5G). In striking contrast,
IFN.gamma. expression was essentially unaffected by HF treatment
(FIG. 5F, 5G). Therefore, HF selectively limits IL-17 expression in
both human and mouse T cells.
[0662] Th17 differentiation is synergistically regulated by
TGF.beta. and the pro-inflammatory cytokines IL-6 and IL-21.
Although reports had indicated that HF can attenuate TGF.beta.
signaling at high concentrations (>50 nM) (Gnainsky et al., Cell
Tiss. Res. 328:153, 200; Flanders, Int. J. Exp. Pathol. 85:47,
2004), it was discovered that low dose HF inhibited neither
TGF.beta.-induced Smad phosphorylation nor a variety of other
lymphocyte responses to TGF.beta. (Li et al., Ann. Rev. Immunol.
24:99, 2006; Glimcher et al., Nat. Rev. Immunol. 4:900, 2004; van
Vlasselaer et al., J. Immunol. 148:2062, 1992), in contrast to the
type 1 TGF.beta. receptor kinase inhibitor SB-431542, which
abrogated all responses to TGF.beta. (FIG. 10). Since STAT3 is the
major transducer of IL-6 and IL-21 signaling, the kinetics of STAT3
phosphorylation in HF-treated T cells were examined. HF did not
interfere with STAT3 activation during the first 6 hours of Th17
differentiation, but rather decreased the maintenance of STAT3
phosphorylation, beginning around 12 hours post activation (FIG.
6A, 6B).
[0663] Next, it was investigated whether inhibition of Th17
differentiation by HF could be restored by transgenic expression of
a hyperactive STAT3 protein (STAT3C) (Bromberg et al., Cell 98:295,
1999). T cells isolated from homozygous mice containing a floxed
stop-STAT3C-IRES-EGFP (STAT3C-GFPfl/fl) or stop-YFP (YFPfl/fl)
cassette inserted into the ROSA26 locus were transduced with a
cell-permeant TAT-Cre fusion protein to delete the stop cassette
and these cells were activated in the presence of TGF.beta. plus
IL-6, with either HF or 9 (MAZ1310). As expected, HF strongly
impaired Th17 differentiation of cells expressing YFP or those not
expressing a transgene (FIG. 6C, top three panels); in contrast, T
cells expressing STAT3C (defined by their concomitant expression of
GFP) remained capable of differentiating into Th17 cells even in
the presence of 10 nM HF (FIG. 6C, bottom panel). Data from a
number of similar experiments are quantified and summarized in FIG.
6D. Collectively, these results suggest that HF inhibits Th17
differentiation through its ability to prevent sustained activation
of STAT3. STAT3 promotes Th17 lineage commitment through the
induction of the orphan nuclear receptors ROR.gamma.t and
ROR.alpha. (Yang et al., J. Biol. Chem. 282:9358, 2007; Ivanov et
al., Cell 126:1121, 2006; Yang et al., Immunity 28:29, 2008).
Consistent with the finding that HF did not affect STAT3
phosphorylation during the first 12 hours of stimulation, HF did
not interfere with the upregulation of ROR.gamma. or ROR.alpha.
during Th17 differentiation (FIG. 11A). Moreover, HF inhibited Th17
differentiation as effectively in T cells retrovirally transduced
with ROR.gamma.t-expressing retroviruses as in those transduced
with empty retroviruses (FIG. 11B, 11C). T cells differentiated in
the presence of HF showed enhanced Foxp3 expression (FIG. 6E), as
expected from the observations that HF inhibits STAT3 signaling and
Th17 differentiation (Yang et al., J. Biol. Chem. 282:9358, 2007).
This result suggested that HF redirects developing Th17 cells to
the iTreg lineage rather than simply blocking their effector
function. However, upregulation of Foxp3 by HF was neither
necessary nor sufficient to inhibit Th17 differentiation;
retroviral expression of FOXP3 in T cells did not decrease IL-17
expression induced by TGF.beta. plus IL-6 (FIG. 12A), though it
markedly reduced IL-2 and IFN.gamma. production in T cells cultured
under non-polarizing conditions. Moreover, HF strongly repressed
IL-17 expression in T cells lacking Foxp3 (FIG. 12B). Therefore,
the inhibitory effects of HF on Th17 differentiation are not
exerted indirectly through the upregulation of Foxp3. Rather, HF
impairs the maintenance of STAT3 phosphorylation in developing Th17
cells, resulting in a reciprocal increase in iTreg cell
development.
[0664] The 12-hour lag period between the addition of HF to T cell
cultures and the ensuing effect on STAT3 phosphorylation strongly
suggested an indirect effect. To identify more proximal cellular
effects of HF treatment, we used DNA microarrays to define the
transcriptional profiles of HF- and MAZ1310-treated T cells
activated in Th17-priming conditions for 3 or 6 hours. Eighty-one
annotated genes that were differentially expressed at both time
points in HF-versus MAZ1310-treated cells were identified, the
majority of which were upregulated following HF treatment (FIG. 7A,
FIG. 15). Among the HF-inducible transcripts, a large number of
genes functionally associated with amino acid synthesis and
transport, as well as protein synthesis, were observed (FIG. 7A,
FIG. 15). Similar gene expression profiles have been observed
during cellular responses to amino acid starvation (Fafournoux et
al., Biochem. J. 351:1, 2000; Peng et al., Mol. Cell Biol. 22:5575,
2002). Insufficient cellular levels of amino acids lead to the
accumulation of uncharged tRNAs that, in turn, activate the amino
acid response (AAR) pathway via the protein kinase GCN2. Activated
GCN2 phosphorylates and inhibits eukaryotic translation initiation
factor 2A (eIF2.alpha.), thereby reducing overall protein
translation, while specifically enhancing translation of the
transcription factor ATF4 (Harding et al., Mol. Cell 11:619, 2003;
Harding et al., Mol. Cell 6:1099, 2000). Indeed, a number of
stress-induced genes reportedly regulated by ATF4 in mouse
embryonic fibroblasts (Harding et al., Mol. Cell 11:619, 2003) were
over-represented among the genes induced by HF treatment in T cells
(FIG. 7B, FIG. 16). These analyses suggest that at least a portion
of the transcriptional response to HF is mediated by ATF4.
Furthermore, quantitative real-time PCR (qPCR) experiments
confirmed that at least three known AAR-associated genes (Asns,
Gpt2, eIF4Ebp1) were induced by HF treatment within 4 hours of T
cell activation (FIG. 7C).
[0665] To directly address whether HF activates the AAR pathway,
eIF2.alpha. phosphorylation and ATF4 protein levels in HF-treated T
cells was examined. HF induced detectable eIF2.alpha.
phosphorylation at 2.5 nM, and this effect plateaued at 5-10 nM HF
(FIG. 7D). ATF4 expression levels were highest in T cells treated
with 5-10 nM HF and were reduced in cells treated with higher
concentrations of HF (20-40 nM) (FIG. 7D), demonstrating a positive
correlation between the concentrations of HF that induce ATF4
expression and those that selectively inhibit Th17 differentiation
(FIG. 5A). In kinetic analyses, eIF2.alpha. phosphorylation in
HF-treated cells reached maximum levels by 2 hours and ATF4 protein
continued to accumulate until 4 hours (FIG. 7E), indicating that HF
activates the AAR pathway before any detectable effects on STAT3
phosphorylation or IL-17 production are observed. AAR activation
was a general consequence of HF treatment; HF induced eIF2a
phosphorylation, not only in T cells activated in Th17-priming
conditions, but also in resting naive T cells and T cells activated
in ThN, Th1, Th2, and iTreg polarizing conditions (FIG. 7F). HF
treatment also increased eIF2.alpha. phosphorylation in cultured
fibroblasts (FIG. 13) and microarray analyses of fibroblasts
revealed that HF induced a pattern of early gene induction similar
to that seen in T cells. These data demonstrate that activation of
the AAR pathway by HF is not a cell type-specific effect. HF
treatment induced ATF4 expression in all differentiated T cells,
but not in naive T cells (FIG. 7F). This result most likely
reflects the low metabolic rate and relatively inefficient
translation capacity of naive T cells (Rathmell et al., Eur. J.
Immunol. 33:2223, 2003). Thus, the rapid activation of the AAR
pathway by HF could underlie both its selective inhibition of Th17
differentiation and its effects on fibroblasts (Pines and Nagler,
Gen. Pharmacol. 30:445, 1998).
[0666] A variety of other cellular stresses (ER stress, oxidative
stress, viral infection) also result in eIF2.alpha. phosphorylation
and ATF4 translation, a phenomenon termed the integrated stress
response (ISR) (Harding et al., Mol. Cell 11:619, 2003; Harding et
al., Mol. Cell 6:1099, 2000). Individual stressors, however, can
also activate stress type-specific pathways. For instance, the
unfolded protein response (UPR), which is activated by ER stress,
results in expression of the transcription factor Xbp-1 through a
mechanism involving IRE-1-dependent splicing, as well as nuclear
translocation of the ER-sequestered transcription factor ATF6 in
addition to eIF2.alpha. phosphorylation catalyzed by the protein
kinase Perk (Ron and Walter, Nat. Rev. Mol. Cell Biol. 8:519, 2007;
Brunsing et al., J. Biol. Chem. 283, 17954, 2008; Lin et al.,
Science 318:944, 2007). Xbp-1 and ATF6, in turn, upregulate ER
chaperones such as GRP78/BiP and calreticulin, whose expression is
specific to the UPR and independent of the eIF2.alpha./ATF4 ISR
pathway (Ron and Walter, Nat. Rev. Mol. Cell Biol. 8:519, 2007; Lee
et al., Mol. Cell Biol. 23: 7448, 2003). However, HF did not induce
the expression of these and other hallmark ER stress response
genes.
[0667] To delineate the stress response pathway activated by HF,
the effects of amino acid starvation with those of tunicamycin (an
inducer of ER stress) or HF treatment during T cell activation were
compared. As expected, cells deprived of cysteine (Cys) and
methionine (Met) displayed eIF2.alpha. phosphorylation, ATF4
expression, and upregulation of AAR-associated genes but did not
induce Xbp-1 splicing (FIG. 8A, FIG. 14A, 14B). In contrast,
tunicamycin treatment induced eIF2.alpha. phosphorylation and ATF4
expression together with Xbp-1 splicing (FIG. 8A), as
characteristic of the UPR. The effects of HF treatment closely
resembled those of amino acid starvation, inducing eIF2.alpha.
phosphorylation without promoting Xbp-1 splicing (FIG. 8A). Taken
together, these data indicate that HF specifically induces an
AAR.
[0668] Next, the effects of amino acid starvation on Th17
differentiation and STAT3 activation were investigated. It was
discovered that the functional consequences of Cys/Met-deprivation
were remarkably similar to those of HF treatment in T cells.
Cys/Met deprivation profoundly and selectively impaired Th17
differentiation in a manner directly related to the concentration
of these amino acids in the culture medium. T cells cultured under
limiting Cys/Met concentrations showed greatly diminished Th17
differentiation but upregulated CD25 expression and differentiated
into Th1, Th2, and iTreg subsets as effectively as T cells cultured
in complete medium (FIG. 8B, FIG. 14C). As shown for HF (FIG. 5E),
inhibition of IL-17 expression by amino acid starvation was
unrelated to cell survival or proliferation (FIG. 14D). Further
similar to the effects of HF, Cys/Met-deprivation did not affect
the early phase of STAT3 phosphorylation but impaired the
maintenance of STAT3 phosphorylation (FIG. 8C, 8D). Moreover,
L-tryptophanol, a tryptophan derivative that competitively inhibits
tryptophanyl-tRNA loading, or limiting concentrations of a
different amino acid, leucine, also impaired IL-17 production (FIG.
8E), suggesting that inhibition of Th17 differentiation is a
general consequence of amino acid starvation. The mammalian target
of rapamycin (mTOR) pathway represents a second, complementary
mechanism through which cells respond to amino acid availability
(Fingar and Blenis, Oncogene 23:3151, 2004). However, early
transcriptional responses induced by HF and the mTOR inhibitor
rapamycin are distinct (Peng et al., Mol. Cell Biol. 22:5575, 2002
and FIG. 15), and HF did not inhibit signaling downstream of mTOR
in fibroblasts.
[0669] To test whether inhibition of IL-17 expression was specific
to stress induced by amino acid starvation, the influence of
tunicamycin on T cell activation and differentiation was tested.
Surprisingly, low concentrations of tunicamycin had little
influence on IL-17 expression in T cells (FIG. 8F, FIG. 14C) but
instead preferentially impaired Th1 and Th2 differentiation (FIG.
8F, FIG. 14C). These data suggest that individual stress response
pathways can regulate distinct aspects of T cell differentiation
and effector function but also indicate that eIF2.alpha.
phosphorylation and ATF4 translation (shared consequences of both
AAR and UPR) are not sufficient to explain the selective regulation
of Th17 differentiation by HF or amino acid deprivation. The impact
of cellular stress on the immune system is complex. Data herein
show here that Th17 differentiation is particularly susceptible to
stress induced by amino acid deprivation, whereas ER stress blunts
Th1 and Th2 differentiation. In addition to these effects on T cell
effector function, eIF2.alpha. phosphorylation induced during ER
stress may have cytoprotective effects in oligodendrocytes and
pancreatic 03 cells during acute inflammation associated with
autoimmune encephalomyelitis and diabetes (Puccetti and Grohmann,
Nat. Rev. Immunol. 7:817, 2007; Lin et al., J. Clin. Invest.
117:448, 2007). Diverse cellular responses to stress may regulate
both T cell function and the downstream cellular targets of
inflammatory cytokine signaling during tissue inflammation.
[0670] The distinctive sensitivity of Th17 cells to AAR pathway
activation may have a role during adaptive immune responses in
vivo. For example, indoleamine 2,3-dioxygenase (IDO), an
IFN.gamma.-induced enzyme that breaks down tryptophan, has been
shown to cause local depletion of tryptophan at sites of
inflammation and activate the AAR pathway in resident T cells
(Puccetti and Grohmann, Nat. Rev. Immunol. 7:817, 2007; Munn et
al., Immunity 22:633, 2005). While local IDO accumulation is most
often associated with proliferative impairment in T cells,
expansion or conversion of Foxp3+ T cells also has been reported
following upregulation of IDO (Puccetti and Grohmann, Nat. Rev.
Immunol. 7:817, 2007; Park et al., Arthritis Res. 10:R11, 2008).
Given the reciprocal relationship between the development of
pro-inflammatory Th17 cells and tissueprotective iTreg cells, it is
postulated that IDO-mediated immune tolerance involves local AAR
mediated inhibition of Th17 differentiation and consequent skewing
of the Th17: iTreg balance in favor of iTreg cells (Romani et al.,
J. Immunol. 180:5157, 2008).
Materials and Methods
Mice
[0671] Mice were housed in specific pathogen-free barrier
facilities and were used in accordance with protocols approved by
the animal care and use committees of the Immune Disease Institute
and Harvard Medical School. Wild-type C57B/6 mice were purchased
from Jackson laboratories (Bar Harbor, Me.) and were used for all
in vitro culture experiments unless otherwise noted.
ROSA26-YFPfl/fl (Srinivas et al., BMC Dev. Biol. 1:4, 2001) and
ROSA26-STAT3C-GFPfl/fl (Mesaros et al., Cell. Metab. 7:236, 2008)
mice have been described. Dr. Alexander Rudensky provided lymphoid
organs from Foxp3gfp and Foxp3ko mice (Gavin et al., Nature
445:771, 2007).
Cell Isolation
[0672] Primary murine T and B cells were purified by cell sorting.
CD4.sup.+ CD25.sup.- T cells were positively selected using CD4
dynabeads and detachabeads (Dynal, Oslo, Norway) per manufacturers
instructions followed by nTreg depletion using a CD25 microbead kit
(Miltenyi biotech, Auburn, Calif.). Naive (CD4.sup.+ CD62Lhi CD44lo
Foxp3gfp- or CD4.sup.+ CD62Lhi CD44lo CD25.sup.-) T cells were
purified from Foxp3gfp or Foxp3ko mice, respectively, by FACS
sorting. CD8.sup.+ T cells or B cells were isolated from CD4.sup.-
fractions using CD8 negative isolation kit (Dynal) or CD43 negative
isolation kit (Miltenyi biotech), respectively. Resting human
CD4.sup.+ T cells were isolated from PBMC of healthy human donors
using Dynal CD4 Positive Isolation Kit (Invitrogen, Carlsbad,
Calif.) as previously described (Sundrud et al., Blood 106:3440,
2005). CD4.sup.+ cells were further purified to obtain memory T
cells by staining with PE-conjugated anti-human CD45RO-PE
antibodies (BD Biosciences), and sorting on a FACSAria cytometer
(BD Biosciences). Following purification, cells were greater than
99% CD4.sup.+ CD45RO.sup.+. CD14.sup.+ monocytes were isolated from
autologous PBMC by MACS sorting using a magnetic separator
(AutoMACS, Miltenyi Biotech) and were more then 99% pure following
isolation.
Cytokines, Antibodies and Cell Culture
[0673] Purified CD4.sup.+ CD25.sup.- T cells were activated in
vitro as previously described (Djuretic et al., Nat. Immunol.
8:145, 2007) using 0.3 .mu.g/ml hamster anti-mouse CD3 (clone
145-2C11) (ATCC, Manassas, Va.) and 0.5 .mu.g/ml hamster anti-mouse
CD28 (BD Pharmingen, San Jose, Calif.). Activated cell cultures
were differentiated using the following combinations of cytokines
and antibodies: iTreg--recombinant human TGF.beta.1 (3
ng/ml--R&D systems, Minneapolis, Minn.), Th17--TGF.beta.1 (3
ng/ml) plus recombinant mouse IL-6 (30 ng/ml--R&D systems). Th1
and Th2 differentiation was performed as previously described
(Djuretic et al., Nat. Immunol. 8:145, 2007). Human IL-2
supernatant (National Cancer Institute) was used in culture at 0.01
U/ml and was added at 48 hours-post activation when T cells were
split into tissue culture wells lacking CD3 and CD28 antibodies,
with the exception of Th17 cultures that were maintained in the
absence of exogenous IL-2. CD8.sup.+ T cells were activated with 1
.mu.g/ml anti-CD3 and 1 .mu.g/ml anti-CD28 and were expanded in 0.1
U/ml IL-2 until day 6 post activation. CD43-depleted B cells were
activated in vitro by culturing with 25 .mu.g/ml LPS (Sigma, St.
Louis, Mo.) for 3-4 days in the presence or absence of TGF.beta..
All reagents (see below) were added at the time of T cell
activation and again at 48 hours post activation unless indicated
otherwise. For some experiments, purified CD4.sup.+ CD25.sup.- T
cells, CD8.sup.+ T cells or B cells were labeled with 1 .mu.M CFSE
(Invitrogen) prior to activation in accordance with manufacturer's
instructions. Human T cell activation was performed by plating
purified monocytes in a 96-well flat bottom plate at a
concentration of 2.times.10.sup.4 cells per well in complete medium
overnight. 10.sup.5 purified human memory T cells were added to
monocyte cultures in the presence of soluble anti-CD3/anti-CD28
beads (Dynabeads, Invitrogen). T cells were expanded in the
presence HF or 9 (MAZ1310) for up to 6 days.
Inhibitors and Amino Acid Starvation
[0674] 1 kg of 10% pure HF was received as a gift from Hangpoon
Chemical Co. (Seoul, Korea), which was further purified via HPLC to
>99% purity and used for experiments. Compound 9 (MAZ1310,
Kamberov, Ph.D. Dissertation, Harvard University, 2008) was
generated by chemical derivatization of halofuginone and was used
at equal concentrations as a negative control. HF and 9 were
prepared as 100 mM stock solutions in DMSO and diluted to the
indicated concentrations. SB-431542 (Inman et al., Mol. Pharmacol.
62:65, 2002) (Tocris Bioscience, Ellisville, Mo.) was prepared as a
10 mM stock solution in DMSO and was used in culture at 10 .mu.M.
L-tryptophanol was prepared as a 20 mM stock solution in 0.1 M
NaOH, pH 7.4 and was used at 0.2 mM. For amino acid starvation
experiments, T cells were activated and differentiated as above in
D-MEM medium without L-cysteine and L-methionine (Invitrogen,
Carlsbad, Calif.), or D-MEM medium without L-leucine. Stocks
containing 20 mM L-cysteine (Sigma, St. Louis, Mo.) plus 10 mM
L-methionine (Sigma), or 400 mM L-leucine (Sigma) were prepared in
ddH2O, pH 1.0 and were added to medium at the indicated
concentrations.
Tat-Cre Transduction
[0675] 6xHis-TAT-NLS-Cre (HTNC--herein called TAT-Cre) was prepared
as previously described (Peitz et al., Proc. Nat. Acad. Sci. USA
99:4489, 2002). Purified T cells where rested in complete medium
for 30 minutes, washed 3 times in ADCF-Mab serum free medium
(Hyclone, Logan, Utah) and resuspended in pre-warmed serum free
medium supplemented with 50 .mu.g/ml of TATCre. Following a 45
minute incubation at 37.degree. C., transduction was stopped using
media containing 10% FCS and T cells were rested for 4-6 hours in
complete medium prior to activation.
Retroviral Transductions
[0676] MIG and MIG.ROR.gamma.t retroviral cDNA were gifts from Dr.
Dan Littman. pRV and pRV.FOXP3 retroviral constructs have been
described previously (Wu et al., Cell 126:375, 2006). Retroviral
particles were generated using the phoenix-Eco system (ATCC).
Supernatants were concentrated by centrifugation and stored at
-80.degree. C. prior to use in culture. Thawed retroviral
supernatants were added to T cell cultures 12 hours after T cell
activation in the presence of 8 g/ml polybrene (American
Bioanalytical, Natick, Mass.) and centrifuged for 1 hour at room
temperature to enhance infections.
Detection of Cytokine Production
[0677] Cytokines secreted into media supernatant were measured
using the mouse Th1/Th2 cytometric bead array (CBA--BD Pharmingen)
in accordance with manufacturers instructions. Briefly, CD4+ CD25-
T cells were activated in anti-CD3/anti-CD28-coated tissue culture
wells (see above) and supernatants were collected at the indicated
times.
[0678] For detection of intracellular cytokines in murine cells,
cultured T or B cells were stimulated with 10 nM PMA (Sigma) and 1
mM ionomycin (Sigma) for 4-5 hours in the presence of 10 mM
brefeldin A (Sigma). Stimulated cells were harvested, washed with
PBS and fixed with PBS plus 4% paraformaldehyde at room temperature
for 20 minutes. Cells were then washed with PBS, permeabilized with
PBS supplemented with 1% BSA and 0.5% saponin (Sigma) at room
temperature for 10 minutes before cytokine-specific antibodies were
added and incubated with cells for an additional 20 minutes at room
temperature. Human T cells were restimulated with PMA (20 ng/ml)
(Sigma) and lonomycin (500 ng/ml) (Sigma) for 6 hours in the
presence of golgi plug (BD Biosciences) and intracellular staining
was performed using cytofix/cytoperm kit (BD Biosciences) per
manufacturers instructions. All stained cells were stored at
4.degree. C. in PBS plus 1% paraformaldehyde prior to FACS
analyses.
FACS Analyses and Sorting
[0679] All cell surface staining was performed in FACS buffer
(PBS/2% FBS/0.1% NaN.sub.3) and antibodies were incubated with
cells on ice for 20-30 minutes. Cells were washed with FACS buffer
and fixed with FACS buffer plus 1% paraformaldehyde prior to data
acquisition. For phospho-STAT3 intracellular staining, stimulated T
cells cultured with or without TGF.beta. plus IL-6 for the
indicated times were harvested on ice and fixed in PBS plus 2%
paraformaldehyde for 10 minutes at 37.degree. C. Fixed cells were
washed twice with staining buffer (PBS/1% BSA/0.1% NaN.sub.3) and
then permeabilized with perm buffer III (BD Pharmingen) on ice for
30 minutes. Cells were then washed twice with staining buffer and
PE-conjugated anti-STAT3 (pY705) (BD Pharmingen) was added per the
manufacturer's instructions and incubated with cells at room
temperature for 45-60 minutes. Cells were then washed and stored in
staining buffer prior to data acquisition. Foxp3 intracellular
staining was performed using a Foxp3 intracellular staining kit
(eBioscience, San Diego, Calif.) in accordance with the
manufacturer's instructions. Fluorescent-conjugated antibodies
purchased from BD Pharmingen were percp-Cy5.5-conjugated anti-CD4,
PE-conjugated anti-CD25, PE-conjugated anti-IL-17, PE-conjugated
anti-phospho-STAT3 and APC-conjugated anti-human IFN.gamma..
Fluorescent conjugated antibodies purchased from eBioscience
include FITC-conjugated anti-CD8, APC-conjugated anti-mouse/rat
Foxp3, PE-conjugated anti-IL-4, APC-conjugated anti-IFN.gamma.,
PE-conjugated anti-granzyme B, APC-conjugated streptavidin,
PE-conjugated anti-IL-6, and PE-conjugated anti-human IL-17.
Biotin-conjugated anti-IgA antibody was purchased from Southern
biotech (Birmingham, Ala.). All FACS data was acquired on a
FACSCalibur flow cytometer (BD Pharmingen) and analyzed using
FlowJo software (Treestar, Inc., Ashland, Oreg.). FACS sorting was
performed on a FACS-Diva cytometer (BD Pharmingen).
Quantitative Real-Time PCR
[0680] T cells were activated as described above, collected at the
indicated times and pellets were flash-frozen in liquid nitrogen.
Total RNA was obtained by RNeasy (Quiagen, Valencia, Calif.) column
purification per manufacturers instructions. ROR.gamma.t expression
was determined after reverse transcription using the message sensor
kit (Ambion--Austin, Tex.) per the manufacturer's instructions and
taqman primers and probe as described elsewhere (Ivanov et al.,
Cell 126:1121, 2006). Sybrgreen quantitative real-time PCR was
performed on T cell RNA samples following reverse transcription via
SuperScript II first-strand cDNA synthesis kit (Invitrogen,
Carlsbad, Calif.). All PCR data was collected on an iCycler thermal
cycler (Biorad, Hercules, Calif.). Primer sequences used for
detecting stress response genes are listed below.
TABLE-US-00004 Asns forward: (SEQ ID NO: 1)
5'-TGACTGCCTTTCCGTGCAGTGTCTGAG-3' Asns reverse: (SEQ ID NO: 2)
5'-ACAGCCAAGCGGTGAAAGCCAAAGCAGC-3' Gpt2 forward: (SEQ ID NO: 3) 5'-
TAGTCACAGCAGCGCTGCAGCCGAAGC-3' Gpt2 reverse: (SEQ ID NO: 4) 5'-
TACTCCACCGCCTTCACCTGCGGGTTC-3' eIF4Ebp1 forward: (SEQ ID NO: 5) 5'-
ACCAGGATTATCTATGACCGGAAATTTC-3' eIF4Ebp1 reverse: (SEQ ID NO: 6)
5'- TGGGAGGCTCATCGCTGGTAGGGCTAG-3' Hprt forward: (SEQ ID NO: 7)
5'-GGGGGCTATAAGTTCTTTGCTGACC-3' Hprt reverse: (SEQ ID NO: 8)
5'-TCCAACACTTCGAGAGGTCCTTTTCAC-3'
Western Blotting
[0681] Whole cell lysates were generated from T cells activated for
the indicated times. For STAT3 and Smad2/3 western blots cells were
harvested, washed in PBS and lysed in 50 mM Tris, pH 7.4, 0.1% SDS,
1% Triton-X100, 140 mM NaCl, 1 mM EDTA, 1 mM EGTA supplemented with
protease inhibitors tablets (Roche, Germany), 1 mM NaF and 1 mM
Na.sub.3VO.sub.4. For eIF2.alpha. and ATF4 western blots, cells
were harvested as above and lysed in 50 mM Tris, pH 7.4, 2% SDS,
20% glycerol and 2 mM EDTA supplemented with protease and
phosphatase inhibitors as above. All lysates were cleared via
centrifugation and 15-30 .mu.g of protein was resolved by SDS-PAGE.
Protein was transferred to nitrocellulose membranes, blocked and
blotted using specific antibodies. Antibodies used for western blot
analysis were anti-phospho-Smad2, anti-STAT3 (pY705), anti-STAT3,
anti-eIF2.alpha..sup.pS51, anti-eIF2.alpha. (all from Cell
Signaling Technology, Danvers, Mass.). Anti-ATF4/CREB2 and
anti-.beta.-actin were purchased from Santa Cruz Biotechnology
(Santa Cruz, Calif.). HRP-conjugated secondary antibodies were all
purchased from Sigma, with the exception of HRP-conjugated
anti-armenian hamster antibody (Jackson Immunoresearch, West Grove,
Pa.).
Microarrays, Data Analyses and Statistics
[0682] RNA prepared from activated T cells treated with 10 nM HF or
MAZ1310 for either 3 or 6 hours, was amplified, biotin-labeled
(MessageAmp II Biotin-Enhanced kit, Ambion, Austin, Tex.), and
purified using the RNeasy Mini Kit (Qiagen, Valencia, Calif.).
Resulting cRNAs were hybridized to M430 2.0 chips (Affymetrix,
Inc.). Raw data were normalized using the RMA algorithm implemented
in the "Expression File Creator" module from the GenePattern
software package (Reich et al., Nat. Gen. 38:500, 2006) (available
on the internet at the following address:
broad.mit.edu/cancer/software/genepattern/). Data were visualized
using the GenePattern "Multiplot" modules. Gene expression
distribution analyses were performed using Chi-squared statistical
tests. For all other statistical comparisons, p values were
generated using one-tailed student T-tests on duplicate or
triplicate samples.
Example 10
Depletion of Amino Acids or tRNA Synthetase Inhibition with
L-Tryptophanol Inhibits Th17 Differentiation
[0683] T cells were cultured in complete medium (complete--200
.mu.M Cys/100 .mu.M Met/4 mM Leu), medium containing 0.1.times.,
0.2.times., or 1.times. cysteine and methionine (Cys/Met), medium
containing 0.1.times. leucine (Leu), or complete medium plus 0.2 mM
L-tryptophanol. Cells were activated in the presence or absence of
TGF.beta. plus IL-6, expanded for 4 days and restimulated with PMA
and ionomycin for intracellular cytokine staining. For
intracellular cytokine staining, fixed cells were washed twice with
staining buffer (PBS/1% BSA/0.1% NaN.sub.3) and permeabilized with
perm buffer III (BD Pharmingen) on ice for 30 minutes. Cells were
then washed and stored in staining buffer prior to data
acquisition. All FACS data were acquired on a FACSCalibur flow
cytometer (BD Pharmingen) and analyzed using FlowJo software
(Treestar, Inc., Ashland Oreg.). FACS sorting was performed on a
FACS-Diva cytometer (BD Pharmingen). The results, depicted in FIG.
17, show that depletion of Cys/Met, depletion of Leu, and treatment
with tryptophanol all inhibited Th17 differentiation.
Example 11
Modulation of Th17-Mediated Effects In Vivo
[0684] The ability of systemic HF treatment to block IL-17
expression and associated autoimmune inflammation in vivo was
examined using two distinct models of experimental autoimmune
encephalomyelitis (EAE). The first model used is referred to as
adjuvant-driven EAE and is actively induced by immunization of
wild-type mice with the immunodominant myelin-derived peptide
antigen MOG.sub.33-55 emulsified in Complete Freund's Adjuvant
(CFA). The second model, a passive model of EAE induction, is
initiated by the transfer of myelin proteolipid protein
(PLP)-reactive T cells into lymphopenic hosts.
[0685] Adjuvant-driven EAE was induced in 8 week-old wild-type B6
mice purchased from Charles River laboratories (Kingston, N.Y.) by
subcutaneous injection of MOG.sub.33-55 peptide emulsified in
Incomplete Freund's Adjuvant (IFA) plus 5 mg/ml heat-killed M.
tuburculosis (BD Biosciences) in both dorsal flanks as described in
Veldhoen et al. (Nat. Immunol. 7(11):1151-1156, 2006).
[0686] Passive EAE was induced by intravenous transfer of purified
CD3.sup.+ splenic T cells isolated from PLP TCR transgenic B10.S
mice into syngeneic RAG2-deficient mice (3.times.10.sup.6
cells/mouse) (Waldner et al., J. Clin. Invest. 113(7):990-997,
2004).
[0687] Mice were injected daily with HF (2 .mu.g/mouse) or vehicle
control (DMSO) i.p. Clinical signs of EAE were assessed according
to the following score: 0, no signs of disease; 1, flaccid tail; 2,
weak gait/hind limb paresis; 3, hind limb paralysis; 4,
tetraplegia; 5, moribund. Cytokine production during EAE was
determined either in peripheral T cells isolated from spleen or
lymph nodes of mice prior to disease onset (day 6-10) or in
mononuclear cells isolated from the brain and spinal cords of mice
with severe disease (clinical score.gtoreq.2) between days 15-20.
Briefly, splenocytes were stained for intracellular cytokines
following erythrocyte lysis with ammonium chloride buffer. T cells
were isolated from brain and spinal cords of mice with active EAE
following perfusion with cold PBS. Minced CNS tissue was digested
with liberase C1 (0.33 mg/ml, Roche Diagnostics) or collagenase D
(10 mg/ml, Roche Diagnostics) at 37.degree. C. for 30-45 minutes.
Cell suspensions were passed through 70 .mu.m cell strainers (VWR)
and fractionated by 70%/30% Percoll gradient centrifugation.
Mononuclear cells were collected from the interphase, washed, and
used for intracellular cytokine analysis.
[0688] The adjuvant-driven EAE model is associated with
infiltration of both IL-17- and IFN.gamma.-expressing CD4.sup.+ T
cells into the CNS (FIG. 18A). Low-dose HF treatment (2 .mu.g HF
daily, .about.0.1 mg/kg) significantly reduced both the severity of
adjuvant-driven EAE disease and frequency of disease onset (FIG.
18B). The second, passive model of EAE induction leads to a
predominant Th response, rather than Th17 response, within CNS
infiltrates (FIG. 18C). In marked contrast to the adjuvant-driven
EAE model, HF-treated mice in the passive EAE model developed
disease symptoms with kinetics and severity similar to control
treated animals (FIG. 18D). The contrasting effects of HF in these
two models of EAE support the notion that HF selectively inhibits
IL-17-associated inflammatory T cell function without inducing
general T cell hyporesponsiveness. Taken together, these data
suggest that HF can modulate autoimmune inflammation associated
with Th17, but not Th1, responses.
[0689] HF-mediated protection from adjuvant-driven EAE was
accompanied by a reduction in T cell-derived IL-17-expression, both
in peripheral lymph nodes prior to disease onset and in CNS tissue
during active disease (FIG. 18E), as well as an overall reduction
in CD4.sup.+ T cell infiltrates into the CNS (FIG. 19). Consistent
with in vitro results, HF impaired IL-17 production but did not
affect IFN.gamma. expression in the same T cell populations.
Moreover, splenocytes isolated ex vivo from HF-injected mice
displayed increased eIF2.alpha. phosphorylation and expression of
AAR-associated transcripts (FIG. 18F). Thus, systemic
administration of low doses of HF activates the AAR, leading to a
selective impairment of Th17 differentiation, and concomitant
blunting of IL-17 associated inflammatory responses in vivo.
[0690] Thus, consistent with in vitro data, it was discovered that
HF protects mice from adjuvant-driven EAE through in vivo
activation of the AAR. HF selectively reduced the number of IL-17
expressing T cells in vivo, but had no effect on the number of
IFN.gamma. T-cells. These data are consistent with reports showing
that adjuvant-driven EAE disease is particularly sensitive to
modulation of IL-17 expression. Notably, HF had no effect on an
independent, passive model of EAE that develops in the absence of a
Th17 response, demonstrating that HF is neither globally
immunosuppressive nor generically protective against CNS
inflammation. Both Th and Th17 cells can drive EAE pathogenesis
when transferred into mice. In the adjuvant-driven EAE model
described above, a roughly equal induction of Th1 and Th17 cells
was observed, whereas in the passive model of EAE, encephalitogenic
T cells were biased towards a Th response. Thus, the lack of an
effect of HF in the passive model, in comparison to the
adjuvant-driven EAE is likely due to the distinctive inflammatory T
cell responses in the two models.
[0691] Two follow-on experiments were performed to further
determine the therapeutic benefit of HF on EAE. In one, HF was
injected into mice immunized to produce EAE as described above,
with the exception that HF was not introduced into the animals
until day 10 following immunization. These data demonstrate that HF
controls autoimmune inflammation even after inflammatory
pathogenesis is evident, providing a more accurate representation
of autoimmune disease in humans. See FIG. 24. In the other
experiment, HF was injected into mice immunized to produce EAE as
described above, with the exception that HF injection was
terminated at day 10 following immunization. These data as shown in
FIG. 25 demonstrate that HF exerts a protective effect that extends
well beyond the time of treatment, consistent with its proposed
role in preventing the differentiation of pro-inflammatory Th17
cells. Therefore, HF has been shown to have a therapeutic benefit
in EAE after the onset of symptoms as well as to prevent the
development of the symptoms of EAE.
Example 12
Synthesis of (2S/R,3R/S)-tert-butyl
2-(3-(7-bromo-6-chloro-4-oxoquinazolin-3(4H)-yl)-2-oxopropyl)-3-hydroxypi-
peridine-1-carboxylate (9)
##STR00202##
[0693] Di-tert-butyldicarbonate (982 mg) in 10 mL DMF was added to
a solution of 1.5 g halofuginone hydrobromide and 1.3 mL
diisopropylethylamine in 100 mL. The reaction mixture was stirred
for 16 h at room temperature. After addition of water the aquous
layer was extracted three times with diethyl ether. The combined
organic layers were dried over sodium sulfate and evaporated to
dryness. The crude product was purified on silica gel with
dichloromethane/methanol to yield the desired Boc protected product
as white solid in quantitative yield.
Example 13
Synthesis of tert-butyl
2-(2-(2-hex-5-ynamidoethoxy)ethoxy)ethylcarbamate
##STR00203##
[0695] PyBOP (2.34 g) in 5 mL dichloromethane were added to a
solution of 476 mg hex-5-ynoic acid and 1.16 g
diisopropylethylamine in 20 mL dichloromethane. After 2 min 700 mg
Boc-2,2'-(ethane-1,2-diylbis(oxy))diethanamine were added and the
reaction mixture was stirred for 3 h at room temperature. The
reaction mixture was washed with citric acid solution and saturated
sodium bicarbonate solution. The organic layer was then dried over
sodium sulfate and evaporated to dryness. The crude product was
purified on silica gel with dichloromethane and methanol to yield
the desired product as colorless oil (820 mg).
Example 14
Synthesis of (2S/R,3R/S)-tert-butyl
2-(3-(6-chloro-7-(2,2-dimethyl-4,15-dioxo-3,8,11-trioxa-5,14-diazaicos-19-
-yn-20-yl)-4-oxoquinazolin-3(4H)-yl)-2-oxopropyl)-3-hydroxypiperidine-1-ca-
rboxylate
##STR00204##
[0697] (2S/R,3R/S)-tert-butyl
2-(3-(7-bromo-6-chloro-4-oxoquinazolin-3(4H)-yl)-2-oxopropyl)-3-hydroxypi-
peridine-1-carboxylate (100 mg) and 130 mg tert-butyl
2-(2-(2-hex-5-ynamidoethoxy)ethoxy)ethylcarbamate were dissolved in
5 mL triethylamine and 5 mL THF. The solution was degassed and 50
mg tetrakis(triphenylphosphine)palladium and 40 mg copper iodide
are added. The reaction mixture was stirred at 50.degree. C. for 16
h. After cooling to room temperature dichloromethane was added and
the organic layer was washed with brine. The organic layer was
dried over sodium sulfate and evaporated to dryness. The crude
product was purified on silica with hexanes and ethyl acetate (5:1)
as eluent to afford 130 mg of the product as off white solid.
Example 15
Synthesis of
N-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-6-(6-chloro-3-(3-((2S/R,3R/S)-3-hydr-
oxypiperidin-2-yl)-2-oxopropyl)-4-oxo-3,4-dihydroquinazolin-7-yl)hex-5-yna-
mide (8)
##STR00205##
[0699] (2S/R,3R/S)-tert-butyl
2-(3-(6-chloro-7-(2,2-dimethyl-4,15-dioxo-3,8,11-trioxa-5,14-diazaicos-19-
-yn-20-yl)-4-oxoquinazolin-3(4H)-yl)-2-oxopropyl)-3-hydroxypiperidine-1-ca-
rboxylate (6 mg) was dissolved in 1 mL dichloromethane, followed by
the addition of 200 uL TFA. The reaction mixture was stirred at
room temperature for 16 hours and the solvent was removed under
reduced pressure. The product was used without further purification
as trifluoroacetate.
Example 16
Synthesis of (2S/R,3R/S)-tert-butyl
2-(3-(6-chloro-4-oxo-7-((trimethylsilyl)ethynyl)quinazolin-3(4H)-yl)-2-ox-
opropyl)-3-hydroxypiperidine-1-carboxylate
##STR00206##
[0701] (2S/R,3R/S)-tert-butyl
2-(3-(7-bromo-6-chloro-4-oxoquinazolin-3(4H)-yl)-2-oxopropyl)-3-hydroxypi-
peridine-1-carboxylate (55 mg) and 24 mg ethynyltrimethylsilane
were dissolved in 5 mL triethylamine. The solution was degassed and
11.5 mg tetrakis(triphenylphosphine)palladium and 8 mg copper
iodide were added. The reaction mixture was stirred at 50.degree.
C. for 16 h. After cooling to room temperature dichloromethane and
water were added and the aqueous layer was extracted twice with
dichloromethane. The organic layer was dried over sodium sulfate
and evaporated to dryness. The crude product was purified on basic
alumina with dichloromethane and methanol as eluent to afford 25 mg
of the desired product as white solid.
Example 17
Synthesis (2S/R,3R/S)-tert-butyl
2-(3-(6-chloro-7-ethynyl-4-oxoquinazolin-3(4H)-yl)-2-oxopropyl)-3-hydroxy-
piperidine-1-carboxylate
##STR00207##
[0703] (2S/R,3R/S)-tert-butyl
2-(3-(6-chloro-4-oxo-7-((trimethylsilyl)ethynyl)quinazolin-3(4H)-yl)-2-ox-
opropyl)-3-hydroxypiperidine-1-carboxylate (11 mg) was dissolved in
500 uL methanol, followed by the addition of 3 mg potassium
carbonate. The reaction mixture was stirred for 2 h at room
temperature. The solvent was removed and the product was purified
on basic alumina with dichloromethane and methanol as eluent to
afford 6 mg of the desired product as white solid.
Example 18
Synthesis of
6-chloro-3-(3-((2S/R,3R/S)-3-hydroxypiperidin-2-yl)-2-oxopropyl)-7-((trim-
ethylsilyl)ethynyl)quinazolin-4(3H)-one (5)
##STR00208##
[0705] (2S/R,3R/S)-tert-butyl
2-(3-(6-chloro-4-oxo-7-((trimethylsilyl)ethynyl)quinazolin-3(4H)-yl)-2-ox-
opropyl)-3-hydroxypiperidine-1-carboxylate (10 mg) was dissolved in
500 uL dichloromethane, followed by the addition of 50 uL TFA. The
reaction mixture was stirred at room temperature for 5 h and the
solvent was removed under reduced pressure. The product was used
without further purification as the trifluoroacetate salt.
Example 19
6-chloro-7-ethynyl-3-(3-((2S/R,3R/S)-3-hydroxypiperidin-2-yl)-2-oxopropyl)-
quinazolin-4(3H)-one (7)
##STR00209##
[0707] (2S/R,3R/S)-tert-butyl
2-(3-(6-chloro-7-ethynyl-4-oxoquinazolin-3(4H)-yl)-2-oxopropyl)-3-hydroxy-
piperidine-1-carboxylate (6 mg) was dissolved in 500 .mu.L
dichloromethane, followed by the addition of 50 .mu.L TFA. The
reaction mixture was stirred at room temperature for 5 h and the
solvent was removed under reduced pressure. The product was
purified by HPLC (water/MeCN) to yield 3 mg of the desired product
as white solid.
Example 20
(2S/R,3R/S)-tert-butyl
2-((Z)-3-(7-bromo-6-chloro-4-oxoquinazolin-3(4H)-yl)-2-hydrazonopropyl)-3-
-hydroxypiperidine-1-carboxylate
##STR00210##
[0709] (2S/R,3R/S)-tert-butyl
2-(3-(7-bromo-6-chloro-4-oxoquinazolin-3(4H)-yl)-2-oxopropyl)-3-hydroxypi-
peridine-1-carboxylate (40 mg) and 40 uL hydrazine were dissolved
in 2 mL absolute ethanol and heated for 16 h. The solvent was
evaporated and the crude product was purified on silica with
dichloromethane and methanol to yield 15 mg of the desired
hydrazone as white solid.
Example 21
Synthesis of
7-bromo-6-chloro-3-((Z)-2-hydrazono-3-((2S/R,3R/S)-3-hydroxypiperidin-2-y-
l)propyl)quinazolin-4(3H)-one
##STR00211##
[0711] Halofuginone trifluoroacetate (24 mg) and 40 uL hydrazine
were dissolved in 2 mL absolute ethanol. The reaction mixture was
stirred for 5 h at room temperature whereby the desired product
precipitated as white solid. The product was filtered off and
washed twice with ethanol to yield 16 mg of the desired hydrazone
without further purification.
Example 22
Synthesis of
(Z)--N'-(1-(7-bromo-6-chloro-4-oxoquinazolin-3(4H)-yl)-3-((2S/R,3R/S)-3-h-
ydroxypiperidin-2-yl)propan-2-ylidene)acetohydrazide (14)
##STR00212##
[0713] Halofuginone trifluoroacetate (24 mg) and 14 mg
acetohydrazide were dissolved in 2 mL absolute ethanol. The
reaction mixture was stirred for 1 h at room temperature followed
by the removal of the solvent under reduced pressure. The crude
product was purified by HPLC to yield the desired product as
colorless oil (16 mg).
Example 23
Synthesis of methyl
6-((Z)-2-(1-(7-bromo-6-chloro-4-oxoquinazolin-3(4H)-yl)-3-((2S/R,3R/S)-3--
hydroxypiperidin-2-yl)propan-2-ylidene)hydrazinyl)-6-oxohexanoate
(12)
##STR00213##
[0715] Halofuginone trifluoroacetate (50 mg) and 50 mg methyl
6-hydrazinyl-6-oxohexanoate were dissolved in 2 mL absolute
ethanol. The reaction mixture was stirred for 24 h at room
temperature followed by the removal of the solvent under reduced
pressure.
Example 24
Synthesis of
N-(6-((Z)-2-(1-(7-bromo-6-chloro-4-oxoquinazolin-3(4H)-yl)-3-((2S/R,3R/S)-
-3-hydroxypiperidin-2-yl)propan-2-ylidene)hydrazinyl)-6-oxohexyl)-6-(5-(2--
oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)hexanamide
(13)
##STR00214##
[0717] Halofuginone trifluoroacetate (25 mg) and 25 mg
N-(6-hydrazinyl-6-oxohexyl)-6-(5-(2-oxohexahydro-1H-thieno[3,4-d]imidazol-
-4-yl)pentanamido)hexanamide were dissolved in 2 mL absolute
ethanol. The reaction mixture was stirred for 48 h at 65.degree. C.
followed by the removal of the solvent under reduced pressure. The
crude product was purified by HPLC to yield the desired product as
colorless oil (30 mg).
Example 25
Synthesis of (2S/R,3R/S)-tert-butyl
2-(3-(7-bromo-6-chloro-4-oxoquinazolin-3(4H)-yl)-2-oxopropyl)-3-(diethoxy-
phosphoryloxy)piperidine-1-carboxylate
##STR00215##
[0719] (2S/R,3R/S)-tert-butyl
2-(3-(7-bromo-6-chloro-4-oxoquinazolin-3(4H)-yl)-2-oxopropyl)-3-hydroxypi-
peridine-1-carboxylate (52 mg), 15 mg triethylamine and 17.2 mg
diethyl phosphorochloridate were dissolved in 2 mL dichloromethane
followed by the addition of 40 uL titaniumtetraisopropoxide. After
16 h stirring at room temperature additional 0.5 eq phosphochloride
and 0.2 eq titaniumtetraisopropoxide were added and the reaction
mixture was stirred for additional 24 hours. After addition of few
drops water and methanol the solvent was removed under reduced
pressure and the crude product was purified by HPLC to yield the
desired phosphate as white solid (10 mg).
Example 26
Synthesis of
(2S/R,3R/S)-2-(3-(7-bromo-6-chloro-4-oxoquinazolin-3(4H)-yl)-2-oxopropyl)-
piperidin-3-yl diethyl phosphate (16)
##STR00216##
[0721] (2S/R,3R/S)-tert-butyl
2-(3-(7-bromo-6-chloro-4-oxoquinazolin-3(4H)-yl)-2-oxopropyl)-3-(diethoxy-
phosphoryloxy)piperidine-1-carboxylate (6 mg) was dissolved in 2 mL
dichloromethane followed by the addition of 0.1 mL trifluoroacetic
acid. The reaction mixture was stirred for 2 h followed by the
removal of the solvent under reduced pressure. The product was used
without further purification.
Example 27
MAZ1320 and MAZ1686 Inhibit Th17 Differentiation
[0722] Two novel derivatives of halofuginone (MAZ1320 and MAZ 1686,
shown below) selectively inhibit the differentiation of Th17 cells
without inhibiting Th differentiation. Methods for determing Th17
differentiation and FACS analysis are described above. Shown in
FIG. 20 is FACS analysis of CD4+ CD25- T cells analyzed according
to IL17 expression (marking Th17 differentiation, y-axis) and Th1
(IFNgamma, x-axis) following differentiation in the presence of the
indicated concentrations of HF or derivatives thereof (MAZ1320,
MAZ1685, MAZ1686).
##STR00217##
Example 28
HF Inhibits the Incorporation of Proline into tRNA
[0723] Rabbit reticulocyte lysate was incubated with total bovine
tRNA, 0.1 .mu.M puromycin (to prevent translation), and
.sup.14C-labeled proline or .sup.35S-labeled methionine for minutes
in the presence or absence of HF or MAZ1310 (an inactive HF
derivative). Total RNA was extracted with acidic phenol-chloroform
and tRNA was isolated using miRVANA microRNA isolation kit.
Radioisotope incorporation into tRNA was measured in a
scintillation counter. FIG. 21 shows data normalized to control (no
HF or MAZ1310 addition) material. HF is an inhibitor of prolyl-tRNA
synthetase (EPRS) but not a different tRNA synthetase (methionyl
tRNA synthetase) in a crude in vitro translation system.
Example 29
HF Inhibits Purified EPRS
[0724] Prolyl-tRNA synthetase (EPRS) purified from rabbit liver was
tested for its ability to incorporate .sup.14C-labeled proline into
tRNA in the presence of the indicated concentration of HF or its
inactive derivative, MAZ1310. Purified enzyme was incubated with
100 .mu.g/ml bovine tRNA, 50 .mu.M .sup.14C-labeled proline, 5 mM
ATP, and 10 mM MgCl.sub.2 for 20 minutes. Charged tRNA was isolated
by precipitation on Whatman filter paper and washing with cold 5%
TCA. TCA precipitable counts were assayed by scintillation
counting. FIG. 22 shows the TCA preciptiable counts for 10 .mu.M
HF, 4 .mu.M HF, 1 .mu.M HF, and MAZ1310. HF directly inhibits
purified mammalian EPRS.
##STR00218##
Example 30
A tRNA Synthetase Inhibitor Structurally Unrelated to HF
Selectively Inhibits Th17 Differentiation
[0725] Borrelidin, a threonyl tRNA synthetase inhibitor
structurally unrelated to HF was tested for its ability to alter
T-cell differentiation. Methods for determining Th17
differentiation and FACS analysis are described above. Shown in
FIG. 23 is FACS analysis of CD4+CD25-T cells analyzed according to
IL17 expression (marking Th17 differentiation), IFNgamma expression
(marking Th1 differentiation), FoxP3 expression (marking Tref
differentiation), or ILA (marking Th2 differentiation) following
differentiation under conditions for polarization of each effector
T-cell subtype in the presence of the indicated concentrations of
HF (10 nM) or borrelidin. Borrelidin inhibits Th17 differentiation
without affecting Th1, Th2, or Treg differentiation or cell number,
a selectivity identical to that of HF. tRNA synthetase inhibition
therefore provides a general approach to the selective inhibition
of Th17 differentiation without generalized immunosuppression.
OTHER EMBODIMENTS
[0726] The foregoing has been a description of certain non-limiting
preferred embodiments of the invention. Those of ordinary skill in
the art will appreciate that various changes and modifications to
this description may be made without departing from the spirit or
scope of the present invention, as defined in the following claims.
Sequence CWU 1
1
8127DNAArtificial sequenceSynthetic Polynucleotide 1tgactgcctt
tccgtgcagt gtctgag 27228DNAArtificial sequenceSynthetic
Polynucleotide 2acagccaagc ggtgaaagcc aaagcagc 28327DNAArtificial
sequenceSynthetic Polynucleotide 3tagtcacagc agcgctgcag ccgaagc
27427DNAArtificial sequenceSynthetic Polynucleotide 4tactccaccg
ccttcacctg cgggttc 27528DNAArtificial sequenceSynthetic
Polynucleotide 5accaggatta tctatgaccg gaaatttc 28627DNAArtificial
sequenceSynthetic Polynucleotide 6tgggaggctc atcgctggta gggctag
27725DNAArtificial sequenceSynthetic Polynucleotide 7gggggctata
agttctttgc tgacc 25827DNAArtificial sequenceSynthetic
Polynucleotide 8tccaacactt cgagaggtcc ttttcac 27
* * * * *