U.S. patent application number 15/652788 was filed with the patent office on 2018-06-14 for prmt5 inhibitors containing a dihydro- or tetrahydroisoquinoline and uses thereof.
This patent application is currently assigned to Epizyme, Inc.. The applicant listed for this patent is Epizyme, Inc.. Invention is credited to Paula Ann Boriack-Sjodin, Richard Chesworth, Kenneth W. Duncan, Lei Jin, Michael John Munchhof.
Application Number | 20180162847 15/652788 |
Document ID | / |
Family ID | 50029211 |
Filed Date | 2018-06-14 |
United States Patent
Application |
20180162847 |
Kind Code |
A1 |
Duncan; Kenneth W. ; et
al. |
June 14, 2018 |
PRMT5 INHIBITORS CONTAINING A DIHYDRO- OR TETRAHYDROISOQUINOLINE
AND USES THEREOF
Abstract
Described herein are compounds of Formula (A), pharmaceutically
acceptable salts thereof, and pharmaceutical compositions thereof.
Compounds of the present invention are useful for inhibiting PRMT5
activity. Methods of using the compounds for treating
PRMT5-mediated disorders are also described. ##STR00001##
Inventors: |
Duncan; Kenneth W.;
(Westwood, MA) ; Chesworth; Richard; (Concord,
MA) ; Boriack-Sjodin; Paula Ann; (Lexington, MA)
; Munchhof; Michael John; (Salem, CT) ; Jin;
Lei; (Wellesley, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Epizyme, Inc. |
Cambridge |
MA |
US |
|
|
Assignee: |
Epizyme, Inc.
Cambridge
MA
|
Family ID: |
50029211 |
Appl. No.: |
15/652788 |
Filed: |
July 18, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14654226 |
Jun 19, 2015 |
9745291 |
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PCT/US2013/077250 |
Dec 20, 2013 |
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15652788 |
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61790928 |
Mar 15, 2013 |
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61745490 |
Dec 21, 2012 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/502 20130101;
A61P 35/00 20180101; C07D 217/04 20130101; A61P 7/06 20180101; A61K
31/538 20130101; A61P 43/00 20180101; C07D 405/14 20130101; A61K
31/498 20130101; A61K 31/472 20130101; A61K 31/5377 20130101; A61K
31/4725 20130101; C07D 413/12 20130101; A61K 31/497 20130101; C07D
401/12 20130101; C07D 405/12 20130101; A61P 3/04 20180101; C07D
217/08 20130101; C07D 401/14 20130101; C07D 217/16 20130101; A61P
3/00 20180101; A61K 31/517 20130101; C07D 417/12 20130101; A61P
3/10 20180101; A61P 7/00 20180101 |
International
Class: |
C07D 413/12 20060101
C07D413/12; C07D 217/08 20060101 C07D217/08; C07D 401/14 20060101
C07D401/14; C07D 217/16 20060101 C07D217/16; C07D 401/12 20060101
C07D401/12; C07D 405/12 20060101 C07D405/12; C07D 417/12 20060101
C07D417/12; C07D 217/04 20060101 C07D217/04; A61K 31/5377 20060101
A61K031/5377; A61K 31/517 20060101 A61K031/517; A61K 31/502
20060101 A61K031/502; A61K 31/498 20060101 A61K031/498; A61K 31/497
20060101 A61K031/497; A61K 31/4725 20060101 A61K031/4725; A61K
31/472 20060101 A61K031/472; A61K 31/538 20060101 A61K031/538; C07D
405/14 20060101 C07D405/14 |
Claims
1.-101. (canceled)
102. A kit or packaged pharmaceutical comprising a compound of
Formula (A), or a pharmaceutically acceptable salt thereof, and
instructions for use thereof: ##STR00613## or a pharmaceutically
acceptable salt thereof, wherein: represents a single or double
bond: R.sup.12 is halogen or optionally substituted C.sub.1-3alkyl,
and R.sup.13 is hydrogen, halogen, --NR.sup.A1R.sup.A2, optionally
substituted C.sub.1-3alkyl or --OR.sup.1; or R.sup.12 is hydrogen,
halogen or optionally substituted C.sub.1-3alkyl, and R.sup.13 is
hydrogen, halogen, --NR.sup.A1R.sup.A2, or optionally substituted
C.sub.1-3alkyl; R.sup.A1 and R.sup.A2 are each independently
hydrogen, optionally substituted C.sub.1-3 alkyl, a nitrogen
protecting group, or R.sup.A1 and R.sup.A2 are taken together with
the intervening nitrogen atom to form an optionally substituted 3-6
membered heterocyclic ring; R.sup.1 is hydrogen, R.sup.z, or
--C(O)R.sup.z, wherein R.sup.z is optionally substituted C.sub.1-6
alkyl; X is a bond, --O--, --N(R)--, --CR.sup.4R.sup.5--,
--O--CR.sup.4R.sup.5, --N(R)--CR.sup.4R.sup.5--,
--O--CR.sup.4R.sup.5--O--, --N(R)--CR.sup.4R.sup.5--O,
--N(R)--CR.sup.4R.sup.5--N(R)--, --O--CR.sup.4R.sup.5--N(R)--,
--CR.sup.4R.sup.5--O--, --CR.sup.4R.sup.5--N(R)--,
--O--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--N(R)--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--O--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--N(R)--, or
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--; each R is independently
hydrogen or optionally substituted C.sub.1-6 aliphatic; R.sup.2 and
R.sup.3 are each independently selected from the group consisting
of hydrogen, halo, --CN, --NO.sub.2, optionally substituted
aliphatic, optionally substituted carbocyclyl, optionally
substituted phenyl, optionally substituted heterocyclyl, optionally
substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A,
--C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
R.sup.4 and R.sup.5 are each independently selected from the group
consisting of hydrogen, halo, --CN, --NO.sub.2, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted phenyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.4 and R.sup.5 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
R.sup.6 and R.sup.7 are each independently selected from the group
consisting of hydrogen, halo, --CN, --NO.sub.2, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted phenyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.6 and R.sup.7 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
each R.sup.A is independently selected from the group consisting of
hydrogen, optionally substituted aliphatic, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, and optionally substituted heteroaryl; each
R.sup.B is independently selected from the group consisting of
hydrogen, optionally substituted aliphatic, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, and optionally substituted heteroaryl, or two
R.sup.B groups are taken together with their intervening atoms to
form an optionally substituted heterocyclic ring; R.sup.8, R.sup.9,
R.sup.10, and R.sup.11 are independently hydrogen, halo, or
optionally substituted aliphatic; Cy is a monocyclic or bicyclic,
saturated, partially unsaturated, or aromatic ring having 0-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur, wherein Cy is substituted with 0, 1, 2, 3, or 4 R.sup.y
groups; each R.sup.y is independently selected from the group
consisting of halo, --CN, --NO.sub.2, optionally substituted
aliphatic, optionally substituted carbocyclyl, optionally
substituted aryl, optionally substituted heterocyclyl, optionally
substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A,
--C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or an
R.sup.y group may be optionally taken together with R.sup.2 or
R.sup.3 to form an optionally substituted 5- to 6-membered
carbocyclic or heterocyclic ring fused to Cy; each R.sup.x is
independently selected from the group consisting of halo, --CN,
optionally substituted aliphatic, --OR', and --N(R'').sub.2; each
R' is independently hydrogen or optionally substituted aliphatic;
each R'' is independently hydrogen or optionally substituted
aliphatic, or two R'' are taken together with their intervening
atoms to form an optionally substituted heterocyclic ring having
1-2 heteroatoms independently selected from nitrogen, oxygen, and
sulfur; and n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency
permits; wherein each instance of aliphatic is independently an
alkyl, alkenyl, alkynyl, cycloalkyl, or cycloalkenyl group; and
each instance of optionally substituted is independently
unsubstituted or substituted, and each instance of substituted is,
when on a carbon atom, halogen, --CN, --NO.sub.2, --N.sub.3,
--SO.sub.2H, --SO.sub.3H, --OH, --OR.sup.aa, --ON(R.sup.bb).sub.2,
--N(R.sup.bb).sub.2, --N(R.sup.bb).sub.3.sup.+X.sup.-,
--N(OR.sup.cc)R.sup.bb, --SH, --SR.sup.aa, --SSR.sup.cc,
--C(.dbd.O)R.sup.aa, --CO.sub.2H, --CHO, --C(OR.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --OC(.dbd.O)R.sup.aa, --OCO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.2, --OC(.dbd.O)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.O)R.sup.aa, --NR.sup.bbCO.sub.2R.sup.aa,
--NR.sup.bbC(.dbd.O)N(R.sup.bb).sub.2, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --OC(.dbd.NR.sup.bb)R.sup.aa,
--OC(.dbd.NR.sup.bb)OR.sup.aa,
--C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--OC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--C(.dbd.O)NR.sup.bbSO.sub.2R.sup.aa, --NR.sup.bbSO.sub.2R.sup.aa,
--SO.sub.2N(R.sup.bb).sub.2, --SO.sub.2R.sup.aa,
--SO.sub.2OR.sup.aa, --OSO.sub.2R.sup.aa, --S(.dbd.O)R.sup.aa,
--OS(.dbd.O)R.sup.aa, --Si(R.sup.aa).sub.3,
--OSi(R.sup.aa).sub.3--C(.dbd.S)N(R.sup.bb).sub.2,
--C(.dbd.O)SR.sup.aa, --C(.dbd.S)SR.sup.aa, --SC(.dbd.S)SR.sup.aa,
--SC(.dbd.O)SR.sup.aa, --OC(.dbd.O)SR.sup.aa,
--SC(.dbd.O)OR.sup.aa, --SC(.dbd.O)R.sup.aa,
--P(.dbd.O)(R.sup.aa).sub.2, --OP(.dbd.O)(R.sup.aa).sub.2,
--OP(.dbd.O)(OR.sup.cc).sub.2, --P(.dbd.O).sub.2N(R.sup.bb).sub.2,
--OP(.dbd.O).sub.2N(R.sup.bb).sub.2, --P(.dbd.O)(NR.sup.bb).sub.2,
--OP(.dbd.O)(NR.sup.bb).sub.2,
--NR.sup.bbP(.dbd.O)(OR.sup.cc).sub.2,
--NR.sup.bbP(.dbd.O)(NR.sup.bb).sub.2, --P(R.sup.cc).sub.2,
--OP(R.sup.cc).sub.2, --OP(R.sup.cc).sub.3, --B(R.sup.aa).sub.2,
--B(OR.sup.cc).sub.2, --BR(OR.sup.cc), C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups, or two
geminal hydrogens on a carbon atom are replaced with the group
.dbd.O, .dbd.S, .dbd.NN(R.sup.bb).sub.2,
.dbd.NNR.sup.bbC(.dbd.O)R.sup.aa,
.dbd.NNR.sup.bbC(.dbd.O)OR.sup.aa,
.dbd.NNR.sup.bbS(.dbd.O).sub.2R.sup.aa, .dbd.NR.sup.bb, or
.dbd.NOR.sup.cc; each instance of substituted is, when on a
nitrogen, independently --OH, --OR.sup.aa, --N(R.sup.cc).sub.2,
--CN, --C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.bb)R.sup.aa, --C(.dbd.NR.sup.cc)OR.sup.aa,
--C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2, --SO.sub.2N(R.sup.cc).sub.2,
--SO.sub.2R.sup.cc, --SO.sub.2OR.sup.cc, --SOR.sup.aa,
--C(.dbd.S)N(R.sup.cc).sub.2, --C(.dbd.O)SR.sup.cc,
--C(.dbd.S)SR.sup.cc, --P(.dbd.O)(R.sup.aa).sub.2,
--P(.dbd.O)(NR.sup.cc).sub.2, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, or two R.sup.cc groups attached to a nitrogen
atom are joined to form a 3-14 membered heterocyclyl or 5-14
membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups; each instance
of R.sup.aa is, independently, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, or two R.sup.aa groups are joined to form a
3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,
wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 R.sup.dd groups; each instance of R.sup.bb is,
independently, hydrogen, --OH, --OR.sup.aa, --N(R.sup.cc).sub.2,
--CN, --C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.cc)OR.sup.aa, --C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2,
--SO.sub.2N(R.sup.cc).sub.2, --SO.sub.2R.sup.cc,
--SO.sub.2OR.sup.cc, --SOR.sup.aa, --C(.dbd.S)N(R.sup.cc).sub.2,
--C(.dbd.O)SR.sup.cc, --C(.dbd.S)SR.sup.cc,
--P(.dbd.O)(R.sup.aa).sub.2, --P(.dbd.O).sub.2N(R.sup.cc).sub.2,
C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, or 5-14 membered heteroaryl, or two
R.sup.bb groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups;
each instance of R.sup.cc is, independently, hydrogen, C.sub.1-10
alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10
alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered heterocyclyl,
C.sub.6-14 aryl, or 5-14 membered heteroaryl, or two R.sup.cc
groups are joined to form a 3-14 membered heterocyclyl or 5-14
membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups; each instance
of R.sup.dd is, independently, halogen, --CN, --NO.sub.2,
--N.sub.3, --SO.sub.2H, --SO.sub.3H, --OH, --OR.sup.ee,
--ON(R.sup.ff).sub.2, --N(R.sup.ff).sub.2,
--N(R.sup.ff).sub.3.sup.+X.sup.-, --N(OR.sup.ee)R.sup.ff, --SH,
--SR.sup.ee, --SSR.sup.ee, --C(.dbd.O)R.sup.ee, --CO.sub.2H,
--CO.sub.2R.sup.ee, --OC(.dbd.O)R.sup.ee, --OCO.sub.2R.sup.ee,
--C(.dbd.O)N(R.sup.ff).sub.2, --OC(.dbd.O)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.O)R.sup.ee, --NR.sup.ffCO.sub.2R.sup.ee,
--NR.sup.ffC(.dbd.O)N(R.sup.ff).sub.2,
--C(.dbd.NR.sup.ff)OR.sup.ee, --OC(.dbd.NR.sup.ff)R.sup.ee,
--OC(.dbd.NR.sup.ff)OR.sup.ee,
--C(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--OC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffSO.sub.2R.sup.ee, --SO.sub.2N(R.sup.ff).sub.2,
--SO.sub.2R.sup.ee, --SO.sub.2OR.sup.ee, --OSO.sub.2R.sup.ee,
--S(.dbd.O)R.sup.ee, --Si(R.sup.ee).sub.3, --OSi(R.sup.ee).sub.3,
--C(.dbd.S)N(R.sup.ff).sub.2, --C(.dbd.O)SR.sup.ee,
--C(.dbd.S)SR.sup.ee, --SC(.dbd.S)SR.sup.ee,
--P(.dbd.O)(R.sup.ee).sub.2, --OP(.dbd.O)(R.sup.ee).sub.2,
--OP(.dbd.O)(OR.sup.ee).sub.2, C.sub.1-6 alkyl, C.sub.1-6
perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-10
carbocyclyl, 3-10 membered heterocyclyl, C.sub.6-10 aryl, or 5-10
membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups, or two
geminal R.sup.dd substituents are joined to form .dbd.O or .dbd.S;
each instance of R.sup.ee is, independently, C.sub.1-6 alkyl,
C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl,
C.sub.3-10 carbocyclyl, C.sub.6-10 aryl, 3-10 membered
heterocyclyl, or 3-10 membered heteroaryl, wherein each alkyl,
alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl
is independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg
groups; each instance of R.sup.ff is, independently, hydrogen,
C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered
heterocyclyl, C.sub.6-10 aryl, or 5-10 membered heteroaryl, or two
R.sup.ff groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R
.sup.gg groups; and each instance of R.sup.gg is, independently,
halogen, --CN, --NO.sub.2, --N.sub.3, --SO.sub.2H, --SO.sub.3H,
--OH, --OC.sub.1-6 alkyl, --ON(C.sub.1-6 alkyl).sub.2,
--N(C.sub.1-6 alkyl).sub.2, --N(C.sub.1-6
alkyl).sub.3.sup.+X.sup.-, --NH(C.sub.1-6
alkyl).sub.2.sup.+X.sup.-, --NH.sub.2(C.sub.1-6
alkyl).sup.+X.sup.-, --NH.sub.3.sup.+X.sup.-, --N(OC.sub.1-6
alkyl)(C.sub.1-6 alkyl), --N(OH)(C.sub.1-6 alkyl), --NH(OH), --SH,
--SC.sub.1-6 alkyl, --SS(C.sub.1-6 alkyl), --C(.dbd.O)(C.sub.1-6
alkyl), --CO.sub.2H, --CO.sub.2(C.sub.1-6 alkyl),
--OC(.dbd.O)(C.sub.1-6 alkyl), --OCO.sub.2(C.sub.1-6 alkyl),
--C(.dbd.O)NH.sub.2, --C(.dbd.O)N(C.sub.1-6 alkyl).sub.2,
--OC(.dbd.O)NH(C.sub.1-6 alkyl), --NHC(.dbd.O)(C.sub.1-6 alkyl),
--N(C.sub.1-6 alkyl)C(.dbd.O)(C.sub.1-6 alkyl),
--NHCO.sub.2(C.sub.1-6 alkyl), --NHC(.dbd.O)N(C.sub.1-6
alkyl).sub.2, --NHC(.dbd.O)NH(C.sub.1-6 alkyl),
--NHC(.dbd.O)NH.sub.2, --C(.dbd.NH)O(C.sub.1-6 alkyl),
--OC(.dbd.NH)(C.sub.1-6 alkyl), --OC(.dbd.NH)OC.sub.1-6 alkyl,
--C(.dbd.NH)N(C.sub.1-6 alkyl).sub.2, --C(.dbd.NH)NH(C.sub.1-6
alkyl), --C(.dbd.NH)NH.sub.2, --OC(.dbd.NH)N(C.sub.1-6
alkyl).sub.2, --OC(NH)NH(C.sub.1-6 alkyl), --OC(NH)NH.sub.2,
--NHC(NH)N(C.sub.1-6 alkyl).sub.2, --NHC(.dbd.NH)NH.sub.2,
--NHSO.sub.2(C.sub.1-6 alkyl), --SO.sub.2N(C.sub.1-6 alkyl).sub.2,
--SO.sub.2NH(C.sub.1-6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2C.sub.1-6 alkyl, --SO.sub.2OC.sub.1-6 alkyl,
--OSO.sub.2C.sub.1-6 alkyl, --SOC.sub.1-6 alkyl, --Si(C.sub.1-6
alkyl).sub.3, --OSi(C.sub.1-6 alkyl).sub.3-C(.dbd.S)N(C.sub.1-6
alkyl).sub.2, C(.dbd.S)NH(C.sub.1-6 alkyl), C(.dbd.S)NH.sub.2,
--C(.dbd.O)S(C.sub.1-6 alkyl), --C(.dbd.S)SC.sub.1-6 alkyl,
--SC(.dbd.S)SC.sub.1-6 alkyl, --P(.dbd.O)(C.sub.1-6 alkyl).sub.2,
--OP(.dbd.O)(C.sub.1-6 alkyl).sub.2, --OP(.dbd.O)(OC.sub.1-6
alkyl).sub.2, C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6
alkenyl, C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, C.sub.6-10
aryl, 3-10 membered heterocyclyl, or 5-10 membered heteroaryl, or
two geminal R.sup.gg substituents are joined to form .dbd.O or
.dbd.S; wherein X.sup.- is a counterion.
103. A method of inhibiting PRMT5 comprising contacting a cell with
an effective amount of a compound of Formula (A), or a
pharmaceutically acceptable salt thereof; ##STR00614## or a
pharmaceutically acceptable salt thereof, wherein: represents a
single or double bond; R.sup.12 is halogen or optionally
substituted C.sub.1-3alkyl, and R.sup.13 is hydrogen, halogen,
--NR.sup.A1R.sup.A2, optionally substituted C.sub.1-3alkyl or
--OR.sup.1; or R.sup.12 is hydrogen, halogen or optionally
substituted C.sub.1-3alkyl, and R.sup.13 is hydrogen, halogen,
--NR.sup.A1R.sup.2, or optionally substituted C.sub.1-3alkyl;
R.sup.A1 and R.sup.A2 are each independently hydrogen, optionally
substituted C.sub.1-3 alkyl, a nitrogen protecting group, or
R.sup.A1 and R.sup.A2 are taken together with the intervening
nitrogen atom to form an optionally substituted 3-6 membered
heterocyclic ring; R.sup.1 is hydrogen, R.sup.z, or --C(O)R.sup.z,
wherein R.sup.z is optionally substituted C.sub.1-6 alkyl; X is a
bond, --O--, --N(R)--, --CR.sup.4R.sup.5--, --O--CR.sup.4R.sup.5,
--N(R)--CR.sup.4R.sup.5--, --O--CR.sup.4R.sup.5--O--,
--N(R)--CR.sup.4R.sup.5--O, --N(R)--CR.sup.4R.sup.5--N(R)--,
--O--CR.sup.4R.sup.5--N(R)--, --CR.sup.4R.sup.5--O--,
--CR.sup.4R.sup.5--N(R)--, --O--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--N(R)--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--O--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--N(R)--, or
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--; each R is independently
hydrogen or optionally substituted C.sub.1-6 aliphatic; R.sup.2 and
R.sup.3 are each independently selected from the group consisting
of hydrogen, halo, --CN, --NO.sub.2, optionally substituted
aliphatic, optionally substituted carbocyclyl, optionally
substituted phenyl, optionally substituted heterocyclyl, optionally
substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A,
--C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
R.sup.4 and R.sup.5 are each independently selected from the group
consisting of hydrogen, halo, --CN, --NO.sub.2, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted phenyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.4 and R.sup.5 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
R.sup.6 and R.sup.7 are each independently selected from the group
consisting of hydrogen, halo, --CN, --NO.sub.2, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted phenyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.6 and R.sup.7 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
each R.sup.A is independently selected from the group consisting of
hydrogen, optionally substituted aliphatic, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, and optionally substituted heteroaryl; each
R.sup.B is independently selected from the group consisting of
hydrogen, optionally substituted aliphatic, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, and optionally substituted heteroaryl, or two
R.sup.B groups are taken together with their intervening atoms to
form an optionally substituted heterocyclic ring; R.sup.8, R.sup.9,
R.sup.10, and R.sup.11 are independently hydrogen, halo, or
optionally substituted aliphatic; Cy is a monocyclic or bicyclic,
saturated, partially unsaturated, or aromatic ring having 0-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur, wherein Cy is substituted with 0, 1, 2, 3, or 4 R.sup.y
groups; each R.sup.y is independently selected from the group
consisting of halo, --CN, --NO.sub.2, optionally substituted
aliphatic, optionally substituted carbocyclyl, optionally
substituted aryl, optionally substituted heterocyclyl, optionally
substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A,
--C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or an
R.sup.y group may be optionally taken together with R.sup.2 or
R.sup.3 to form an optionally substituted 5- to 6-membered
carbocyclic or heterocyclic ring fused to Cy; each R.sup.x is
independently selected from the group consisting of halo, --CN,
optionally substituted aliphatic, --OR', and --N(R'').sub.2; each
R' is independently hydrogen or optionally substituted aliphatic;
each R'' is independently hydrogen or optionally substituted
aliphatic, or two R'' are taken together with their intervening
atoms to form an optionally substituted heterocyclic ring having
1-2 heteroatoms independently selected from nitrogen, oxygen, and
sulfur; and n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency
permits; wherein each instance of aliphatic is independently an
alkyl, alkenyl, alkynyl, cycloalkyl, or cycloalkenyl group; and
each instance of optionally substituted is independently
unsubstituted or substituted, and each instance of substituted is,
when on a carbon atom, halogen, --CN, --NO.sub.2, --N.sub.3,
--SO.sub.2H, --SO.sub.3H, --OH, --OR.sup.aa, --ON(R.sup.bb).sub.2,
--N(R.sup.bb).sub.2, --N(R.sup.bb).sub.3.sup.+X.sup.-,
--N(OR.sup.cc)R.sup.bb, --SH, --SR.sup.aa, --SSR.sup.cc,
--C(.dbd.O)R.sup.aa, --CO.sub.2H, --CHO, --C(OR.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --OC(.dbd.O)R.sup.aa, --OCO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.2, --OC(.dbd.O)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.O)R.sup.aa, --NR.sup.bbCO.sub.2R.sup.aa,
--NR.sup.bbC(.dbd.O)N(R.sup.bb).sub.2, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --OC(.dbd.NR.sup.bb)R.sup.aa,
--OC(.dbd.NR.sup.bb)OR.sup.aa,
--C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--OC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--C(.dbd.O)NR.sup.bbSO.sub.2R.sup.aa, --NR.sup.bbSO.sub.2R.sup.aa,
--SO.sub.2N(R.sup.bb).sub.2, --SO.sub.2R.sup.aa,
--SO.sub.2OR.sup.aa, --OSO.sub.2R.sup.aa, --S(.dbd.O)R.sup.aa,
--OS(.dbd.O)R.sup.aa, --Si(R.sup.aa).sub.3,
--OSi(R.sup.aa).sub.3--C(.dbd.S)N(R.sup.bb).sub.2,
--C(.dbd.O)SR.sup.aa, --C(.dbd.S)SR.sup.aa, --SC(.dbd.S)SR.sup.aa,
--SC(.dbd.O)SR.sup.aa, --OC(.dbd.O)SR.sup.aa,
--SC(.dbd.O)OR.sup.aa, --SC(.dbd.O)R.sup.aa,
--P(.dbd.O)(R.sup.aa).sub.2, --OP(.dbd.O)(R.sup.aa).sub.2,
--OP(.dbd.O)(OR.sup.cc).sub.2, --P(.dbd.O).sub.2N(R.sup.bb).sub.2,
--OP(.dbd.O).sub.2N(R.sup.bb).sub.2, --P(.dbd.O)(NR.sup.bb).sub.2,
--OP(.dbd.O)(NR.sup.bb).sub.2,
--NR.sup.bbP(.dbd.O)(OR.sup.cc).sub.2,
--NR.sup.bbP(.dbd.O)(NR.sup.bb).sub.2, --P(R.sup.cc).sub.2,
--OP(R.sup.cc).sub.2, --OP(R.sup.cc).sub.3, --B(R.sup.aa).sub.2,
--B(OR.sup.cc).sub.2, --BR(OR.sup.cc), C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups, or two
geminal hydrogens on a carbon atom are replaced with the group
.dbd.O, .dbd.S, .dbd.NN(R.sup.bb).sub.2,
.dbd.NNR.sup.bbC(.dbd.O)R.sup.aa,
.dbd.NNR.sup.bbC(.dbd.O)OR.sup.aa,
.dbd.NNR.sup.bbS(.dbd.O).sub.2R.sup.aa, .dbd.NR.sup.bb, or
.dbd.NOR.sup.cc; each instance of substituted is, when on a
nitrogen, independently --OH, --OR.sup.aa, --N(R.sup.cc).sub.2,
--CN, --C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.bb)R.sup.aa, --C(.dbd.NR.sup.cc)OR.sup.aa,
--C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2, --SO.sub.2N(R.sup.cc).sub.2,
--SO.sub.2R.sup.aa, --SO.sub.2OR.sup.cc, --SOR.sup.aa,
--C(.dbd.S)N(R.sup.cc).sub.2, --C(.dbd.O)SR.sup.cc,
--C(.dbd.S)SR.sup.cc, --P(.dbd.O)(R.sup.aa).sub.2,
--P(.dbd.O)(NR.sup.cc).sub.2, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, or two R.sup.cc groups attached to a nitrogen
atom are joined to form a 3-14 membered heterocyclyl or 5-14
membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups; each instance
of R.sup.aa is, independently, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, or two R.sup.aa groups are joined to form a
3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,
wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 R.sup.dd groups; each instance of R.sup.bb is,
independently, hydrogen, --OH, --OR.sup.aa, --N(R.sup.cc).sub.2,
--CN, --C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.cc)OR.sup.aa, --C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2,
--SO.sub.2N(R.sup.cc).sub.2, --SO.sub.2R.sup.cc,
--SO.sub.2OR.sup.cc, --SOR.sup.aa, --C(.dbd.S)N(R.sup.cc).sub.2,
--C(.dbd.O)SR.sup.cc, --C(.dbd.S)SR.sup.cc,
--P(.dbd.O)(R.sup.aa).sub.2, --P(.dbd.O).sub.2N(R.sup.cc).sub.2,
C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, or 5-14 membered heteroaryl, or two
R.sup.bb groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups;
each instance of R.sup.cc is, independently, hydrogen, C.sub.1-10
alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10
alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered heterocyclyl,
C.sub.6-14 aryl, or 5-14 membered heteroaryl, or two R.sup.cc
groups are joined to form a 3-14 membered heterocyclyl or 5-14
membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups; each instance
of R.sup.dd is, independently, halogen, --CN, --NO.sub.2,
--N.sub.3, --SO.sub.2H, --SO.sub.3H, --OH, --OR.sup.ee,
--ON(R.sup.ff).sub.2, --N(R.sup.ff).sub.2,
--N(R.sup.ff).sub.3.sup.+X.sup.-, --N(OR.sup.ee)R.sup.ff, --SH,
--SR.sup.ee, --SSR.sup.ee, --C(.dbd.O)R.sup.ee, --CO.sub.2H,
--CO.sub.2R.sup.ee, --OC(.dbd.O)R.sup.ee, --OCO.sub.2R.sup.ee,
--C(.dbd.O)N(R.sup.ff).sub.2, --OC(.dbd.O)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.O)R.sup.ee, --NR.sup.ffCO.sub.2R.sup.ee,
--NR.sup.ffC(.dbd.O)N(R.sup.ff).sub.2,
--C(.dbd.NR.sup.ff)OR.sup.ee, --OC(.dbd.NR.sup.ff)R.sup.ee,
--OC(.dbd.NR.sup.ff)OR.sup.ee,
--C(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--OC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffSO.sub.2R.sup.ee, --SO.sub.2N(R.sup.ff).sub.2,
--SO.sub.2R.sup.ee, --SO.sub.2OR.sup.ee, --OSO.sub.2R.sup.ee,
--S(.dbd.O)R.sup.ee, --Si(R.sup.ee).sub.3, --OSi(R.sup.ee).sub.3,
--C(.dbd.S)N(R.sup.ff).sub.2, --C(.dbd.O)SR.sup.ee,
--C(.dbd.S)SR.sup.ee, --SC(.dbd.S)SR.sup.ee,
--P(.dbd.O)(R.sup.ee).sub.2, --OP(.dbd.O)(R.sup.ee).sub.2,
--OP(.dbd.O)(OR.sup.ee).sub.2, C.sub.1-6 alkyl, C.sub.1-6
perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-10
carbocyclyl, 3-10 membered heterocyclyl, C.sub.6-10 aryl, or 5-10
membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups, or two
geminal R.sup.dd substituents are joined to form .dbd.O or .dbd.S;
each instance of R.sup.ee is, independently, C.sub.1-6 alkyl,
C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl,
C.sub.3-10 carbocyclyl, C.sub.6-10 aryl, 3-10 membered
heterocyclyl, or 3-10 membered heteroaryl, wherein each alkyl,
alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl
is independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg
groups; each instance of R.sup.ff is, independently, hydrogen,
C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered
heterocyclyl, C.sub.6-10 aryl, or 5-10 membered heteroaryl, or two
R.sup.ff
groups are joined to form a 3-14 membered heterocyclyl or 5-14
membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups; and each
instance of R.sup.gg is, independently, halogen, --CN, --NO.sub.2,
--N.sub.3, --SO.sub.2H, --SO.sub.3H, --OH, --OC.sub.1-6 alkyl,
--ON(C.sub.1-6 alkyl).sub.2, --N(C.sub.1-6 alkyl).sub.2,
--N(C.sub.1-6 alkyl).sub.3.sup.+X.sup.-, --NH(C.sub.1-6
alkyl).sub.2.sup.+X.sup.-, --NH.sub.2(C.sub.1-6
alkyl).sup.+X.sup.-, --NH.sub.3.sup.+X.sup.-, --N(OC.sub.1-6
alkyl)(C.sub.1-6 alkyl), --N(OH)(C.sub.1-6 alkyl), --NH(OH), --SH,
--SC.sub.1-6 alkyl, --SS(C.sub.1-6 alkyl), --C(.dbd.O)(C.sub.1-6
alkyl), --CO.sub.2H, --CO.sub.2(C.sub.1-6 alkyl),
--OC(.dbd.O)(C.sub.1-6 alkyl), --OCO.sub.2(C.sub.1-6 alkyl),
--C(.dbd.O)NH.sub.2, --C(.dbd.O)N(C.sub.1-6 alkyl).sub.2,
--OC(.dbd.O)NH(C.sub.1-6 alkyl), --NHC(.dbd.O)(C.sub.1-6 alkyl),
--N(C.sub.1-6 alkyl)C(.dbd.O)(C.sub.1-6 alkyl),
--NHCO.sub.2(C.sub.1-6 alkyl), --NHC(.dbd.O)N(C.sub.1-6
alkyl).sub.2, --NHC(.dbd.O)NH(C.sub.1-6 alkyl),
--NHC(.dbd.O)NH.sub.2, --C(.dbd.NH)O(C.sub.1-6 alkyl),
--OC(.dbd.NH)(C.sub.1-6 alkyl), --OC(.dbd.NH)OC.sub.1-6 alkyl,
--C(.dbd.NH)N(C.sub.1-6 alkyl).sub.2, --C(.dbd.NH)NH(C.sub.1-6
alkyl), --C(.dbd.NH)NH.sub.2, --OC(.dbd.NH)N(C.sub.1-6
alkyl).sub.2, --OC(NH)NH(C.sub.1-6 alkyl), --OC(NH)NH.sub.2,
--NHC(NH)N(C.sub.1-6 alkyl).sub.2, --NHC(.dbd.NH)NH.sub.2,
--NHSO.sub.2(C.sub.1-6 alkyl), --SO.sub.2N(C.sub.1-6 alkyl).sub.2,
--SO.sub.2NH(C.sub.1-6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2C.sub.1-6 alkyl, --SO.sub.2OC.sub.1-6 alkyl,
--OSO.sub.2C.sub.1-6 alkyl, --SOC.sub.1-6 alkyl, --Si(C.sub.1-6
alkyl).sub.3, --OSi(C.sub.1-6 alkyl).sub.3-C(.dbd.S)N(C.sub.1-6
alkyl).sub.2, C(.dbd.S)NH(C.sub.1-6 alkyl), C(.dbd.S)NH.sub.2,
--C(.dbd.O)S(C.sub.1-6 alkyl), --C(.dbd.S)SC.sub.1-6 alkyl,
--SC(.dbd.S)SC.sub.1-6 alkyl, --P(.dbd.O)(C.sub.1-6 alkyl).sub.2,
--OP(.dbd.O)(C.sub.1-6 alkyl).sub.2, --OP(.dbd.O)(OC.sub.1-6
alkyl).sub.2, C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6
alkenyl, C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, C.sub.6-10
aryl, 3-10 membered heterocyclyl, or 5-10 membered heteroaryl, or
two geminal R.sup.gg substituents are joined to form .dbd.O or
.dbd.S; wherein X.sup.- is a counterion.
104. A method of altering gene expression comprising contacting a
cell with an effective amount of a compound of Formula (A), or a
pharmaceutically acceptable salt thereof: ##STR00615## or a
pharmaceutically acceptable salt thereof, wherein: represents a
single or double bond; R.sup.12 is halogen or optionally
substituted C.sub.1-3alkyl, and R.sup.13 is hydrogen, halogen,
--NR.sup.A1R.sup.A2, optionally substituted C.sub.1-3alkyl or
--OR.sup.1; or R.sup.12 is hydrogen, halogen or optionally
substituted C.sub.1-3alkyl, and R.sup.13 is hydrogen, halogen,
--NR.sup.A1R.sup.A2, or optionally substituted C.sub.1-3alkyl;
R.sup.A1 and R.sup.A2 are each independently hydrogen, optionally
substituted C.sub.1-3 alkyl, a nitrogen protecting group, or
R.sup.A1 and R.sup.A2 are taken together with the intervening
nitrogen atom to form an optionally substituted 3-6 membered
heterocyclic ring; R.sup.1 is hydrogen, R.sup.z, or --C(O)R.sup.z,
wherein R.sup.z is optionally substituted C.sub.1-6 alkyl; X is a
bond, --O--, --N(R)--, --CR.sup.4R.sup.5--, --O--CR.sup.4R.sup.5,
--N(R)--CR.sup.4R.sup.5--, --O--CR.sup.4R.sup.5--O--,
--N(R)--CR.sup.4R.sup.5--O, --N(R)--CR.sup.4R.sup.5--N(R)--,
--O--CR.sup.4R.sup.5--N(R)--, --CR.sup.4R.sup.5--O--,
--CR.sup.4R.sup.5--N(R)--, --O--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--N(R)--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--O--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--N(R)--, or
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--; each R is independently
hydrogen or optionally substituted C.sub.1-6 aliphatic; R.sup.2 and
R.sup.3 are each independently selected from the group consisting
of hydrogen, halo, --CN, --NO.sub.2, optionally substituted
aliphatic, optionally substituted carbocyclyl, optionally
substituted phenyl, optionally substituted heterocyclyl, optionally
substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A,
--C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
R.sup.4 and R.sup.5 are each independently selected from the group
consisting of hydrogen, halo, --CN, --NO.sub.2, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted phenyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.4 and R.sup.5 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
R.sup.6 and R.sup.7 are each independently selected from the group
consisting of hydrogen, halo, --CN, --NO.sub.2, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted phenyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.6 and R.sup.7 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
each R.sup.A is independently selected from the group consisting of
hydrogen, optionally substituted aliphatic, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, and optionally substituted heteroaryl; each
R.sup.B is independently selected from the group consisting of
hydrogen, optionally substituted aliphatic, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, and optionally substituted heteroaryl, or two
R.sup.B groups are taken together with their intervening atoms to
form an optionally substituted heterocyclic ring; R.sup.8, R.sup.9,
R.sup.10, and R.sup.11 are independently hydrogen, halo, or
optionally substituted aliphatic; Cy is a monocyclic or bicyclic,
saturated, partially unsaturated, or aromatic ring having 0-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur, wherein Cy is substituted with 0, 1, 2, 3, or 4 R.sup.y
groups; each R.sup.y is independently selected from the group
consisting of halo, --CN, --NO.sub.2, optionally substituted
aliphatic, optionally substituted carbocyclyl, optionally
substituted aryl, optionally substituted heterocyclyl, optionally
substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A,
--C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or an
R.sup.y group may be optionally taken together with R.sup.2 or
R.sup.3 to form an optionally substituted 5- to 6-membered
carbocyclic or heterocyclic ring fused to Cy; each R.sup.x is
independently selected from the group consisting of halo, --CN,
optionally substituted aliphatic, --OR', and --N(R'').sub.2; each
R' is independently hydrogen or optionally substituted aliphatic;
each R'' is independently hydrogen or optionally substituted
aliphatic, or two R'' are taken together with their intervening
atoms to form an optionally substituted heterocyclic ring having
1-2 heteroatoms independently selected from nitrogen, oxygen, and
sulfur; and n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency
permits; wherein each instance of aliphatic is independently an
alkyl, alkenyl, alkynyl, cycloalkyl, or cycloalkenyl group; and
each instance of optionally substituted is independently
unsubstituted or substituted, and each instance of substituted is,
when on a carbon atom, halogen, --CN, --NO.sub.2, --N.sub.3,
--SO.sub.2H, --SO.sub.3H, --OH, --OR.sup.aa, --ON(R.sup.bb).sub.2,
--N(R.sup.bb).sub.2, --N(R.sup.bb).sub.3.sup.+X.sup.-,
--N(OR.sup.cc)R.sup.bb, --SH, --SR.sup.aa, --SSR.sup.cc,
--C(.dbd.O)R.sup.aa, --CO.sub.2H, --CHO, --C(OR.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --OC(.dbd.O)R.sup.aa, --OCO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.2, --OC(.dbd.O)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.O)R.sup.aa, --NR.sup.bbCO.sub.2R.sup.aa,
--NR.sup.bbC(.dbd.O)N(R.sup.bb).sub.2, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --OC(.dbd.NR.sup.bb)R.sup.aa,
--OC(.dbd.NR.sup.bb)OR.sup.aa,
--C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--OC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--C(.dbd.O)NR.sup.bbSO.sub.2R.sup.aa, --NR.sup.bbSO.sub.2R.sup.aa,
--SO.sub.2N(R.sup.bb).sub.2, --SO.sub.2R.sup.aa,
--SO.sub.2OR.sup.aa, --OSO.sub.2R.sup.aa, --S(.dbd.O)R.sup.aa,
--OS(.dbd.O)R.sup.aa, --Si(R.sup.aa).sub.3,
--OSi(R.sup.aa).sub.3--C(.dbd.S)N(R.sup.bb).sub.2,
--C(.dbd.O)SR.sup.aa, --C(.dbd.S)SR.sup.aa, --SC(.dbd.S)SR.sup.aa,
--SC(.dbd.O)SR.sup.aa, --OC(.dbd.O)SR.sup.aa,
--SC(.dbd.O)OR.sup.aa, --SC(.dbd.O)R.sup.aa,
--P(.dbd.O)(R.sup.aa).sub.2, --OP(.dbd.O)(R.sup.aa).sub.2,
--OP(.dbd.O)(OR.sup.cc).sub.2, --P(.dbd.O).sub.2N(R.sup.bb).sub.2,
--OP(.dbd.O).sub.2N(R.sup.bb).sub.2, --P(.dbd.O)(NR.sup.bb).sub.2,
--OP(.dbd.O)(NR.sup.bb).sub.2,
--NR.sup.bbP(.dbd.O)(OR.sup.cc).sub.2,
--NR.sup.bbP(.dbd.O)(NR.sup.bb).sub.2, --P(R.sup.cc).sub.2,
--OP(R.sup.cc).sub.2, --OP(R.sup.cc).sub.3, --B(R.sup.aa).sub.2,
--B(OR.sup.cc).sub.2, --BR(OR.sup.cc), C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups, or two
geminal hydrogens on a carbon atom are replaced with the group
.dbd.O, .dbd.S, .dbd.NN(R.sup.bb).sub.2,
.dbd.NNR.sup.bbC(.dbd.O)R.sup.aa,
.dbd.NNR.sup.bbC(.dbd.O)OR.sup.aa,
.dbd.NNR.sup.bbS(.dbd.O).sub.2R.sup.aa, .dbd.NR.sup.bb, or
.dbd.NOR.sup.cc; each instance of substituted is, when on a
nitrogen, independently --OH, --OR.sup.aa, --N(R.sup.cc).sub.2,
--CN, --C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.bb)R.sup.aa, --C(.dbd.NR.sup.cc)OR.sup.aa,
--C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2, --SO.sub.2N(R.sup.cc).sub.2,
--SO.sub.2R.sup.cc, --SO.sub.2OR.sup.aa, --SOR.sup.aa,
--C(.dbd.S)N(R.sup.cc).sub.2, --C(.dbd.O)SR.sup.cc,
--C(.dbd.S)SR.sup.cc, --P(.dbd.O)(R.sup.aa).sub.2,
--P(.dbd.O)(NR.sup.cc).sub.2, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, or two R.sup.cc groups attached to a nitrogen
atom are joined to form a 3-14 membered heterocyclyl or 5-14
membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups; each instance
of R.sup.aa is, independently, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, or two R.sup.aa groups are joined to form a
3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,
wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 R.sup.dd groups; each instance of R.sup.bb is,
independently, hydrogen, --OH, --OR.sup.aa, --N(R.sup.cc).sub.2,
--CN, --C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.cc)OR.sup.aa, --C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2,
--SO.sub.2N(R.sup.cc).sub.2, --SO.sub.2R.sup.cc,
--SO.sub.2OR.sup.cc, --SOR.sup.aa, --C(.dbd.S)N(R.sup.cc).sub.2,
--C(.dbd.O)SR.sup.cc, --C(.dbd.S)SR.sup.cc,
--P(.dbd.O)(R.sup.aa).sub.2, --P(.dbd.O).sub.2N(R.sup.cc).sub.2,
C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, or 5-14 membered heteroaryl, or two
R.sup.bb groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups;
each instance of R.sup.cc is, independently, hydrogen, C.sub.1-10
alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10
alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered heterocyclyl,
C.sub.6-4 aryl, or 5-14 membered heteroaryl, or two R.sup.cc groups
are joined to form a 3-14 membered heterocyclyl or 5-14 membered
heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl,
heterocyclyl, aryl, and heteroaryl is independently substituted
with 0, 1, 2, 3, 4, or 5 R.sup.dd groups; each instance of R.sup.dd
is, independently, halogen, --CN, --NO.sub.2, --N.sub.3,
--SO.sub.2H, --SO.sub.3H, --OH, --OR.sup.ee, --ON(R.sup.ff).sub.2,
--N(R.sup.ff).sub.2, --N(R.sup.ff).sub.3.sup.+X.sup.-,
--N(OR.sup.ee)R.sup.ff, --SH, --SR.sup.ee, --SSR.sup.ee,
--C(.dbd.O)R.sup.ee, --CO.sub.2H, --CO.sub.2R.sup.ee,
--OC(.dbd.O)R.sup.ee, --OCO.sub.2R.sup.ee,
--C(.dbd.O)N(R.sup.ff).sub.2, --OC(.dbd.O)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.O)R.sup.ee, --NR.sup.ffCO.sub.2R.sup.ee,
--NR.sup.ffC(.dbd.O)N(R.sup.ff).sub.2,
--C(.dbd.NR.sup.ff)OR.sup.ee, --OC(.dbd.NR.sup.ff)R.sup.ee,
--OC(.dbd.NR.sup.ff)OR.sup.ee,
--C(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--OC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffSO.sub.2R.sup.ee, --SO.sub.2N(R.sup.ff).sub.2,
--SO.sub.2R.sup.ee, --SO.sub.2OR.sup.ee, --OSO.sub.2R.sup.ee,
--S(.dbd.O)R.sup.ee, --Si(R.sup.ee).sub.3, --OSi(R.sup.ee).sub.3,
--C(.dbd.S)N(R.sup.ff).sub.2, --C(.dbd.O)SR.sup.ee,
--C(.dbd.S)SR.sup.ee, --SC(.dbd.S)SR.sup.ee,
--P(.dbd.O)(R.sup.ee).sub.2, --OP(.dbd.O)(R.sup.ee).sub.2,
--OP(.dbd.O)(OR.sup.ee).sub.2, C.sub.1-6 alkyl, C.sub.1-6
perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-10
carbocyclyl, 3-10 membered heterocyclyl, C.sub.6-10 aryl, or 5-10
membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups, or two
geminal R.sup.dd substituents are joined to form .dbd.O or .dbd.S;
each instance of R.sup.ee is, independently, C.sub.1-6 alkyl,
C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl,
C.sub.3-10 carbocyclyl, C.sub.6-10 aryl, 3-10 membered
heterocyclyl, or 3-10 membered heteroaryl, wherein each alkyl,
alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl
is independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg
groups; each instance of R.sup.ff is, independently, hydrogen,
C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered
heterocyclyl, C.sub.6-10 aryl, or 5-10 membered heteroaryl, or two
R
.sup.ff groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups;
and each instance of R.sup.gg is, independently, halogen, --CN,
--NO.sub.2, --N.sub.3, --SO.sub.2H, --SO.sub.3H, --OH, --OC.sub.1-6
alkyl, --ON(C.sub.1-6 alkyl).sub.2, --N(C.sub.1-6 alkyl).sub.2,
--N(C.sub.1-6 alkyl).sub.3.sup.+X.sup.-, --NH(C.sub.1-6
alkyl).sub.2.sup.+X.sup.-, --NH.sub.2(C.sub.1-6
alkyl).sup.+X.sup.-, --NH.sub.3.sup.+X.sup.-, --N(OC.sub.1-6
alkyl)(C.sub.1-6 alkyl), --N(OH)(C.sub.1-6 alkyl), --NH(OH), --SH,
--SC.sub.1-6 alkyl, --SS(C.sub.1-6 alkyl), --C(.dbd.O)(C.sub.1-6
alkyl), --CO.sub.2H, --CO.sub.2(C.sub.1-6 alkyl),
--OC(.dbd.O)(C.sub.1-6 alkyl), --OCO.sub.2(C.sub.1-6 alkyl),
--C(.dbd.O)NH.sub.2, --C(.dbd.O)N(C.sub.1-6 alkyl).sub.2,
--OC(.dbd.O)NH(C.sub.1-6 alkyl), --NHC(.dbd.O)(C.sub.1-6 alkyl),
--N(C.sub.1-6 alkyl)C(.dbd.O)(C.sub.1-6 alkyl),
--NHCO.sub.2(C.sub.1-6 alkyl), --NHC(.dbd.O)N(C.sub.1-6
alkyl).sub.2, --NHC(.dbd.O)NH(C.sub.1-6 alkyl),
--NHC(.dbd.O)NH.sub.2, --C(.dbd.NH)O(C.sub.1-6 alkyl),
--OC(.dbd.NH)(C.sub.1-6 alkyl), --OC(.dbd.NH)OC.sub.1-6 alkyl,
--C(.dbd.NH)N(C.sub.1-6 alkyl).sub.2, --C(.dbd.NH)NH(C.sub.1-6
alkyl), --C(.dbd.NH)NH.sub.2, --OC(.dbd.NH)N(C.sub.1-6
alkyl).sub.2, --OC(NH)NH(C.sub.1-6 alkyl), --OC(NH)NH.sub.2,
--NHC(NH)N(C.sub.1-6 alkyl).sub.2, --NHC(.dbd.NH)NH.sub.2,
--NHSO.sub.2(C.sub.1-6 alkyl), --SO.sub.2N(C.sub.1-6 alkyl).sub.2,
--SO.sub.2NH(C.sub.1-6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2C.sub.1-6 alkyl, --SO.sub.2OC.sub.1-6 alkyl,
--OSO.sub.2C.sub.1-6 alkyl, --SOC.sub.1-6 alkyl, --Si(C.sub.1-6
alkyl).sub.3, --OSi(C.sub.1-6 alkyl).sub.3-C(.dbd.S)N(C.sub.1-6
alkyl).sub.2, C(.dbd.S)NH(C.sub.1-6 alkyl), C(.dbd.S)NH.sub.2,
--C(.dbd.O)S(C.sub.1-6 alkyl), --C(.dbd.S)SC.sub.1-6 alkyl,
--SC(.dbd.S)SC.sub.1-6 alkyl, --P(.dbd.O)(C.sub.1-6 alkyl).sub.2,
--OP(.dbd.O)(C.sub.1-6 alkyl).sub.2, --OP(.dbd.O)(OC.sub.1-6
alkyl).sub.2, C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6
alkenyl, C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, C.sub.6-10
aryl, 3-10 membered heterocyclyl, or 5-10 membered heteroaryl, or
two geminal R.sup.gg substituents are joined to form .dbd.O or
.dbd.S; wherein X.sup.- is a counterion.
105. A method of altering transcription comprising contacting a
cell with an effective amount of a compound of Formula (A), or a
pharmaceutically acceptable salt thereof; ##STR00616## or a
pharmaceutically acceptable salt thereof, wherein: represents a
single or double bond; R.sup.12 is halogen or optionally
substituted C.sub.1-3alkyl, and R.sup.13 is hydrogen, halogen,
--NR.sup.A1R.sup.A2, optionally substituted C.sub.1-3alkyl or
--OR.sup.1; or R.sup.12 is hydrogen, halogen or optionally
substituted C.sub.1-3alkyl, and R.sup.13 is hydrogen, halogen,
--NR.sup.A1R.sup.2, or optionally substituted C.sub.1-3alkyl;
R.sup.A1 and R.sup.A2 are each independently hydrogen, optionally
substituted C.sub.1-3 alkyl, a nitrogen protecting group, or
R.sup.A1 and R.sup.A2 are taken together with the intervening
nitrogen atom to form an optionally substituted 3-6 membered
heterocyclic ring; R.sup.1 is hydrogen, R.sup.z, or --C(O)R.sup.z,
wherein R.sup.z is optionally substituted C.sub.1-6 alkyl; X is a
bond, --O--, --N(R)--, --CR.sup.4R.sup.5--, --O--CR.sup.4R.sup.5,
--N(R)--CR.sup.4R.sup.5--, --O--CR.sup.4R.sup.5--O--,
--N(R)--CR.sup.4R.sup.5--O, --N(R)--CR.sup.4R.sup.5--N(R)--,
--O--CR.sup.4R.sup.5--N(R)--, --CR.sup.4R.sup.5--O--,
--CR.sup.4R.sup.5--N(R)--, --O--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--N(R)--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--O--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--N(R)--, or
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--; each R is independently
hydrogen or optionally substituted C.sub.1-6 aliphatic; R.sup.2 and
R.sup.3 are each independently selected from the group consisting
of hydrogen, halo, --CN, --NO.sub.2, optionally substituted
aliphatic, optionally substituted carbocyclyl, optionally
substituted phenyl, optionally substituted heterocyclyl, optionally
substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A,
--C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
R.sup.4 and R.sup.5 are each independently selected from the group
consisting of hydrogen, halo, --CN, --NO.sub.2, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted phenyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.4 and R.sup.5 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
R.sup.6 and R.sup.7 are each independently selected from the group
consisting of hydrogen, halo, --CN, --NO.sub.2, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted phenyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.6 and R.sup.7 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
each R.sup.A is independently selected from the group consisting of
hydrogen, optionally substituted aliphatic, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, and optionally substituted heteroaryl; each
R.sup.B is independently selected from the group consisting of
hydrogen, optionally substituted aliphatic, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, and optionally substituted heteroaryl, or two
R.sup.B groups are taken together with their intervening atoms to
form an optionally substituted heterocyclic ring; R.sup.8, R.sup.9,
R.sup.10, and R.sup.11 are independently hydrogen, halo, or
optionally substituted aliphatic; Cy is a monocyclic or bicyclic,
saturated, partially unsaturated, or aromatic ring having 0-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur, wherein Cy is substituted with 0, 1, 2, 3, or 4 R.sup.y
groups; each R.sup.y is independently selected from the group
consisting of halo, --CN, --NO.sub.2, optionally substituted
aliphatic, optionally substituted carbocyclyl, optionally
substituted aryl, optionally substituted heterocyclyl, optionally
substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A,
--C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or an
R.sup.y group may be optionally taken together with R.sup.2 or
R.sup.3 to form an optionally substituted 5- to 6-membered
carbocyclic or heterocyclic ring fused to Cy; each R.sup.x is
independently selected from the group consisting of halo, --CN,
optionally substituted aliphatic, --OR', and --N(R'').sub.2; each
R' is independently hydrogen or optionally substituted aliphatic;
each R'' is independently hydrogen or optionally substituted
aliphatic, or two R'' are taken together with their intervening
atoms to form an optionally substituted heterocyclic ring having
1-2 heteroatoms independently selected from nitrogen, oxygen, and
sulfur; and n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency
permits; wherein each instance of aliphatic is independently an
alkyl, alkenyl, alkynyl, cycloalkyl, or cycloalkenyl group; and
each instance of optionally substituted is independently
unsubstituted or substituted, and each instance of substituted is,
when on a carbon atom, halogen, --CN, --NO.sub.2, --N.sub.3,
--SO.sub.2H, --SO.sub.3H, --OH, --OR.sup.aa, --ON(R.sup.bb).sub.2,
--N(R.sup.bb).sub.2, --N(R.sup.bb).sub.3.sup.+X.sup.-,
--N(OR.sup.cc)R.sup.bb, --SH, --SR.sup.aa, --SSR.sup.cc,
--C(.dbd.O)R.sup.aa, --CO.sub.2H, --CHO, --C(OR.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --OC(.dbd.O)R.sup.aa, --OCO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.2, --OC(.dbd.O)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.O)R.sup.aa, --NR.sup.bbCO.sub.2R.sup.aa,
--NR.sup.bbC(.dbd.O)N(R.sup.bb).sub.2, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --OC(.dbd.NR.sup.bb)R.sup.aa,
--OC(.dbd.NR.sup.bb)OR.sup.aa,
--C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--OC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--C(.dbd.O)NR.sup.bbSO.sub.2R.sup.aa, --NR.sup.bbSO.sub.2R.sup.aa,
--SO.sub.2N(R.sup.bb).sub.2, --SO.sub.2R.sup.aa,
--SO.sub.2OR.sup.aa, --OSO.sub.2R.sup.aa, --S(.dbd.O)R.sup.aa,
--OS(.dbd.O)R.sup.aa, --Si(R.sup.aa).sub.3,
--OSi(R.sup.aa).sub.3--C(.dbd.S)N(R.sup.bb).sub.2,
--C(.dbd.O)SR.sup.aa, --C(.dbd.S)SR.sup.aa, --SC(.dbd.S)SR.sup.aa,
--SC(.dbd.O)SR.sup.aa, --OC(.dbd.O)SR.sup.aa,
--SC(.dbd.O)OR.sup.aa, --SC(.dbd.O)R.sup.aa,
--P(.dbd.O)(R.sup.aa).sub.2, --OP(.dbd.O)(R.sup.aa).sub.2,
--OP(.dbd.O)(OR.sup.cc).sub.2, --P(.dbd.O).sub.2N(R.sup.bb).sub.2,
--OP(.dbd.O).sub.2N(R.sup.bb).sub.2, --P(.dbd.O)(NR.sup.bb).sub.2,
--OP(.dbd.O)(NR.sup.bb).sub.2,
--NR.sup.bbP(.dbd.O)(OR.sup.cc).sub.2,
--NR.sup.bbP(.dbd.O)(NR.sup.bb).sub.2, --P(R.sup.cc).sub.2,
--OP(R.sup.cc).sub.2, --OP(R.sup.cc).sub.3, --B(R.sup.aa).sub.2,
--B(OR.sup.cc).sub.2, --BR(OR.sup.cc), C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups, or two
geminal hydrogens on a carbon atom are replaced with the group
.dbd.O, .dbd.S, .dbd.NN(R.sup.bb).sub.2,
.dbd.NNR.sup.bbC(.dbd.O)R.sup.aa,
.dbd.NNR.sup.bbC(.dbd.O)OR.sup.aa,
.dbd.NNR.sup.bbS(.dbd.O).sub.2R.sup.aa, .dbd.NR.sup.bb, or
=NOR.sup.cc; each instance of substituted is, when on a nitrogen,
independently --OH, --OR.sup.aa, --N(R.sup.cc).sub.2, --CN,
--C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.bb)R.sup.aa, --C(.dbd.NR.sup.cc)OR.sup.aa,
--C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2, --SO.sub.2N(R.sup.cc).sub.2,
--SO.sub.2R.sup.aa, --SO.sub.2OR.sup.cc, --SOR.sup.aa,
--C(.dbd.S)N(R.sup.cc).sub.2, --C(.dbd.O)SR.sup.cc,
--C(.dbd.S)SR.sup.cc, --P(.dbd.O)(R.sup.aa).sub.2,
--P(.dbd.O)(NR.sup.cc).sub.2, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, or two R.sup.cc groups attached to a nitrogen
atom are joined to form a 3-14 membered heterocyclyl or 5-14
membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups; each instance
of R.sup.aa is, independently, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, or two R.sup.aa groups are joined to form a
3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,
wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 R.sup.dd groups; each instance of R.sup.bb is,
independently, hydrogen, --OH, --OR.sup.aa, --N(R.sup.cc).sub.2,
--CN, --C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.cc)OR.sup.aa, --C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2,
--SO.sub.2N(R.sup.cc).sub.2, --SO.sub.2R.sup.cc,
--SO.sub.2OR.sup.cc, --SOR.sup.aa, --C(.dbd.S)N(R.sup.cc).sub.2,
--C(.dbd.O)SR.sup.cc, --C(.dbd.S)SR.sup.cc,
--P(.dbd.O)(R.sup.aa).sub.2, --P(.dbd.O).sub.2N(R.sup.cc).sub.2,
C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, or 5-14 membered heteroaryl, or two
R.sup.bb groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups;
each instance of R.sup.cc is, independently, hydrogen, C.sub.1-10
alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10
alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered heterocyclyl,
C.sub.6-4 aryl, or 5-14 membered heteroaryl, or two R.sup.cc groups
are joined to form a 3-14 membered heterocyclyl or 5-14 membered
heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl,
heterocyclyl, aryl, and heteroaryl is independently substituted
with 0, 1, 2, 3, 4, or 5 R.sup.dd groups; each instance of R.sup.dd
is, independently, halogen, --CN, --NO.sub.2, --N.sub.3,
--SO.sub.2H, --SO.sub.3H, --OH, --OR.sup.ee, --ON(R.sup.ff).sub.2,
--N(R.sup.ff).sub.2, --N(R.sup.ff).sub.3.sup.+X.sup.-,
--N(OR.sup.ee)R.sup.ff, --SH, --SR.sup.ee, --SSR.sup.ee,
--C(.dbd.O)R.sup.ee, --CO.sub.2H, --CO.sub.2R.sup.ee,
--OC(.dbd.O)R.sup.ee, --OCO.sub.2R.sup.ee,
--C(.dbd.O)N(R.sup.ff).sub.2, --OC(.dbd.O)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.O)R.sup.ee, --NR.sup.ffCO.sub.2R.sup.ee,
--NR.sup.ffC(.dbd.O)N(R.sup.ff).sub.2,
--C(.dbd.NR.sup.ff)OR.sup.ee, --OC(.dbd.NR.sup.ff)R.sup.ee,
--OC(.dbd.NR.sup.ff)OR.sup.ee,
--C(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--OC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffSO.sub.2R.sup.ee, --SO.sub.2N(R.sup.ff).sub.2,
--SO.sub.2R.sup.ee, --SO.sub.2R.sup.ee, --OSO.sub.2R.sup.ee,
--S(.dbd.O)R.sup.ee, --Si(R.sup.ee).sub.3, --OSi(R.sup.ee).sub.3,
--C(.dbd.S)N(R.sup.ff).sub.2, --C(.dbd.O)SR.sup.ee,
--C(.dbd.S)SR.sup.ee, --SC(.dbd.S)SR.sup.ee,
--P(.dbd.O)(R.sup.ee).sub.2, --OP(.dbd.O)(R.sup.ee).sub.2,
--OP(.dbd.O)(OR.sup.ee).sub.2, C.sub.1-6 alkyl, C.sub.1-6
perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-10
carbocyclyl, 3-10 membered heterocyclyl, C.sub.6-10 aryl, or 5-10
membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups, or two
geminal R.sup.dd substituents are joined to form .dbd.O or .dbd.S;
each instance of R.sup.ee is, independently, C.sub.1-6 alkyl,
C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl,
C.sub.3-10 carbocyclyl, C.sub.6-10 aryl, 3-10 membered
heterocyclyl, or 3-10 membered heteroaryl, wherein each alkyl,
alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl
is independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg
groups; each instance of R.sup.ff is, independently, hydrogen,
C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered
heterocyclyl, C.sub.6-10 aryl, or 5-10 membered heteroaryl, or two
R.sup.ff
groups are joined to form a 3-14 membered heterocyclyl or 5-14
membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups; and each
instance of R.sup.gg is, independently, halogen, --CN, --NO.sub.2,
--N.sub.3, --SO.sub.2H, --SO.sub.3H, --OH, --OC.sub.1-6 alkyl,
--ON(C.sub.1-6 alkyl).sub.2, --N(C.sub.1-6 alkyl).sub.2,
--N(C.sub.1-6 alkyl).sub.3.sup.+X.sup.-, --NH(C.sub.1-6
alkyl).sub.2.sup.+X.sup.-, --NH.sub.2(C.sub.1-6
alkyl).sup.+X.sup.-, --NH.sub.3.sup.+X.sup.-, --N(OC.sub.1-6
alkyl)(C.sub.1-6 alkyl), --N(OH)(C.sub.1-6 alkyl), --NH(OH), --SH,
--SC.sub.1-6 alkyl, --SS(C.sub.1-6 alkyl), --C(.dbd.O)(C.sub.1-6
alkyl), --CO.sub.2H, --CO.sub.2(C.sub.1-6 alkyl),
--OC(.dbd.O)(C.sub.1-6 alkyl), --OCO.sub.2(C.sub.1-6 alkyl),
--C(.dbd.O)NH.sub.2, --C(.dbd.O)N(C.sub.1-6 alkyl).sub.2,
--OC(.dbd.O)NH(C.sub.1-6 alkyl), --NHC(.dbd.O)(C.sub.1-6 alkyl),
--N(C.sub.1-6 alkyl)C(.dbd.O)(C.sub.1-6 alkyl),
--NHCO.sub.2(C.sub.1-6 alkyl), --NHC(.dbd.O)N(C.sub.1-6
alkyl).sub.2, --NHC(.dbd.O)NH(C.sub.1-6 alkyl),
--NHC(.dbd.O)NH.sub.2, --C(.dbd.NH)O(C.sub.1-6 alkyl),
--OC(.dbd.NH)(C.sub.1-6 alkyl), --OC(.dbd.NH)OC.sub.1-6 alkyl,
--C(.dbd.NH)N(C.sub.1-6 alkyl).sub.2, --C(.dbd.NH)NH(C.sub.1-6
alkyl), --C(.dbd.NH)NH.sub.2, --OC(.dbd.NH)N(C.sub.1-6
alkyl).sub.2, --OC(NH)NH(C.sub.1-6 alkyl), --OC(NH)NH.sub.2,
--NHC(NH)N(C.sub.1-6 alkyl).sub.2, --NHC(.dbd.NH)NH.sub.2,
--NHSO.sub.2(C.sub.1-6 alkyl), --SO.sub.2N(C.sub.1-6 alkyl).sub.2,
--SO.sub.2NH(C.sub.1-6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2C.sub.1-6 alkyl, --SO.sub.2OC.sub.1-6 alkyl,
--OSO.sub.2C.sub.1-6 alkyl, --SOC.sub.1-6 alkyl, --Si(C.sub.1-6
alkyl).sub.3, --OSi(C.sub.1-6 alkyl).sub.3-C(.dbd.S)N(C.sub.1-6
alkyl).sub.2, C(.dbd.S)NH(C.sub.1-6 alkyl), C(.dbd.S)NH.sub.2,
--C(.dbd.O)S(C.sub.1-6 alkyl), --C(.dbd.S)SC.sub.1-6 alkyl,
--SC(.dbd.S)SC.sub.1-6 alkyl, --P(.dbd.O)(C.sub.1-6 alkyl).sub.2,
--OP(.dbd.O)(C.sub.1-6 alkyl).sub.2, --OP(.dbd.O)(OC.sub.1-6
alkyl).sub.2, C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6
alkenyl, C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, C.sub.6-10
aryl, 3-10 membered heterocyclyl, or 5-10 membered heteroaryl, or
two geminal R.sup.gg substituents are joined to form .dbd.0 or
.dbd.S; wherein X.sup.- is a counterion.
106. (canceled)
107. (canceled)
108. A method of treating a PRMT5-mediated disorder, comprising
administering to a subject in need thereof a therapeutically
effective amount of a compound of Formula (A), or a
pharmaceutically acceptable salt thereof, or a pharmaceutical
composition comprising a compound of Formula (A), or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable excipient: ##STR00617## or a pharmaceutically acceptable
salt thereof, wherein: represents a single or double bond; R.sup.12
is halogen or optionally substituted C.sub.1-3alkyl, and R.sup.13
is hydrogen, halogen, --NR.sup.A1R.sup.A2, optionally substituted
C.sub.1-3alkyl or --OR.sup.1; or R.sup.12 is hydrogen, halogen or
optionally substituted C.sub.1-3alkyl, and R.sup.13 is hydrogen,
halogen, --NR.sup.A1R.sup.A2, or optionally substituted
C.sub.1-3alkyl; R.sup.A1 and R.sup.A2 are each independently
hydrogen, optionally substituted C.sub.1-3 alkyl, a nitrogen
protecting group, or R.sup.A1 and R.sup.A2 are taken together with
the intervening nitrogen atom to form an optionally substituted 3-6
membered heterocyclic ring; R.sup.1 is hydrogen, R.sup.z, or
--C(O)R.sup.z, wherein R.sup.z is optionally substituted C.sub.1-6
alkyl; X is a bond, --O--, --N(R)--, --CR.sup.4R.sup.5--,
--O--CR.sup.4R.sup.5, --N(R)--CR.sup.4R.sup.5--,
--O--CR.sup.4R.sup.5--O--, --N(R)--CR.sup.4R.sup.5--O,
--N(R)--CR.sup.4R.sup.5--N(R)--, --O--CR.sup.4R.sup.5--N(R)--,
--CR.sup.4R.sup.5--O--, --CR.sup.4R.sup.5--N(R)--,
--O--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--N(R)--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--O--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--N(R)--, or
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--; each R is independently
hydrogen or optionally substituted C.sub.1-6 aliphatic; R.sup.2 and
R.sup.3 are each independently selected from the group consisting
of hydrogen, halo, --CN, --NO.sub.2, optionally substituted
aliphatic, optionally substituted carbocyclyl, optionally
substituted phenyl, optionally substituted heterocyclyl, optionally
substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A,
--C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
R.sup.4 and R.sup.5 are each independently selected from the group
consisting of hydrogen, halo, --CN, --NO.sub.2, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted phenyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.4 and R.sup.5 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
R.sup.6 and R.sup.7 are each independently selected from the group
consisting of hydrogen, halo, --CN, --NO.sub.2, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted phenyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.6 and R.sup.7 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
each R.sup.A is independently selected from the group consisting of
hydrogen, optionally substituted aliphatic, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, and optionally substituted heteroaryl; each
R.sup.B is independently selected from the group consisting of
hydrogen, optionally substituted aliphatic, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, and optionally substituted heteroaryl, or two
R.sup.B groups are taken together with their intervening atoms to
form an optionally substituted heterocyclic ring; R.sup.8, R.sup.9,
R.sup.10, and R.sup.11 are independently hydrogen, halo, or
optionally substituted aliphatic; Cy is a monocyclic or bicyclic,
saturated, partially unsaturated, or aromatic ring having 0-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur, wherein Cy is substituted with 0, 1, 2, 3, or 4 R.sup.y
groups; each R.sup.y is independently selected from the group
consisting of halo, --CN, --NO.sub.2, optionally substituted
aliphatic, optionally substituted carbocyclyl, optionally
substituted aryl, optionally substituted heterocyclyl, optionally
substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A,
--C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or an
R.sup.y group may be optionally taken together with R.sup.2 or
R.sup.3 to form an optionally substituted 5- to 6-membered
carbocyclic or heterocyclic ring fused to Cy; each R.sup.x is
independently selected from the group consisting of halo, --CN,
optionally substituted aliphatic, --OR', and --N(R'').sub.2; each
R' is independently hydrogen or optionally substituted aliphatic;
each R'' is independently hydrogen or optionally substituted
aliphatic, or two R'' are taken together with their intervening
atoms to form an optionally substituted heterocyclic ring having
1-2 heteroatoms independently selected from nitrogen, oxygen, and
sulfur; and n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency
permits; wherein each instance of aliphatic is independently an
alkyl, alkenyl, alkynyl, cycloalkyl, or cycloalkenyl group; and
each instance of optionally substituted is independently
unsubstituted or substituted, and each instance of substituted is,
when on a carbon atom, halogen, --CN, --NO.sub.2, --N.sub.3,
--SO.sub.2H, --SO.sub.3H, --OH, --OR.sup.aa, --ON(R.sup.bb).sub.2,
--N(R.sup.bb).sub.2, --N(R.sup.bb).sub.3.sup.+X.sup.-,
--N(OR.sup.cc)R.sup.bb, --SH, --SR.sup.aa, --SSR.sup.cc,
--C(.dbd.O)R.sup.aa, --CO.sub.2H, --CHO, --C(OR.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --OC(.dbd.O)R.sup.aa, --OCO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.2, --OC(.dbd.O)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.O)R.sup.aa, --NR.sup.bbCO.sub.2R.sup.aa,
--NR.sup.bbC(.dbd.O)N(R.sup.bb).sub.2, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --OC(.dbd.NR.sup.bb)R.sup.aa,
--OC(.dbd.NR.sup.bb)OR.sup.aa,
--C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--OC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--C(.dbd.O)NR.sup.bbSO.sub.2R.sup.aa, --NR.sup.bbSO.sub.2R.sup.aa,
--SO.sub.2N(R.sup.bb).sub.2, --SO.sub.2R.sup.aa,
--SO.sub.2OR.sup.aa, --OSO.sub.2R.sup.aa, --S(.dbd.O)R.sup.aa,
--OS(.dbd.O)R.sup.aa, --Si(R.sup.aa).sub.3,
--OSi(R.sup.aa).sub.3--C(.dbd.S)N(R.sup.bb).sub.2,
--C(.dbd.O)SR.sup.aa, --C(.dbd.S)SR.sup.aa, --SC(.dbd.S)SR.sup.aa,
--SC(.dbd.O)SR.sup.aa, --OC(.dbd.O)SR.sup.aa,
--SC(.dbd.O)OR.sup.aa, --SC(.dbd.O)R.sup.aa,
--P(.dbd.O)(R.sup.aa).sub.2, --OP(.dbd.O)(R.sup.aa).sub.2,
--OP(.dbd.O)(OR.sup.cc).sub.2, --P(.dbd.O).sub.2N(R.sup.bb).sub.2,
--OP(.dbd.O).sub.2N(R.sup.bb).sub.2, --P(.dbd.O)(NR.sup.bb).sub.2,
--OP(.dbd.O)(NR.sup.bb).sub.2,
--NR.sup.bbP(.dbd.O)(OR.sup.cc).sub.2,
--NR.sup.bbP(.dbd.O)(NR.sup.bb).sub.2, --P(R.sup.cc).sub.2,
--OP(R.sup.cc).sub.2, --OP(R.sup.cc).sub.3, --B(R.sup.aa).sub.2,
--B(OR.sup.cc).sub.2, --BR(OR.sup.cc), C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 Rd.sup.d groups, or two
geminal hydrogens on a carbon atom are replaced with the group
.dbd.O, .dbd.S, .dbd.NN(R.sup.bb).sub.2,
.dbd.NNR.sup.bbC(.dbd.O)R.sup.aa,
.dbd.NNR.sup.bbC(.dbd.O)OR.sup.aa,
.dbd.NNR.sup.bbS(.dbd.O).sub.2R.sup.aa, .dbd.NR.sup.bb, or
.dbd.NOR.sup.cc; each instance of substituted is, when on a
nitrogen, independently --OH, --OR.sup.aa, --N(R.sup.cc).sub.2,
--CN, --C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.bb)R.sup.aa, --C(.dbd.NR.sup.cc)OR.sup.aa,
--C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2, --SO.sub.2N(R.sup.cc).sub.2,
--SO.sub.2R.sup.cc, --SO.sub.2OR.sup.cc, --SOR.sup.aa,
--C(.dbd.S)N(R.sup.cc).sub.2, --C(.dbd.O)SR.sup.cc,
--C(.dbd.S)SR.sup.cc, --P(.dbd.O)(R.sup.aa).sub.2,
--P(.dbd.O)(NR.sup.cc).sub.2, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, or two R.sup.cc groups attached to a nitrogen
atom are joined to form a 3-14 membered heterocyclyl or 5-14
membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups; each instance
of R.sup.aa is, independently, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, or 5-14
membered heteroaryl, or two R.sup.aa groups are joined to form a
3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,
wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 R.sup.dd groups; each instance of R.sup.bb is,
independently, hydrogen, --OH, --OR.sup.aa, --N(R.sup.cc).sub.2,
--CN, --C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.cc)OR.sup.aa, --C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2,
--SO.sub.2N(R.sup.cc).sub.2, --SO.sub.2R.sup.cc,
--SO.sub.2OR.sup.cc, --SOR.sup.aa, --C(.dbd.S)N(R.sup.cc).sub.2,
--C(.dbd.O)SR.sup.cc, --C(.dbd.S)SR.sup.cc,
--P(.dbd.O)(R.sup.aa).sub.2, --P(.dbd.O).sub.2N(R.sup.cc).sub.2,
C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, or 5-14 membered heteroaryl, or two
R.sup.bb groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups;
each instance of R.sup.cc is, independently, hydrogen, C.sub.1-10
alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10
alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered heterocyclyl,
C.sub.6-14 aryl, or 5-14 membered heteroaryl, or two R.sup.cc
groups are joined to form a 3-14 membered heterocyclyl or 5-14
membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups; each instance
of R.sup.dd is, independently, halogen, --CN, --NO.sub.2,
--N.sub.3, --SO.sub.2H, --SO.sub.3H, --OH, --OR.sup.ee,
--ON(R.sup.ff).sub.2, --N(R.sup.ff).sub.2,
--N(R.sup.ff).sub.3.sup.+X.sup.-, --N(OR.sup.ee)R.sup.ff, --SH,
--SR.sup.ee, --SSR.sup.ee, --C(.dbd.O)R.sup.ee, --CO.sub.2H,
--CO.sub.2R.sup.ee, --OC(.dbd.O)R.sup.ee, --OCO.sub.2R.sup.ee,
--C(.dbd.O)N(R.sup.ff).sub.2, --OC(.dbd.O)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.O)R.sup.ee, --NR.sup.ffCO.sub.2R.sup.ee,
--NR.sup.ffC(.dbd.O)N(R.sup.ff).sub.2,
--C(.dbd.NR.sup.ff)OR.sup.ee, --OC(.dbd.NR.sup.ff)R.sup.ee,
--OC(.dbd.NR.sup.ff)OR.sup.ee,
--C(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--OC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffSO.sub.2R.sup.ee, --SO.sub.2N(R.sup.ff).sub.2,
--SO.sub.2R.sup.ee, --SO.sub.2OR.sup.ee, --OSO.sub.2R.sup.ee,
--S(.dbd.O)R.sup.ee, --Si(R.sup.ee).sub.3, --OSi(R.sup.ee).sub.3,
--C(.dbd.S)N(R.sup.ff).sub.2, --C(.dbd.O)SR.sup.ee,
--C(.dbd.S)SR.sup.ee, --SC(.dbd.S)SR.sup.ee,
--P(.dbd.O)(R.sup.ee).sub.2, --OP(.dbd.O)(R.sup.ee).sub.2,
--OP(.dbd.O)(OR.sup.ee).sub.2, C.sub.1-6 alkyl, C.sub.1-6
perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-10
carbocyclyl, 3-10 membered heterocyclyl, C.sub.6-10 aryl, or 5-10
membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups, or two
geminal R.sup.dd substituents are joined to form .dbd.O or .dbd.S;
each instance of R.sup.ee is, independently, C.sub.1-6 alkyl,
C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl,
C.sub.3-10 carbocyclyl, C.sub.6-10 aryl, 3-10 membered
heterocyclyl, or 3-10 membered heteroaryl, wherein each alkyl,
alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl
is independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg
groups; each instance of R.sup.ff is, independently, hydrogen,
C
.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered
heterocyclyl, C.sub.6-10 aryl, or 5-10 membered heteroaryl, or two
R.sup.ff groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups;
and each instance of R.sup.gg is, independently, halogen, --CN,
--NO.sub.2, --N.sub.3, --SO.sub.2H, --SO.sub.3H, --OH, --OC.sub.1-6
alkyl, --ON(C.sub.1-6 alkyl).sub.2, --N(C.sub.1-6 alkyl).sub.2,
--N(C.sub.1-6 alkyl).sub.3.sup.+X.sup.-, --NH(C.sub.1-6
alkyl).sub.2.sup.+X.sup.-, --NH.sub.2(C.sub.1-6
alkyl).sup.+X.sup.-, --NH.sub.3.sup.+X.sup.-, --N(OC.sub.1-6
alkyl)(C.sub.1-6 alkyl), --N(OH)(C.sub.1-6 alkyl), --NH(OH), --SH,
--SC.sub.1-6 alkyl, --SS(C.sub.1-6 alkyl), --C(.dbd.O)(C.sub.1-6
alkyl), --CO.sub.2H, --CO.sub.2(C.sub.1-6 alkyl),
--OC(.dbd.O)(C.sub.1-6 alkyl), --OCO.sub.2(C.sub.1-6 alkyl),
--C(.dbd.O)NH.sub.2, --C(.dbd.O)N(C.sub.1-6 alkyl).sub.2,
--OC(.dbd.O)NH(C.sub.1-6 alkyl), --NHC(.dbd.O)(C.sub.1-6 alkyl),
--N(C.sub.1-6 alkyl)C(.dbd.O)(C.sub.1-6 alkyl),
--NHCO.sub.2(C.sub.1-6 alkyl), --NHC(.dbd.O)N(C.sub.1-6
alkyl).sub.2, --NHC(.dbd.O)NH(C.sub.1-6 alkyl),
--NHC(.dbd.O)NH.sub.2, --C(.dbd.NH)O(C.sub.1-6 alkyl),
--OC(.dbd.NH)(C.sub.1-6 alkyl), --OC(.dbd.NH)OC.sub.1-6 alkyl,
--C(.dbd.NH)N(C.sub.1-6 alkyl).sub.2, --C(.dbd.NH)NH(C.sub.1-6
alkyl), --C(.dbd.NH)NH.sub.2, --OC(.dbd.NH)N(C.sub.1-6
alkyl).sub.2, --OC(NH)NH(C.sub.1-6 alkyl), --OC(NH)NH.sub.2,
--NHC(NH)N(C.sub.1-6 alkyl).sub.2, --NHC(.dbd.NH)NH.sub.2,
--NHSO.sub.2(C.sub.1-6 alkyl), --SO.sub.2N(C.sub.1-6 alkyl).sub.2,
--SO.sub.2NH(C.sub.1-6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2C.sub.1-6 alkyl, --SO.sub.2OC.sub.1-6 alkyl,
--OSO.sub.2C.sub.1-6 alkyl, --SOC.sub.1-6 alkyl, --Si(C.sub.1-6
alkyl).sub.3, --OSi(C.sub.1-6 alkyl).sub.3-C(.dbd.S)N(C.sub.1-6
alkyl).sub.2, C(.dbd.S)NH(C.sub.1-6 alkyl), C(.dbd.S)NH.sub.2,
--C(.dbd.O)S(C.sub.1-6 alkyl), --C(.dbd.S)SC.sub.1-6 alkyl,
--SC(.dbd.S)SC.sub.1-6 alkyl, --P(.dbd.O)(C.sub.1-6 alkyl).sub.2,
--OP(.dbd.O)(C.sub.1-6 alkyl).sub.2, --OP(.dbd.O)(OC.sub.1-6
alkyl).sub.2, C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6
alkenyl, C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, C.sub.6-10
aryl, 3-10 membered heterocyclyl, or 5-10 membered heteroaryl, or
two geminal R.sup.gg substituents are joined to form .dbd.O or
.dbd.S; wherein X.sup.- is a counterion.
109. The method of claim 108, wherein the disorder is a
proliferative disorder.
110. The method of claim 109, wherein the disorder is cancer, a
metabolic disorder, or a blood disorder.
111. The method of claim 110, wherein the cancer is hematopoietic
cancer, lung cancer, prostate cancer, melanoma, or pancreatic
cancer.
112. (canceled)
113. The method of claim 110, wherein the metabolic disorder is
diabetes or obesity.
114. (canceled)
115. (canceled)
116. The method of claim 110, wherein the disorder is a
hemoglobinopathy.
117. The method of claim 116, wherein the disorder is sickle cell
anemia or .beta.-thalessemia.
118. (canceled)
119. The method of claim 108, wherein the compound is of Formula
(A-5): ##STR00618## or a pharmaceutically acceptable salt
thereof.
120. The method of claim 108, wherein the compound is of Formula
(A-9): ##STR00619## or a pharmaceutically acceptable salt
thereof.
121. The method of claim 108, wherein R.sup.12 is hydrogen, and
R.sup.13 is --NR.sup.A1R.sup.A2.
122. The method of claim 108, wherein R.sup.12 is hydrogen, and
R.sup.13 is optionally substituted C.sub.1-3alkyl.
123. The method of claim 108, wherein n is 0.
124. The method of claim 108, wherein R.sup.2 and R.sup.3 are each
hydrogen.
125. The method of claim 108, wherein Cy is phenyl substituted with
0, 1, or 2 R.sup.y groups.
126. The method of claim 108, wherein Cy is a bicyclic saturated,
partially unsaturated, or aromatic ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur, wherein
Cy is substituted with 0, 1, 2, 3, or 4 R.sup.y groups.
127. The method of claim 108, wherein Cy is substituted with 1 or 2
R.sup.y groups, and at least one R.sup.y is halo, --CN, --NO.sub.2,
optionally substituted aliphatic, optionally substituted
carbocyclyl, optionally substituted aryl, optionally substituted
heterocyclyl, optionally substituted heteroaryl, --OR.sup.A,
--N(R.sup.B).sub.2, --C(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--S(O)R.sup.A, --NR.sup.BSO.sub.2R.sup.A, or
--SO.sub.2N(R.sup.B).sub.2.
128. The method of claim 108, wherein the compound is selected from
the group consisting of: ##STR00620## ##STR00621## ##STR00622##
##STR00623## and pharmaceutically acceptable salts thereof.
Description
RELATED APPLICATIONS
[0001] The present application is a Continuation application of
U.S. patent application U.S. Ser. No. 14/654,226, filed Jun. 19,
2015, which is a national stage filing under 35 U.S.C. .sctn. 371
of International PCT application PCT/US2013/077250, filed Dec. 20,
2013 which claims priority under 35 U.S.C. .sctn. 119(e) to U.S.
provisional patent applications, U.S. Ser. No. 61/745,490, filed
Dec. 21, 2012, and U.S. Ser. No. 61/790,928, filed Mar. 15, 2013,
the entire contents of each of which are incorporated herein by
reference.
BACKGROUND OF THE INVENTION
[0002] Epigenetic regulation of gene expression is an important
biological determinant of protein production and cellular
differentiation and plays a significant pathogenic role in a number
of human diseases.
[0003] Epigenetic regulation involves heritable modification of
genetic material without changing its nucleotide sequence.
Typically, epigenetic regulation is mediated by selective and
reversible modification (e.g., methylation) of DNA and proteins
(e.g., histones) that control the conformational transition between
transcriptionally active and inactive states of chromatin. These
covalent modifications can be controlled by enzymes such as
methyltransferases (e.g., PRMT5), many of which are associated with
specific genetic alterations that can cause human disease.
[0004] Disease-associated chromatin-modifying enzymes (e.g., PRMT5)
play a role in diseases such as proliferative disorders, metabolic
disorders, and blood disorders. Thus, there is a need for the
development of small molecules that are capable of inhibiting the
activity of PRMT5.
DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS
[0005] Protein arginine methyltransferase 5 (PRMT5) catalyzes the
addition of two methyl groups to the two .omega.-guanidino nitrogen
atoms of arginine, resulting in .omega.-NG, N'G symmetric
dimethylation of arginine (sDMA) of the target protein. PRMT5
functions in the nucleus as well as in the cytoplasm, and its
substrates include histones, spliceosomal proteins, transcription
factors (See e.g., Sun et al., 20011, PNAS 108: 20538-20543). PRMT5
generally functions as part of a molecule weight protein complex.
While the protein complexes of PRMT5 can have a variety of
components, they generally include the protein MEP50 (methylosome
protein 50). In addition, PRMT5 acts in conjunction with cofactor
SAM (S-adenosyl methionine).
[0006] PRMT5 is an attractive target for modulation given its role
in the regulation of diverse biological processes. It has now been
found that compounds described herein, and pharmaceutically
acceptable salts and compositions thereof, are effective as
inhibitors of PRMT5. Such compounds have the general Formula
(A):
##STR00002##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.2,
R.sup.3, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13,
R.sup.x, n, X, and Cy are as defined herein.
[0007] In some embodiments, the inhibitors of PRMT5 have the
general Formula (I):
##STR00003##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.1,
R.sup.2, R.sup.3, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.x, n,
X, and Cy are as described herein.
[0008] In some embodiments, pharmaceutical compositions are
provided which comprise a compound described herein (e.g., a
compound of Formula (A), e.g., Formula (I)), or a pharmaceutically
acceptable salt thereof, and optionally a pharmaceutically
acceptable excipient.
[0009] In certain embodiments, compounds described herein inhibit
activity of PRMT5. In certain embodiments, methods of inhibiting
PRMT5 are provided which comprise contacting PRMT5 with an
effective amount of a compound of Formula (A), e.g., Formula (I),
or a pharmaceutically acceptable salt thereof. The PRMT5 may be
purified or crude, and may be present in a cell, tissue, or a
subject. Thus, such methods encompass inhibition of PRMT5 activity
both in vitro and in vivo. In certain embodiments, the PRMT5 is
wild-type PRMT5. In certain embodiments, the PRMT5 is
overexpressed. In certain embodiments, the PRMT5 is a mutant. In
certain embodiments, the PRMT5 is in a cell. In certain
embodiments, the PRMT5 is in an animal, e.g., a human. In some
embodiments, the PRMT5 is in a subject that is susceptible to
normal levels of PRMT5 activity due to one or more mutations
associated with a PRMT5 substrate. In some embodiments, the PRMT5
is in a subject known or identified as having abnormal PRMT5
activity (e.g., overexpression). In some embodiments, a provided
compound is selective for PRMT5 over other methyltransferases. In
certain embodiments, a provided compound is at least about 10-fold
selective, at least about 20-fold selective, at least about 30-fold
selective, at least about 40-fold selective, at least about 50-fold
selective, at least about 60-fold selective, at least about 70-fold
selective, at least about 80-fold selective, at least about 90-fold
selective, or at least about 100-fold selective relative to one or
more other methyltransferases.
[0010] In certain embodiments, methods of altering gene expression
in a cell are provided which comprise contacting a cell with an
effective amount of a compound of Formula (A), or a
pharmaceutically acceptable salt thereof, or a pharmaceutical
composition thereof. In certain embodiments, the cell in culture in
vitro. In certain embodiments, cell is in an animal, e.g., a
human.
[0011] In certain embodiments, methods of altering transcription in
a cell are provided which comprise contacting a cell with an
effective amount of a compound of Formula (A), e.g., Formula (I),
or a pharmaceutically acceptable salt thereof, or a pharmaceutical
composition thereof. In certain embodiments, the cell in culture in
vitro. In certain embodiments, the cell is in an animal, e.g., a
human.
[0012] In some embodiments, methods of treating a PRMT5-mediated
disorder are provided which comprise administering to a subject
suffering from a PRMT5-mediated disorder an effective amount of a
compound described herein (e.g., a compound of Formula (A), e.g.,
Formula (I)), or a pharmaceutically acceptable salt thereof, or a
pharmaceutical composition thereof. In certain embodiments, the
PRMT5-mediated disorder is a proliferative disorder, a metabolic
disorder, or a blood disorder. In certain embodiments, compounds
described herein are useful for treating cancer. In certain
embodiments, compounds described herein are useful for treating
hematopoietic cancer, lung cancer, prostate cancer, melanoma, or
pancreatic cancer. In certain embodiments, compounds described
herein are useful for treating a hemoglobinopathy. In certain
embodiments, compounds described herein are useful for treating
sickle cell anemia. In certain embodiments, compounds described
herein are useful for treating diabetes or obesity. In certain
embodiments, a provided compound is useful in treating inflammatory
and autoimmune disease.
[0013] Compounds described herein are also useful for the study of
PRMT5 in biological and pathological phenomena, the study of
intracellular signal transduction pathways mediated by PRMT5, and
the comparative evaluation of new PRMT5 inhibitors.
[0014] This application refers to various issued patent, published
patent applications, journal articles, and other publications, all
of which are incorporated herein by reference.
[0015] Definitions of specific functional groups and chemical terms
are described in more detail below. The chemical elements are
identified in accordance with the Periodic Table of the Elements,
CAS version, Handbook of Chemistry and Physics, 75.+-.Ed., inside
cover, and specific functional groups are generally defined as
described therein. Additionally, general principles of organic
chemistry, as well as specific functional moieties and reactivity,
are described in Thomas Sorrell, Organic Chemistry, 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; and Carruthers, Some Modern
Methods of Organic Synthesis, 3.sup.rd Edition, Cambridge
University Press, Cambridge, 1987.
[0016] Compounds described herein can comprise one or more
asymmetric centers, and thus can exist in various isomeric forms,
e.g., enantiomers and/or diastereomers. For example, the compounds
described herein can be in the form of an individual enantiomer,
diastereomer or geometric isomer, or can be in the form of a
mixture of stereoisomers, including racemic mixtures and mixtures
enriched in one or more stereoisomer. Isomers can be isolated from
mixtures by methods known to those skilled in the art, including
chiral high pressure liquid chromatography (HPLC) and the formation
and crystallization of chiral salts; or preferred isomers can be
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); and
Wilen, Tables of Resolving Agents and Optical Resolutions p. 268
(E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind.
1972). The present disclosure additionally encompasses compounds
described herein as individual isomers substantially free of other
isomers, and alternatively, as mixtures of various isomers.
[0017] It is to be understood that the compounds of the present
invention may be depicted as different tautomers. It should also be
understood that when compounds have tautomeric forms, all
tautomeric forms are intended to be included in the scope of the
present invention, and the naming of any compound described herein
does not exclude any tautomer form.
##STR00004##
[0018] Unless otherwise stated, structures depicted herein are also
meant to include compounds that differ only in the presence of one
or more isotopically enriched atoms. For example, compounds having
the present structures except for the replacement of hydrogen by
deuterium or tritium, replacement of .sup.19F with .sup.18F, or the
replacement of a carbon by a .sup.13C- or .sup.14C-enriched carbon
are within the scope of the disclosure. Such compounds are useful,
for example, as analytical tools or probes in biological
assays.
[0019] 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. In some embodiments, an aliphatic group is 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 alkyl, alkenyl, alkynyl, cycloalkyl, and
cycloalkenyl moieties.
[0020] When a range of values is listed, it is intended to
encompass each value and sub-range within the range. For example
"C.sub.1-6 alkyl" is intended to encompass, C.sub.1, C.sub.2,
C.sub.3, C.sub.4, C.sub.5, C.sub.6, C.sub.1-6, C.sub.1-5,
C.sub.1-4, C.sub.1-3, C.sub.1-2, C.sub.2-6, C.sub.2-5, C.sub.2-4,
C.sub.2-3, C.sub.3-6, C.sub.3-5, C.sub.3-4, C.sub.4-6, C.sub.4-5,
and C.sub.5-6 alkyl.
[0021] "Alkyl" refers to a radical of a straight-chain or branched
saturated hydrocarbon group having from 1 to 20 carbon atoms
("C.sub.1-20 alkyl"). In some embodiments, an alkyl group has 1 to
10 carbon atoms ("C.sub.1-10 alkyl"). In some embodiments, an alkyl
group has 1 to 9 carbon atoms ("C.sub.1-9 alkyl"). In some
embodiments, an alkyl group has 1 to 8 carbon atoms ("C.sub.1-8
alkyl"). In some embodiments, an alkyl group has 1 to 7 carbon
atoms ("C.sub.1-7 alkyl"). In some embodiments, an alkyl group has
1 to 6 carbon atoms ("C.sub.1-6 alkyl"). In some embodiments, an
alkyl group has 1 to 5 carbon atoms ("C.sub.1-5 alkyl"). In some
embodiments, an alkyl group has 1 to 4 carbon atoms ("C.sub.1-4
alkyl"). In some embodiments, an alkyl group has 1 to 3 carbon
atoms ("C.sub.1-3 alkyl"). In some embodiments, an alkyl group has
1 to 2 carbon atoms ("C.sub.1-2 alkyl"). In some embodiments, an
alkyl group has 1 carbon atom ("C.sub.1 alkyl"). In some
embodiments, an alkyl group has 2 to 6 carbon atoms ("C.sub.2-6
alkyl"). Examples of C.sub.1-6 alkyl groups include methyl
(C.sub.1), ethyl (C.sub.2), n-propyl (C.sub.3), isopropyl
(C.sub.3), n-butyl (C.sub.4), tert-butyl (C.sub.4), sec-butyl
(C.sub.4), iso-butyl (C.sub.4), n-pentyl (C.sub.5), 3-pentanyl
(C.sub.5), amyl (C.sub.5), neopentyl (C.sub.5), 3-methyl-2-butanyl
(C.sub.5), tertiary amyl (C.sub.5), and n-hexyl (C.sub.6).
Additional examples of alkyl groups include n-heptyl (C.sub.7),
n-octyl (C.sub.5) and the like. In certain embodiments, each
instance of an alkyl group is independently optionally substituted,
e.g., unsubstituted (an "unsubstituted alkyl") or substituted (a
"substituted alkyl") with one or more substituents. In certain
embodiments, the alkyl group is unsubstituted C.sub.1-10 alkyl
(e.g., --CH.sub.3). In certain embodiments, the alkyl group is
substituted C.sub.1-10 alkyl.
[0022] In some embodiments, an alkyl group is substituted with one
or more halogens. "Perhaloalkyl" is a substituted alkyl group as
defined herein wherein all of the hydrogen atoms are independently
replaced by a halogen, e.g., fluoro, bromo, chloro, or iodo. In
some embodiments, the alkyl moiety has 1 to 8 carbon atoms
("C.sub.1-8 perhaloalkyl"). In some embodiments, the alkyl moiety
has 1 to 6 carbon atoms ("C.sub.1-6 perhaloalkyl"). In some
embodiments, the alkyl moiety has 1 to 4 carbon atoms ("C.sub.1-4
perhaloalkyl"). In some embodiments, the alkyl moiety has 1 to 3
carbon atoms ("C.sub.1-3 perhaloalkyl"). In some embodiments, the
alkyl moiety has 1 to 2 carbon atoms ("C.sub.1-2 perhaloalkyl"). In
some embodiments, all of the hydrogen atoms are replaced with
fluoro. In some embodiments, all of the hydrogen atoms are replaced
with chloro. Examples of perhaloalkyl groups include --CF.sub.3,
--CF.sub.2CF.sub.3, --CF.sub.2CF.sub.2CF.sub.3, --CCl.sub.3,
--CFCl.sub.2, --CF.sub.2Cl, and the like.
[0023] "Alkenyl" refers to a radical of a straight-chain or
branched hydrocarbon group having from 2 to 20 carbon atoms, one or
more carbon-carbon double bonds, and no triple bonds ("C.sub.2-20
alkenyl"). In some embodiments, an alkenyl group has 2 to 10 carbon
atoms ("C.sub.2-10 alkenyl"). In some embodiments, an alkenyl group
has 2 to 9 carbon atoms ("C.sub.2-9 alkenyl"). In some embodiments,
an alkenyl group has 2 to 8 carbon atoms ("C.sub.2-8 alkenyl"). In
some embodiments, an alkenyl group has 2 to 7 carbon atoms
("C.sub.2-7 alkenyl"). In some embodiments, an alkenyl group has 2
to 6 carbon atoms ("C.sub.2-6 alkenyl"). In some embodiments, an
alkenyl group has 2 to 5 carbon atoms ("C.sub.2-5 alkenyl"). In
some embodiments, an alkenyl group has 2 to 4 carbon atoms
("C.sub.2-4 alkenyl"). In some embodiments, an alkenyl group has 2
to 3 carbon atoms ("C.sub.2-3 alkenyl"). In some embodiments, an
alkenyl group has 2 carbon atoms ("C.sub.2 alkenyl"). The one or
more carbon-carbon double bonds can be internal (such as in
2-butenyl) or terminal (such as in 1-butenyl). Examples of
C.sub.2-4 alkenyl groups include ethenyl (C.sub.2), 1-propenyl
(C.sub.3), 2-propenyl (C.sub.3), 1-butenyl (C.sub.4), 2-butenyl
(C.sub.4), butadienyl (C.sub.4), and the like. Examples of
C.sub.2-6 alkenyl groups include the aforementioned C.sub.2-4
alkenyl groups as well as pentenyl (C.sub.5), pentadienyl
(C.sub.5), hexenyl (C.sub.6), and the like. Additional examples of
alkenyl include heptenyl (C.sub.7), octenyl (C.sub.8), octatrienyl
(C.sub.8), and the like. In certain embodiments, each instance of
an alkenyl group is independently optionally substituted, e.g.,
unsubstituted (an "unsubstituted alkenyl") or substituted (a
"substituted alkenyl") with one or more substituents. In certain
embodiments, the alkenyl group is unsubstituted C.sub.2-10 alkenyl.
In certain embodiments, the alkenyl group is substituted C.sub.2-10
alkenyl.
[0024] "Alkynyl" refers to a radical of a straight-chain or
branched hydrocarbon group having from 2 to 20 carbon atoms, one or
more carbon-carbon triple bonds, and optionally one or more double
bonds ("C.sub.2-20 alkynyl"). In some embodiments, an alkynyl group
has 2 to 10 carbon atoms ("C.sub.2-10 alkynyl"). In some
embodiments, an alkynyl group has 2 to 9 carbon atoms ("C.sub.2-9
alkynyl"). In some embodiments, an alkynyl group has 2 to 8 carbon
atoms ("C.sub.2-8 alkynyl"). In some embodiments, an alkynyl group
has 2 to 7 carbon atoms ("C.sub.2-7 alkynyl"). In some embodiments,
an alkynyl group has 2 to 6 carbon atoms ("C.sub.2-6 alkynyl"). In
some embodiments, an alkynyl group has 2 to 5 carbon atoms
("C.sub.2-5 alkynyl"). In some embodiments, an alkynyl group has 2
to 4 carbon atoms ("C.sub.2-4 alkynyl"). In some embodiments, an
alkynyl group has 2 to 3 carbon atoms ("C.sub.2-3 alkynyl"). In
some embodiments, an alkynyl group has 2 carbon atoms ("C.sub.2
alkynyl"). The one or more carbon-carbon triple bonds can be
internal (such as in 2-butynyl) or terminal (such as in 1-butynyl).
Examples of C.sub.2-4 alkynyl groups include, without limitation,
ethynyl (C.sub.2), 1-propynyl (C.sub.3), 2-propynyl (C.sub.3),
1-butynyl (C.sub.4), 2-butynyl (C.sub.4), and the like. Examples of
C.sub.2-6 alkenyl groups include the aforementioned C.sub.2-4
alkynyl groups as well as pentynyl (C.sub.5), hexynyl (C.sub.6),
and the like. Additional examples of alkynyl include heptynyl
(C.sub.7), octynyl (C.sub.8), and the like. In certain embodiments,
each instance of an alkynyl group is independently optionally
substituted, e.g., unsubstituted (an "unsubstituted alkynyl") or
substituted (a "substituted alkynyl") with one or more
substituents. In certain embodiments, the alkynyl group is
unsubstituted C.sub.2-10 alkynyl. In certain embodiments, the
alkynyl group is substituted C.sub.2-10 alkynyl.
[0025] "Carbocyclyl" or "carbocyclic" refers to a radical of a
non-aromatic cyclic hydrocarbon group having from 3 to 14 ring
carbon atoms ("C.sub.3-14 carbocyclyl") and zero heteroatoms in the
non-aromatic ring system. In some embodiments, a carbocyclyl group
has 3 to 10 ring carbon atoms ("C.sub.3-10 carbocyclyl").
"Carbocyclyl" or "carbocyclic" refers to a radical of a
non-aromatic cyclic hydrocarbon group having from 3 to 14 ring
carbon atoms ("C.sub.3-14 carbocyclyl") and zero heteroatoms in the
non-aromatic ring system. In some embodiments, a carbocyclyl group
has 3 to 10 ring carbon atoms ("C.sub.3-10 carbocyclyl"). In some
embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms
("C.sub.3-8 carbocyclyl"). In some embodiments, a carbocyclyl group
has 3 to 6 ring carbon atoms ("C.sub.3-6 carbocyclyl"). In some
embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms
("C.sub.5-10 carbocyclyl"). Exemplary C.sub.3-6 carbocyclyl groups
include, without limitation, cyclopropyl (C.sub.3), cyclopropenyl
(C.sub.3), cyclobutyl (C.sub.4), cyclobutenyl (C.sub.4),
cyclopentyl (C.sub.5), cyclopentenyl (C.sub.5), cyclohexyl
(C.sub.6), cyclohexenyl (C.sub.6), cyclohexadienyl (C.sub.6), and
the like. Exemplary C.sub.3-8 carbocyclyl groups include, without
limitation, the aforementioned C.sub.3-6 carbocyclyl groups as well
as cycloheptyl (C.sub.7), cycloheptenyl (C.sub.7), cycloheptadienyl
(C.sub.7), cycloheptatrienyl (C.sub.7), cyclooctyl (C.sub.8),
cyclooctenyl (C.sub.8), bicyclo[2.2.1]heptanyl (C.sub.7),
bicyclo[2.2.2]octanyl (C.sub.8), and the like. Exemplary C.sub.3-10
carbocyclyl groups include, without limitation, the aforementioned
C.sub.3-8 carbocyclyl groups as well as cyclononyl (C.sub.9),
cyclononenyl (C.sub.9), cyclodecyl (C.sub.10), cyclodecenyl
(C.sub.10), octahydro-1H-indenyl (C.sub.9), decahydronaphthalenyl
(C.sub.10), spiro[4.5]decanyl (C.sub.10), and the like. As the
foregoing examples illustrate, in certain embodiments, the
carbocyclyl group is either monocyclic ("monocyclic carbocyclyl")
or contain a fused, bridged or spiro ring system such as a bicyclic
system ("bicyclic carbocyclyl") and can be saturated or can be
partially unsaturated. "Carbocyclyl" also includes ring systems
wherein the carbocyclyl ring, as defined above, is fused with one
or more aryl or heteroaryl groups wherein the point of attachment
is on the carbocyclyl ring, and in such instances, the number of
carbons continue to designate the number of carbons in the
carbocyclic ring system. In certain embodiments, each instance of a
carbocyclyl group is independently optionally substituted, e.g.,
unsubstituted (an "unsubstituted carbocyclyl") or substituted (a
"substituted carbocyclyl") with one or more substituents. In
certain embodiments, the carbocyclyl group is unsubstituted
C.sub.3-10 carbocyclyl. In certain embodiments, the carbocyclyl
group is a substituted C.sub.3-10 carbocyclyl.
[0026] In some embodiments, "carbocyclyl" is a monocyclic,
saturated carbocyclyl group having from 3 to 14 ring carbon atoms
("C.sub.3-14 cycloalkyl"). In some embodiments, a cycloalkyl group
has 3 to 10 ring carbon atoms ("C.sub.3-10 cycloalkyl"). In some
embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms
("C.sub.3-8 cycloalkyl"). In some embodiments, a cycloalkyl group
has 3 to 6 ring carbon atoms ("C.sub.3-6 cycloalkyl"). In some
embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms
("C.sub.5-6 cycloalkyl"). In some embodiments, a cycloalkyl group
has 5 to 10 ring carbon atoms ("C.sub.5-10 cycloalkyl"). Examples
of C.sub.5-6 cycloalkyl groups include cyclopentyl (C.sub.5) and
cyclohexyl (C.sub.5). Examples of C.sub.3-6 cycloalkyl groups
include the aforementioned C.sub.5-6 cycloalkyl groups as well as
cyclopropyl (C.sub.3) and cyclobutyl (C.sub.4). Examples of
C.sub.3-8 cycloalkyl groups include the aforementioned C.sub.3-6
cycloalkyl groups as well as cycloheptyl (C.sub.7) and cyclooctyl
(C.sub.8). In certain embodiments, each instance of a cycloalkyl
group is independently unsubstituted (an "unsubstituted
cycloalkyl") or substituted (a "substituted cycloalkyl") with one
or more substituents. In certain embodiments, the cycloalkyl group
is unsubstituted C.sub.3-10 cycloalkyl. In certain embodiments, the
cycloalkyl group is substituted C.sub.3-10 cycloalkyl.
[0027] "Heterocyclyl" or "heterocyclic" refers to a radical of a 3-
to 14-membered non-aromatic ring system having ring carbon atoms
and 1 to 4 ring heteroatoms, wherein each heteroatom is
independently selected from nitrogen, oxygen, and sulfur ("3-14
membered heterocyclyl"). In certain embodiments, heterocyclyl or
heterocyclic refers to a radical of a 3-10 membered non-aromatic
ring system having ring carbon atoms and 1-4 ring heteroatoms,
wherein each heteroatom is independently selected from nitrogen,
oxygen, and sulfur ("3-10 membered heterocyclyl"). In heterocyclyl
groups that contain one or more nitrogen atoms, the point of
attachment can be a carbon or nitrogen atom, as valency permits. A
heterocyclyl group can either be monocyclic ("monocyclic
heterocyclyl") or a fused, bridged or spiro ring system such as a
bicyclic system ("bicyclic heterocyclyl"), and can be saturated or
can be partially unsaturated. Heterocyclyl bicyclic ring systems
can include one or more heteroatoms in one or both rings.
"Heterocyclyl" also includes ring systems wherein the heterocyclyl
ring, as defined above, is fused with one or more carbocyclyl
groups wherein the point of attachment is either on the carbocyclyl
or heterocyclyl ring, or ring systems wherein the heterocyclyl
ring, as defined above, is fused with one or more aryl or
heteroaryl groups, wherein the point of attachment is on the
heterocyclyl ring, and in such instances, the number of ring
members continue to designate the number of ring members in the
heterocyclyl ring system. In certain embodiments, each instance of
heterocyclyl is independently optionally substituted, e.g.,
unsubstituted (an "unsubstituted heterocyclyl") or substituted (a
"substituted heterocyclyl") with one or more substituents. In
certain embodiments, the heterocyclyl group is unsubstituted 3-10
membered heterocyclyl. In certain embodiments, the heterocyclyl
group is substituted 3-10 membered heterocyclyl.
[0028] In some embodiments, a heterocyclyl group is a 5-10 membered
non-aromatic ring system having ring carbon atoms and 1-4 ring
heteroatoms, wherein each heteroatom is independently selected from
nitrogen, oxygen, and sulfur ("5-10 membered heterocyclyl"). In
some embodiments, a heterocyclyl group is a 5-8 membered
non-aromatic ring system having ring carbon atoms and 1-4 ring
heteroatoms, wherein each heteroatom is independently selected from
nitrogen, oxygen, and sulfur ("5-8 membered heterocyclyl"). In some
embodiments, a heterocyclyl group is a 5-6 membered non-aromatic
ring system having ring carbon atoms and 1-4 ring heteroatoms,
wherein each heteroatom is independently selected from nitrogen,
oxygen, and sulfur ("5-6 membered heterocyclyl"). In some
embodiments, the 5-6 membered heterocyclyl has 1-3 ring heteroatoms
independently selected from nitrogen, oxygen, and sulfur. In some
embodiments, the 5-6 membered heterocyclyl has 1-2 ring heteroatoms
independently selected from nitrogen, oxygen, and sulfur. In some
embodiments, the 5-6 membered heterocyclyl has one ring heteroatom
selected from nitrogen, oxygen, and sulfur.
[0029] Exemplary 3-membered heterocyclyl groups containing one
heteroatom include, without limitation, azirdinyl, oxiranyl, and
thiorenyl. Exemplary 4-membered heterocyclyl groups containing one
heteroatom include, without limitation, azetidinyl, oxetanyl, and
thietanyl. Exemplary 5-membered heterocyclyl groups containing one
heteroatom include, without limitation, tetrahydrofuranyl,
dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl,
pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary
5-membered heterocyclyl groups containing two heteroatoms include,
without limitation, dioxolanyl, oxasulfuranyl, disulfuranyl, and
oxazolidin-2-one. Exemplary 5-membered heterocyclyl groups
containing three heteroatoms include, without limitation,
triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary
6-membered heterocyclyl groups containing one heteroatom include,
without limitation, piperidinyl, tetrahydropyranyl,
dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclyl
groups containing two heteroatoms include, without limitation,
piperazinyl, morpholinyl, dithianyl, and dioxanyl. Exemplary
6-membered heterocyclyl groups containing two heteroatoms include,
without limitation, triazinanyl. Exemplary 7-membered heterocyclyl
groups containing one heteroatom include, without limitation,
azepanyl, oxepanyl and thiepanyl. Exemplary 8-membered heterocyclyl
groups containing one heteroatom include, without limitation,
azocanyl, oxecanyl, and thiocanyl. Exemplary 5-membered
heterocyclyl groups fused to a C.sub.6 aryl ring (also referred to
herein as a 5,6-bicyclic heterocyclic ring) include, without
limitation, indolinyl, isoindolinyl, dihydrobenzofuranyl,
dihydrobenzothienyl, benzoxazolinonyl, and the like. Exemplary
6-membered heterocyclyl groups fused to an aryl ring (also referred
to herein as a 6,6-bicyclic heterocyclic ring) include, without
limitation, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the
like.
[0030] "Aryl" refers to a radical of a monocyclic or polycyclic
(e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g.,
having 6, 10, or 14 r electrons shared in a cyclic array) having
6-14 ring carbon atoms and zero heteroatoms provided in the
aromatic ring system ("C.sub.6-14 aryl"). In some embodiments, an
aryl group has six ring carbon atoms ("C.sub.6 aryl"; e.g.,
phenyl). In some embodiments, an aryl group has ten ring carbon
atoms ("C.sub.10 aryl"; e.g., naphthyl such as 1-naphthyl and
2-naphthyl). In some embodiments, an aryl group has fourteen ring
carbon atoms ("C.sub.14 aryl"; e.g., anthracyl). "Aryl" also
includes ring systems wherein the aryl ring, as defined above, is
fused with one or more carbocyclyl or heterocyclyl groups wherein
the radical or point of attachment is on the aryl ring, and in such
instances, the number of carbon atoms continue to designate the
number of carbon atoms in the aryl ring system. In certain
embodiments, each instance of an aryl group is independently
optionally substituted, e.g., unsubstituted (an "unsubstituted
aryl") or substituted (a "substituted aryl") with one or more
substituents. In certain embodiments, the aryl group is
unsubstituted C.sub.6-14 aryl. In certain embodiments, the aryl
group is substituted C.sub.6-14 aryl.
[0031] "Heteroaryl" refers to a radical of a 5-14 membered
monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2
aromatic ring system (e.g., having 6 or 10 .pi. electrons shared in
a cyclic array) having ring carbon atoms and 1-4 ring heteroatoms
provided in the aromatic ring system, wherein each heteroatom is
independently selected from nitrogen, oxygen and sulfur ("5-14
membered heteroaryl"). In certain embodiments, heteroaryl refers to
a radical of a 5-10 membered monocyclic or bicyclic 4n+2 aromatic
ring system having ring carbon atoms and 1-4 ring heteroatoms
provided in the aromatic ring system, wherein each heteroatom is
independently selected from nitrogen, oxygen and sulfur ("5-10
membered heteroaryl"). In heteroaryl groups that contain one or
more nitrogen atoms, the point of attachment can be a carbon or
nitrogen atom, as valency permits. Heteroaryl bicyclic ring systems
can include one or more heteroatoms in one or both rings.
"Heteroaryl" includes ring systems wherein the heteroaryl ring, as
defined above, is fused with one or more carbocyclyl or
heterocyclyl groups wherein the point of attachment is on the
heteroaryl ring, and in such instances, the number of ring members
continue to designate the number of ring members in the heteroaryl
ring system. "Heteroaryl" also includes ring systems wherein the
heteroaryl ring, as defined above, is fused with one or more aryl
groups wherein the point of attachment is either on the aryl or
heteroaryl ring, and in such instances, the number of ring members
designates the number of ring members in the fused
(aryl/heteroaryl) ring system. Bicyclic heteroaryl groups wherein
one ring does not contain a heteroatom (e.g., indolyl, quinolinyl,
carbazolyl, and the like) the point of attachment can be on either
ring, e.g., either the ring bearing a heteroatom (e.g., 2-indolyl)
or the ring that does not contain a heteroatom (e.g.,
5-indolyl).
[0032] In some embodiments, a heteroaryl group is a 5-14 membered
aromatic ring system having ring carbon atoms and 1-4 ring
heteroatoms provided in the aromatic ring system, wherein each
heteroatom is independently selected from nitrogen, oxygen, and
sulfur ("5-14 membered heteroaryl"). In some embodiments, a
heteroaryl group is a 5-10 membered aromatic ring system having
ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic
ring system, wherein each heteroatom is independently selected from
nitrogen, oxygen, and sulfur ("5-10 membered heteroaryl"). In some
embodiments, a heteroaryl group is a 5-8 membered aromatic ring
system having ring carbon atoms and 1-4 ring heteroatoms provided
in the aromatic ring system, wherein each heteroatom is
independently selected from nitrogen, oxygen, and sulfur ("5-8
membered heteroaryl"). In some embodiments, a heteroaryl group is a
5-6 membered aromatic ring system having ring carbon atoms and 1-4
ring heteroatoms provided in the aromatic ring system, wherein each
heteroatom is independently selected from nitrogen, oxygen, and
sulfur ("5-6 membered heteroaryl"). In some embodiments, the 5-6
membered heteroaryl has 1-3 ring heteroatoms independently selected
from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6
membered heteroaryl has 1-2 ring heteroatoms independently selected
from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6
membered heteroaryl has 1 ring heteroatom selected from nitrogen,
oxygen, and sulfur. In certain embodiments, each instance of a
heteroaryl group is independently optionally substituted, e.g.,
unsubstituted ("unsubstituted heteroaryl") or substituted
("substituted heteroaryl") with one or more substituents. In
certain embodiments, the heteroaryl group is unsubstituted 5-14
membered heteroaryl. In certain embodiments, the heteroaryl group
is substituted 5-14 membered heteroaryl.
[0033] Exemplary 5-membered heteroaryl groups containing one
heteroatom include, without limitation, pyrrolyl, furanyl and
thiophenyl. Exemplary 5-membered heteroaryl groups containing two
heteroatoms include, without limitation, imidazolyl, pyrazolyl,
oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary
5-membered heteroaryl groups containing three heteroatoms include,
without limitation, triazolyl, oxadiazolyl, and thiadiazolyl.
Exemplary 5-membered heteroaryl groups containing four heteroatoms
include, without limitation, tetrazolyl. Exemplary 6-membered
heteroaryl groups containing one heteroatom include, without
limitation, pyridinyl. Exemplary 6-membered heteroaryl groups
containing two heteroatoms include, without limitation,
pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered
heteroaryl groups containing three or four heteroatoms include,
without limitation, triazinyl and tetrazinyl, respectively.
Exemplary 7-membered heteroaryl groups containing one heteroatom
include, without limitation, azepinyl, oxepinyl, and thiepinyl.
Exemplary 5,6-bicyclic heteroaryl groups include, without
limitation, indolyl, isoindolyl, indazolyl, benzotriazolyl,
benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl,
benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl,
benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and
purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, without
limitation, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl,
cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.
[0034] "Fused" or "ortho-fused" are used interchangeably herein,
and refer to two rings that have two atoms and one bond in common,
e.g.,
##STR00005##
[0035] "Bridged" refers to a ring system containing (1) a
bridgehead atom or group of atoms which connect two or more
non-adjacent positions of the same ring; or (2) a bridgehead atom
or group of atoms which connect two or more positions of different
rings of a ring system and does not thereby form an ortho-fused
ring, e.g.,
##STR00006##
[0036] "Spiro" or "Spiro-fused" refers to a group of atoms which
connect to the same atom of a carbocyclic or heterocyclic ring
system (geminal attachment), thereby forming a ring, e.g.,
##STR00007##
Spiro-fusion at a bridgehead atom is also contemplated.
[0037] "Partially unsaturated" refers to a group that includes at
least one double or triple bond. The term "partially unsaturated"
is intended to encompass rings having multiple sites of
unsaturation, but is not intended to include aromatic groups (e.g.,
aryl or heteroaryl groups) as herein defined. Likewise, "saturated"
refers to a group that does not contain a double or triple bond,
i.e., contains all single bonds.
[0038] In some embodiments, aliphatic, alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl groups, as defined
herein, are optionally substituted (e.g., "substituted" or
"unsubstituted" aliphatic, "substituted" or "unsubstituted" alkyl,
"substituted" or "unsubstituted" alkenyl, "substituted" or
"unsubstituted" alkynyl, "substituted" or "unsubstituted"
carbocyclyl, "substituted" or "unsubstituted" heterocyclyl,
"substituted" or "unsubstituted" aryl or "substituted" or
"unsubstituted" heteroaryl group). In general, the term
"substituted", whether preceded by the term "optionally" or not,
means that at least one hydrogen present on a group (e.g., a carbon
or nitrogen atom) is replaced with a permissible substituent, e.g.,
a substituent which upon substitution results in a stable compound,
e.g., a compound which does not spontaneously undergo
transformation such as by rearrangement, cyclization, elimination,
or other reaction. Unless otherwise indicated, a "substituted"
group has a substituent at one or more substitutable positions of
the group, and when more than one position in any given structure
is substituted, the substituent is either the same or different at
each position. The term "substituted" is contemplated to include
substitution with all permissible substituents of organic
compounds, including any of the substituents described herein that
results in the formation of a stable compound. The present
disclosure contemplates any and all such combinations in order to
arrive at a stable compound. For purposes of this disclosure,
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.
[0039] Exemplary carbon atom substituents include, but are not
limited to, halogen, --CN, --NO.sub.2, --N.sub.3, --SO.sub.2H,
--SO.sub.3H, --OH, --OR.sup.aa, --ON(R.sup.bb).sub.2,
--N(R.sup.bb).sub.2, --N(R.sup.bb).sub.3.sup.+X.sup.-,
--N(OR.sup.cc)R.sup.bb, --SH, --SR.sup.aa, --SSR.sup.cc,
--C(.dbd.O)R.sup.aa, --CO.sub.2H, --CHO, --C(OR.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --OC(.dbd.O)R.sup.aa, --OCO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.2, --OC(.dbd.O)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.O)R.sup.aa, --NR.sup.bbCO.sub.2R.sup.aa,
--NR.sup.bbC(.dbd.O)N(R.sup.bb).sub.2, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --OC(.dbd.NR.sup.bb)R.sup.aa,
--OC(.dbd.NR.sup.bb)OR.sup.aa,
--C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--OC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--C(.dbd.O)NR.sup.bbSO.sub.2R.sup.aa, --NR.sup.bbSO.sub.2R.sup.aa,
--SO.sub.2N(R.sup.bb).sub.2, --SO.sub.2R.sup.aa,
--SO.sub.2OR.sup.aa, --OSO.sub.2R.sup.aa, --S(.dbd.O)R.sup.aa,
--OS(.dbd.O)R.sup.aa, --Si(R.sup.aa).sub.3,
--OSi(R.sup.aa).sub.3--C(.dbd.S)N(R.sup.bb).sub.2,
--C(.dbd.O)SR.sup.aa, --C(.dbd.S)SR.sup.aa, --SC(.dbd.S)SR.sup.aa,
--SC(.dbd.O)SR.sup.aa, --OC(.dbd.O)SR.sup.aa,
--SC(.dbd.O)OR.sup.aa, --SC(.dbd.O)R.sup.aa,
--P(.dbd.O).sub.2R.sup.aa, --OP(.dbd.O).sub.2R.sup.aa,
--P(.dbd.O)(R.sup.aa).sub.2, --OP(.dbd.O)(R.sup.aa).sub.2,
--OP(.dbd.O)(OR.sup.cc).sub.2, --P(.dbd.O).sub.2N(R.sup.bb).sub.2,
--OP(.dbd.O).sub.2N(R.sup.bb).sub.2, --P(.dbd.O)(NR.sup.bb).sub.2,
--OP(.dbd.O)(NR.sup.bb).sub.2,
--NR.sup.bbP(.dbd.O)(OR.sup.cc).sub.2,
--NR.sup.bbP(.dbd.O)(NR.sup.bb).sub.2, --P(R.sup.cc).sub.2,
--P(R.sup.cc).sub.3, --OP(R.sup.cc).sub.2, --OP(R.sup.cc).sub.3,
--B(R.sup.aa).sub.2, --B(OR.sup.cc).sub.2, --BR.sup.aa(OR.sup.cc),
C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl,
wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 R.sup.dd groups; [0040] or two geminal hydrogens on a
carbon atom are replaced with the group .dbd.O, .dbd.S,
.dbd.NN(R.sup.bb).sub.2, .dbd.NNR.sup.bbC(.dbd.O)R.sup.aa,
.dbd.NNR.sup.bbC(.dbd.O)OR.sup.aa,
.dbd.NNR.sup.bbS(.dbd.O).sub.2R.sup.aa, .dbd.NR.sup.bb, or
.dbd.NOR.sup.cc; [0041] each instance of R.sup.aa is,
independently, selected from C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, and 5-14
membered heteroaryl, or two R.sup.aa groups are joined to form a
3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,
wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 R.sup.dd groups; [0042] each instance of R.sup.bb is,
independently, selected from hydrogen, --OH, --OR.sup.aa,
--N(R.sup.cc).sub.2, --CN, --C(.dbd.O)R.sup.aa,
--C(.dbd.O)N(R.sup.cc).sub.2, --CO.sub.2R.sup.aa,
--SO.sub.2R.sup.aa, --C(.dbd.NR.sup.cc)OR.sup.aa,
--C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2, --SO.sub.2N(R.sup.cc).sub.2,
--SO.sub.2R.sup.cc, --SO.sub.2OR.sup.cc, --SOR.sup.aa,
--C(.dbd.S)N(R.sup.cc).sub.2, --C(.dbd.O)SR.sup.cc,
--C(.dbd.S)SR.sup.cc, --P(.dbd.O).sub.2R.sup.aa,
--P(.dbd.O)(R.sup.aa).sub.2, --P(.dbd.O).sub.2N(R.sup.cc).sub.2,
--P(.dbd.O)(NR.sup.cc).sub.2, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, and 5-14
membered heteroaryl, or two R.sup.bb groups are joined to form a
3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,
wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 R.sup.dd groups; [0043] each instance of R.sup.cc is,
independently, selected from hydrogen, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, and 5-14
membered heteroaryl, or two R.sup.cc groups are joined to form a
3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,
wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 R.sup.dd groups; [0044] each instance of R.sup.dd is,
independently, selected from halogen, --CN, --NO.sub.2, --N.sub.3,
--SO.sub.2H, --SO.sub.3H, --OH, --OR.sup.ee, --ON(R.sup.ff).sub.2,
--N(R.sup.ff).sub.2, --N(R.sup.ff).sub.3.sup.+X.sup.-,
--N(OR.sup.ee)R.sup.ff, --SH, --SR.sup.ee, --SSR.sup.ee,
--C(.dbd.O)R.sup.ee, --CO.sub.2H, --CO.sub.2R.sup.ee,
--OC(.dbd.O)R.sup.ee, --OCO.sub.2R.sup.ee,
--C(.dbd.O)N(R.sup.ff).sub.2, --OC(.dbd.O)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.O)R.sup.ee, --NR.sup.ffCO.sub.2R.sup.ee,
--NR.sup.ffC(.dbd.O)N(R.sup.ff).sub.2,
--C(.dbd.NR.sup.ff)OR.sup.ee, --OC(.dbd.NR.sup.ff)R.sup.ee,
--OC(.dbd.NR.sup.ff)OR.sup.ee,
--C(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--OC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffSO.sub.2R.sup.ee, --SO.sub.2N(R.sup.ff).sub.2,
--SO.sub.2R.sup.ee, --SO.sub.2OR.sup.ee, --OSO.sub.2R.sup.ee,
--S(.dbd.O)R.sup.ee, --Si(R.sup.ee).sub.3, --OSi(R.sup.ee).sub.3,
--C(.dbd.S)N(R.sup.ff).sub.2, --C(.dbd.O)SR.sup.ee,
--C(.dbd.S)SR.sup.ee, --SC(.dbd.S)SR.sup.ee,
--P(.dbd.O).sub.2R.sup.ee, --P(.dbd.O)(R.sup.ee).sub.2,
--OP(.dbd.O)(R.sup.ee).sub.2, --OP(.dbd.O)(OR.sup.ee).sub.2,
C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered
heterocyclyl, C.sub.6-10 aryl, 5-10 membered heteroaryl, wherein
each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and
heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5
R.sup.gg groups, or two geminal R.sup.dd substituents can be joined
to form .dbd.O or .dbd.S; [0045] each instance of R.sup.ee is,
independently, selected from C.sub.1-6 alkyl, C.sub.1-6
perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-10
carbocyclyl, C.sub.6-10 aryl, 3-10 membered heterocyclyl, and 3-10
membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently
substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups; [0046] each
instance of R.sup.ff is, independently, selected from hydrogen,
C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered
heterocyclyl, C.sub.6-10 aryl and 5-10 membered heteroaryl, or two
R.sup.ff groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups;
and [0047] each instance of R.sup.gg is, independently, halogen,
--CN, --NO.sub.2, --N.sub.3, --SO.sub.2H, --SO.sub.3H, --OH,
--OC.sub.1-6 alkyl, --ON(C.sub.1-6 alkyl).sub.2, --N(C.sub.1-6
alkyl).sub.2, --N(C.sub.1-6 alkyl).sub.3.sup.+X.sup.-,
--NH(C.sub.1-6 alkyl).sub.2.sup.+X.sup.-, --NH.sub.2(C.sub.1-6
alkyl).sup.+X.sup.-, --NH.sub.3.sup.+X.sup.-, --N(OC.sub.1-6
alkyl)(C.sub.1-6 alkyl), --N(OH)(C.sub.1-6 alkyl), --NH(OH), --SH,
--SC.sub.1-6 alkyl, --SS(C.sub.1-10 alkyl), --C(.dbd.O)(C.sub.1-6
alkyl), --CO.sub.2H, --CO.sub.2(C.sub.1-6 alkyl),
--OC(.dbd.O)(C.sub.1-6 alkyl), --OCO.sub.2(C.sub.1-6 alkyl),
--C(.dbd.O)NH.sub.2, --C(.dbd.O)N(C.sub.1-6 alkyl).sub.2,
--OC(.dbd.O)NH(C.sub.1-6 alkyl), --NHC(.dbd.O)(C.sub.1-6 alkyl),
--N(C.sub.1-6 alkyl)C(.dbd.O)(C.sub.1-6 alkyl),
--NHCO.sub.2(C.sub.1-6 alkyl), --NHC(.dbd.O)N(C.sub.1-6
alkyl).sub.2, --NHC(.dbd.O)NH(C.sub.1-6 alkyl),
--NHC(.dbd.O)NH.sub.2, --C(.dbd.NH)O(C.sub.1-6 alkyl),
--OC(.dbd.NH)(C.sub.1-6 alkyl), --OC(.dbd.NH)OC.sub.1-6 alkyl,
--C(.dbd.NH)N(C.sub.1-6 alkyl).sub.2, --C(.dbd.NH)NH(C.sub.1-6
alkyl), --C(.dbd.NH)NH.sub.2, --OC(.dbd.NH)N(C.sub.1-6
alkyl).sub.2, --OC(NH)NH(C.sub.1-6 alkyl), --OC(NH)NH.sub.2,
--NHC(NH)N(C.sub.1-6 alkyl).sub.2, --NHC(.dbd.NH)NH.sub.2,
--NHSO.sub.2(C.sub.1-6 alkyl), --SO.sub.2N(C.sub.1-6 alkyl).sub.2,
--SO.sub.2NH(C.sub.1-6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2C.sub.1-6 alkyl, --SO.sub.2OC.sub.1-6 alkyl,
--OSO.sub.2C.sub.1-6 alkyl, --SOC.sub.1-6 alkyl, --Si(C.sub.1-4
alkyl).sub.3, --OSi(C.sub.1-6 alkyl).sub.3--C(.dbd.S)N(C.sub.1-6
alkyl).sub.2, C(.dbd.S)NH(C.sub.1-6 alkyl), C(.dbd.S)NH.sub.2,
--C(.dbd.O)S(C.sub.1-6 alkyl), --C(.dbd.S)SC.sub.1-6 alkyl,
--SC(.dbd.S)SC.sub.1-6 alkyl, --P(.dbd.O).sub.2(C.sub.1-6 alkyl),
--P(.dbd.O)(C.sub.1-6 alkyl).sub.2, --OP(.dbd.O)(C.sub.1-6
alkyl).sub.2, --OP(.dbd.O)(OC.sub.1-6 alkyl).sub.2, C.sub.1-6
alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6
alkynyl, C.sub.3-10 carbocyclyl, C.sub.6-10 aryl, 3-10 membered
heterocyclyl, 5-10 membered heteroaryl; or two geminal R.sup.gg
substituents can be joined to form .dbd.O or .dbd.S; wherein
X.sup.- is a counterion.
[0048] A "counterion" or "anionic counterion" is a negatively
charged group associated with a cationic quaternary amino group in
order to maintain electronic neutrality. Exemplary counterions
include halide ions (e.g., F.sup.-, Cl.sup.-, Br.sup.-, I.sup.-),
NO.sub.3.sup.-, ClO.sub.4.sup.-, OH.sup.-, H.sub.2PO.sub.4.sup.-,
HSO.sub.4.sup.-, sulfonate ions (e.g., methansulfonate,
trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate,
10-camphor sulfonate, naphthalene-2-sulfonate,
naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonic
acid-2-sulfonate, and the like), and carboxylate ions (e.g.,
acetate, ethanoate, propanoate, benzoate, glycerate, lactate,
tartrate, glycolate, and the like).
[0049] "Halo" or "halogen" refers to fluorine (fluoro, --F),
chlorine (chloro, --Cl), bromine (bromo, --Br), or iodine (iodo,
--I).
[0050] Nitrogen atoms can be substituted or unsubstituted as
valency permits, and include primary, secondary, tertiary, and
quarternary nitrogen atoms. Exemplary nitrogen atom substitutents
include, but are not limited to, hydrogen, --OH, --OR.sup.aa,
--N(R.sup.cc).sub.2, --CN, --C(.dbd.O)R.sup.aa,
--C(.dbd.O)N(R.sup.cc).sub.2, --CO.sub.2R.sup.aa,
--SO.sub.2R.sup.aa, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.cc)OR.sup.aa, --C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2,
--SO.sub.2N(R.sup.cc).sub.2, --SO.sub.2R.sup.cc,
--SO.sub.2OR.sup.cc, --SOR.sup.aa, --C(.dbd.S)N(R.sup.cc).sub.2,
--C(.dbd.O)SR.sup.cc, --C(.dbd.S)SR.sup.cc,
--P(.dbd.O).sub.2R.sup.aa, --P(.dbd.O)(R.sup.aa).sub.2,
--P(.dbd.O).sub.2N(R.sup.cc).sub.2, --P(.dbd.O)(NR.sup.cc).sub.2,
C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6 14 aryl, and 5-14 membered heteroaryl, or two
R.sup.cc groups attached to a nitrogen atom are joined to form a
3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,
wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 R.sup.dd groups, and wherein R.sup.aa, R.sup.bb, R.sup.cc
and R.sup.dd are as defined above.
[0051] In certain embodiments, the substituent present on a
nitrogen atom is a nitrogen protecting group (also referred to as
an amino protecting group). Nitrogen protecting groups include, but
are not limited to, --OH, --OR.sup.aa, --N(R.sup.cc).sub.2,
--C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.cc)R.sup.aa, --C(.dbd.NR.sup.cc)OR.sup.aa,
--C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2, --SO.sub.2N(R.sup.cc).sub.2,
--SO.sub.2R.sup.cc, --SO.sub.2OR.sup.cc, --SOR.sup.aa,
--C(.dbd.S)N(R.sup.cc).sub.2, --C(.dbd.O)SR.sup.cc,
--C(.dbd.S)SR.sup.cc, C.sub.1-10 alkyl (e.g., aralkyl,
heteroaralkyl), C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, and 5-14
membered heteroaryl groups, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aralkyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups,
and wherein R.sup.aa, R.sup.bb, R.sup.cc, and R.sup.dd are as
defined herein. Nitrogen protecting groups are 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, incorporated herein by
reference.
[0052] Amide nitrogen protecting groups (e.g., --C(.dbd.O)R.sup.aa)
include, but are not limited to, formamide, acetamide,
chloroacetamide, trichloroacetamide, trifluoroacetamide,
phenylacetamide, 3-phenylpropanamide, picolinamide,
3-pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide,
p-phenylbenzamide, o-nitophenylacetamide, o-nitrophenoxyacetamide,
acetoacetamide, (N'-dithiobenzyloxyacylamino)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,
o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.
[0053] Carbamate nitrogen protecting groups (e.g.,
--C(.dbd.O)OR.sup.aa) include, but are not limited to, 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-10,10,10,10,10-tetrahydrothioxanthyl)]meth-
yl 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, t-amyl carbamate, S-benzyl
thiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate,
cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl
carbamate, p-decyloxybenzyl carbamate, 2,2-dimethoxyacylvinyl
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, and 2,4,6-trimethylbenzyl carbamate.
[0054] Sulfonamide nitrogen protecting groups (e.g.,
--S(.dbd.O).sub.2R.sup.aa) include, but are not limited to,
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), J3-trimethylsilylethanesulfonamide (SES),
9-anthracenesulfonamide,
4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),
benzylsulfonamide, trifluoromethylsulfonamide, and
phenacylsulfonamide.
[0055] Other nitrogen protecting groups include, but are not
limited to, phenothiazinyl-(10)-acyl derivative,
N'-p-toluenesulfonylaminoacyl derivative, N'-phenylaminothioacyl
derivative, N-benzoylphenylalanyl derivative, N-acetylmethionine
derivative, 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(pentaacylchromium- or tungsten)acyl]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,
and 3-nitropyridinesulfenamide (Npys).
[0056] In certain embodiments, the substituent present on an oxygen
atom is an oxygen protecting group (also referred to as a hydroxyl
protecting group). Oxygen protecting groups include, but are not
limited to, --R.sup.aa, --N(R.sup.bb).sub.2, --C(.dbd.O)SR.sup.aa,
--C(.dbd.O)R.sup.aa, --CO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.2, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--S(.dbd.O)R.sup.aa, --SO.sub.2R.sup.aa, --Si(R.sup.aa).sub.3,
--P(R.sup.cc).sub.2, --P(R.sup.cc).sub.3,
--P(.dbd.O).sub.2R.sup.aa, --P(.dbd.O)(R.sup.aa).sub.2,
--P(.dbd.O)(OR.sup.cc).sub.2, --P(.dbd.O).sub.2N(R.sup.bb).sub.2,
and --P(.dbd.O)(NR.sup.bb).sub.2, wherein R.sup.aa, R.sup.bb, and
R.sup.cc are as defined herein. Oxygen protecting groups are 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, incorporated
herein by reference.
[0057] Exemplary oxygen protecting groups include, but are not
limited to, 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 (Bn), 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-benzodisulfuran-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), t-butyl carbonate (BOC), 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-(methoxyacyl)benzoate, a-naphthoate, nitrate, alkyl
N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate,
borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate,
sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate
(Ts).
[0058] In certain embodiments, the substituent present on a sulfur
atom is a sulfur protecting group (also referred to as a thiol
protecting group). Sulfur protecting groups include, but are not
limited to, --R.sup.aa, --N(R.sup.bb).sub.2, --C(.dbd.O)SR.sup.aa,
--C(.dbd.O)R.sup.aa, --CO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.2, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--S(.dbd.O)R.sup.aa, --SO.sub.2R.sup.aa, --Si(R.sup.aa).sub.3,
--P(R.sup.cc).sub.2, --P(R.sup.cc).sub.3,
--P(.dbd.O).sub.2R.sup.aa, --P(.dbd.O)(R.sup.aa).sub.2,
--P(.dbd.O)(OR.sup.cc).sub.2, --P(.dbd.O).sub.2N(R.sup.bb).sub.2,
and --P(.dbd.O)(NR.sup.bb).sub.2, wherein R.sup.aa, R.sup.bb, and
R.sup.cc are as defined herein. Sulfur protecting groups are 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, incorporated
herein by reference.
[0059] As used herein, a "leaving group", or "LG", is a term
understood in the art to refere to a molecular fragment that
departs with a pair of electrons upon heterolytic bond cleavage,
wherein the molecular fragment is an anion or neutral molecule.
See, for example, Smith, March Advanced Organic Chemistry 6th ed.
(501-502). Examples of suitable leaving groups include, but are not
limited to, halides (such as chloride, bromide, or iodide),
alkoxycarbonyloxy, aryloxycarbonyloxy, alkanesulfonyloxy,
arenesulfonyloxy, alkyl-carbonyloxy (e.g., acetoxy),
arylcarbonyloxy, aryloxy, methoxy, N,O-dimethylhydroxylamino,
pixyl, haloformates, --NO.sub.2, trialkylammonium, and aryliodonium
salts. In some embodiments, the leaving group is a sulfonic acid
ester. In some embodiments, the sulfonic acid ester comprises the
formula --OSO.sub.2R.sup.LG1 wherein R.sup.LG1 is selected from the
group consisting alkyl optionally, alkenyl optionally substituted,
heteroalkyl optionally substituted, aryl optionally substituted,
heteroaryl optionally substituted, arylalkyl optionally
substituted, and heterarylalkyl optionally substituted. In some
embodiments, R.sup.LG1 is substituted or unsubstituted
C.sub.1-C.sub.6 alkyl. In some embodiments, R.sup.LG1 is methyl. In
some embodiments, R.sup.LG1 is --CF.sub.3. In some embodiments,
R.sup.LG1 is substituted or unsubstituted aryl. In some
embodiments, R.sup.LG1 is substituted or unsubstituted phenyl. In
some embodiments R.sup.LG1 is:
##STR00008##
[0060] These and other exemplary substituents are described in more
detail in the Detailed Description, Examples, and claims. The
present disclosure is not intended to be limited in any manner by
the above exemplary listing of substituents.
[0061] "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 other 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.
Pharmaceutically acceptable salts of the compounds describe herein
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,
quaternary salts.
[0062] A "subject" to which administration is contemplated
includes, but is not limited to, humans (e.g., a male or female of
any age group, e.g., a pediatric subject (e.g, infant, child,
adolescent) or adult subject (e.g., young adult, middle-aged adult
or senior adult)) and/or other non-human animals, for example,
non-human mammals (e.g., primates (e.g., cynomolgus monkeys, rhesus
monkeys); commercially relevant mammals such as cattle, pigs,
horses, sheep, goats, cats, and/or dogs), birds (e.g., commercially
relevant birds such as chickens, ducks, geese, and/or turkeys),
rodents (e.g., rats and/or mice), reptiles, amphibians, and fish.
In certain embodiments, the non-human animal is a mammal. The
non-human animal may be a male or female at any stage of
development. A non-human animal may be a transgenic animal.
[0063] "Condition," "disease," and "disorder" are used
interchangeably herein.
[0064] "Treat," "treating" and "treatment" encompasses an action
that occurs while a subject is suffering from a condition which
reduces the severity of the condition or retards or slows the
progression of the condition ("therapeutic treatment"). "Treat,"
"treating" and "treatment" also encompasses an action that occurs
before a subject begins to suffer from the condition and which
inhibits or reduces the severity of the condition ("prophylactic
treatment").
[0065] An "effective amount" of a compound refers to an amount
sufficient to elicit the desired biological response, e.g., treat
the condition. As will be appreciated by those of ordinary skill in
this art, the effective amount of a compound described herein may
vary depending on such factors as the desired biological endpoint,
the pharmacokinetics of the compound, the condition being treated,
the mode of administration, and the age and health of the subject.
An effective amount encompasses therapeutic and prophylactic
treatment.
[0066] A "therapeutically effective amount" of a compound is an
amount sufficient to provide a therapeutic benefit in the treatment
of a condition or to delay or minimize one or more symptoms
associated with the condition. A therapeutically effective amount
of a compound means an amount of therapeutic agent, alone or in
combination with other therapies, which provides a therapeutic
benefit in the treatment of the condition. The term
"therapeutically effective amount" can encompass an amount that
improves overall therapy, reduces or avoids symptoms or causes of
the condition, or enhances the therapeutic efficacy of another
therapeutic agent.
[0067] A "prophylactically effective amount" of a compound is an
amount sufficient to prevent a condition, or one or more symptoms
associated with the condition or prevent its recurrence. A
prophylactically effective amount of a compound means an amount of
a therapeutic agent, alone or in combination with other agents,
which provides a prophylactic benefit in the prevention of the
condition. The term "prophylactically effective amount" can
encompass an amount that improves overall prophylaxis or enhances
the prophylactic efficacy of another prophylactic agent.
[0068] As used herein, the term "methyltransferase" represents
transferase class enzymes that are able to transfer a methyl group
from a donor molecule to an acceptor molecule, e.g., an amino acid
residue of a protein or a nucleic base of a DNA molecule.
Methytransferases typically use a reactive methyl group bound to
sulfur in S-adenosyl methionine (SAM) as the methyl donor. In some
embodiments, a methyltransferase described herein is a protein
methyltransferase. In some embodiments, a methyltransferase
described herein is a histone methyltransferase. Histone
methyltransferases (HMT) are histone-modifying enzymes, (including
histone-lysine N-methyltransferase and histone-arginine
N-methyltransferase), that catalyze the transfer of one or more
methyl groups to lysine and arginine residues of histone proteins.
In certain embodiments, a methyltransferase described herein is a
histone-arginine N-methyltransferase.
[0069] As generally described above, provided herein are compounds
useful as PRMT5 inhibitors. In certain embodiments, the present
disclosure provides a compound of Formula (A):
##STR00009##
or a pharmaceutically acceptable salt thereof, wherein: [0070]
represents a single or double bond; [0071] R.sup.12 is hydrogen,
halogen, or optionally substituted C.sub.1-3alkyl; [0072] R.sup.13
is hydrogen, halogen, --NR.sup.A1R.sup.A2, optionally substituted
C.sub.1-3alkyl or --OR.sup.1; [0073] R.sup.A1 and R.sup.A2 are each
independently hydrogen, optionally substituted C.sub.1-3 alkyl, a
nitrogen protecting group, or R.sup.A1 and R.sup.A2 are taken
together with the intervening nitrogen atom to form an optionally
substituted 3-6 membered heterocyclic ring; [0074] R.sup.1 is
hydrogen, R.sup.z, or --C(O)R.sup.z, wherein R.sup.z is optionally
substituted C.sub.1-6 alkyl; [0075] X is a bond, --O--, --N(R)--,
--CR.sup.4R.sup.5--, --O--CR.sup.4R.sup.5,
--N(R)--CR.sup.4R.sup.5--, --O--CR.sup.4R.sup.5--O--,
--N(R)--CR.sup.4R.sup.5--O, --N(R)--CR.sup.4R.sup.5--N(R)--,
--O--CR.sup.4R.sup.5--N(R)--, --CR.sup.4R.sup.5--O--,
--CR.sup.4R.sup.5--N(R)--, --O--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--N(R)--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--O--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--N(R)--, or
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--; [0076] each R is
independently hydrogen or optionally substituted C.sub.1-6
aliphatic; [0077] R.sup.2 and R.sup.3 are each independently
selected from the group consisting of hydrogen, halo, --CN,
--NO.sub.2, optionally substituted aliphatic, optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, optionally substituted heteroaryl,
--OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A, --C(.dbd.O)R.sup.A,
--C(O)OR.sup.A, --C(O)SR.sup.A, --C(O)N(R.sup.B).sub.2,
--C(O)N(R.sup.B)N(R.sup.B).sub.2, --OC(O)R.sup.A,
--OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
[0078] R.sup.4 and R.sup.5 are each independently selected from the
group consisting of hydrogen, halo, --CN, --NO.sub.2, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted phenyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.4 and R.sup.5 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
[0079] R.sup.6 and R.sup.7 are each independently selected from the
group consisting of hydrogen, halo, --CN, --NO.sub.2, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted phenyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.6 and R.sup.7 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring;
[0080] each R.sup.A is independently selected from the group
consisting of hydrogen, optionally substituted aliphatic,
optionally substituted carbocyclyl, optionally substituted
heterocyclyl, optionally substituted aryl, and optionally
substituted heteroaryl; [0081] each R.sup.B is independently
selected from the group consisting of hydrogen, optionally
substituted aliphatic, optionally substituted carbocyclyl,
optionally substituted heterocyclyl, optionally substituted aryl,
and optionally substituted heteroaryl, or two R.sup.B groups are
taken together with their intervening atoms to form an optionally
substituted heterocyclic ring; [0082] R.sup.8, R.sup.9, R.sup.10,
and R.sup.11 are independently hydrogen, halo, or optionally
substituted aliphatic; Cy is a monocyclic or bicyclic, saturated,
partially unsaturated, or aromatic ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur, wherein
Cy is substituted with 0, 1, 2, 3, or 4 R.sup.y groups; [0083] each
R.sup.y is independently selected from the group consisting of
halo, --CN, --NO.sub.2, optionally substituted aliphatic,
optionally substituted carbocyclyl, optionally substituted aryl,
optionally substituted heterocyclyl, optionally substituted
heteroaryl, --OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A,
C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --C(O)N(R.sup.B)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or an
R.sup.y group may be optionally taken together with R.sup.2 or
R.sup.3 to form an optionally substituted 5- to 6-membered
carbocyclic or heterocyclic ring fused to Cy; [0084] each R.sup.x
is independently selected from the group consisting of halo, --CN,
optionally substituted aliphatic, --OR', and --N(R'').sub.2; [0085]
R' is hydrogen or optionally substituted aliphatic; [0086] each R''
is independently hydrogen or optionally substituted aliphatic, or
two R'' are taken together with their intervening atoms to form an
optionally substituted heterocyclic ring having 1-2 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; and
[0087] n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency
permits.
[0088] In some embodiments, the carbon attached to R.sup.12 has
(S)-stereochemistry. In some embodiments, the carbon attached to
R.sup.12 has (R)-stereochemistry. In some embodiments, the carbon
attached to R.sup.13 has (S)-stereochemistry. In some embodiments,
the carbon attached to R.sup.13 has (R)-stereochemistry. In some
embodiments, R.sup.12 is hydrogen. In some embodiments, R.sup.13 is
hydrogen. In some embodiments, both R.sup.12 and R.sup.13 are
hydrogen. In some embodiments, R.sup.12 is optionally substituted
C.sub.1-3alkyl. In some embodiments, R.sup.13 is optionally
substituted C.sub.1-3alkyl. In some embodiments, both R.sup.12 and
R.sup.13 are optionally substituted C.sub.1-3alkyl. In some
embodiments, R.sup.12 is halogen e.g., fluoro, bromo, chloro, or
iodo, provided that R.sup.13 is not --OR.sup.1. In some
embodiments, R.sup.13 is halogen e.g., fluoro, bromo, chloro, or
iodo. In some embodiments, both R.sup.12 and R.sup.13 are halogen
e.g., fluoro, bromo, chloro, or iodo. In some embodiments, R.sup.12
is halogen e.g., fluoro, bromo, chloro, or iodo and R.sup.13 is
optionally substituted C.sub.1-3alkyl. In some embodiments,
R.sup.12 is optionally substituted C.sub.1-3alkyl and R.sup.13 is
halogen e.g., fluoro, bromo, chloro, or iodo. In some embodiments,
R.sup.13 is --OR.sup.1. In some embodiments, R.sup.12 is optionally
substituted C.sub.1-3alkyl and R.sup.13 is --OR.sup.1. In some
embodiments, R.sup.12 is hydrogen and R.sup.13 is --OR.sup.1. In
some embodiments, R.sup.12 is hydrogen and R.sup.13 optionally
substituted C.sub.1-3alkyl. In some embodiments, R.sup.12 is
optionally substituted C.sub.1-3alkyl and R.sup.13 is hydrogen. In
some embodiments, R.sup.12 is halogen e.g., fluoro, bromo, chloro,
or iodo and R.sup.13 is hydrogen. In some embodiments, R.sup.12 is
hydrogen and R.sup.13 is halogen e.g., fluoro, bromo, chloro, or
iodo.
[0089] As generally defined above, R.sup.12 is hydrogen, halogen,
or optionally substituted C.sub.1-3alkyl. In certain embodiments,
R.sup.12 is hydrogen. In certain embodiments, R.sup.12 is
optionally substituted C.sub.1-3alkyl, e.g., optionally substituted
with halogen. In certain embodiments, R.sup.12 is optionally
substituted C.sub.1alkyl, e.g., methyl or trifluoromethyl. In
certain embodiments, R.sup.12 is optionally substituted C.sub.2
alkyl, e.g., ethyl. In certain embodiments, R.sup.12 is optionally
substituted C.sub.3 alkyl, e.g., propyl. In certain embodiments,
R.sup.12 is fluoro, provided that R.sup.13 is not --OR.sup.1. In
certain embodiments, R.sup.12 is chloro, provided that R.sup.13 is
not --OR.sup.1. In certain embodiments, R.sup.12 is bromo, provided
that R.sup.13 is not --OR.sup.1. In certain embodiments, R.sup.12
is iodo, provided that R.sup.13 is not --OR.sup.1.
[0090] As generally defined above, R.sup.13 is hydrogen, halogen,
optionally substituted C.sub.1-3alkyl, --NR.sup.A1R.sup.A2, or
--OR.sup.1. In certain embodiments, R.sup.13 is hydrogen. In
certain embodiments, R.sup.13 is optionally substituted
C.sub.1-3alkyl, e.g., optionally substituted with halogen. In
certain embodiments, R.sup.13 is optionally substituted
C.sub.1alkyl, e.g., methyl or trifluoromethyl. In certain
embodiments, R.sup.13 is optionally substituted C.sub.2 alkyl,
e.g., ethyl. In certain embodiments, R.sup.13 is optionally
substituted C.sub.3 alkyl, e.g., propyl. In certain embodiments,
R.sup.13 is fluoro. In certain embodiments, R.sup.13 is chloro. In
certain embodiments, R.sup.13 is bromo. In certain embodiments,
R.sup.13 is iodo.
[0091] For example, in some embodiments of Formula (A), wherein
R.sup.13 is hydrogen, the present disclosure provides a compound of
Formula (A-1):
##STR00010##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.2,
R.sup.3, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.x,
n, X, and Cy are as described herein.
[0092] In some embodiments of Formula (A), wherein R.sup.12 is
hydrogen, the present disclosure provides a compound of Formula
(A-2):
##STR00011##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.2,
R.sup.3, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.13, R.sup.x,
n, X, and Cy are as described herein.
[0093] In some embodiments of Formula (A), wherein both R.sup.12
and R.sup.13 are hydrogen, the present disclosure provides a
compound of Formula (A-3):
##STR00012##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.2,
R.sup.3, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.x, n, X, and
Cy are as described herein.
[0094] In some embodiments of Formula (A), wherein R.sup.13 is
--OR.sup.1, the present disclosure provides a compound of Formula
(A-4):
##STR00013##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.1,
R.sup.2, R.sup.3, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12,
R.sup.x, n, X, and Cy are as described herein.
[0095] In some embodiments of Formula (A), wherein R.sup.13 is
--NR.sup.A1R.sup.A2, the present disclosure provides a compound of
Formula (A-5):
##STR00014##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.2,
R.sup.3, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.x,
R.sup.A1, R.sup.A2, n, X, and Cy are as described herein.
[0096] In some embodiments of Formula (A), wherein R.sup.8,
R.sup.9, R.sup.10, and R.sup.11 are hydrogen, the present
disclosure provides a compound of Formula (A-6):
##STR00015##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.2,
R.sup.3, R.sup.12, R.sup.13, R.sup.x, R.sup.A1, R.sup.A2, n, X, and
Cy are as described herein.
[0097] In some embodiments of Formula (A), wherein R.sup.8,
R.sup.9, R.sup.10, and R.sup.11 are hydrogen and R.sup.13 is
--OR.sup.1, the present disclosure provides a compound of Formula
(A-7):
##STR00016##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.1,
R.sup.2, R.sup.3, R.sup.12, R.sup.x, R.sup.A1, R.sup.A2, n, X, and
Cy are as described herein.
[0098] In some embodiments of Formula (A), wherein R.sup.12 is
hydrogen and R.sup.13 is --OR.sup.1, the present disclosure
provides a compound of Formula (I):
##STR00017##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.1,
R.sup.2, R.sup.3, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.x, n,
X, and Cy are as described herein.
[0099] In certain embodiments, a provided compound is of Formula
(I-a):
##STR00018##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.1,
R.sup.2, R.sup.3, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.x, n,
X, and Cy are as described herein.
[0100] In certain embodiments, a provided compound is of Formula
(I-b):
##STR00019##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.1,
R.sup.2, R.sup.3, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.x, n,
X, and Cy are as described herein.
[0101] In certain embodiments, a provided compound is of Formula
(I-c):
##STR00020##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.1,
R.sup.2, R.sup.3, R.sup.x, n, X, and Cy are as described
herein.
[0102] In certain embodiments, a provided compound is of Formula
(I'):
##STR00021##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.1,
R.sup.2, R.sup.3, R.sup.x, n, X, and Cy are as described
herein.
[0103] In certain embodiments, a provided compound is of Formula
(I'-a):
##STR00022##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.1,
R.sup.2, R.sup.3, R.sup.x, n, X, and Cy are as described
herein.
[0104] In certain embodiments, a provided compound is of Formula
(I'-b):
##STR00023##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.1,
R.sup.2, R.sup.3, R.sup.x, n, X, and Cy are as described
herein.
[0105] In certain embodiments, a provided compound is of Formula
(A-8):
##STR00024##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.1,
R.sup.2, R.sup.3, R.sup.x, R.sup.12, R.sup.13, n, X, and Cy are as
described herein.
[0106] In certain embodiments, a provided compound is of Formula
(II):
##STR00025##
or a pharmaceutically acceptable salt thereof, wherein R.sup.2,
R.sup.3, R.sup.x, n, and Cy are as described herein.
[0107] In certain embodiments, a provided compound is of Formula
(II-a):
##STR00026##
or a pharmaceutically acceptable salt thereof, wherein R.sup.2,
R.sup.3, R.sup.x, n, and Cy are as described herein.
[0108] In certain embodiments, a provided compound is of Formula
(II-b):
##STR00027##
or a pharmaceutically acceptable salt thereof, wherein R.sup.2,
R.sup.3, R.sup.x, n, and Cy are as described herein.
[0109] In certain embodiments, a provided compound is of Formula
(A-9):
##STR00028##
or a pharmaceutically acceptable salt thereof, wherein R.sup.2,
R.sup.3, R.sup.x, R.sup.12, R.sup.13, n, and Cy are as described
herein.
[0110] In certain embodiments, a provided compound is of Formula
(III):
##STR00029##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.2,
R.sup.3, R.sup.x, n, and Cy are as described herein.
[0111] In certain embodiments, a provided compound is of Formula
(III-a):
##STR00030##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.2,
R.sup.3, R.sup.x, n, and Cy are as described herein.
[0112] In certain embodiments, a provided compound is of Formula
(III-b):
##STR00031##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.2,
R.sup.3, R.sup.x, n, and Cy are as described herein.
[0113] In certain embodiments, a provided compound is of Formula
(A-10):
##STR00032##
or a pharmaceutically acceptable salt thereof, wherein R, R.sup.2,
R.sup.3, R.sup.x, R.sup.12, R.sup.13, n, and Cy are as described
herein.
[0114] In certain embodiments, a provided compound is of Formula
(IV):
##STR00033##
or a pharmaceutically acceptable salt thereof, wherein R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.x, n, and Cy are as described
herein.
[0115] In certain embodiments, a provided compound is of Formula
(IV-a):
##STR00034##
or a pharmaceutically acceptable salt thereof, wherein R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.x, n, and Cy are as described
herein.
[0116] In certain embodiments, a provided compound is of Formula
(IV-b):
##STR00035##
or a pharmaceutically acceptable salt thereof, wherein R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.x, n, and Cy are as described
herein.
[0117] In certain embodiments, a provided compound is of Formula
(A-11):
##STR00036##
or a pharmaceutically acceptable salt thereof, wherein R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.x, R.sup.12, R.sup.13, n, and Cy
are as described herein.
[0118] In certain embodiments, a provided compound is of Formula
(A-12):
##STR00037##
or a pharmaceutically acceptable salt thereof, wherein R.sup.12 and
R.sup.13 are described herein, Cy is a 5,6-fused bicyclic
heteroaryl as described herein, and Cy is substituted with 0, 1, 2,
3, 4, or 5 R.sup.y groups, as valency permits.
[0119] In certain embodiments, a provided compound is of Formula
(VII):
##STR00038##
or a pharmaceutically acceptable salt thereof, wherein Cy is a
5,6-fused bicyclic heteroaryl as described herein, and Cy is
substituted with 0, 1, 2, 3, 4, or 5 R.sup.y groups, as valency
permits.
[0120] In certain embodiments, a provided compound is of Formula
(VII-a):
##STR00039##
or a pharmaceutically acceptable salt thereof, wherein Cy is a
5,6-fused bicyclic heteroaryl as described herein, and Cy is
substituted with 0, 1, 2, 3, 4, or 5 R.sup.y groups, as valency
permits.
[0121] In certain embodiments, a provided compound is of Formula
(VII-b):
##STR00040##
or a pharmaceutically acceptable salt thereof, wherein Cy is a
5,6-fused bicyclic heteroaryl as described herein, and Cy is
substituted with 0, 1, 2, 3, 4, or 5 R.sup.y groups, as valency
permits.
[0122] In certain embodiments, a provided compound is of Formula
(A-13):
##STR00041##
or a pharmaceutically acceptable salt thereof, wherein R.sup.12 and
R.sup.13 are described herein, Cy is a 5,6-fused bicyclic
heteroaryl as described herein, and Cy is substituted with 0, 1, 2,
3, 4, or 5 R.sup.y groups, as valency permits.
[0123] In certain embodiments, a provided compound is of Formula
(V):
##STR00042##
or a pharmaceutically acceptable salt thereof, wherein R.sup.2,
R.sup.3, R.sup.x, n, and Cy are as described herein.
[0124] In certain embodiments, a provided compound is of Formula
(V-a):
##STR00043##
or a pharmaceutically acceptable salt thereof, wherein R.sup.2,
R.sup.3, R.sup.x, n, and Cy are as described herein.
[0125] In certain embodiments, a provided compound is of Formula
(V-b):
##STR00044##
or a pharmaceutically acceptable salt thereof, wherein R.sup.2,
R.sup.3, R.sup.x, n, and Cy are as described herein.
[0126] In certain embodiments, a provided compound is of Formula
(A-14):
##STR00045##
or a pharmaceutically acceptable salt thereof, wherein R.sup.2,
R.sup.3, R.sup.x, R.sup.12, R.sup.13, n, and Cy are as described
herein.
[0127] In some embodiments, represents a single bond. In some
embodiments, represents a double bond.
[0128] As defined generally above, R.sup.1 is hydrogen, R.sup.z, or
--C(O)R.sup.z, wherein R.sup.z is optionally substituted C.sub.1-6
alkyl. In certain embodiments, R.sup.1 is hydrogen. In some
embodiments, R.sup.1 is optionally substituted C.sub.1-6 alkyl. In
certain embodiments, R.sup.1 is unsubstituted C.sub.1-6 alkyl. In
certain embodiments, R.sup.1 is methyl, ethyl, or propyl. In some
embodiments, R.sup.1 is --C(O)R.sup.z, wherein R.sup.z is
optionally substituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.1 is --C(O)R.sup.z, wherein R.sup.z is unsubstituted
C.sub.1-6 alkyl. In certain embodiments, R.sup.1 is acetyl.
[0129] As defined generally above, X is a bond, --O--, --N(R)--,
--CR.sup.4R.sup.5--, --O--CR.sup.4R.sup.5, N(R)--CR.sup.4R.sup.5--,
--O--CR.sup.4R.sup.5--O--, --N(R)--CR.sup.4R.sup.5--O,
--N(R)--CR.sup.4R.sup.5--N(R)--, --O--CR.sup.4R.sup.5--N(R)--,
--CR.sup.4R.sup.5--O--, --CR.sup.4R.sup.5--N(R)--,
--O--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--N(R)--CR.sup.4R.sup.5--CR.sup.6R.sup.7--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--O--,
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--N(R)--, or
--CR.sup.6R.sup.7--CR.sup.4R.sup.5--. In certain embodiments, X is
a bond, --O--, --N(R)-- or --CR.sup.4R.sup.5--, wherein R, R.sup.4,
and R.sup.5 are as described herein. In certain embodiments, X is a
bond. In certain embodiments, X is --O--. In some embodiments, X is
--N(R)--. In certain embodiments, X is --NH--. In certain
embodiments, X is --N(R)--, wherein R is optionally substituted
C.sub.1-6 aliphatic. In certain embodiments, X is --N(R)--, wherein
R is optionally substituted C.sub.1-6 alkyl. In certain
embodiments, X is --N(R)--, wherein R is unsubstituted C.sub.1-6
alkyl. In certain embodiments, X is --N(Me)-. In some embodiments,
X is --CR.sup.4R.sup.5--. In certain embodiments, X is
--CH.sub.2--. In certain embodiments, X is --CH.sub.2--O--.
[0130] As defined generally above, each R is independently hydrogen
or optionally substituted C.sub.1-6 aliphatic. In certain
embodiments, R is hydrogen. In some embodiments, R is optionally
substituted C.sub.1-6 aliphatic. In some embodiments, R is
substituted C.sub.1-6 aliphatic. In some embodiments, R is
unsubstituted C.sub.1-6 aliphatic. In some embodiments, R is
optionally substituted C.sub.1-6 alkyl. In some embodiments, R is
substituted C.sub.1-6 alkyl. In some embodiments, R is
unsubstituted C.sub.1-6 alkyl. In some embodiments, R is methyl,
ethyl, or propyl.
[0131] As defined generally above, R.sup.2 and R.sup.3 are each
independently selected from the group consisting of hydrogen, halo,
--CN, --NO.sub.2, optionally substituted aliphatic, optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, optionally substituted heteroaryl,
--OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A, --C(.dbd.O)R.sup.A,
--C(O)OR.sup.A, --C(O)SR.sup.A, C(O)N(R.sup.B).sub.2,
--C(O)N(R.sup.B)N(R.sup.B).sub.2, --OC(O)R.sup.A,
--OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring.
In certain embodiments, R.sup.2 and R.sup.3 are each independently
selected from the group consisting of hydrogen, halo, --CN,
--NO.sub.2, optionally substituted aliphatic, optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, optionally substituted heteroaryl,
--OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A, --C(.dbd.O)R.sup.A,
--C(O)OR.sup.A, --C(O)SR.sup.A, --C(O)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2, --SC(O)R.sup.A,
--C(.dbd.NR.sup.B)R.sup.A, --C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --SO.sub.2R.sup.A, --NR.sup.BSO.sub.2R.sup.A, and
--SO.sub.2N(R.sup.B).sub.2; or R.sup.2 and R.sup.3 are taken
together with their intervening atoms to form an optionally
substituted carbocyclic or heterocyclic ring.
[0132] In certain embodiments, R.sup.2 is hydrogen. In some
embodiments, R.sup.2 is not hydrogen. In some embodiments, R.sup.2
is halo. In certain embodiments, R.sup.2 is fluoro. In some
embodiments, R.sup.2 is optionally substituted aliphatic. In
certain embodiments, R.sup.2 is optionally substituted C.sub.1-6
aliphatic. In certain embodiments, R.sup.2 is optionally
substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.2 is
substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.2 is
--CF.sub.3, CHF.sub.2, or CH.sub.2F. In certain embodiments,
R.sup.2 is unsubstituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.2 is methyl, ethyl, or propyl. In some embodiments, R.sup.2
is --CN or --NO.sub.2. In some embodiments, R.sup.2 is optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, or optionally substituted heteroaryl. In
some embodiments, R.sup.2 is --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --OC(O)R.sup.A, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2, --SC(O)R.sup.A,
--C(.dbd.NR.sup.B)R.sup.A, --C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --SO.sub.2R.sup.A, --NR.sup.BSO.sub.2R.sup.A, or
--SO.sub.2N(R.sup.B).sub.2. In certain embodiments, R.sup.2 is
--N(R.sup.B).sub.2. In certain embodiments, R.sup.2 is --NHR.sup.B.
In certain embodiments, R.sup.2 is --NH.sub.2. In certain
embodiments, R.sup.2 is --OR.sup.A. In certain embodiments, R.sup.2
is --OH.
[0133] In certain embodiments, R.sup.3 is hydrogen. In some
embodiments, R.sup.3 is not hydrogen. In some embodiments, R.sup.3
is halo. In certain embodiments, R.sup.3 is fluoro. In some
embodiments, R.sup.3 is optionally substituted aliphatic. In
certain embodiments, R.sup.3 is optionally substituted C.sub.1-6
aliphatic. In certain embodiments, R.sup.3 is optionally
substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.3 is
substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.3 is
--CF.sub.3, CHF.sub.2, or CH.sub.2F. In certain embodiments,
R.sup.3 is unsubstituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.3 is methyl, ethyl, or propyl. In some embodiments, R.sup.3
is --CN or --NO.sub.2. In some embodiments, R.sup.3 is optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, or optionally substituted heteroaryl. In
some embodiments, R.sup.3 is --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --OC(O)R.sup.A, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2, --SC(O)R.sup.A,
--C(.dbd.NR.sup.B)R.sup.A, --C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --SO.sub.2R.sup.A, --NR.sup.BSO.sub.2R.sup.A, or
--SO.sub.2N(R.sup.B).sub.2. In certain embodiments, R.sup.3 is
--N(R.sup.B).sub.2. In certain embodiments, R.sup.3 is --NHR.sup.B.
In certain embodiments, R.sup.3 is --NH.sub.2. In certain
embodiments, R.sup.3 is --OR.sup.A. In certain embodiments, R.sup.3
is --OH.
[0134] In some embodiments, R.sup.2 and R.sup.3 are the same. In
some embodiments, R.sup.2 and R.sup.3 are different. In some
embodiments, R.sup.2 and R.sup.3 are each hydrogen. In some
embodiments, R.sup.2 is hydrogen and R.sup.3 is not hydrogen. In
some embodiments, R.sup.2 is hydrogen and R.sup.3 is optionally
substituted aliphatic. In some embodiments, R.sup.2 is hydrogen and
R.sup.3 is C.sub.1-6 alkyl. In some embodiments, R.sup.2 is
hydrogen and R.sup.3 is methyl. In some embodiments, R.sup.2 is
hydrogen and R.sup.3 is ethyl or propyl. In some embodiments,
R.sup.2 is hydrogen and R.sup.3 is --CF.sub.3, CHF.sub.2, or
CH.sub.2F. In some embodiments, R.sup.2 is hydrogen and R.sup.3 is
--N(R.sup.B).sub.2 or --OR.sup.A. In some embodiments, R.sup.2 is
hydrogen and R.sup.3 is --NH.sub.2. In some embodiments, R.sup.2 is
hydrogen and R.sup.3 is --OH. In some embodiments, R.sup.2 and
R.sup.3 are not hydrogen. In some embodiments, R.sup.2 and R.sup.3
are independently optionally substituted aliphatic. In some
embodiments, R.sup.2 and R.sup.3 are methyl. In some embodiments,
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic
ring.
[0135] As defined generally above, R.sup.4 and R.sup.5 are
independently selected from the group consisting of hydrogen, halo,
--CN, --NO.sub.2, optionally substituted aliphatic, optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, optionally substituted heteroaryl,
--OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A, --C(.dbd.O)R.sup.A,
--C(O)OR.sup.A, --C(O)SR.sup.A, C(O)N(R.sup.B).sub.2,
--C(O)N(R.sup.B)N(R.sup.B).sub.2, --OC(O)R.sup.A,
--OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.4 and R.sup.5 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring.
In certain embodiments, R.sup.4 and R.sup.5 are independently
selected from the group consisting of hydrogen, halo, --CN,
--NO.sub.2, optionally substituted aliphatic, optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, optionally substituted heteroaryl,
--OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A, --C(.dbd.O)R.sup.A,
--C(O)OR.sup.A, --C(O)SR.sup.A, --C(O)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2, --SC(O)R.sup.A,
--C(.dbd.NR.sup.B)R.sup.A, --C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --SO.sub.2R.sup.A, --NR.sup.BSO.sub.2R.sup.A, and
--SO.sub.2N(R.sup.B).sub.2; or R.sup.4 and R.sup.5 are taken
together with their intervening atoms to form an optionally
substituted carbocyclic or heterocyclic ring.
[0136] In certain embodiments, R.sup.4 is hydrogen. In some
embodiments, R.sup.4 is not hydrogen. In some embodiments, R.sup.4
is halo. In certain embodiments, R.sup.4 is fluoro. In some
embodiments, R.sup.4 is optionally substituted aliphatic. In
certain embodiments, R.sup.4 is optionally substituted C.sub.1-6
aliphatic. In certain embodiments, R.sup.4 is optionally
substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.4 is
substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.4 is
--CF.sub.3, CHF.sub.2, or CH.sub.2F. In certain embodiments,
R.sup.4 is unsubstituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.4 is methyl, ethyl, or propyl. In some embodiments, R.sup.4
is --CN or --NO.sub.2. In some embodiments, R.sup.4 is optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, or optionally substituted heteroaryl. In
some embodiments, R.sup.4 is --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --OC(O)R.sup.A, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2, --SC(O)R.sup.A,
--C(.dbd.NR.sup.B)R.sup.A, --C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --SO.sub.2R.sup.A, --NR.sup.BSO.sub.2R.sup.A, or
--SO.sub.2N(R.sup.B).sub.2. In certain embodiments, R.sup.4 is
--N(R.sup.B).sub.2. In certain embodiments, R.sup.4 is --NHR.sup.B.
In certain embodiments, R.sup.4 is --NH.sub.2. In certain
embodiments, R.sup.4 is --OR.sup.A. In certain embodiments, R.sup.4
is --OH.
[0137] In certain embodiments, R.sup.5 is hydrogen. In some
embodiments, R.sup.5 is not hydrogen. In some embodiments, R.sup.5
is halo. In certain embodiments, R.sup.5 is fluoro. In some
embodiments, R.sup.5 is optionally substituted aliphatic. In
certain embodiments, R.sup.5 is optionally substituted C.sub.1-6
aliphatic. In certain embodiments, R.sup.5 is optionally
substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.5 is
substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.5 is
--CF.sub.3, CHF.sub.2, or CH.sub.2F. In certain embodiments,
R.sup.5 is unsubstituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.5 is methyl, ethyl, or propyl. In some embodiments, R.sup.5
is --CN or --NO.sub.2. In some embodiments, R.sup.5 is optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, or optionally substituted heteroaryl. In
some embodiments, R.sup.5 is --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --OC(O)R.sup.A, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2, --SC(O)R.sup.A,
--C(.dbd.NR.sup.B)R.sup.A, --C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --SO.sub.2R.sup.A, --NR.sup.BSO.sub.2R.sup.A, or
--SO.sub.2N(R.sup.B).sub.2. In certain embodiments, R.sup.5 is
--N(R.sup.B).sub.2. In certain embodiments, R.sup.5 is --NHR.sup.B.
In certain embodiments, R.sup.5 is --NH.sub.2. In certain
embodiments, R.sup.5 is --OR.sup.A. In certain embodiments, R.sup.5
is --OH.
[0138] In some embodiments, R.sup.4 and R.sup.5 are the same. In
some embodiments, R.sup.4 and R.sup.5 are different. In some
embodiments, R.sup.4 and R.sup.5 are each hydrogen. In some
embodiments, R.sup.4 is hydrogen and R.sup.5 is not hydrogen. In
some embodiments, R.sup.4 is hydrogen and R.sup.5 is optionally
substituted aliphatic. In some embodiments, R.sup.4 is hydrogen and
R.sup.5 is C.sub.1-6 alkyl. In some embodiments, R.sup.4 is
hydrogen and R.sup.5 is methyl. In some embodiments, R.sup.4 is
hydrogen and R.sup.5 is ethyl or propyl. In certain embodiments,
R.sup.4 and hydrogen and R.sup.5 is --CF.sub.3, CHF.sub.2, or
CH.sub.2F. In some embodiments, R.sup.4 is hydrogen and R.sup.5 is
--N(R.sup.B).sub.2 or --OR.sup.A. In some embodiments, R.sup.4 is
hydrogen and R.sup.5 is --NH.sub.2. In some embodiments, R.sup.4 is
hydrogen and R.sup.5 is --OH. In some embodiments, R.sup.4 and
R.sup.5 are not hydrogen. In some embodiments, R.sup.4 and R.sup.5
are independently optionally substituted aliphatic. In some
embodiments, R.sup.4 and R.sup.5 are methyl. In some embodiments,
R.sup.4 and R.sup.5 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic
ring.
[0139] As defined generally above, R.sup.6 and R.sup.7 are
independently selected from the group consisting of hydrogen, halo,
--CN, --NO.sub.2, optionally substituted aliphatic, optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, optionally substituted heteroaryl,
--OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A, --C(.dbd.O)R.sup.A,
--C(O)OR.sup.A, --C(O)SR.sup.A, C(O)N(R.sup.B).sub.2,
--C(O)N(R.sup.B)N(R.sup.B).sub.2, --OC(O)R.sup.A,
--OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2; or
R.sup.6 and R.sup.7 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic ring.
In certain embodiments, R.sup.6 and R.sup.7 are independently
selected from the group consisting of hydrogen, halo, --CN,
--NO.sub.2, optionally substituted aliphatic, optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, optionally substituted heteroaryl,
--OR.sup.A, --N(R.sup.B).sub.2, --SR.sup.A, --C(.dbd.O)R.sup.A,
--C(O)OR.sup.A, --C(O)SR.sup.A, --C(O)N(R.sup.B).sub.2,
--OC(O)R.sup.A, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2, --SC(O)R.sup.A,
--C(.dbd.NR.sup.B)R.sup.A, --C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --SO.sub.2R.sup.A, --NR.sup.BSO.sub.2R.sup.A, and
--SO.sub.2N(R.sup.B).sub.2; or R.sup.6 and R.sup.7 are taken
together with their intervening atoms to form an optionally
substituted carbocyclic or heterocyclic ring.
[0140] In certain embodiments, R.sup.6 is hydrogen. In some
embodiments, R.sup.6 is not hydrogen. In some embodiments, R.sup.6
is halo. In certain embodiments, R.sup.6 is fluoro. In some
embodiments, R.sup.6 is optionally substituted aliphatic. In
certain embodiments, R.sup.6 is optionally substituted C.sub.1-6
aliphatic. In certain embodiments, R.sup.6 is optionally
substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.6 is
substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.6 is
--CF.sub.3, CHF.sub.2, or CH.sub.2F. In certain embodiments,
R.sup.6 is unsubstituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.6 is methyl, ethyl, or propyl. In some embodiments, R.sup.6
is --CN or --NO.sub.2. In some embodiments, R.sup.6 is optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, or optionally substituted heteroaryl. In
some embodiments, R.sup.6 is --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --OC(O)R.sup.A, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2, --SC(O)R.sup.A,
--C(.dbd.NR.sup.B)R.sup.A, --C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --SO.sub.2R.sup.A, --NR.sup.BSO.sub.2R.sup.A, or
--SO.sub.2N(R.sup.B).sub.2. In certain embodiments, R.sup.6 is
--N(R.sup.B).sub.2. In certain embodiments, R.sup.6 is --NHR.sup.B.
In certain embodiments, R.sup.6 is --NH.sub.2. In certain
embodiments, R.sup.6 is --OR.sup.A. In certain embodiments, R.sup.6
is --OH.
[0141] In certain embodiments, R.sup.7 is hydrogen. In some
embodiments, R.sup.7 is not hydrogen. In some embodiments, R.sup.7
is halo. In certain embodiments, R.sup.7 is fluoro. In some
embodiments, R.sup.7 is optionally substituted aliphatic. In
certain embodiments, R.sup.7 is optionally substituted C.sub.1-6
aliphatic. In certain embodiments, R.sup.7 is optionally
substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.7 is
substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.7 is
--CF.sub.3, CHF.sub.2, or CH.sub.2F. In certain embodiments,
R.sup.7 is unsubstituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.7 is methyl, ethyl, or propyl. In some embodiments, R.sup.7
is --CN or --NO.sub.2. In some embodiments, R.sup.7 is optionally
substituted carbocyclyl, optionally substituted phenyl, optionally
substituted heterocyclyl, or optionally substituted heteroaryl. In
some embodiments, R.sup.7 is --OR.sup.A, --N(R.sup.B).sub.2,
--SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A, --C(O)SR.sup.A,
--C(O)N(R.sup.B).sub.2, --OC(O)R.sup.A, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2, --SC(O)R.sup.A,
--C(.dbd.NR.sup.B)R.sup.A, --C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
--C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --SO.sub.2R.sup.A, --NR.sup.BSO.sub.2R.sup.A, or
--SO.sub.2N(R.sup.B).sub.2. In certain embodiments, R.sup.7 is
--N(R.sup.B).sub.2. In certain embodiments, R.sup.7 is --NHR.sup.B.
In certain embodiments, R.sup.7 is --NH.sub.2. In certain
embodiments, R.sup.7 is --OR.sup.A. In certain embodiments, R.sup.7
is --OH.
[0142] In some embodiments, R.sup.6 and R.sup.7 are the same. In
some embodiments, R.sup.6 and R.sup.7 are different. In some
embodiments, R.sup.6 and R.sup.7 are each hydrogen. In some
embodiments, R.sup.6 is hydrogen and R.sup.7 is not hydrogen. In
some embodiments, R.sup.6 is hydrogen and R.sup.7 is optionally
substituted aliphatic. In some embodiments, R.sup.6 is hydrogen and
R.sup.7 is C.sub.1-6 alkyl. In some embodiments, R.sup.6 is
hydrogen and R.sup.7 is methyl. In some embodiments, R.sup.6 is
hydrogen and R.sup.7 is ethyl or propyl. In certain embodiments,
R.sup.6 and hydrogen and R.sup.7 is --CF.sub.3, CHF.sub.2, or
CH.sub.2F. In some embodiments, R.sup.6 is hydrogen and R.sup.7 is
--N(R.sup.B).sub.2 or --OR.sup.A. In some embodiments, R.sup.6 is
hydrogen and R.sup.7 is --NH.sub.2. In some embodiments, R.sup.6 is
hydrogen and R.sup.7 is --OH. In some embodiments, R.sup.6 and
R.sup.7 are not hydrogen. In some embodiments, R.sup.6 and R.sup.7
are independently optionally substituted aliphatic. In some
embodiments, R.sup.6 and R.sup.7 are methyl. In some embodiments,
R.sup.6 and R.sup.7 are taken together with their intervening atoms
to form an optionally substituted carbocyclic or heterocyclic
ring.
[0143] As defined generally above, R.sup.8, R.sup.9, R.sup.10, and
R.sup.11 are each independently hydrogen, halo, or optionally
substituted aliphatic. In some embodiments, R.sup.8, R.sup.9,
R.sup.10, and R.sup.11 are hydrogen. In some embodiments, R.sup.9,
R.sup.10, and R.sup.11 are hydrogen, and R.sup.8 is optionally
substituted aliphatic. In some embodiments, R.sup.9, R.sup.10, and
R.sup.11 are hydrogen, and R.sup.8 is optionally substituted
C.sub.1-6 aliphatic. In some embodiments, R.sup.9, R.sup.10, and
R.sup.11 are hydrogen, and R.sup.8 is optionally substituted
C.sub.1-3 aliphatic. In some embodiments, R.sup.9, R.sup.10, and
R.sup.11 are hydrogen, and R.sup.8 is methyl. In some embodiments,
R.sup.8, R.sup.9, and R.sup.10 are hydrogen, and R.sup.11 is
optionally substituted aliphatic. In some embodiments, R.sup.8,
R.sup.9, and R.sup.10 are hydrogen, and R.sup.11 is optionally
substituted C.sub.1-6 aliphatic. In some embodiments, R.sup.8,
R.sup.9, and R.sup.10 are hydrogen, and R.sup.11 is optionally
substituted C.sub.1-3 aliphatic. In some embodiments, R.sup.8,
R.sup.9, and R.sup.10 are hydrogen, and R.sup.11 is methyl. In some
embodiments, R.sup.8 is hydrogen. In some embodiments, R.sup.8 is
halo. In certain embodiments, R.sup.8 is fluoro. In some
embodiments, R.sup.8 is optionally substituted C.sub.1-6 aliphatic.
In some embodiments, R.sup.8 is optionally substituted C.sub.1-3
alkyl. In certain embodiments, R.sup.8 is methyl. In some
embodiments, R.sup.9 is hydrogen. In some embodiments, R.sup.9 is
halo. In certain embodiments, R.sup.9 is fluoro. In some
embodiments, R.sup.9 is optionally substituted C.sub.1-6 aliphatic.
In some embodiments, R.sup.9 is optionally substituted C.sub.1-3
alkyl. In certain embodiments, R.sup.9 is methyl. In some
embodiments, R.sup.10 is hydrogen. In some embodiments, R.sup.10 is
halo. In certain embodiments, R.sup.10 is fluoro. In some
embodiments, R.sup.10 is optionally substituted C.sub.1-6
aliphatic. In some embodiments, R.sup.10 is optionally substituted
C.sub.1-3 alkyl. In certain embodiments, R.sup.10 is methyl. In
some embodiments, R.sup.11 is hydrogen. In some embodiments,
R.sup.11 is halo. In certain embodiments, R.sup.11 is fluoro. In
some embodiments, R.sup.11 is optionally substituted C.sub.1-6
aliphatic. In some embodiments, R.sup.11 is optionally substituted
C.sub.1-3 alkyl. In certain embodiments, R.sup.11 is methyl.
[0144] As generally defined above, R.sup.12 is hydrogen, halogen,
or optionally substituted C.sub.1-3alkyl. In certain embodiments,
R.sup.12 is hydrogen. In certain embodiments, R.sup.12 is
optionally substituted C.sub.1-3alkyl, e.g., optionally substituted
with halogen. In certain embodiments, R.sup.12 is optionally
substituted C.sub.1alkyl, e.g., methyl or trifluoromethyl. In
certain embodiments, R.sup.12 is optionally substituted C.sub.2
alkyl, e.g., ethyl. In certain embodiments, R.sup.12 is optionally
substituted C.sub.3 alkyl, e.g., propyl. In certain embodiments,
R.sup.12 is fluoro, provided that R.sup.13 is not --OR.sup.1. In
certain embodiments, R.sup.12 is chloro, provided that R.sup.13 is
not --OR.sup.1. In certain embodiments, R.sup.12 is bromo, provided
that R.sup.13 is not --OR.sup.1. In certain embodiments, R.sup.12
is iodo, provided that R.sup.13 is not --OR.sup.1.
[0145] As generally defined above, R.sup.13 is hydrogen, halogen,
optionally substituted C.sub.1-3alkyl, --NR.sup.A1R.sup.A2, or
--OR.sup.1. In certain embodiments, R.sup.13 is hydrogen. In
certain embodiments, R.sup.13 is optionally substituted
C.sub.1-3alkyl, e.g., optionally substituted with halogen. In
certain embodiments, R.sup.13 is optionally substituted
C.sub.1alkyl, e.g., methyl or trifluoromethyl. In certain
embodiments, R.sup.13 is optionally substituted C.sub.2 alkyl,
e.g., ethyl. In certain embodiments, R.sup.13 is optionally
substituted C.sub.3 alkyl, e.g., propyl. In certain embodiments,
R.sup.13 is fluoro. In certain embodiments, R.sup.13 is chloro. In
certain embodiments, R.sup.13 is bromo. In certain embodiments,
R.sup.13 is iodo. In certain embodiments, R.sup.13 is
--NR.sup.A1R.sup.A2.
[0146] As defined generally above, Cy is a monocyclic or bicyclic,
saturated, partially unsaturated, or aromatic ring having 0-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur, wherein Cy is substituted with 0, 1, 2, 3, or 4 R.sup.y
groups. In certain embodiments, Cy is unsubstituted. In certain
embodiments, Cy is substituted with one or two R.sup.y groups. In
certain embodiments, Cy is substituted with one R.sup.y group. In
certain embodiments, Cy is substituted with two R.sup.y groups. In
certain embodiments, Cy is substituted with three R.sup.y groups.
In certain embodiments, Cy is substituted with four R.sup.y
groups.
[0147] In certain embodiments, Cy is phenyl substituted with 0, 1,
2, 3, or 4 R.sup.y groups. In certain embodiments, Cy is phenyl
substituted with one or two R.sup.y groups. In certain embodiments,
Cy is unsubstituted phenyl. In certain embodiments, Cy is phenyl
substituted with one R.sup.y group. In certain embodiments, Cy is
phenyl substituted with two R.sup.y groups. In certain embodiments,
Cy is phenyl substituted with three R.sup.y groups. In certain
embodiments, Cy is phenyl substituted with four R.sup.y groups.
[0148] In certain embodiments, Cy is
##STR00046##
In certain embodiments, Cy is selected from the group consisting
of
##STR00047##
[0149] In certain embodiments, Cy is
##STR00048##
[0150] In certain embodiments, Cy is
##STR00049##
In certain embodiments, Cy is
##STR00050##
In certain embodiments, Cy is selected from the group consisting
of
##STR00051##
[0151] In certain embodiments, Cy is
##STR00052##
In certain embodiments, Cy is
##STR00053##
In certain embodiments, Cy is
##STR00054##
In certain embodiments, Cy is selected from the group consisting
of
##STR00055##
[0152] In certain embodiments, Cy is
##STR00056##
In certain embodiments, Cy is
##STR00057##
In certain embodiments, Cy is
##STR00058##
In certain embodiments, Cy is
##STR00059##
In certain embodiments, Cy is
##STR00060##
[0153] In certain embodiments, Cy is a 5- to 6-membered heteroaryl
having 1-3 heteroatoms independently selected from nitrogen,
oxygen, and sulfur, and is substituted with 0, 1, 2, 3, or 4
R.sup.y groups. In certain embodiments, Cy is an unsubstituted 5-
to 6-membered heteroaryl having 1-3 heteroatoms independently
selected from nitrogen, oxygen, and sulfur. In certain embodiments,
Cy is a 5- to 6-membered heteroaryl having 1-3 heteroatoms
independently selected from nitrogen, oxygen, and sulfur, and is
substituted with one or two R.sup.y groups. In certain embodiments,
Cy is a 5- to 6-membered heteroaryl having 1-3 heteroatoms
independently selected from nitrogen, oxygen, and sulfur, and is
substituted with one R.sup.y group. In certain embodiments, Cy is a
5-membered heteroaryl having 1-3 heteroatoms independently selected
from nitrogen, oxygen, and sulfur (e.g., furanyl, thienyl,
pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyrazolyl,
isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl), and is
substituted with 0, 1, 2, 3, or 4 R.sup.y groups. In certain
embodiments, Cy is a 6-membered heteroaryl having 1-3 nitrogens
(e.g., pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, triazinyl), and
is substituted with 0, 1, 2, 3, or 4 R.sup.y groups.
[0154] In certain embodiments, Cy is a bicyclic saturated,
partially unsaturated, or aromatic ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur, wherein
Cy is substituted with 0, 1, 2, 3, or 4 R.sup.y groups. In certain
embodiments, Cy is an 8- to 12-membered bicyclic saturated,
partially unsaturated, or aromatic ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur, wherein
Cy is substituted with 0, 1, 2, 3, or 4 R.sup.y groups. In certain
embodiments, Cy is an unsubstituted bicyclic saturated, partially
unsaturated, or aromatic ring having 0-4 heteroatoms independently
selected from nitrogen, oxygen, and sulfur. In certain embodiments,
Cy is a bicyclic saturated, partially unsaturated, or aromatic ring
having 0-4 heteroatoms independently selected from nitrogen,
oxygen, and sulfur, wherein Cy is substituted with one or two
R.sup.y groups. In certain embodiments, Cy is a bicyclic saturated,
partially unsaturated, or aromatic ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur, wherein
Cy is substituted with one R.sup.y group. In certain embodiments,
Cy is a bicyclic saturated, partially unsaturated, or aromatic ring
having 0-4 heteroatoms independently selected from nitrogen,
oxygen, and sulfur, wherein Cy is substituted with two R.sup.y
groups. In certain embodiments, Cy is a bicyclic saturated,
partially unsaturated, or aromatic ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur, wherein
Cy is substituted with three R.sup.y groups. In certain
embodiments, Cy is a bicyclic saturated, partially unsaturated, or
aromatic ring having 0-4 heteroatoms independently selected from
nitrogen, oxygen, and sulfur, wherein Cy is substituted with four
R.sup.y groups.
[0155] In certain embodiments, Cy is an 8- to 10-membered bicyclic
heteroaryl having 1-4 heteroatoms independently selected from
nitrogen, oxygen, and sulfur, wherein Cy is substituted with 0, 1,
2, 3, or 4 R.sup.y groups. In certain embodiments, Cy is a
9-membered bicyclic heteroaryl having 1-3 heteroatoms independently
selected from nitrogen, oxygen, and sulfur (e.g., indolyl,
isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl,
isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl,
benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl,
benzisothiazolyl, benzthiadiazolyl, indolizinyl), wherein Cy is
substituted with 0, 1, 2, 3, or 4 R.sup.y groups. In certain
embodiments, Cy is a 10-membered bicyclic heteroaryl having 1-3
heteroatoms independently selected from nitrogen, oxygen, and
sulfur (e.g., naphthyridinyl, quinolinyl, isoquinolinyl,
quinoxalinyl, quinazolinyl), wherein Cy is substituted with 0, 1,
2, 3, or 4 R.sup.y groups. In certain embodiments, Cy is selected
from the group consisting of quinoline, benzimidazole,
benzopyrazole, quinoxaline, tetrahydroquinoline,
tetrahydroisoquinoline, naphthalene, tetrahydronaphthalene,
2,3-dihydrobenzo[b][1,4]dioxine, isoindole,
2H-benzo[b][1,4]oxazin-3(4H)-one,
3,4-dihydro-2H-benzo[b][1,4]oxazine, and quinoxalin-2(1H)-one,
wherein Cy is substituted with 0, 1, 2, 3, or 4 R.sup.y groups.
[0156] In certain embodiments, Cy is a monocyclic or bicyclic
heterocyclic ring having 1-4 heteroatoms independently selected
from nitrogen, oxygen, and sulfur, wherein Cy is substituted with
0, 1, 2, 3, 4, or 5 R.sup.y groups, as valency permits. In certain
embodiments, Cy is a monocyclic heterocyclic ring, e.g., a
monocyclic 5-membered or 6-membered heterocyclic ring substituted
with 0, 1, 2, 3, 4, or 5 R.sup.y groups, as valency permits. In
certain embodiments, Cy is a bicyclic heterocyclic ring, e.g., a
6,6-bicyclic or 5,6-bicyclic heterocyclic ring substituted with 0,
1, 2, 3, 4, or 5 R.sup.y groups, as valency permits. In certain
embodiments, Cy is a 5,6-bicyclic heterocyclic ring wherein the
point of attachment is on the 6-membered ring. In certain
embodiments, wherein Cy is a 5,6-bicyclic heterocyclic ring, Cy is
an optionally substituted dihydroimidazo pyrimidinyl ring.
[0157] As defined generally above, each R.sup.y is independently
selected from the group consisting of halo, --CN, --NO.sub.2,
optionally substituted aliphatic, optionally substituted
carbocyclyl, optionally substituted aryl, optionally substituted
heterocyclyl, optionally substituted heteroaryl, --OR.sup.A,
--N(R.sup.B).sub.2, --SR.sup.A, --C(.dbd.O)R.sup.A, --C(O)OR.sup.A,
--C(O)SR.sup.A, C(O)N(R.sup.B).sub.2,
--C(O)N(R.sup.B)N(R.sup.B).sub.2, --OC(O)R.sup.A,
--OC(O)N(R.sup.B).sub.2, --NR.sup.BC(O)R.sup.A,
--NR.sup.BC(O)N(R.sup.B).sub.2,
--NR.sup.BC(O)N(R.sup.B)N(R.sup.B).sub.2, --NR.sup.BC(O)OR.sup.A,
--SC(O)R.sup.A, --C(.dbd.NR.sup.B)R.sup.A,
--C(.dbd.NNR.sup.B)R.sup.A, --C(.dbd.NOR.sup.A)R.sup.A,
--C(.dbd.NR.sup.B)N(R.sup.B).sub.2,
--NR.sup.BC(.dbd.NR.sup.B)R.sup.B, --C(.dbd.S)R.sup.A,
C(.dbd.S)N(R.sup.B).sub.2, --NR.sup.BC(.dbd.S)R.sup.A,
--S(O)R.sup.A, --OS(O).sub.2R.sup.A, --SO.sub.2R.sup.A,
--NR.sup.BSO.sub.2R.sup.A, and --SO.sub.2N(R.sup.B).sub.2, wherein
R.sup.A and R.sup.B are described herein; or an R.sup.y group may
be optionally taken together with R.sup.2 or R.sup.3 to form an
optionally substituted 5- to 6-membered carbocyclic or heterocyclic
ring fused to Cy.
[0158] In some embodiments, at least one R.sup.y is halo. In
certain embodiments, at least one R.sup.y is fluoro. In certain
embodiments, at least one R.sup.y is chloro. In some embodiments,
at least one R.sup.y is --CN. In some embodiments, at least one
R.sup.y is --OR.sup.A, wherein R.sup.A is optionally substituted
aliphatic. In some embodiments, at least one R.sup.y is --OR.sup.A,
wherein R.sup.A is unsubstituted C.sub.1-6 alkyl. In certain
embodiments, at least one R.sup.y is methoxy, ethoxy, or propoxy.
In certain embodiments, at least one R.sup.y is methoxy. In some
embodiments, at least one R.sup.y is --OR.sup.A, wherein R.sup.A is
substituted C.sub.1-6 alkyl. In certain embodiments, at least one
R.sup.y is --OCH.sub.2CH.sub.2N(CH.sub.3).sub.2. In some
embodiments, at least one R.sup.y is --N(R.sup.B).sub.2. In some
embodiments, at least one R.sup.y is --N(R.sup.B).sub.2, wherein
each R.sup.B is independently selected from hydrogen or C.sub.1-6
alkyl. In some embodiments, at least one R.sup.y is --NHR.sup.B. In
some embodiments, at least one R.sup.y is --N(C.sub.1-6
alkyl).sub.2, --NH(C.sub.1-6 alkyl), or --NH.sub.2. In certain
embodiments, at least one R.sup.y is --NH.sub.2. In certain
embodiments, at least one R.sup.y is --NHCH.sub.3. In certain
embodiments, at least one R.sup.y is --N(CH.sub.3).sub.2.
[0159] In some embodiments, at least one R.sup.y is optionally
substituted aliphatic. In certain embodiments, at least one R.sup.y
is substituted aliphatic. In certain embodiments, at least one
R.sup.y is unsubstituted aliphatic. In some embodiments, at least
one R.sup.y is optionally substituted C.sub.1-6 alkyl. In certain
embodiments, at least one R.sup.y is unsubstituted C.sub.1-6 alkyl.
In certain embodiments, at least one R.sup.y is substituted
C.sub.1-6 alkyl. In certain embodiments, at least one R.sup.y is
methyl, ethyl, or propyl. In certain embodiments, at least one
R.sup.y is methyl. In certain embodiments, at least one R.sup.y is
--CF.sub.3, CHF.sub.2, or CH.sub.2F. In certain embodiments, at
least one R.sup.y is C.sub.1-6 alkyl substituted with aryl,
heteroaryl, or heterocyclyl. In certain embodiments, at least one
R.sup.y is benzyl. In certain embodiments, at least one R.sup.y is
--(C.sub.1-6 alkyl)-heteroaryl. In certain embodiments, at least
one R.sup.y is --(C.sub.1-6 alkyl)-heterocyclyl. In certain
embodiments, at least one R.sup.y is --CH.sub.2-heteroaryl. In
certain embodiments, at least one R.sup.y is --CH.sub.2--
heterocyclyl.
[0160] In some embodiments, at least one R.sup.y is
--C(O)N(R.sup.B).sub.2. In certain embodiments, at least one
R.sup.y is --C(O)NHR.sup.B. In certain embodiments, at least one
R.sup.y is --C(O)NH.sub.2. In certain embodiments, at least one
R.sup.y is --C(O)N(R.sup.B).sub.2, wherein the R.sup.B groups are
taken together with their intervening atoms to form an optionally
substituted 5- to 6-membered heterocyclyl. In certain embodiments,
at least one R.sup.y is --C(O)N(R.sup.B).sub.2, wherein the R.sup.B
groups are taken together with their intervening atoms to form an
optionally substituted morpholinyl.
[0161] In some embodiments, at least one R.sup.y is
--SO.sub.2N(R.sup.B).sub.2. In certain embodiments, at least one
R.sup.y is --SO.sub.2NHR.sup.B. In certain embodiments, at least
one R.sup.y is --SO.sub.2NH.sub.2. In certain embodiments, at least
one R.sup.y is --SO.sub.2N(R.sup.B).sub.2, wherein neither R.sup.B
is hydrogen. In certain embodiments, at least one R.sup.y is
--SO.sub.2NH(C.sub.1-6 alkyl) or --SO.sub.2N(C.sub.1-6
alkyl).sub.2. In certain embodiments, at least one R.sup.y is
--SO.sub.2N(CH.sub.3).sub.2. In certain embodiments, at least one
R.sup.y is --SO.sub.2N(R.sup.B).sub.2, wherein the R.sup.B groups
are taken together with their intervening atoms to form an
optionally substituted 5- to 6-membered heterocyclyl. In certain
embodiments, at least one R.sup.y is --SO.sub.2-morpholinyl. In
certain embodiments, at least one R.sup.y is
--SO.sub.2-piperidinyl, --SO.sub.2-piperazinyl, or
--SO.sub.2-piperidinyl.
[0162] In some embodiments, at least one R.sup.y is
--SO.sub.2R.sup.A. In some embodiments, at least one R.sup.y is
--SO.sub.2R.sup.A, wherein R.sup.A is optionally substituted
aliphatic. In some embodiments, at least one R.sup.y is
--SO.sub.2(C.sub.1-6 alkyl). In some embodiments, at least one
R.sup.y is --SO.sub.2CH.sub.3. In some embodiments, at least one
R.sup.y is --C(O)R.sup.A. In some embodiments, at least one R.sup.y
is --C(O)R.sup.A, wherein R.sup.A is optionally substituted
aliphatic. In some embodiments, at least one R.sup.y is
--C(O)(C.sub.1-6 alkyl). In some embodiments, at least one R.sup.y
is --C(O)CH.sub.3.
[0163] In some embodiments, at least one R.sup.y is
--N(R.sup.B)C(O)R.sup.A. In certain embodiments, at least one
R.sup.y is --NHC(O)R.sup.A. In certain embodiments, at least one
R.sup.y is --NHC(O)(C.sub.1-6 alkyl). In certain embodiments, at
least one R.sup.y is --NHC(O)CH.sub.3.
[0164] In some embodiments, at least one R.sup.y is
--N(R.sup.B)SO.sub.2R.sup.A. In some embodiments, at least one
R.sup.y is --NHSO.sub.2R.sup.A. In some embodiments, at least one
R.sup.y is --N(C.sub.1-6 alkyl)SO.sub.2R.sup.A. In certain
embodiments, at least one R.sup.y is --NHSO.sub.2(C.sub.1-6 alkyl)
or --N(C.sub.1-6 alkyl)SO.sub.2(C.sub.1-6 alkyl). In certain
embodiments, at least one R.sup.y is --NHSO.sub.2CH.sub.3. In
certain embodiments, at least one R.sup.y is
--N(CH.sub.3)SO.sub.2CH.sub.3.
[0165] In some embodiments, at least one R.sup.y is optionally
substituted heterocyclyl, optionally substituted carbocyclyl,
optionally substituted aryl, or optionally substituted heteroaryl.
In certain embodiments, at least one R.sup.y is optionally
substituted 5- to 6-membered heterocyclyl having 1-2 heteroatoms
independently selected from nitrogen, oxygen, and sulfur. In
certain embodiments, at least one R.sup.y is optionally substituted
5-membered heterocyclyl having one heteroatom selected from
nitrogen, oxygen, and sulfur. In certain embodiments, at least one
R.sup.y is optionally substituted pyrrolidinyl. In certain
embodiments, at least one R.sup.y is pyrroldinyl,
hydroxypyrrolidinyl, or methylpyrrolidinyl. In certain embodiments,
at least one R.sup.y is optionally substituted 6-membered
heterocyclyl having 1-2 heteroatoms independently selected from
nitrogen, oxygen, and sulfur. In certain embodiments, at least one
R.sup.y is optionally substituted 6-membered heterocyclyl having
one heteroatom selected from nitrogen, oxygen, and sulfur. In
certain embodiments, at least one R.sup.y is optionally substituted
piperidinyl. In certain embodiments, at least one R.sup.y is
optionally substituted 6-membered heterocyclyl having two
heteroatoms each independently selected from nitrogen, oxygen, and
sulfur. In certain embodiments, at least one R.sup.y is optionally
substituted piperdinyl, optionally substituted piperazinyl, or
optionally substituted morpholinyl. In certain embodiments, at
least one R.sup.y is morpholinyl, tetrahydropyranyl, piperidinyl,
methylpiperidinyl, piperazinyl, methylpiperazinyl,
acetylpiperazinyl, methylsulfonylpiperazinyl, aziridinyl, or
methylaziridinyl. In some embodiments, at least one R.sup.y is
optionally substituted 5- to 6-membered heteroaryl having 1-3
heteroatoms independently selected from nitrogen, oxygen, and
sulfur. In certain embodiments, at least one R.sup.y is optionally
substituted 5-membered heteroaryl having 1-3 heteroatoms each
independently selected from nitrogen, oxygen, and sulfur. In
certain embodiments, at least one R.sup.y is optionally substituted
5-membered heteroaryl having one heteroatom selected from nitrogen,
oxygen, and sulfur. In certain embodiments, at least one R.sup.y is
optionally substituted 5-membered heteroaryl having two heteroatoms
each independently selected from nitrogen, oxygen, and sulfur. In
certain embodiments, at least one R.sup.y is optionally substituted
6-membered heteroaryl having 1-3 nitrogens. In certain embodiments,
at least one R.sup.y is optionally substituted pyrazolyl. In
certain embodiments, at least one R.sup.y is optionally substituted
imidazolyl. In certain embodiments, at least one R.sup.y is
optionally substituted pyridyl. In certain embodiments, at least
one R.sup.y is optionally substituted pyrimidyl. In certain
embodiments, at least one R.sup.y is pyrazolyl, methylpyrazolyl,
imidazolyl, or methylimidazolyl.
[0166] In some embodiments, R.sup.y is --OR.sup.A. In some
embodiments, R.sup.y is --OR.sup.A, wherein R.sup.A is optionally
substituted heterocyclyl. In some embodiments, R.sup.y is
--OR.sup.A, wherein R.sup.A is optionally substituted heteroaryl.
In some embodiments, R.sup.y is --OR.sup.A, wherein R.sup.A is
optionally substituted cycloalkyl. In some embodiments, R.sup.y is
--N(R.sup.B).sub.2. In some embodiments, R.sup.y is --NHR.sup.B. In
some embodiments, R.sup.y is --NHR.sup.B, wherein R.sup.B is
optionally substituted heterocyclyl. In some embodiments, R.sup.y
is --NHR.sup.B, wherein R.sup.B is optionally substituted
heteroaryl. In some embodiments, R.sup.y is --NHR.sup.B, wherein
R.sup.B is optionally substituted cycloalkyl. In some embodiments,
R.sup.y is --N(R.sup.B).sub.2, wherein one R.sup.B is optionally
substituted heterocyclyl, and the other R.sup.B is C.sub.1-4 alkyl.
In some embodiments, R.sup.y is --N(R.sup.B).sub.2, wherein one
R.sup.B is optionally substituted heteroaryl, and the other R.sup.B
is C.sub.1-4 alkyl. In some embodiments, R.sup.y is
--N(R.sup.B).sub.2, wherein one R.sup.B is optionally substituted
cycloalkyl, and the other R.sup.B is C.sub.1-4 alkyl.
[0167] In some embodiments, an R.sup.y group is taken together with
R.sup.2 or R.sup.3 and their intervening atoms to form a 5- to
6-membered carbocyclic or heterocyclic ring fused to Cy.
[0168] In certain embodiments, Cy is selected from the group
consisting of:
##STR00061## ##STR00062## ##STR00063## ##STR00064##
##STR00065##
[0169] In certain embodiments, Cy is selected from the group
consisting of:
##STR00066## ##STR00067## ##STR00068## ##STR00069## ##STR00070##
##STR00071## ##STR00072## ##STR00073## ##STR00074## ##STR00075##
##STR00076## ##STR00077## ##STR00078## ##STR00079## ##STR00080##
##STR00081## ##STR00082## ##STR00083## ##STR00084## ##STR00085##
##STR00086## ##STR00087## ##STR00088## ##STR00089##
[0170] In certain embodiments, Cy is selected from the group
consisting of:
##STR00090## ##STR00091## ##STR00092## ##STR00093## ##STR00094##
##STR00095## ##STR00096## ##STR00097## ##STR00098## ##STR00099##
##STR00100## ##STR00101## ##STR00102## ##STR00103## ##STR00104##
##STR00105## ##STR00106## ##STR00107## ##STR00108## ##STR00109##
##STR00110## ##STR00111## ##STR00112## ##STR00113## ##STR00114##
##STR00115## ##STR00116## ##STR00117## ##STR00118## ##STR00119##
##STR00120## ##STR00121## ##STR00122##
[0171] In certain embodiments, Cy is selected from the group
consisting of:
##STR00123## ##STR00124## ##STR00125## ##STR00126## ##STR00127##
##STR00128##
[0172] In some embodiments, Cy is selected from the group
consisting of:
##STR00129##
[0173] In some embodiments, Cy is selected from the group
consisting of:
##STR00130## ##STR00131## ##STR00132## ##STR00133## ##STR00134##
##STR00135## ##STR00136##
[0174] In some embodiments, Cy is selected from the group
consisting of:
##STR00137## ##STR00138##
[0175] In certain embodiments, Cy is a 5,6-fused bicyclic
heteroaryl ring system such as one of the following:
##STR00139## ##STR00140## ##STR00141## ##STR00142## ##STR00143##
##STR00144##
wherein the point of attachment can be any carbon or nitrogen atom,
as valency permits, and the ring may be substituted with 0, 1, 2,
3, 4, or 5 R.sup.y groups, as valency permits.
[0176] In some embodiments, Cy is selected from the group
consisting of:
##STR00145## ##STR00146## ##STR00147## ##STR00148## ##STR00149##
##STR00150## ##STR00151## ##STR00152## ##STR00153##
##STR00154##
each of which may be optionally substituted with 1, 2, 3, 4, or 5
R.sup.y groups as valency permits.
[0177] In certain embodiments, Cy is selected from the group
consisting of:
##STR00155##
[0178] As defined generally above, each R.sup.x is independently
selected from the group consisting of halo, --CN, optionally
substituted aliphatic, --OR', and --N(R'').sub.2. In certain
embodiments, at least one R.sup.x is halo. In certain embodiments,
at least one R.sup.x is fluoro. In certain embodiments, at least
one R.sup.x is --CN. In certain embodiments, at least one R.sup.x
is optionally substituted aliphatic. In certain embodiments, at
least one R.sup.x is optionally substituted C.sub.1-6 alkyl. In
certain embodiments, at least one R.sup.x is methyl. In certain
embodiments, at least one R.sup.x is --CF.sub.3. In certain
embodiments, at least one R.sup.x is --OR' or --N(R'').sub.2. In
certain embodiments, R.sup.x is not --OR' or --N(R'').sub.2. In
certain embodiments, at least one R.sup.x is --OCH.sub.3. In
certain embodiments, R.sup.x is not --OCH.sub.3.
[0179] As is generally understood from the above disclosure, the
ring system:
##STR00156##
is a fused bicyclic ring system, i.e., a phenyl ring fused to a
nitrogen containing ring, wherein the point of attachment to the
parent moiety is on the nitrogen, and wherein the fused bicyclic
system is optionally substituted with (R.sup.X)n, wherein n and
R.sup.x are as defined herein. As is generally understood, each of
the atoms of the phenyl ring and the nitrogen-containing ring can
be independently optionally substituted with R.sup.x, as valency
permits.
[0180] In certain embodiments, the fused bicyclic ring system is
optionally substituted with one or more R.sup.x, with the proviso
that when the nitrogen-containing ring is substituted at one of the
positions alpha to the nitrogen, R.sup.x is not-C(.dbd.O)R.sup.x1,
wherein R.sup.x1 is optionally substituted aliphatic, optionally
substituted carbocyclyl, optionally substituted aryl, optionally
substituted heterocyclyl, optionally substituted heteroaryl,
--OR.sup.A, --N(R.sup.B).sub.2, or --SR.sup.A, wherein R.sup.A and
R.sup.B are as generally defined herein. In certain embodiments,
the nitrogen-containing ring does not comprise an R.sup.x
substituent. In certain embodiments, only atoms of the phenyl ring
are optionally substituted with one or more R.sup.x.
[0181] In certain embodiments, the nitrogen-containing ring is
optionally substituted and the fused bicyclic ring system is of the
formula:
##STR00157##
wherein R.sup.x is as defined above, and n1 is 0, 1, 2, 3, or
4.
[0182] Thus, one of ordinary skill in the art will appreciate that
an R.sup.x group can be attached anywhere on the
tetrahydroisoquinoline or dihydroisoquinoline ring. In certain
embodiments, an R.sup.x group is attached to the benzene portion of
the tetrahydroisoquinoline or dihydroisoquinoline ring. In certain
embodiments, an R.sup.x group is attached to the tetrahydropyridine
or dihydropyridine portion of the tetrahydroisoquinoline or
dihydroisoquinoline ring. In certain embodiments, R.sup.x groups
are attached to both the benzene portion and the tetrahydropyridine
(or dihydropyridine) portion of the tetrahydroisoquinoline (or
dihydroisoquinoline) ring. See, for example, the structures shown
below:
##STR00158##
[0183] For example, in certain embodiments, a provided compound is
of Formula (VI):
##STR00159##
or a pharmaceutically acceptable salt thereof.
[0184] As generally defined above, R.sup.A1 and R.sup.A2 are
independently hydrogen, substituted or unsubstituted C.sub.1-3
alkyl, substituted or unsubstituted acyl, or a nitrogen protecting
group. In some embodiments, R.sup.A1 is hydrogen. In some
embodiments, R.sup.A1 is substituted or unsubstituted C.sub.1-3
alkyl. In some embodiments, R.sup.A1 is unsubstituted C.sub.1-3
alkyl. In some embodiments, R.sup.A1 is methyl, ethyl, n-propyl, or
isopropyl. In some embodiments, R.sup.A1 is substituted C.sub.1-3
alkyl. In some embodiments, R.sup.A1 is --CF.sub.3, --CHF.sub.2,
--CH.sub.2F, or --CH(CF.sub.3)CH.sub.3. In some embodiments,
R.sup.A1 is substituted or unsubstituted acyl. In some embodiments,
R.sup.A1 is acetyl. In some embodiments, R.sup.A1 is a nitrogen
protecting group. In some embodiments, R.sup.A1 is
CH.sub.3SO.sub.2--. In some embodiments, R.sup.A2 is hydrogen. In
some embodiments, R.sup.A2 is substituted or unsubstituted
C.sub.1-3 alkyl. In some embodiments, R.sup.A2 is unsubstituted
C.sub.1-3 alkyl. In some embodiments, R.sup.A2 is methyl, ethyl,
n-propyl, or isopropyl. In some embodiments, R.sup.A2 is
substituted C.sub.1-3 alkyl. In some embodiments, R.sup.A2 is
--CF.sub.3, --CHF.sub.2, --CH.sub.2F, or --CH(CF.sub.3)CH.sub.3. In
some embodiments, R.sup.A2 is substituted or unsubstituted acyl. In
some embodiments, R.sup.A2 is acetyl. In some embodiments, R.sup.A2
is a nitrogen protecting group. In some embodiments, R.sup.A2 is
CH.sub.3SO.sub.2--. In some embodiments, R.sup.A1 is hydrogen, and
R.sup.A2 is hydrogen. In some embodiments, R.sup.A1 is hydrogen,
and R.sup.A2 is substituted or unsubstituted C.sub.1-3 alkyl. In
some embodiments, R.sup.A1 is hydrogen, and R.sup.A2 is methyl,
ethyl, n-propyl, or isopropyl. In some embodiments, R.sup.A1 is
hydrogen, and R.sup.A2 is --CF.sub.3, --CHF.sub.2, --CH.sub.2F, or
--CH(CF.sub.3)CH.sub.3. In some embodiments, R.sup.A1 is hydrogen,
and R.sup.A2 is substituted or unsubstituted acyl. In some
embodiments, R.sup.A1 is hydrogen, and R.sup.A2 is acetyl. In some
embodiments, R.sup.A1 is hydrogen, and R.sup.A2 is a nitrogen
protecting group. In some embodiments, R.sup.A1 is hydrogen and
R.sup.A2 is CH.sub.3SO.sub.2--. In some embodiments, R.sup.A1 is
substituted or unsubstituted C.sub.1-3 alkyl, and R.sup.A2 is
substituted or unsubstituted C.sub.1-3 alkyl. In some embodiments,
R.sup.A1 is substituted or unsubstituted C.sub.1-3 alkyl, and
R.sup.A2 is methyl. In some embodiments, R.sup.A1 is substituted or
unsubstituted C.sub.1-3 alkyl, and R.sup.A2 is ethyl. In some
embodiments, R.sup.A1 is substituted or unsubstituted C.sub.1-3
alkyl, and R.sup.A2 is n-propyl. In some embodiments, R.sup.A1 is
substituted or unsubstituted C.sub.1-3 alkyl, and R.sup.A2 is
isopropyl. In some embodiments, R.sup.A1 is substituted or
unsubstituted C.sub.1-3 alkyl, and R.sup.A2 is substituted or
unsubstituted acyl. In some embodiments, R.sup.A1 is substituted or
unsubstituted C.sub.1-3 alkyl, and R.sup.A2 is a nitrogen
protecting group. In some embodiments, R.sup.A1 is methyl, and
R.sup.A2 is substituted or unsubstituted C.sub.1-3 alkyl. In some
embodiments, R.sup.A1 is methyl, and R.sup.A2 is methyl. In some
embodiments, R.sup.A1 is methyl, and R.sup.A2 is ethyl. In some
embodiments, R.sup.A1 is methyl, and R.sup.A2 is n-propyl. In some
embodiments, R.sup.A1 is methyl, and R.sup.A2 is isopropyl. In some
embodiments, R.sup.A1 is methyl, and R.sup.A2 is substituted or
unsubstituted acyl. In some embodiments, R.sup.A1 is methyl, and
R.sup.A2 is a nitrogen protecting group. In some embodiments,
R.sup.A1 is ethyl, and R.sup.A2 is substituted or unsubstituted
C.sub.1-3 alkyl. In some embodiments, R.sup.A1 is ethyl, and
R.sup.A2 is methyl. In some embodiments, R.sup.A1 is ethyl, and
R.sup.A2 is ethyl. In some embodiments, R.sup.A1 is ethyl, and
R.sup.A2 is n-propyl. In some embodiments, R.sup.A1 is ethyl, and
R.sup.A2 is isopropyl. In some embodiments, R.sup.A1 is ethyl, and
R.sup.A2 is substituted or unsubstituted acyl. In some embodiments,
R.sup.A1 is ethyl, and R.sup.A2 is a nitrogen protecting group. In
some embodiments, R.sup.A1 is n-propyl, and R.sup.A2 is substituted
or unsubstituted C.sub.1-3 alkyl. In some embodiments, R.sup.A1 is
n-propyl, and R.sup.A2 is methyl. In some embodiments, R.sup.A1 is
n-propyl, and R.sup.A2 is ethyl. In some embodiments, R.sup.A1 is
n-propyl, and R.sup.A2 is n-propyl. In some embodiments, R.sup.A1
is n-propyl and R.sup.A2 is isopropyl. In some embodiments,
R.sup.A1 is n-propyl, and R.sup.A2 is substituted or unsubstituted
acyl. In some embodiments, R.sup.A1 is n-propyl and R.sup.A2 is a
nitrogen protecting group. In some embodiments, R.sup.A1 is
isopropyl and R.sup.A2 is substituted or unsubstituted C.sub.1-3
alkyl. In some embodiments, R.sup.A1 is isopropyl and R.sup.A2 is
methyl. In some embodiments, R.sup.A1 is isopropyl and R.sup.A2 is
ethyl. In some embodiments, R.sup.A1 is isopropyl, and R.sup.A2 is
n-propyl. In some embodiments, R.sup.A1 is isopropyl, and R.sup.A2
is isopropyl. In some embodiments, R.sup.A1 is isopropyl, and
R.sup.A2 is substituted or unsubstituted acyl. In some embodiments,
R.sup.A1 is isopropyl, and R.sup.A2 is a nitrogen protecting group.
In some embodiments, R.sup.A1 is substituted or unsubstituted acyl,
and R.sup.A2 is substituted or unsubstituted C.sub.1-3 alkyl. In
some embodiments, R.sup.A1 is a nitrogen protecting group, and
R.sup.A2 is substituted or unsubstituted C.sub.1-3 alkyl. In some
embodiments, R.sup.A1 is a nitrogen protecting group and R.sup.A2
is methyl. In some embodiments, R.sup.A1 is a nitrogen protecting
group, and R.sup.A2 is ethyl. In some embodiments, R.sup.A1 is a
nitrogen protecting group, and R.sup.A2 is n-propyl. In some
embodiments, R.sup.A1 is a nitrogen protecting group, and R.sup.A2
is isopropyl. In some embodiments, R.sup.A1 is a nitrogen
protecting group, and R.sup.A2 is a nitrogen protecting group.
[0185] As generally defined above, R.sup.A1 and R.sup.A2 can be
taken together with the intervening nitrogen atom to form a
substituted or unsubstituted 3-6 membered heterocyclic ring. In
certain embodiments, R.sup.A1 and R.sup.A2 can be taken together
with the intervening nitrogen atom to form a substituted or
unsubstituted azetidine. In certain embodiments, R.sup.A1 and
R.sup.A2 can be taken together with the intervening nitrogen atom
to form a substituted or unsubstituted pyrrolidine. In certain
embodiments, R.sup.A1 and R.sup.A2 can be taken together with the
intervening nitrogen atom to form a substituted or unsubstituted
piperidine. In certain embodiments, R.sup.A1 and R.sup.A2 can be
taken together with the intervening nitrogen atom to form a
substituted or unsubstituted piperazine. In certain embodiments,
R.sup.A1 and R.sup.A2 can be taken together with the intervening
nitrogen atom to form a substituted or unsubstituted
morpholine.
[0186] As defined generally above, n is 0, 1, 2, 3, 4, 5, 6, 7, 8,
9, or 10. In certain embodiments, n is 0. In certain embodiments, n
is 1. In certain embodiments, n is 2.
[0187] In some embodiments, e.g. for Formula (A), Formula (I), or
any subgenera thereof, the provided compound is of a free base
form. In some embodiments, e.g. for Formula (A), Formula (I), or
any subgenera thereof, the provided compound is in the form of a
pharmaceutically acceptable salt as generally defined herein. In
some embodiments, the provided compound is a hydrochloride salt
thereof. In some embodiments, the provided compound is a tartrate
salt thereof. In some embodiments, the provided compound is a
monotartrate salt thereof. In some embodiments, the provided
compound is a bitartrate salt thereof.
[0188] In certain embodiments, a provided compound is a compound
listed in Table 1A, or a pharmaceutically acceptable salt
thereof.
TABLE-US-00001 TABLE 1A Exemplary Compounds Cmpd LC-MS m/z No
Structure Exact mass (M + H) 1 ##STR00160## 340.1787 341.2 2
##STR00161## 338.1994 339.2 3 ##STR00162## 352.2151 353.2 4
##STR00163## 485.2678 486.2 5 ##STR00164## 354.1943 355.1 6
##STR00165## 391.1896 392.2 7 ##STR00166## 358.1693 359.1 8
##STR00167## 365.1739 366.1 9 ##STR00168## 354.1943 355.2 10
##STR00169## 358.1693 359.2 11 ##STR00170## 365.1739 366.2 12
##STR00171## 370.1893 371.2 13 ##STR00172## 383.1845 384.2 14
##STR00173## 418.1562 419.2 15 ##STR00174## 354.1943 355.1 16
##STR00175## 358.1693 359.1 17 ##STR00176## 365.1739 366.1 18
##STR00177## 339.1947 340.1 19 ##STR00178## 354.1943 355.2 20
##STR00179## 370.1893 371.1 21 ##STR00180## 397.2002 398.2 22
##STR00181## 370.1893 371.1 23 ##STR00182## 394.2005 395.1 24
##STR00183## 346.2256 347.2 25 ##STR00184## 368.21 369.2 26
##STR00185## 354.1943 355.2 27 ##STR00186## 383.2209 384.2 28
##STR00187## 433.1671 434.1 29 ##STR00188## 383.1845 384.2 30
##STR00189## 394.2005 395.1 31 ##STR00190## 397.2002 398.1 32
##STR00191## 340.1787 341.2 33 ##STR00192## 340.1787 341.2 34
##STR00193## 383.1845 384.1 35 ##STR00194## 383.2209 384.2 36
##STR00195## 397.2002 398.1 37 ##STR00196## 418.1562 419.1 38
##STR00197## 383.2209 384.2 39 ##STR00198## 405.2052 406.2 40
##STR00199## 422.2569 423.2 41 ##STR00200## 348.2049 349.2 42
##STR00201## 433.1671 434.1 43 ##STR00202## 419.2209 420.2 44
##STR00203## 391.1896 392.1 45 ##STR00204## 391.1896 392.1 46
##STR00205## 394.2005 395.2 47 ##STR00206## 418.1562 419.1 48
##STR00207## 476.2424 477.2 49 ##STR00208## 425.2315 426.2 50
##STR00209## 406.2005 407.2 51 ##STR00210## 325.179 326.1 52
##STR00211## 419.1515 420.1 53 ##STR00212## 433.1671 434.1 54
##STR00213## 404.2212 405.2 55 ##STR00214## 324.1838 325.1 56
##STR00215## 330.2307 331.2 57 ##STR00216## 324.1838 325.1 58
##STR00217## 330.2307 331.2 59 ##STR00218## 339.1947 340.2 60
##STR00219## 375.1947 376.1 61 ##STR00220## 339.1947 340.1 62
##STR00221## 339.1947 340.1 63 ##STR00222## 339.1947 340.1 64
##STR00223## 394.2256 395.1 65 ##STR00224## 419.1515 420.1 66
##STR00225## 390.1943 391.2 67 ##STR00226## 406.2005 407.2 68
##STR00227## 406.2005 407.2 69 ##STR00228## 393.2165 394.2 70
##STR00229## 463.2583 464.2 71 ##STR00230## 477.274 478.3 72
##STR00231## 405.2052 406.2 73 ##STR00232## 405.2052 406.1 74
##STR00233## 405.2052 406.1 75 ##STR00234## 439.2471 440.2 76
##STR00235## 405.2052 406.2 77 ##STR00236## 425.2315 426.2 78
##STR00237## 425.2315 426.2 79 ##STR00238## 424.2474 425.2 80
##STR00239## 425.1951 426.2 81 ##STR00240## 411.2158 412.1 82
##STR00241## 394.2005 395.2 83 ##STR00242## 453.2264 454.2 84
##STR00243## 438.2631 439.3 85 ##STR00244## 438.2631 439.3 86
##STR00245## 469.2577 470.2 87 ##STR00246## 469.2577 470.2 88
##STR00247## 427.2471 428.1 89 ##STR00248## 427.2471 428.2 90
##STR00249## 466.258 467.2 91 ##STR00250## 395.2209 396.1 92
##STR00251## 392.1848 393.2 93 ##STR00252## 439.2471 440.2 94
##STR00253## 490.258 491.2 95 ##STR00254## 449.2427 450.2 96
##STR00255## 463.2583 464.2 97 ##STR00256## 421.2002 422.2 98
##STR00257## 421.2002 422.2 99 ##STR00258## 409.2365 410.2 100
##STR00259## 398.1842 399.1 101 ##STR00260## 437.2315 438.2 102
##STR00261## 473.1984 474.2 103 ##STR00262## 423.2522 424.3 104
##STR00263## 434.2318 435.2 105 ##STR00264## 434.2318 435.2 106
##STR00265## 502.225 503.2 107 ##STR00266## 488.2093 489.2 108
##STR00267## 502.225 503.2 109 ##STR00268## 447.1828 448.1 110
##STR00269## 473.1984 474.1 111 ##STR00270## 489.1934 490.1 112
##STR00271## 397.2002 398.2 113 ##STR00272## 411.2158 412.2 114
##STR00273## 432.1719 433.1 115 ##STR00274## 406.2005 407.1 116
##STR00275## 409.2365 410.2 117 ##STR00276## 423.2522 424.2 118
##STR00277## 394.2005 395.2 119 ##STR00278## 395.2209 396.1 120
##STR00279## 409.2365 410.2 121 ##STR00280## 440.2424 441.2 122
##STR00281## 530.2199 531.2
123 ##STR00282## 433.1671 434.1 124 ##STR00283## 411.1794 412.2 125
##STR00284## 408.1798 409.2 126 ##STR00285## 433.1671 434.1 127
##STR00286## 384.2161 385.1 128 ##STR00287## 398.1954 399.1 129
##STR00288## 408.1798 409.1 130 ##STR00289## 437.2678 438.3 131
##STR00290## 419.1515 420.1 132 ##STR00291## 395.2209 396.2 133
##STR00292## 384.2161 385.2 134 ##STR00293## 437.2678 438.3 135
##STR00294## 409.2365 410.3 136 ##STR00295## 423.2522 424.2 137
##STR00296## 406.2005 407.1 138 ##STR00297## 420.2161 421.1 139
##STR00298## 434.1624 435.1 140 ##STR00299## 434.1624 435.1 141
##STR00300## 455.242 456.2 142 ##STR00301## 447.1828 448.2 143
##STR00302## 384.2161 385.2 144 ##STR00303## 427.1707 428.2 145
##STR00304## 356.1848 357.1 146 ##STR00305## 407.2209 408.2 147
##STR00306## 398.1954 399.2 148 ##STR00307## 398.1954 399.2 149
##STR00308## 489.1934 490.1 150 ##STR00309## 395.2209 396.2 151
##STR00310## 409.2365 410.2 152 ##STR00311## 355.1896 356.2 153
##STR00312## 420.2161 421.2 154 ##STR00313## 392.1848 393.2 155
##STR00314## 447.1828 448.1 156 ##STR00315## 420.2161 421.2 157
##STR00316## 447.1828 448.1 158 ##STR00317## 408.2161 409.2 159
##STR00318## 394.2005 395.2 160 ##STR00319## 394.2005 395.1 161
##STR00320## 454.2216 455.2 162 ##STR00321## 398.1954 399.2 163
##STR00322## 407.2209 408.2 164 ##STR00323## 407.2209 408.2 165
##STR00324## 340.1787 341.1 166 ##STR00325## 348.2049 349.1 167
##STR00326## 377.2315 378.2 168 ##STR00327## 407.1957 408.2 169
##STR00328## 462.1937 463.1 170 ##STR00329## 433.1671 434.1 171
##STR00330## 334.1893 335.1 172 ##STR00331## 440.2424 441.2 173
##STR00332## 354.1943 355.1 174 ##STR00333## 368.1736 369.2 175
##STR00334## 378.2056 379.1 176 ##STR00335## 390.2631 391.1 177
##STR00336## 351.1947 352.1 178 ##STR00337## 364.2151 365.1 179
##STR00338## 364.2151 365.1 180 ##STR00339## 364.2151 365.1 181
##STR00340## 392.1848 393.2 182 ##STR00341## 363.1947 364.1 183
##STR00342## 368.1736 369.2 184 ##STR00343## 357.2416 358.1 185
##STR00344## 365.2103 366.1 186 ##STR00345## 406.2005 407.1 187
##STR00346## 420.2161 421.1 188 ##STR00347## 496.2686 497.3 189
##STR00348## 477.2376 478.3 190 ##STR00349## 438.2631 439.3 191
##STR00350## 504.2406 505.1 192 ##STR00351## 378.2056 379.1 193
##STR00352## 395.1667 396.2
[0189] In certain embodiments, a provided compound is a compound
listed in Table IB, or a pharmaceutically acceptable salt
thereof.
TABLE-US-00002 TABLE 1B Exemplary Compounds LC-MS Cmpd m/z No
Structure Exact mass (M + H) 194 ##STR00353## 406.2005 407.1 195
##STR00354## 406.2005 407.2 196 ##STR00355## 470.2529 471.3 197
##STR00356## 424.2474 425.3 198 ##STR00357## 476.2093 477.1 199
##STR00358## 480.2737 481.3 200 ##STR00359## 379.1896 380.2 201
##STR00360## 490.225 491.1 202 ##STR00361## 502.225 503.2 203
##STR00362## 516.2406 517.2 204 ##STR00363## 437.2678 438.3 205
##STR00364## 437.2678 438.2 206 ##STR00365## 378.2056 379.2 207
##STR00366## 392.2212 393.3 208 ##STR00367## 412.1666 413.2 209
##STR00368## 392.2212 393.1 210 ##STR00369## 502.225 503.3 211
##STR00370## 516.2406 517.3 212 ##STR00371## 380.1848 381.2 213
##STR00372## 396.1962 397.2 214 ##STR00373## 544.2719 545.3 215
##STR00374## 504.2043 505.3 216 ##STR00375## 532.2356 533.3 217
##STR00376## 518.2199 519.3 218 ##STR00377## 491.2533 492.3 219
##STR00378## 516.2406 517.3 220 ##STR00379## 516.2406 517.3 221
##STR00380## 488.2093 489.3 222 ##STR00381## 392.2212 393.3 223
##STR00382## 380.1848 381.2 224 ##STR00383## 392.2212 393.3 225
##STR00384## 396.1962 397.1 226 ##STR00385## 412.1666 413.0 227
##STR00386## 412.1666 413.3 228 ##STR00387## 392.2212 393.3 229
##STR00388## 378.2056 379.2 230 ##STR00389## 477.2376 478.3 231
##STR00390## 491.2533 492.2 232 ##STR00391## 502.225 503.2 233
##STR00392## 532.2356 533.1 234 ##STR00393## 420.2161 421.2 235
##STR00394## 423.2522 424.2 236 ##STR00395## 423.2522 424.2 237
##STR00396## 439.2471 440.3 238 ##STR00397## 530.2563 531.2 239
##STR00398## 379.2008 380.0 240 ##STR00399## 328.1899 329.0 241
##STR00400## 396.1962 397.1 242 ##STR00401## 408.2161 409.1 243
##STR00402## 404.2212 405.2 244 ##STR00403## 484.2686 485.2 245
##STR00404## 491.2533 492.1 246 ##STR00405## 518.2199 519.2 247
##STR00406## 530.2563 531.3 248 ##STR00407## 378.2056 379.1 249
##STR00408## 408.2161 409.1 250 ##STR00409## 392.2212 393.1 251
##STR00410## 382.2307 383.4 252 ##STR00411## 531.2515 532.2 253
##STR00412## 423.2522 424.2 254 ##STR00413## 437.2678 438.1 255
##STR00414## 420.2161 421.1 256 ##STR00415## 418.2369 419.1 257
##STR00416## 516.2406 517.2 258 ##STR00417## 437.2678 438.3 259
##STR00418## 342.2056 343.1 260 ##STR00419## 532.2719 533.1 261
##STR00420## 423.2522 424.2 262 ##STR00421## 423.2522 424.2 263
##STR00422## 423.2522 424.2 264 ##STR00423## 423.2522 424.2 265
##STR00424## 516.2406 517.2 266 ##STR00425## 558.2876 559.3 267
##STR00426## 477.2376 478.3 268 ##STR00427## 392.2212 393.3 269
##STR00428## 395.21 396.9 270 ##STR00429## 424.2474 425.2 271
##STR00430## 424.2474 425.2 272 ##STR00431## 530.679 531.3 273
##STR00432## 516.2406 517.3 274 ##STR00433## 530.2563 531.2 275
##STR00434## 545.2672 546.3 276 ##STR00435## 545.2672 546.2 277
##STR00436## 559.2828 560.3 278 ##STR00437## 571.2828 572.3
[0190] In certain embodiments, a provided compound is a compound
listed in Table IC, or a pharmaceutically acceptable salt
thereof.
TABLE-US-00003 TABLE 1C Exemplary Compounds Cmpd LC-MS m/z No
Structure Exact mass (M + H) 279 ##STR00438## 390.2056 -- 280
##STR00439## 404.2212 -- 281 ##STR00440## 418.2369 -- 282
##STR00441## 432.2161 -- 283 ##STR00442## 468.1831 -- 284
##STR00443## 443.1821 -- 285 ##STR00444## 486.2243 -- 286
##STR00445## 458.2682 -- 287 ##STR00446## 459.2634 -- 288
##STR00447## 460.2474 -- 289 ##STR00448## 430.2369 -- 290
##STR00449## 444.2525 -- 291 ##STR00450## 544.2832 -- 292
##STR00451## 558.2988 -- 293 ##STR00452## 572.3145 -- 294
##STR00453## 586.2937 -- 295 ##STR00454## 622.2607 -- 296
##STR00455## 597.2597 -- 297 ##STR00456## 640.3019 -- 298
##STR00457## 612.3458 -- 299 ##STR00458## 613.3410 -- 300
##STR00459## 614.3250 -- 301 ##STR00460## 584.3145 -- 302
##STR00461## 598.3301 --
[0191] In certain embodiments, a provided compound inhibits PRMT5.
In certain embodiments, a provided compound inhibits wild-type
PRMT5. In certain embodiments, a provided compound inhibits a
mutant PRMT5. In certain embodiments, a provided compound inhibits
PRMT5, e.g., as measured in an assay described herein. In certain
embodiments, the PRMT5 is from a human. In certain embodiments, a
provided compound inhibits PRMT5 at an IC.sub.50 less than or equal
to 10 .mu.M. In certain embodiments, a provided compound inhibits
PRMT5 at an IC.sub.50 less than or equal to 1 .mu.M. In certain
embodiments, a provided compound inhibits PRMT5 at an IC.sub.50
less than or equal to 0.1 M. In certain embodiments, a provided
compound inhibits PRMT5 in a cell at an EC.sub.50 less than or
equal to 10 .mu.M. In certain embodiments, a provided compound
inhibits PRMT5 in a cell at an EC.sub.50 less than or equal to 1
.mu.M. In certain embodiments, a provided compound inhibits PRMT5
in a cell at an EC.sub.50 less than or equal to 0.1 .mu.M. In
certain embodiments, a provided compound inhibits cell
proliferation at an EC.sub.50 less than or equal to 10 .mu.M. In
certain embodiments, a provided compound inhibits cell
proliferation at an EC.sub.50 less than or equal to 1 .mu.M. In
certain embodiments, a provided compound inhibits cell
proliferation at an EC.sub.50 less than or equal to 0.1 .mu.M. In
some embodiments, a provided compound is selective for PRMT5 over
other methyltransferases. In certain embodiments, a provided
compound is at least about 10-fold selective, at least about
20-fold selective, at least about 30-fold selective, at least about
40-fold selective, at least about 50-fold selective, at least about
60-fold selective, at least about 70-fold selective, at least about
80-fold selective, at least about 90-fold selective, or at least
about 100-fold selective for PRMT5 relative to one or more other
methyltransferases.
[0192] It will be understood by one of ordinary skill in the art
that the PRMT5 can be wild-type PRMT5, or any mutant or variant of
PRMT5.
[0193] In some embodiments embodiment, the mutant or variant of
PRMT5 contains one or more mutations (e.g., conservative
substitutions). In some embodiments, provided herein is a PRMT5
point mutant. In some embodiments, the PRMT point mutant has an
amino acid sequence that a degree of homology to the amino acid
sequence of SEQ ID NO: 1 of at least about 80%, e.g., at least
about 85%, at least about 90%, at least about 95%, or at least
about 97%. Further provided is a protein that has a degree of
homology to the amino acid sequence of SEQ ID NO: 2 of at least
about 80%, e.g., at least about 85%, at least about 90%, at least
about 95%, or at least about 97%.
[0194] In certain embodiments, the PRMT5 is isoform A (GenBank
accession no. NP006100) (SEQ ID NO.:1):
TABLE-US-00004 MAAMAVGGAG GSRVSSGRDL NCVPEIADTL GAVAKQGFDF
LCMPVFHPRF KREFIQEPAK NRPGPQTRSD LLLSGRDWNT LIVGKLSPWI RPDSKVEKIR
RNSEAAMLQE LNFGAYLGLP AFLLPLNQED NTNLARVLTN HIHTGHHSSM FWMRVPLVAP
EDLRDDIIEN APTTHTEEYS GEEKTWMWWH NFRTLCDYSK RIAVALEIGA DLPSNHVIDR
WLGEPIKAAI LPTSIFLTNK KGFPVLSKMH QRLIFRLLKL EVQFIITGTN HHSEKEFCSY
LQYLEYLSQN RPPPNAYELF AKGYEDYLQS PLQPLMDNLE SQTYEVFEKD PIKYSQYQQA
IYKCLLDRVP EEEKDTNVQV LMVLGAGRGP LVNASLRAAK QADRRIKLYA VEKNPNAVVT
LENWQFEEWG SQVTVVSSDM REWVAPEKAD IIVSELLGSF ADNELSPECL DGAQHFLKDD
GVSIPGEYTS FLAPISSSKL YNEVRACREK DRDPEAQFEM PYVVRLHNFH QLSAPQPCFT
FSHPNRDPMI DNNRYCTLEF PVEVNTVLHG FAGYFETVLY QDITLSIRPE THSPGMFSWF
PILFPIKQPI TVREGQTICV RFWRCSNSKK VWYEWAVTAP VCSAIHNPTG RSYTIGL
[0195] In certain embodiments, the PRMT5 is isoform B (GenBank
accession no. NP001034708) (SEQ ID NO.:2):
TABLE-US-00005 MRGPNSGTEK GRLVIPEKQG FDFLCMPVFH PRFKREFIQE
PAKNRPGPQT RSDLLLSGRD WNTLIVGKLS PWIRPDSKVE KIRRNSEAAM LQELNFGAYL
GLPAFLLPLN QEDNTNLARV LTNHIHTGHH SSMFWMRVPL VAPEDLRDDI IENAPTTHTE
EYSGEEKTWM WWHNFRTLCD YSKRIAVALE IGADLPSNHV IDRWLGEPIK AAILPTSIFL
TNKKGFPVLS KMHQRLIFRL LKLEVQFIIT GTNHHSEKEF CSYLQYLEYL SQNRPPPNAY
ELFAKGYEDY LQSPLQPLMD NLESQTYEVF EKDPIKYSQY QQAIYKCLLD RVPEEEKDTN
VQVLMVLGAG RGPLVNASLR AAKQADRRIK LYAVEKNPNA VVTLENWQFE EWGSQVTVVS
SDMREWVAPE KADIIVSELL GSFADNELSP ECLDGAQHFL KDDGVSIPGE YTSFLAPISS
SKLYNEVRAC REKDRDPEAQ FEMPYVVRLH NFHQLSAPQP CFTFSHPNRD PMIDNNRYCT
LEFPVEVNTV LHGFAGYFET VLYQDITLSI RPETHSPGMF SWFPILFPIK QPITVREGQT
ICVRFWRCSN SKKVWYEWAV TAPVCSAIHN PTGRSYTIGL
[0196] In certain embodiments, the PRMT5 is transcript variant 1
(GenBank accession no. NM_006109).
[0197] The present disclosure provides pharmaceutical compositions
comprising a compound described herein, e.g., a compound of Formula
(A), e.g., Formula (I), or a pharmaceutically acceptable salt
thereof, as described herein, and optionally a pharmaceutically
acceptable excipient. It will be understood by one of ordinary
skill in the art that the compounds described herein, or salts
thereof, may be present in various forms, such as amorphous,
hydrates, solvates, or polymorphs. In certain embodiments, a
provided composition comprises two or more compounds described
herein. In certain embodiments, a compound described herein, or a
pharmaceutically acceptable salt thereof, is provided in an
effective amount in the pharmaceutical composition. In certain
embodiments, the effective amount is a therapeutically effective
amount. In certain embodiments, the effective amount is an amount
effective for inhibiting PRMT5. In certain embodiments, the
effective amount is an amount effective for treating a
PRMT5-mediated disorder. In certain embodiments, the effective
amount is a prophylactically effective amount. In certain
embodiments, the effective amount is an amount effective to prevent
a PRMT5-mediated disorder.
[0198] In certain embodiments, the provided pharmaceutical
compositions comprise a compound described herein, e.g., a compound
of Formula (A), e.g., Formula (I), or any subgenera thereof, and
optionally a pharmaceutically acceptable excipient, wherein the
compound is of a free base form. In certain embodiments, the
provided pharmaceutical compositions comprise a compound described
herein, e.g., a compound of Formula (A), e.g., Formula (I), or any
subgenera thereof, and optionally a pharmaceutically acceptable
excipient, wherein the compound is in the form of a
pharmaceutically acceptable salt as generally defined herein. In
certain embodiments, the provided pharmaceutical compositions
comprise a hydrochloride salt of a compound described herein and
optionally a pharmaceutically acceptable excipient. In certain
embodiments, the provided pharmaceutical compositions comprise a
tartrate salt of a compound described herein and optionally a
pharmaceutically acceptable excipient. In certain embodiments, the
provided pharmaceutical compositions comprise a monotartrate salt
of a compound described herein and optionally a pharmaceutically
acceptable excipient. In certain embodiments, the provided
pharmaceutical compositions comprise a bitartrate salt of a
compound described herein and optionally a pharmaceutically
acceptable excipient. In certain embodiments, the provided
pharmaceutical compositions comprise a monotartrate salt and a
bitartrate salt of a compound described herein and optionally a
pharmaceutically acceptable excipient. In certain embodiments, the
provided pharmaceutical compositions comprise a compound described
herein in a form of free base, and a pharmaceutically acceptable
salt thereof, and optionally a pharmaceutically acceptable
excipient.
[0199] Pharmaceutically acceptable excipients include any and all
solvents, diluents, or other liquid vehicles, dispersions,
suspension aids, surface active agents, isotonic agents, thickening
or emulsifying agents, preservatives, solid binders, lubricants,
and the like, as suited to the particular dosage form desired.
General considerations in formulation and/or manufacture of
pharmaceutical compositions agents can be found, for example, in
Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W.
Martin (Mack Publishing Co., Easton, Pa., 1980), and Remington: The
Science and Practice of Pharmacy, 21st Edition (Lippincott Williams
& Wilkins, 2005).
[0200] Pharmaceutical compositions described herein can be prepared
by any method known in the art of pharmacology. In general, such
preparatory methods include the steps of bringing a compound
described herein (the "active ingredient") into association with a
carrier and/or one or more other accessory ingredients, and then,
if necessary and/or desirable, shaping and/or packaging the product
into a desired single- or multi-dose unit.
[0201] Pharmaceutical compositions can be prepared, packaged,
and/or sold in bulk, as a single unit dose, and/or as a plurality
of single unit doses. As used herein, a "unit dose" is discrete
amount of the pharmaceutical composition comprising a predetermined
amount of the active ingredient. The amount of the active
ingredient is generally equal to the dosage of the active
ingredient which would be administered to a subject and/or a
convenient fraction of such a dosage such as, for example, one-half
or one-third of such a dosage.
[0202] Relative amounts of the active ingredient, the
pharmaceutically acceptable excipient, and/or any additional
ingredients in a pharmaceutical composition of the present
disclosure will vary, depending upon the identity, size, and/or
condition of the subject treated and further depending upon the
route by which the composition is to be administered. By way of
example, the composition may comprise between 0.1% and 100% (w/w)
active ingredient.
[0203] Pharmaceutically acceptable excipients used in the
manufacture of provided pharmaceutical compositions include inert
diluents, dispersing and/or granulating agents, surface active
agents and/or emulsifiers, disintegrating agents, binding agents,
preservatives, buffering agents, lubricating agents, and/or oils.
Excipients such as cocoa butter and suppository waxes, coloring
agents, coating agents, sweetening, flavoring, and perfuming agents
may also be present in the composition.
[0204] Exemplary diluents include calcium carbonate, sodium
carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate,
calcium hydrogen phosphate, sodium phosphate lactose, sucrose,
cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol,
inositol, sodium chloride, dry starch, cornstarch, powdered sugar,
and mixtures thereof.
[0205] Exemplary granulating and/or dispersing agents include
potato starch, corn starch, tapioca starch, sodium starch
glycolate, clays, alginic acid, guar gum, citrus pulp, agar,
bentonite, cellulose and wood products, natural sponge,
cation-exchange resins, calcium carbonate, silicates, sodium
carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone),
sodium carboxymethyl starch (sodium starch glycolate),
carboxymethyl cellulose, cross-linked sodium carboxymethyl
cellulose (croscarmellose), methylcellulose, pregelatinized starch
(starch 1500), microcrystalline starch, water insoluble starch,
calcium carboxymethyl cellulose, magnesium aluminum silicate
(Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and
mixtures thereof.
[0206] Exemplary surface active agents and/or emulsifiers include
natural emulsifiers (e.g., acacia, agar, alginic acid, sodium
alginate, tragacanth, chondrux, cholesterol, xanthan, pectin,
gelatin, egg yolk, casein, wool fat, cholesterol, wax, and
lecithin), colloidal clays (e.g., bentonite (aluminum silicate) and
Veegum (magnesium aluminum silicate)), long chain amino acid
derivatives, high molecular weight alcohols (e.g., stearyl alcohol,
cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene
glycol distearate, glyceryl monostearate, and propylene glycol
monostearate, polyvinyl alcohol), carbomers (e.g., carboxy
polymethylene, polyacrylic acid, acrylic acid polymer, and
carboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g.,
carboxymethylcellulose sodium, powdered cellulose, hydroxymethyl
cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose,
methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene
sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan (Tween
60), polyoxyethylene sorbitan monooleate (Tween 80), sorbitan
monopalmitate (Span 40), sorbitan monostearate (Span 60], sorbitan
tristearate (Span 65), glyceryl monooleate, sorbitan monooleate
(Span 80)), polyoxyethylene esters (e.g., polyoxyethylene
monostearate (Myrj 45), polyoxyethylene hydrogenated castor oil,
polyethoxylated castor oil, polyoxymethylene stearate, and
Solutol), sucrose fatty acid esters, polyethylene glycol fatty acid
esters (e.g., Cremophor.TM.), polyoxyethylene ethers, (e.g.,
polyoxyethylene lauryl ether (Brij 30)), poly(vinyl-pyrrolidone),
diethylene glycol monolaurate, triethanolamine oleate, sodium
oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate,
sodium lauryl sulfate, Pluronic F68, Poloxamer 188, cetrimonium
bromide, cetylpyridinium chloride, benzalkonium chloride, docusate
sodium, and/or mixtures thereof.
[0207] Exemplary binding agents include starch (e.g., cornstarch
and starch paste), gelatin, sugars (e.g., sucrose, glucose,
dextrose, dextrin, molasses, lactose, lactitol, mannitol, etc.),
natural and synthetic gums (e.g., acacia, sodium alginate, extract
of Irish moss, panwar gum, ghatti gum, mucilage of isapol husks,
carboxymethylcellulose, methylcellulose, ethylcellulose,
hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropyl
methylcellulose, microcrystalline cellulose, cellulose acetate,
poly(vinyl-pyrrolidone), magnesium aluminum silicate (Veegum), and
larch arabogalactan), alginates, polyethylene oxide, polyethylene
glycol, inorganic calcium salts, silicic acid, polymethacrylates,
waxes, water, alcohol, and/or mixtures thereof.
[0208] Exemplary preservatives include antioxidants, chelating
agents, antimicrobial preservatives, antifungal preservatives,
alcohol preservatives, acidic preservatives, and other
preservatives.
[0209] Exemplary antioxidants include alpha tocopherol, ascorbic
acid, acorbyl palmitate, butylated hydroxyanisole, butylated
hydroxytoluene, monothioglycerol, potassium metabisulfite,
propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite,
sodium metabisulfite, and sodium sulfite.
[0210] Exemplary chelating agents include
ethylenediaminetetraacetic acid (EDTA) and salts and hydrates
thereof (e.g., sodium edetate, disodium edetate, trisodium edetate,
calcium disodium edetate, dipotassium edetate, and the like),
citric acid and salts and hydrates thereof (e.g., citric acid
monohydrate), fumaric acid and salts and hydrates thereof, malic
acid and salts and hydrates thereof, phosphoric acid and salts and
hydrates thereof, and tartaric acid and salts and hydrates thereof.
Exemplary antimicrobial preservatives include benzalkonium
chloride, benzethonium chloride, benzyl alcohol, bronopol,
cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol,
chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin,
hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol,
phenylmercuric nitrate, propylene glycol, and thimerosal.
[0211] Exemplary antifungal preservatives include butyl paraben,
methyl paraben, ethyl paraben, propyl paraben, benzoic acid,
hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium
benzoate, sodium propionate, and sorbic acid.
[0212] Exemplary alcohol preservatives include ethanol,
polyethylene glycol, phenol, phenolic compounds, bisphenol,
chlorobutanol, hydroxybenzoate, and phenylethyl alcohol. Exemplary
acidic preservatives include vitamin A, vitamin C, vitamin E,
beta-carotene, citric acid, acetic acid, dehydroacetic acid,
ascorbic acid, sorbic acid, and phytic acid.
[0213] Other preservatives include tocopherol, tocopherol acetate,
deteroxime mesylate, cetrimide, butylated hydroxyanisol (BHA),
butylated hydroxytoluened (BHT), ethylenediamine, sodium lauryl
sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium
bisulfite, sodium metabisulfite, potassium sulfite, potassium
metabisulfite, Glydant Plus, Phenonip, methylparaben, Germall 115,
Germaben II, Neolone, Kathon, and Euxyl. In certain embodiments,
the preservative is an anti-oxidant. In other embodiments, the
preservative is a chelating agent.
[0214] Exemplary buffering agents include citrate buffer solutions,
acetate buffer solutions, phosphate buffer solutions, ammonium
chloride, calcium carbonate, calcium chloride, calcium citrate,
calcium glubionate, calcium gluceptate, calcium gluconate,
D-gluconic acid, calcium glycerophosphate, calcium lactate,
propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium
phosphate, phosphoric acid, tribasic calcium phosphate, calcium
hydroxide phosphate, potassium acetate, potassium chloride,
potassium gluconate, potassium mixtures, dibasic potassium
phosphate, monobasic potassium phosphate, potassium phosphate
mixtures, sodium acetate, sodium bicarbonate, sodium chloride,
sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic
sodium phosphate, sodium phosphate mixtures, tromethamine,
magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free
water, isotonic saline, Ringer's solution, ethyl alcohol, and
mixtures thereof.
[0215] Exemplary lubricating agents include magnesium stearate,
calcium stearate, stearic acid, silica, talc, malt, glyceryl
behanate, hydrogenated vegetable oils, polyethylene glycol, sodium
benzoate, sodium acetate, sodium chloride, leucine, magnesium
lauryl sulfate, sodium lauryl sulfate, and mixtures thereof.
[0216] Exemplary natural oils include almond, apricot kernel,
avocado, babassu, bergamot, black current seed, borage, cade,
camomile, canola, caraway, carnauba, castor, cinnamon, cocoa
butter, coconut, cod liver, coffee, corn, cotton seed, emu,
eucalyptus, evening primrose, fish, flaxseed, geraniol, gourd,
grape seed, hazel nut, hyssop, isopropyl myristate, jojoba, kukui
nut, lavandin, lavender, lemon, litsea cubeba, macademia nut,
mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange,
orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed,
pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood,
sasquana, savoury, sea buckthorn, sesame, shea butter, silicone,
soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut,
and wheat germ oils. Exemplary synthetic oils include, but are not
limited to, butyl stearate, caprylic triglyceride, capric
triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360,
isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol,
silicone oil, and mixtures thereof.
[0217] Liquid dosage forms for oral and parenteral administration
include pharmaceutically acceptable emulsions, microemulsions,
solutions, suspensions, syrups and elixirs. In addition to the
active ingredients, the liquid dosage forms may comprise 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 (e.g., cottonseed, groundnut, corn, germ,
olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl
alcohol, polyethylene glycols and fatty acid esters of sorbitan,
and mixtures thereof. Besides inert diluents, the oral compositions
can include adjuvants such as wetting agents, emulsifying and
suspending agents, sweetening, flavoring, and perfuming agents. In
certain embodiments for parenteral administration, the compounds
described herein are mixed with solubilizing agents such as
Cremophor.TM., alcohols, oils, modified oils, glycols,
polysorbates, cyclodextrins, polymers, and mixtures thereof.
[0218] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions can be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation can 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 can 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.
[0219] The injectable formulations can be sterilized, for example,
by filtration through a bacterial-retaining filter, or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium prior to use.
[0220] 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 can 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.
[0221] Compositions for rectal or vaginal administration are
typically suppositories which can be prepared by mixing the
compounds described herein 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 ingredient.
[0222] Solid dosage forms for oral administration include capsules,
tablets, pills, powders, and granules. In such solid dosage forms,
the active ingredient 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, 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 comprise buffering agents.
[0223] Solid compositions of a similar type can 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 comprise opacifying agents and can 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 can 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.
[0224] The active ingredient can 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 ingredient can be admixed with at least one inert diluent
such as sucrose, lactose, or starch. Such dosage forms may
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 comprise
buffering agents. They may optionally comprise opacifying agents
and can 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.
[0225] Dosage forms for topical and/or transdermal administration
of a provided compound may include ointments, pastes, creams,
lotions, gels, powders, solutions, sprays, inhalants and/or
patches. Generally, the active ingredient is admixed under sterile
conditions with a pharmaceutically acceptable carrier and/or any
desired preservatives and/or buffers as can be required.
Additionally, the present disclosure encompasses the use of
transdermal patches, which often have the added advantage of
providing controlled delivery of an active ingredient to the body.
Such dosage forms can be prepared, for example, by dissolving
and/or dispensing the active ingredient in the proper medium.
Alternatively or additionally, the rate can be controlled by either
providing a rate controlling membrane and/or by dispersing the
active ingredient in a polymer matrix and/or gel.
[0226] Suitable devices for use in delivering intradermal
pharmaceutical compositions described herein include short needle
devices such as those described in U.S. Pat. Nos. 4,886,499;
5,190,521; 5,328,483; 5,527,288; 4,270,537; 5,015,235; 5,141,496;
and 5,417,662. Intradermal compositions can be administered by
devices which limit the effective penetration length of a needle
into the skin, such as those described in PCT publication WO
99/34850 and functional equivalents thereof. Jet injection devices
which deliver liquid vaccines to the dermis via a liquid jet
injector and/or via a needle which pierces the stratum corneum and
produces a jet which reaches the dermis are suitable. Jet injection
devices are described, for example, in U.S. Pat. Nos. 5,480,381;
5,599,302; 5,334,144; 5,993,412; 5,649,912; 5,569,189; 5,704,911;
5,383,851; 5,893,397; 5,466,220; 5,339,163; 5,312,335; 5,503,627;
5,064,413; 5,520,639; 4,596,556; 4,790,824; 4,941,880; 4,940,460;
and PCT publications WO 97/37705 and WO 97/13537. Ballistic
powder/particle delivery devices which use compressed gas to
accelerate vaccine in powder form through the outer layers of the
skin to the dermis are suitable. Alternatively or additionally,
conventional syringes can be used in the classical mantoux method
of intradermal administration.
[0227] Formulations suitable for topical administration include,
but are not limited to, liquid and/or semi liquid preparations such
as liniments, lotions, oil in water and/or water in oil emulsions
such as creams, ointments and/or pastes, and/or solutions and/or
suspensions. Topically-administrable formulations may, for example,
comprise from about 1% to about 10% (w/w) active ingredient,
although the concentration of the active ingredient can be as high
as the solubility limit of the active ingredient in the solvent.
Formulations for topical administration may further comprise one or
more of the additional ingredients described herein.
[0228] A provided pharmaceutical composition can be prepared,
packaged, and/or sold in a formulation suitable for pulmonary
administration via the buccal cavity. Such a formulation may
comprise dry particles which comprise the active ingredient and
which have a diameter in the range from about 0.5 to about 7
nanometers or from about 1 to about 6 nanometers. Such compositions
are conveniently in the form of dry powders for administration
using a device comprising a dry powder reservoir to which a stream
of propellant can be directed to disperse the powder and/or using a
self propelling solvent/powder dispensing container such as a
device comprising the active ingredient dissolved and/or suspended
in a low-boiling propellant in a sealed container. Such powders
comprise particles wherein at least 98% of the particles by weight
have a diameter greater than 0.5 nanometers and at least 95% of the
particles by number have a diameter less than 7 nanometers.
Alternatively, at least 95% of the particles by weight have a
diameter greater than 1 nanometer and at least 90% of the particles
by number have a diameter less than 6 nanometers. Dry powder
compositions may include a solid fine powder diluent such as sugar
and are conveniently provided in a unit dose form.
[0229] Low boiling propellants generally include liquid propellants
having a boiling point of below 65.degree. F. at atmospheric
pressure. Generally the propellant may constitute 50 to 99.9% (w/w)
of the composition, and the active ingredient may constitute 0.1 to
20% (w/w) of the composition. The propellant may further comprise
additional ingredients such as a liquid non-ionic and/or solid
anionic surfactant and/or a solid diluent (which may have a
particle size of the same order as particles comprising the active
ingredient).
[0230] Pharmaceutical compositions formulated for pulmonary
delivery may provide the active ingredient in the form of droplets
of a solution and/or suspension. Such formulations can be prepared,
packaged, and/or sold as aqueous and/or dilute alcoholic solutions
and/or suspensions, optionally sterile, comprising the active
ingredient, and may conveniently be administered using any
nebulization and/or atomization device. Such formulations may
further comprise one or more additional ingredients including, but
not limited to, a flavoring agent such as saccharin sodium, a
volatile oil, a buffering agent, a surface active agent, and/or a
preservative such as methylhydroxybenzoate. The droplets provided
by this route of administration may have an average diameter in the
range from about 0.1 to about 200 nanometers.
[0231] Formulations described herein as being useful for pulmonary
delivery are useful for intranasal delivery of a pharmaceutical
composition. Another formulation suitable for intranasal
administration is a coarse powder comprising the active ingredient
and having an average particle from about 0.2 to 500 micrometers.
Such a formulation is administered by rapid inhalation through the
nasal passage from a container of the powder held close to the
nares.
[0232] Formulations for nasal administration may, for example,
comprise from about as little as 0.1% (w/w) and as much as 100%
(w/w) of the active ingredient, and may comprise one or more of the
additional ingredients described herein. A provided pharmaceutical
composition can be prepared, packaged, and/or sold in a formulation
for buccal administration. Such formulations may, for example, be
in the form of tablets and/or lozenges made using conventional
methods, and may contain, for example, 0.1 to 20% (w/w) active
ingredient, the balance comprising an orally dissolvable and/or
degradable composition and, optionally, one or more of the
additional ingredients described herein. Alternately, formulations
for buccal administration may comprise a powder and/or an
aerosolized and/or atomized solution and/or suspension comprising
the active ingredient. Such powdered, aerosolized, and/or
aerosolized formulations, when dispersed, may have an average
particle and/or droplet size in the range from about 0.1 to about
200 nanometers, and may further comprise one or more of the
additional ingredients described herein.
[0233] A provided pharmaceutical composition can be prepared,
packaged, and/or sold in a formulation for ophthalmic
administration. Such formulations may, for example, be in the form
of eye drops including, for example, a 0.1/1.0% (w/w) solution
and/or suspension of the active ingredient in an aqueous or oily
liquid carrier. Such drops may further comprise buffering agents,
salts, and/or one or more other of the additional ingredients
described herein. Other opthalmically-administrable formulations
which are useful include those which comprise the active ingredient
in microcrystalline form and/or in a liposomal preparation. Ear
drops and/or eye drops are contemplated as being within the scope
of this disclosure.
[0234] Although the descriptions of pharmaceutical compositions
provided herein are principally directed to pharmaceutical
compositions which are suitable for administration to humans, it
will be understood by the skilled artisan that such compositions
are generally suitable for administration to animals of all sorts.
Modification of pharmaceutical compositions suitable for
administration to humans in order to render the compositions
suitable for administration to various animals is well understood,
and the ordinarily skilled veterinary pharmacologist can design
and/or perform such modification with ordinary experimentation.
[0235] Compounds provided herein are typically formulated in dosage
unit form for ease of administration and uniformity of dosage. It
will be understood, however, that the total daily usage of provided
compositions will be decided by the attending physician within the
scope of sound medical judgment. The specific therapeutically
effective dose level for any particular subject or organism will
depend upon a variety of factors including the disease, disorder,
or condition being treated and the severity of the disorder; the
activity of the specific active ingredient employed; the specific
composition employed; the age, body weight, general health, sex and
diet of the subject; the time of administration, route of
administration, and rate of excretion of the specific active
ingredient employed; the duration of the treatment; drugs used in
combination or coincidental with the specific active ingredient
employed; and like factors well known in the medical arts.
[0236] The compounds and compositions provided herein can be
administered by any route, including enteral (e.g., oral),
parenteral, intravenous, intramuscular, intra-arterial,
intramedullary, intrathecal, subcutaneous, intraventricular,
transdermal, interdermal, rectal, intravaginal, intraperitoneal,
topical (as by powders, ointments, creams, and/or drops), mucosal,
nasal, bucal, sublingual; by intratracheal instillation, bronchial
instillation, and/or inhalation; and/or as an oral spray, nasal
spray, and/or aerosol. Specifically contemplated routes are oral
administration, intravenous administration (e.g., systemic
intravenous injection), regional administration via blood and/or
lymph supply, and/or direct administration to an affected site. In
general the most appropriate route of administration will depend
upon a variety of factors including the nature of the agent (e.g.,
its stability in the environment of the gastrointestinal tract),
and/or the condition of the subject (e.g., whether the subject is
able to tolerate oral administration).
[0237] The exact amount of a compound required to achieve an
effective amount will vary from subject to subject, depending, for
example, on species, age, and general condition of a subject,
severity of the side effects or disorder, identity of the
particular compound(s), mode of administration, and the like. The
desired dosage can 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 can be delivered using multiple
administrations (e.g., two, three, four, five, six, seven, eight,
nine, ten, eleven, twelve, thirteen, fourteen, or more
administrations).
[0238] In certain embodiments, an effective amount of a compound
for administration one or more times a day to a 70 kg adult human
may comprise about 0.0001 mg to about 3000 mg, about 0.0001 mg to
about 2000 mg, about 0.0001 mg to about 1000 mg, about 0.001 mg to
about 1000 mg, about 0.01 mg to about 1000 mg, about 0.1 mg to
about 1000 mg, about 1 mg to about 1000 mg, about 1 mg to about 100
mg, about 10 mg to about 1000 mg, or about 100 mg to about 1000 mg,
of a compound per unit dosage form.
[0239] In certain embodiments, a compound described herein may be
administered at dosage levels sufficient to deliver from about
0.001 mg/kg to about 1000 mg/kg, from about 0.01 mg/kg to about
mg/kg, from about 0.1 mg/kg to about 40 mg/kg, 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, or 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.
[0240] In some embodiments, a compound described herein is
administered one or more times per day, for multiple days. In some
embodiments, the dosing regimen is continued for days, weeks,
months, or years.
[0241] It will be appreciated that dose ranges as described herein
provide guidance for the administration of provided pharmaceutical
compositions to an adult. The amount to be administered to, for
example, a child or an adolescent can be determined by a medical
practitioner or person skilled in the art and can be lower or the
same as that administered to an adult.
[0242] It will be also appreciated that a compound or composition,
as described herein, can be administered in combination with one or
more additional therapeutically active agents. In certain
embodiments, a compound or composition provided herein is
administered in combination with one or more additional
therapeutically active agents that improve its bioavailability,
reduce and/or modify its metabolism, inhibit its excretion, and/or
modify its distribution within the body. It will also be
appreciated that the therapy employed may achieve a desired effect
for the same disorder, and/or it may achieve different effects.
[0243] The compound or composition can be administered concurrently
with, prior to, or subsequent to, one or more additional
therapeutically active agents. In certain embodiments, the
additional therapeutically active agent is a compound of Formula
(A), e.g., Formula (I). In certain embodiments, the additional
therapeutically active agent is not a compound of Formula (A),
e.g., Formula (I). In general, each agent will be administered at a
dose and/or on a time schedule determined for that agent. In will
further be appreciated that the additional therapeutically active
agent utilized in this combination can be administered together in
a single composition or administered separately in different
compositions. The particular combination to employ in a regimen
will take into account compatibility of a provided compound with
the additional therapeutically active agent and/or the desired
therapeutic effect to be achieved. In general, it is expected that
additional therapeutically active agents utilized in combination be
utilized at levels that do not exceed the levels at which they are
utilized individually. In some embodiments, the levels utilized in
combination will be lower than those utilized individually.
[0244] Exemplary additional therapeutically active agents include,
but are not limited to, small organic molecules such as drug
compounds (e.g., compounds approved by the U.S. Food and Drug
Administration as provided in the Code of Federal Regulations
(CFR)), peptides, proteins, carbohydrates, monosaccharides,
oligosaccharides, polysaccharides, nucleoproteins, mucoproteins,
lipoproteins, synthetic polypeptides or proteins, small molecules
linked to proteins, glycoproteins, steroids, nucleic acids, DNAs,
RNAs, nucleotides, nucleosides, oligonucleotides, antisense
oligonucleotides, lipids, hormones, vitamins, and cells.
[0245] Also encompassed by the present disclosure are kits (e.g.,
pharmaceutical packs). The kits provided may comprise a provided
pharmaceutical composition or compound and a container (e.g., a
vial, ampule, bottle, syringe, and/or dispenser package, or other
suitable container). In some embodiments, provided kits may
optionally further include a second container comprising a
pharmaceutical excipient for dilution or suspension of a provided
pharmaceutical composition or compound. In some embodiments, a
provided pharmaceutical composition or compound provided in the
container and the second container are combined to form one unit
dosage form. In some embodiments, a provided kits further includes
instructions for use.
[0246] Compounds and compositions described herein are generally
useful for the inhibition of PRMT5. In some embodiments, methods of
treating PRMT5-mediated disorder in a subject are provided which
comprise administering an effective amount of a compound described
herein (e.g., a compound of Formula (A), e.g., Formula (I)), or a
pharmaceutically acceptable salt thereof), to a subject in need of
treatment. In certain embodiments, the effective amount is a
therapeutically effective amount. In certain embodiments, the
effective amount is a prophylactically effective amount. In certain
embodiments, the subject is suffering from a PRMT5-mediated
disorder. In certain embodiments, the subject is susceptible to a
PRMT5-mediated disorder.
[0247] As used herein, the term "PRMT5-mediated disorder" means any
disease, disorder, or other pathological condition in which PRMT5
is known to play a role. Accordingly, in some embodiments, the
present disclosure relates to treating or lessening the severity of
one or more diseases in which PRMT5 is known to play a role.
[0248] In some embodiments, the present disclosure provides a
method of inhibiting PRMT5 comprising contacting PRMT5 with an
effective amount of a compound described herein (e.g., a compound
of Formula (A), e.g., Formula (I)), or a pharmaceutically
acceptable salt thereof. The PRMT5 may be purified or crude, and
may be present in a cell, tissue, or subject. Thus, such methods
encompass both inhibition of in vitro and in vivo PRMT5 activity.
In certain embodiments, the method is an in vitro method, e.g.,
such as an assay method. It will be understood by one of ordinary
skill in the art that inhibition of PRMT5 does not necessarily
require that all of the PRMT5 be occupied by an inhibitor at once.
Exemplary levels of inhibition of PRMT5 include at least 10%
inhibition, about 10% to about 25% inhibition, about 25% to about
50% inhibition, about 50% to about 75% inhibition, at least 50%
inhibition, at least 75% inhibition, about 80% inhibition, about
90% inhibition, and greater than 90% inhibition.
[0249] In some embodiments, provided is a method of inhibiting
PRMT5 activity in a subject in need thereof comprising
administering to the subject an effective amount of a compound
described herein (e.g., a compound of Formula (A), e.g., Formula
(I)), or a pharmaceutically acceptable salt thereof, or a
pharmaceutical composition thereof.
[0250] In certain embodiments, provided is a method of altering
gene expression in a cell which comprises contacting a cell with an
effective amount of a compound of Formula (A), e.g., Formula (I),
or a pharmaceutically acceptable salt thereof. In certain
embodiments, the cell in culture in vitro. In certain embodiments,
the cell is in an animal, e.g., a human. In certain embodiments,
the cell is in a subject in need of treatment.
[0251] In certain embodiments, provided is a method of altering
transcription in a cell which comprises contacting a cell with an
effective amount of a compound of Formula (A), e.g., Formula (I),
or a pharmaceutically acceptable salt thereof. In certain
embodiments, the cell in culture in vitro. In certain embodiments,
the cell is in an animal, e.g., a human. In certain embodiments,
the cell is in a subject in need of treatment.
[0252] In certain embodiments, a method is provided of selecting a
therapy for a subject having a disease associated with
PRMT5-mediated disorder or mutation comprising the steps of
determining the presence of PRMT5-mediated disorder or gene
mutation in the PRMT5 gene or and selecting, based on the presence
of PRMT5-mediated disorder a gene mutation in the PRMT5 gene a
therapy that includes the administration of a provided compound. In
certain embodiments, the disease is cancer.
[0253] In certain embodiments, a method of treatment is provided
for a subject in need thereof comprising the steps of determining
the presence of PRMT5-mediated disorder or a gene mutation in the
PRMT5 gene and treating the subject in need thereof, based on the
presence of a PRMT5-mediated disorder or gene mutation in the PRMT5
gene with a therapy that includes the administration of a provided
compound. In certain embodiments, the subject is a cancer
patient.
[0254] In some embodiments, a provided compound is useful in
treating a proliferative disorder, such as cancer, a benign
neoplasm, an autoimmune disease, or an inflammatory disease. For
example, while not being bound to any particular mechanism, PRMT5
has been shown to be involved in cyclin D1 dysregulated cancers.
Increased PRMT5 activity mediates key events associated with cyclin
D1-dependent neoplastic growth including CUL4 repression, CDT1
overexpression, and DNA re-replication. Further, human cancers
harboring mutations in Fbx4, the cyclin D1 E3 ligase, exhibit
nuclear cyclin D1 accumulation and increased PRMT5 activity. See,
e.g., Aggarwal et al., Cancer Cell. (2010) 18(4):329-40.
Additionally, PRMT5 has also been implicated in accelerating cell
cycle progression through G1 phase and modulating regulators of G1;
for example, PRMT5 may upregulate cyclin-dependent kinase (CDK) 4,
CDK6, and cyclins D1, D2 and E1. Moreover, PRMT5 may activate
phosphoinositide 3-kinase (PI3K)/AKT signaling. See, e.g., Wei et
al., Cancer Sci. (2012) 103(9):1640-50. PRMT5 has been reported to
play a role in apoptosis through methylation of E2F-1. See, e.g.,
Cho et al., EMBO J. (2012) 31:1785-1797; Zheng et al., Mol. Cell.
(2013) 52:37-51. PRMT5 has been reported to be an essential
regulator of splicing and affect the alternative splicing of
`sensor` mRNAs that can then lead to defects in downstream events
such as apoptosis. See, e.g., Bezzi et al., Genes Dev. (2013)
27:1903-1916. PRMT5 has been reported to play a role in the RAS-ERK
pathway. See, e.g., Andrew-Perez et al., Sci Signal. (2011) Sep.
13; 4(190)ra58 doi: 10.1126/scisignal.2001936. PRMT5 has been
reported to affect C/EBPb target genes through interaction with the
Mediator complex and hence affect cellular differentiation and
inflammatory response. See, e.g., Tsutsui et al., J. Biol. Chem.
(2013) 288:20955-20965. PRMT5 has been shown to methylate HOXA9
essential for ELAM expression during the EC inflammatory response.
See, e.g., Bandyopadhyay et al., Mol. Cell. Biol. (2012)
32:1202-1203. Thus in some embodiments, the inhibition of PRMT5 by
a provided compound is useful in treating the following
non-limiting list of cancers: breast cancer, esophageal cancer,
bladder cancer, lung cancer, hematopoietic cancer, lymphoma,
medulloblastoma, rectum adenocarcinoma, colon adenocarcinoma,
gastric cancer, pancreatic cancer, liver cancer, adenoid cystic
carcinoma, lung adenocarcinoma, head and neck squamous cell
carcinoma, brain tumors, hepatocellular carcinoma, renal cell
carcinoma, melanoma, oligodendroglioma, ovarian clear cell
carcinoma, and ovarian serous cystadenocarcinoma. See, e.g., Pal et
al., EMBO J. (2007) 26:3558-3569 (mantle cell lymphoma); Wang et
al., Mol. Cell Biol. (2008) 28:6262-77 (chronic lymphocytic
leukemia (CLL)); and Tae et al., Nucleic Acids Res. (2011)
39:5424-5438.
[0255] In some embodiments, the inhibition of PRMT5 by a provided
compound is useful in treating prostate cancer and lung cancer, in
which PRMT5 has been shown to play a role. See, e.g., Gu et al.,
PLoS One 2012; 7(8):e44033; Gu et al., Biochem. J. (2012)
446:235-241. In some embodiments, a provided compound is useful to
delay the onset of, slow the progression of, or ameliorate the
symptoms of cancer. In some embodiments, a provided compound is
administered in combination with other compounds, drugs, or
therapeutics to treat cancer.
[0256] In some embodiments, compounds described herein are useful
for treating a cancer including, but not limited to, acoustic
neuroma, adenocarcinoma, adrenal gland cancer, anal cancer,
angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma,
hemangiosarcoma), appendix cancer, benign monoclonal gammopathy,
biliary cancer (e.g., cholangiocarcinoma), bladder cancer, breast
cancer (e.g., adenocarcinoma of the breast, papillary carcinoma of
the breast, mammary cancer, medullary carcinoma of the breast),
brain cancer (e.g., meningioma; glioma, e.g., astrocytoma,
oligodendroglioma; medulloblastoma), bronchus cancer, carcinoid
tumor, cervical cancer (e.g., cervical adenocarcinoma),
choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer
(e.g., colon cancer, rectal cancer, colorectal adenocarcinoma),
epithelial carcinoma, ependymoma, endotheliosarcoma (e.g., Kaposi's
sarcoma, multiple idiopathic hemorrhagic sarcoma), endometrial
cancer (e.g., uterine cancer, uterine sarcoma), esophageal cancer
(e.g., adenocarcinoma of the esophagus, Barrett's adenocarinoma),
Ewing sarcoma, eye cancer (e.g., intraocular melanoma,
retinoblastoma), familiar hypereosinophilia, gall bladder cancer,
gastric cancer (e.g., stomach adenocarcinoma), gastrointestinal
stromal tumor (GIST), head and neck cancer (e.g., head and neck
squamous cell carcinoma, oral cancer (e.g., oral squamous cell
carcinoma (OSCC), throat cancer (e.g., laryngeal cancer, pharyngeal
cancer, nasopharyngeal cancer, oropharyngeal cancer)),
hematopoietic cancers (e.g., leukemia such as acute lymphocytic
leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myelocytic
leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myelocytic
leukemia (CML) (e.g., B-cell CML, T-cell CML), and chronic
lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL); lymphoma
such as Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and
non-Hodgkin lymphoma (NHL) (e.g., B-cell NHL such as diffuse large
cell lymphoma (DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL)),
follicular lymphoma, chronic lymphocytic leukemia/small lymphocytic
lymphoma (CLL/SLL), mantle cell lymphoma (MCL), marginal zone
B-cell lymphomas (e.g., mucosa-associated lymphoid tissue (MALT)
lymphomas, nodal marginal zone B-cell lymphoma, splenic marginal
zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt
lymphoma, lymphoplasmacytic lymphoma (i.e., "Waldenstrim's
macroglobulinemia"), hairy cell leukemia (HCL), immunoblastic large
cell lymphoma, precursor B-lymphoblastic lymphoma and primary
central nervous system (CNS) lymphoma; and T-cell NHL such as
precursor T-lymphoblastic lymphoma/leukemia, peripheral T-cell
lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g.,
mycosis fungiodes, Sezary syndrome), angioimmunoblastic T-cell
lymphoma, extranodal natural killer T-cell lymphoma, enteropathy
type T-cell lymphoma, subcutaneous panniculitis-like T-cell
lymphoma, anaplastic large cell lymphoma); a mixture of one or more
leukemia/lymphoma as described above; and multiple myeloma (MM)),
heavy chain disease (e.g., alpha chain disease, gamma chain
disease, mu chain disease), hemangioblastoma, inflammatory
myofibroblastic tumors, immunocytic amyloidosis, kidney cancer
(e.g., nephroblastoma a.k.a. Wilms' tumor, renal cell carcinoma),
liver cancer (e.g., hepatocellular cancer (HCC), malignant
hepatoma), lung cancer (e.g., bronchogenic carcinoma, small cell
lung cancer (SCLC), non-small cell lung cancer (NSCLC),
adenocarcinoma of the lung), leiomyosarcoma (LMS), mastocytosis
(e.g., systemic mastocytosis), myelodysplastic syndrome (MDS),
mesothelioma, myeloproliferative disorder (MPD) (e.g., polycythemia
Vera (PV), essential thrombocytosis (ET), agnogenic myeloid
metaplasia (AMM) a.k.a. myelofibrosis (MF), chronic idiopathic
myelofibrosis, chronic myelocytic leukemia (CML), chronic
neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES)),
neuroblastoma, neurofibroma (e.g., neurofibromatosis (NF) type 1 or
type 2, schwannomatosis), neuroendocrine cancer (e.g.,
gastroenteropancreatic neuroendoctrine tumor (GEP-NET), carcinoid
tumor), osteosarcoma, ovarian cancer (e.g., cystadenocarcinoma,
ovarian embryonal carcinoma, ovarian adenocarcinoma), papillary
adenocarcinoma, pancreatic cancer (e.g., pancreatic
andenocarcinoma, intraductal papillary mucinous neoplasm (IPMN),
Islet cell tumors), penile cancer (e.g., Paget's disease of the
penis and scrotum), pinealoma, primitive neuroectodermal tumor
(PNT), prostate cancer (e.g., prostate adenocarcinoma), rectal
cancer, rhabdomyosarcoma, salivary gland cancer, skin cancer (e.g.,
squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma,
basal cell carcinoma (BCC)), small bowel cancer (e.g., appendix
cancer), soft tissue sarcoma (e.g., malignant fibrous histiocytoma
(MFH), liposarcoma, malignant peripheral nerve sheath tumor
(MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous
gland carcinoma, sweat gland carcinoma, synovioma, testicular
cancer (e.g., seminoma, testicular embryonal carcinoma), thyroid
cancer (e.g., papillary carcinoma of the thyroid, papillary thyroid
carcinoma (PTC), medullary thyroid cancer), urethral cancer,
vaginal cancer, and vulvar cancer (e.g., Paget's disease of the
vulva).
[0257] In some embodiments, a provided compound is useful in
treating a metabolic disorder, such as diabetes or obesity. For
example, while not being bound to any particular mechanism, a role
for PRMT5 has been recognized in adipogenesis. Inhibition of PRMT5
expression in multiple cell culture models for adipogenesis
prevented the activation of adipogenic genes, while overexpression
of PRMT5 enhanced adipogenic gene expression and differentiation.
See, e.g., LeBlanc et al., Mol Endocrinol. (2012) 26:583-597.
Additionally, it has been shown that adipogenesis plays a pivotal
role in the etiology and progression of diabetes and obesity. See,
e.g., Camp et al., Trends Mol Med. (2002) 8:442-447. Thus in some
embodiments, the inhibition of PRMT5 by a provided compound is
useful in treating diabetes and/or obesity.
[0258] In some embodiments, a provided compound is useful to delay
the onset of, slow the progression of, or ameliorate the symptoms
of, diabetes. In some embodiments, the diabetes is Type 1 diabetes.
In some embodiments, the diabetes is Type 2 diabetes. In some
embodiments, a provided compound is useful to delay the onset of,
slow the progression of, or ameliorate the symptoms of, obesity. In
some embodiments, a provided compound is useful to help a subject
lose weight. In some embodiments, a provided compound could be used
in combination with other compounds, drugs, or therapeutics, such
as metformin and insulin, to treat diabetes and/or obesity.
[0259] In some embodiments, a provided compound is useful in
treating a blood disorder, e.g., a hemoglobinopathy, such as sickle
cell disease or .beta.-thalassemia. For example, while not being
bound to any particular mechanism, PRMT5 is a known repressor of
y-globin gene expression, and increased fetal y-globin (HbF) levels
in adulthood are associated with symptomatic amelioration in sickle
cell disease and P3-thalassemia. See, e.g., Xu et al.,
Haematologica. (2012) 97:1632-1640; Rank et al. Blood. (2010)
116:1585-1592. Thus in some embodiments, the inhibition of PRMT5 by
a provided compound is useful in treating a blood disorder, such as
a hemoglobinopathy such as sickle cell disease or
.beta.-thalassemia.
[0260] In some embodiments, a provided compound is useful to delay
the onset of, slow the progression of, or ameliorate the symptoms
of, sickle cell disease. In some embodiments, a provided compound
is useful to delay the onset of, slow the progression of, or
ameliorate the symptoms of, .beta.-thalassemia. In some
embodiments, a provided compound could be used in combination with
other compounds, drugs, or therapeutics, to treat a
hemoglobinopathy such as sickle cell disease or
.beta.-thalassemia.
[0261] In some embodiments, a provided compound is useful in
treating inflammatory and autoimmune disease. PRMT5 is reported to
activate NFkB signaling pathway through the methylation of p65.
PRMT5 is reported to interact with Death receptor 4 and Death
receptor 5 contributing to TRAIL-induced activation of inhibitor or
kB kinase (IKK) and nuclear factor-kB (NF-kB). See, e.g., Tanaka et
al., Mol. Cancer. Res. (2009) 7:557-569; Wei et al., Proc. Nat'l.
Acad. Sci. USA (2013) 110:13516-21.
[0262] The term "inflammatory disease" refers to those diseases,
disorders or conditions that are characterized by signs of pain
(dolor, from the generation of noxious substances and the
stimulation of nerves), heat (calor, from vasodilatation), redness
(rubor, from vasodilatation and increased blood flow), swelling
(tumor, from excessive inflow or restricted outflow of fluid),
and/or loss of function (functio laesa, which can be partial or
complete, temporary or permanent. Inflammation takes on many forms
and includes, but is not limited to, acute, adhesive, atrophic,
catarrhal, chronic, cirrhotic, diffuse, disseminated, exudative,
fibrinous, fibrosing, focal, granulomatous, hyperplastic,
hypertrophic, interstitial, metastatic, necrotic, obliterative,
parenchymatous, plastic, productive, proliferous, pseudomembranous,
purulent, sclerosing, seroplastic, serous, simple, specific,
subacute, suppurative, toxic, traumatic, and/or ulcerative
inflammation.
[0263] Exemplary inflammatory diseases include, but are not limited
to, inflammation associated with acne, anemia (e.g., aplastic
anemia, haemolytic autoimmune anaemia), asthma, arteritis (e.g.,
polyarteritis, temporal arteritis, periarteritis nodosa, Takayasu's
arteritis), arthritis (e.g., crystalline arthritis, osteoarthritis,
psoriatic arthritis, gouty arthritis, reactive arthritis,
rheumatoid arthritis and Reiter's arthritis), ankylosing
spondylitis, amylosis, amyotrophic lateral sclerosis, autoimmune
diseases, allergies or allergic reactions, atherosclerosis,
bronchitis, bursitis, chronic prostatitis, conjunctivitis, Chagas
disease, chronic obstructive pulmonary disease, cermatomyositis,
diverticulitis, diabetes (e.g., type I diabetes mellitus, type 2
diabetes mellitus), a skin condition (e.g., psoriasis, eczema,
burns, dermatitis, pruritus (itch)), endometriosis, Guillain-Barre
syndrome, infection, ischaemic heart disease, Kawasaki disease,
glomerulonephritis, gingivitis, hypersensitivity, headaches (e.g.,
migraine headaches, tension headaches), ileus (e.g., postoperative
ileus and ileus during sepsis), idiopathic thrombocytopenic
purpura, interstitial cystitis (painful bladder syndrome),
gastrointestinal disorder (e.g., selected from peptic ulcers,
regional enteritis, diverticulitis, gastrointestinal bleeding,
eosinophilic gastrointestinal disorders (e.g., eosinophilic
esophagitis, eosinophilic gastritis, eosinophilic gastroenteritis,
eosinophilic colitis), gastritis, diarrhea, gastroesophageal reflux
disease (GORD, or its synonym GERD), inflammatory bowel disease
(IBD) (e.g., Crohn's disease, ulcerative colitis, collagenous
colitis, lymphocytic colitis, ischaemic colitis, diversion colitis,
Behcet's syndrome, indeterminate colitis) and inflammatory bowel
syndrome (IBS)), lupus, multiple sclerosis, morphea, myeasthenia
gravis, myocardial ischemia, nephrotic syndrome, pemphigus
vulgaris, pernicious aneaemia, peptic ulcers, polymyositis, primary
biliary cirrhosis, neuroinflammation associated with brain
disorders (e.g., Parkinson's disease, Huntington's disease, and
Alzheimer's disease), prostatitis, chronic inflammation associated
with cranial radiation injury, pelvic inflammatory disease,
reperfusion injury, regional enteritis, rheumatic fever, systemic
lupus erythematosus, schleroderma, scierodoma, sarcoidosis,
spondyloarthopathies, Sjogren's syndrome, thyroiditis,
transplantation rejection, tendonitis, trauma or injury (e.g.,
frostbite, chemical irritants, toxins, scarring, burns, physical
injury), vasculitis, vitiligo and Wegener's granulomatosis.
[0264] In certain embodiments, the inflammatory disease is an acute
inflammatory disease (e.g., for example, inflammation resulting
from infection). In certain embodiments, the inflammatory disease
is a chronic inflammatory disease (e.g., conditions resulting from
asthma, arthritis and inflammatory bowel disease). The compounds
may also be useful in treating inflammation associated with trauma
and non-inflammatory myalgia. The compounds may also be useful in
treating inflammation associated with cancer.
[0265] Exemplary autoimmune diseases, include, but are not limited
to, arthritis (including rheumatoid arthritis,
spondyloarthopathies, gouty arthritis, degenerative joint diseases
such as osteoarthritis, systemic lupus erythematosus, Sjogren's
syndrome, ankylosing spondylitis, undifferentiated spondylitis,
Behcet's disease, haemolytic autoimmune anaemias, multiple
sclerosis, amyotrophic lateral sclerosis, amylosis, acute painful
shoulder, psoriatic, and juvenile arthritis), asthma,
atherosclerosis, osteoporosis, bronchitis, tendonitis, bursitis,
skin condition (e.g., psoriasis, eczema, burns, dermatitis,
pruritus (itch)), enuresis, eosinophilic disease, gastrointestinal
disorder (e.g., selected from peptic ulcers, regional enteritis,
diverticulitis, gastrointestinal bleeding, eosinophilic
gastrointestinal disorders (e.g., eosinophilic esophagitis,
eosinophilic gastritis, eosinophilic gastroenteritis, eosinophilic
colitis), gastritis, diarrhea, gastroesophageal reflux disease
(GORD, or its synonym GERD), inflammatory bowel disease (IBD)
(e.g., Crohn's disease, ulcerative colitis, collagenous colitis,
lymphocytic colitis, ischaemic colitis, diversion colitis, Behcet's
syndrome, indeterminate colitis) and inflammatory bowel syndrome
(IBS)), and disorders ameliorated by a gastroprokinetic agent
(e.g., ileus, postoperative ileus and ileus during sepsis;
gastroesophageal reflux disease (GORD, or its synonym GERD);
eosinophilic esophagitis, gastroparesis such as diabetic
gastroparesis; food intolerances and food allergies and other
functional bowel disorders, such as non-ulcerative dyspepsia (NUD)
and non-cardiac chest pain (NCCP, including costo-chondritis)).
[0266] In some embodiments, a provided compound is useful in
somatic cell reprogramming, such as reprogramming somatic cells
into stem cells. See, e.g., Nagamatsu et al., J Biol Chem. (2011)
286:10641-10648. In some embodiments, a provided compound is useful
in germ cell development, and are thus envisioned useful in the
areas of reproductive technology and regenerative medicine. See,
e.g., Ancelin et al., Nat. Cell. Biol. (2006) 8:623-630.
[0267] In some embodiments, compounds described herein can prepared
using methods shown in general Scheme 1. Compound B can be prepared
via ring opening of a chiral or racemic epoxide group. This amino
alcohol intermediate can be coupled to form an amide via normal
amide coupling methodology using a carboxylic acid A wherein Z is
hydrogen or via amination of an ester of intermediate A when Z is
an optionally substituted aliphatic group. Further substitution of
the tetrahydroisoquinoline ring and/or Cy can be carried out before
or after the coupling reaction.
##STR00462##
[0268] For example, exemplary Schemes 2 and 3 show such
couplings.
##STR00463##
##STR00464##
[0269] In some embodiments, an amide coupling step can be used to
provide a key intermediate for further synthesis, as shown, for
example, in exemplary Scheme 4.
##STR00465##
[0270] In some embodiments of the compounds described herein,
R.sup.12 or R.sup.13 is an amine. A non-limiting example of the
synthetic sequence used to prepare such analogs is provided herein
(see Scheme 5). In this example, an alcohol of Formula (Z-1) is
oxidized under suitable conditions S1 to affect transformation into
an intermediate ketone of Formula (Z-2). A ketone of Formula (Z-2)
can be contacted with a primary or secondary amine under suitable
conditions S2 to affect a reductive amination which can afford an
amino compound of Formula (Z-3).
##STR00466##
[0271] In some embodiments, the oxidation reaction S1 is carried
out directly with a stoichiometric oxidant. In some embodiments,
the stoichiometric oxidant is pyridinium chlorochromate. In some
embodiments, the stoichiometric oxidant is pyridinium dichromate.
In some embodiments, the stoichiometric oxidant is Dess-Martin
periodinane. In some embodiments, the stoichiometric oxidant is
prepared in situ. In some embodiments, the stoichiometric oxidant
is prepared in situ using sulfur trioxide pyridine complex and
dimethylsulfoxide. In some embodiments, the stoichiometric oxidant
is prepared in situ using oxallyl chloride and dimethylsulfoxide.
In some embodiments, the stoichiometric oxidant is prepared in situ
using a carbodiimide and dimethylsulfoxide. In some embodiments,
the stoichiometric oxidant is prepared in situ using
N-chlorosuccinimide and dimethylsulfide. In some embodiments, the
oxidation reaction S1 is catalyzed. In some embodiments, the
catalyst is (2,2,6,6-tetramethyl-piperidin-1-yl)oxyl. In some
embodiments, the catalyst is a ruthenium complex. In some
embodiments, the catalyst is a palladium complex. In some
embodiments, the catalyst is a copper complex. For examples of
standard methods and conditions for alcohol oxidation, see Epstein
et al., Chem. Rev. (1967) 67(3):247-260 and B. M. Trost ed.
"Comprehensive Organic Synthesis", (1991), Vol. 7, p 281-305.
[0272] In some embodiments, both the oxidation step S1 and
reductive amination step S2 occur in one pot. In some embodiments,
both the oxidation step S1 and the reductive amination step S2 are
carried out using the same catalyst. In some embodiments, the
catalyst is a rhodium complex. In some embodiments, the catalyst is
a ruthenium complex. In some embodiments, the catalyst is an
iridium complex.
[0273] In some embodiments, the reductive amination reaction S2 is
carried out using a borohydride. In some embodiments, the reductive
amination reaction S2 is carried out using sodium borohydride. In
some embodiments, the reductive amination reaction S2 is carried
out using sodium cyanoborohydride. In some embodiments, the
reductive amination reaction S2 is carried out using sodium
triacetoxyborohydride. In some embodiments, the reductive amination
reaction S2 is carried out using a borane. In some embodiments, the
reductive amination reaction S2 is carried out using a silyl
hydride. In some embodiments, the reductive amination reaction S2
is carried out using hydrogen. In some embodiments, the reductive
amination reaction S2 is carried out in two steps, by first
contacting a ketone of (Z-2) with an amine to form an intermediate
imine, and then reducing the intermediate imine under sufficient
conditions to afford a compound of Formula (Z-3). In some
embodiments, the reaction conditions S2 comprise addition of a
protic acid. In some embodiments, the reaction conditions S2
comprise addition of an aprotic acid. In some embodiments, the
reaction conditions S2 comprise in situ formation of the reducing
agent. In some embodiments, the reaction conditions S2 comprise a
catalyst. In some embodiments, the reaction conditions S2 comprise
a transition metal catalyst. In some embodiments, the reaction
conditions S2 comprise a palladium or nickel catalyst. In some
embodiments, the reductive amination reaction S2 is
stereoselective. In some embodiments, the stereoselective reductive
amination reaction S2 is carried out in the presence of a chiral
catalyst. For examples of standard methods and conditions for
reductive aminations, see Gomez et al., Adv. Synth. Catal. (2002)
344(10):1037-1057 and Abdel-Magid et al., J. Org. Chem. (1996),
61:3849.
[0274] An alterantive non-limiting synthetic sequence leading to
the aforementioned amine analogs is described herein (see Scheme
6). The hydroxyl moiety of a compound of Formula (Z-4) can be
transformed into a leaving group under sufficient conditions S3 to
afford a compound of Formula (Z-5). The leaving group of a compound
of Formula (Z-5) can be displaced with an amine under suitable
conditions S4 to produce an amino compound of Formula (Z-6).
##STR00467##
[0275] In some embodiments, LG of Formula (Z-5) is a halide. In
some embodiments, LG of Formula (Z-5) is bromine. In some
embodiments, LG of Formula (Z-5) is iodine. In some embodiments, LG
of Formula (Z-5) is a substituted or unsubstituted alkyl sulfonate.
In some embodiments, LG of Formula (Z-5) is a substituted or
unsubstituted aryl sulfonate. In some embodiments, LG of Formula
(Z-5) is methyl sulfonate. In some embodiments, LG of Formula (A-5)
is trifluoromethane sulfonate. In some embodiments, LG of Formula
(Z-5) is a toluene sulfonate. In some embodiments, LG of Formula
(Z-5) is a nitrobenzene sulfonate. In some embodiments, when LG of
Formula (Z-5) is halide, conditions S3 comprise a phosphoryl
halide. In some embodiments, when LG of Formula (Z-5) is halide,
conditions S3 comprise a sulfuryl halide. In some embodiments, when
LG of Formula (Z-5) is sulfonate, conditions S3 comprise a sulfonyl
halide. In some embodiments, when LG of Formula (Z-5) is sulfonate,
conditions S3 comprise a sulfonyl anhydride. For examples of
standard methods and conditions for organohalide or sulfonate ester
synthesis, see Lautens et al., Synthesis (2011) 2:342-346 or
Marcotullio et al., Synthesis (2006) 16:2760-2766.
[0276] In some embodiments, conditions S4 are neutral. In some
embodiments, conditions S4 comprise addition of a base. In certain
embodiments of conditions S4, the base is either inorganic or
organic. In certain embodiments of conditions S4, the base is
inorganic. In certain embodiments of conditions S4, the base is
organic. In certain embodiments of conditions S4, the base is a
metal acetate, alkoxide, amide, amidine, carbonate, hydroxide,
phenoxide, or phosphate. In certain embodiments of conditions S4,
the base is sodium, potassium, or caesium carbonate. In certain
embodiments of conditions S4, the base is sodium, potassium, or
caesium bicarbonate. In certain embodiments of conditions S4, the
base is 1,1,3,3-tetramethylguanidine,
1,4-diazabicyclo[2.2.2]octane, 1,8-bis(dimethylamino)naphthalene,
1,8-diazabicycloundec-7-ene, ammonia, diisopropylamine, imidazole,
N,N-diisopropylethylamine, piperidine, pyridine, pyrrolidine, or
triethylamine. In some embodiments of conditions S4, the solvent is
a polar protic solvent. In some embodiments of conditions S4, the
solvent is a polar aprotic solvent. In some embodiments of
conditions S4, the reaction is performed in the absence of solvent.
In some embodiments, conditions S4 comprise a catalyst. In some
embodiments of conditions S4, the catalyst is an iodide salt. In
some embodiments, both step S3 and the displacement step S4 occur
in one pot. In some embodiments, the hydroxyl moiety of a compound
of Formula (Z-4) is converted into a leaving group in situ. In some
embodiments, the hydroxyl moiety of a compound of Formula (Z-4) is
converted into a leaving group in situ using an azodicarboxylate
and an aryl or alkyl phosphine. For examples of standard methods
and conditions for amine syntheses through alkylation reactions,
see Salvatore et. al, Tetrahedron (2001) 57:7785-7811.
Examples
[0277] In order that the invention described herein may be more
fully understood, the following examples are set forth. It should
be understood that these examples are for illustrative purposes
only and are not to be construed as limiting this invention in any
manner.
Synthetic Methods
Intermediate Synthesis
2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline
##STR00468##
[0279] To a solution of 1,2,3,4-tetrahydroisoquinoline (15 g, 0.11
mol) in MeCN (100 mL) was added K.sub.2CO.sub.3 (30.7 g, 0.23 mol)
at 0.degree. C. 2-(bromomethyl)oxirane (17 g, 0.12 mol) was added
to the reaction after 1 h. The solution was stirred at 22.degree.
C. for 16 h at which time the solids were filtered and washed with
MeCN. The solution was concentrated and the residue was used in the
next step without further purification (17 g, Yield 78%). LCMS
(m/z): 190.1 (M+1).
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol
##STR00469##
[0281] To a solution of
2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (17 g, 0.09
mol) in EtOH (300 mL) at -78.degree. C. was slowly bubbled NH.sub.3
(g). The reaction mixture was then sealed and heated at 80.degree.
C. for 3 h. The reaction mixture was concentrated and the crude
product was used in next step without further purification (18 g,
Yield 96%). LCMS (m/z): 207.1 (M+1).
(R)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline
##STR00470##
[0283] To a solution of 1,2,3,4-tetrahydroisoquinoline (10 g, 0.15
mol) in THF (100 mL) at 0.degree. C. was added KF (22 g, 0.3 mmol).
After 1 h, (S)-oxiran-2-ylmethyl 3-nitrobenzenesulfonate (21.4 g,
0.17 mmol) was added and the resulting solution was stirred at
22.degree. C. for 16 h. The solid was removed by filtration and
washed with THF. The solution was concentrated and the crude
compound was used for next step without further purification (15 g,
Yield 53%). LCMS (m/z): 190.1 (M+1).
(S)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol
##STR00471##
[0285] To a solution of
(R)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (15 g,
0.08 mol) in EtOH (100 mL) at -78.degree. C. was slowly bubbled
NH.sub.3 (g). The reaction mixture was then sealed and heated at
80.degree. C. for 3 h. The reaction mixture was concentrated and
the crude product was used in next step without further
purification (15 g, Yield 92%). LCMS (m/z): 207.1 (M+1).
Alternative synthesis of
(R)-2-oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline
##STR00472##
[0287] To a solution of 1,2,3,4-tetrahydroisoquinoline (Ig, 7.52
mmol) in MeOH (40 mL) was added K.sub.2CO.sub.3 (5.19 g, 37.6 mmol)
under 0.degree. C. After stirring for 30 minutes,
(R)-2-(chloromethyl) oxirane (0.692 g, 7.52 mmol) was added the
reaction. The mixture was then stirred at 0.degree. C. overnight
before filtration and washing of the solid by with MeOH. The
solution was concentrated and the residue purified by column
separation to give the title compound as a colorless oil (70%
purity). This crude was used directly in the next step. LCMS (m/z):
190.1 (M+1).
Alternative synthesis of
(S)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol
##STR00473##
[0289] To a solution of
(R)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (200 mg,
5.2 mmol) in EtOH (20 mL) was added NH.sub.4OH (600 mg, 35.2 mmol)
at -78.degree. C. The reaction mixture was then warmed and heated
at 100.degree. C. for 3 h in a seal tube. The reaction mixture was
concentrated and the crude product was used in next step without
further purification. LCMS (m/z): 207.1 (M+1).
(S)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline
##STR00474##
[0291] To a solution of 1,2,3,4-tetrahydroisoquinoline (5 g, 7.52
mmol) in THF (100 mL) was added KF (8.57 g, 150.4 mmol) at
0.degree. C. (R)-oxiran-2-ylmethyl 3-nitrobenzenesulfonate (10.7 g,
41.4 mmol) was added to the reaction in 1 h. The solution was
stirred at room temperature overnight. The solid was removed by
filtration and washed with THF. The solution was then concentrated
and the residue used for next step without further purification
(11.3 g Yield 80%). LCMS (m/z): 190.1 (M+1).
(R)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol
##STR00475##
[0293] To a solution of
(S)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (2.2 g,
0.012 mol) in EtOH (30 mL), NH.sub.3 was bubbled to the solution
under -78.degree. C. The reaction mixture was then sealed and
heated at 80.degree. C. for 3 h. After LCMS indicated completion of
the reaction, the mixture was concentrated and the crude product
was used in next step without further purification (2.2 g, Yield
90%). LCMS (m/z): 207.1 (M+1).
Compound 1
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-phenoxyacetamide
##STR00476##
[0295] To a stirred mixture of 2-phenoxyacetic acid (100 mg, 0.658
mmol) in DCM (10 mL) was added TEA (200 mg, 1.98 mmol),
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (135 mg,
0.658 mmol) and HATU (250 mg, 0.658 mmol). The mixture was stirred
at 25.degree. C. for 16 hours then quenched with water (20 mL) and
extracted with DCM (3.times.20 mL). The combined extracts were
washed with brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4
and concentrated. The residue was then purified by prep-HPLC to
afford the title compound (76 mg, 34% yield). .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 7.31-7.27 (m, 2H), 7.14-7.08 (m, 3H),
7.05-6.98 (m, 2H), 6.93 (d, J=8.0 Hz, 2H), 4.54 (s, 2H), 4.55-4.52
(m, 1H), 4.42 (s, 2H), 4.06-4.00 (m, 1H), 3.71 (s, 2H), 3.48-3.38
(m, 2H), 2.91 (d, J=5.6 Hz, 2H), 2.84 (d, J=5.6 Hz, 2H), 2.60 (d,
J=6.8 Hz, 2H) ppm. LCMS (m/z): 341.2 [M+H].sup.+.
[0296] Compound 6
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-ylox-
y)acetamide
##STR00477##
[0297] Step 1: 2-(quinolin-8-yloxy)acetic acid
##STR00478##
[0299] To a stirred mixture of quinolin-8-ol (500 mg, 3.45 mmol) in
MeCN (5 mL) was added ethyl bromoacetate (687 mg, 4.14 mmol) and
K.sub.2CO.sub.3 (952 mg, 6.90 mmol). The mixture was stirred at
80.degree. C. for 4 hours until TLC analysis showed completion of
the reaction. The mixture was filtered and the filtrate
concentrated. NaOH (276 mg, 6.90 mmol) and water:EtOH (1:1, 10 mL)
was then added to the residue and the resulting mixture stirred at
50.degree. C. for 4 hours. After cooling, the mixture was acidified
by addition of 1M HCl to pH 3 and then extracted with ethyl acetate
(2.times.30 mL). The combined organic layers were washed with brine
(30 mL), dried over anhydrous Na.sub.2SO.sub.4 and concentrated to
yield the crude target product which was used directly for the next
step.
Step 2:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinoli-
n-8-yloxy)acetamide
##STR00479##
[0301] To a stirred mixture of 2-(quinolin-8-yloxy)acetic acid (100
mg, 0.492 mmol) in DMF (5 mL) was added DIEA (95 mg, 0.738 mmol),
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (100 mg,
0.492 mmol) and BOP-Cl (151 mg, 0.591 mmol). The mixture was
stirred at 25.degree. C. for 48 hours then the reaction mixture was
quenched by addition of water (20 mL) and extracted with DCM
(3.times.20 mL). The combined organic extracts were washed with
brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4 and
concentrated. The residue was then purified by prep-HPLC to afford
the desired product (8 mg, Yield: 4%). .sup.1H NMR (400 MHz, MeOD)
.delta.=8.91 (d, J=4.3 Hz, 1H), 8.42 (d, J=8.3 Hz, 1H), 7.69-7.54
(m, 3H), 7.30 (d, J=7.3 Hz, 1H), 7.12-6.96 (m, 4H), 4.78 (s, 2H),
4.18-4.07 (m, 1H), 3.71 (s, 2H), 3.60-3.49 (m, 1H), 3.48-3.40 (m,
1H), 2.89 (d, J=5.8 Hz, 2H), 2.84 (d, J=4.8 Hz, 2H), 2.65-2.61 (m,
2H). LCMS (m/z): 392.2 (M+1).
Compound 7
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-fluorophenoxy-
)acetamide
##STR00480##
[0302] Step 1: 2-(3-fluorophenoxy)acetic acid
##STR00481##
[0304] To a stirred mixture of 3-fluorophenol (100 mg, 0.893 mmol)
in MeCN (5 mL) was added ethyl bromoacetate (222 mg, 1.34 mmol) and
K.sub.2CO.sub.3 (369 mg, 2.68 mmol). The mixture was stirred at
80.degree. C. for 4 hours. The mixture was then filtered and the
filtrate concentrated. NaOH (71 mg, 1.79 mmol) and water:EtOH (1:1,
10 mL) was added to the residue and the mixture stirred at
50.degree. C. for 4 hours. The mixture was acidified by adding 1M
HCl, and then extracted with ethyl acetate (2.times.30 mL). The
combined extracts were washed with brine (30 mL), dried over
anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was
directly for the next step. LCMS (m/z): 171.0 (M+1).
Step 2:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-fluor-
ophenoxy)acetamide
##STR00482##
[0306] To a stirred mixture of 2-(3-fluorophenoxy)acetic acid (252
mg, 0.893 mmol) in DCM (5 mL) was added DIEA (173 mg, 1.34 mmol),
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (183 mg,
0.893 mmol), and Bop-Cl (273 mg, 1.07 mmol). The mixture was
stirred at 25.degree. C. for 16 hours then quenched with water (20
mL), extracted with DCM (3.times.20 mL). The combined extracts were
washed with brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4
and concentrated. The residue was then purified by prep-HPLC to
afford the product (18 mg, Yield 5.6%). .sup.1H NMR (400 MHz,
METHANOL-d.sub.4) .delta.=7.26-7.29 (m, 1H), 7.13-7.02 (m, 4H),
6.76-6.69 (m, 3H), 4.55 (s, 2H), 4.06-4.00 (m, 1H), 3.71 (s, 1H),
3.47-3.31 (m, 2H), 2.91-2.90 (m, 2H), 2.86-2.77 (m, 2H), 2.61 (d,
J=6.0 Hz, 2H). LCMS (m/z): 359.1 (M+1).
Compound 8
2-(3-cyanophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-
acetamide
##STR00483##
[0307] Step 1: 2-(3-cyanophenoxy) acetic acid
##STR00484##
[0309] To a stirred mixture of 3-hydroxybenzonitrile (100 mg, 0.840
mmol) in MeCN (5 mL) was added ethyl bromoacetate (209 mg, 1.26
mmol) and K.sub.2CO.sub.3 (350 mg, 2.52 mmol). The mixture was
stirred at 80.degree. C. for 4 hours until TLC showed completion of
the reaction. The mixture was filtered and the filtrate
concentrated. NaOH (67 mg, 1.68 mmol) and water:EtOH (1:1, 10 mL)
was added to the residue and the mixture stirred at 50.degree. C.
for 4 hours. After cooling, the mixture was acidified by 1M HCl,
extracted with ethyl acetate (2.times.30 mL). The combined extracts
were washed with brine (30 mL), dried over anhydrous
Na.sub.2SO.sub.4 and concentrated. The residue was used directly in
the next step. LCMS (m/z): 178.0 (M+1).
Step 2:
2-(3-cyanophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydrox-
ypropyl)acetamide
##STR00485##
[0311] To a stirred mixture of 2-(3-cyanophenoxy) acetic acid (100
mg, 0.565 mmol) in DCM (5 mL) was added DIEA (109 mg, 0.85 mmol),
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (116 mg,
0.565 mmol) and BopC1 (173 mg, 0.678 mmol). The resulting mixture
was stirred at 25.degree. C. for 16 hours until LCMS showed the
completion of the reaction. The reaction mixture was quenched by
addition of water (20 mL) then extracted with DCM (3.times.20 mL).
The combined organic extracts were washed with brine (20 mL), dried
over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was
purified by prep-HPLC to afford the desired final product (24 mg,
Yield 12%). .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. 7.48
(dd, J1=J.sub.2=7.6 Hz, 1H), 7.37 (d, J=7.6 Hz, 1H), 7.25-7.21 (m,
2H), 7.13-7.03 (m, 4H), 4.60 (s, 2H), 4.04-4.00 (m, 1H), 3.71 (s,
2H), 3.44 (d, J=6.0 Hz, 2H), 2.93-2.80 (m, 4H), 2.62 (d, J=5.6 Hz,
2H). LCMS (m/z): 366.1 (M+1).
Compound 9
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(p-tolyloxy)acet-
amide
##STR00486##
[0312] Step 1: ethyl 2-(p-tolyloxy)acetate
##STR00487##
[0314] To a mixture of p-cresol (500 mg, 4.63 mmol) and ethyl
2-bromoacetate (928 mg, 5.56 mmol) in CH.sub.3CN (10 mL) was added
K.sub.2CO.sub.3 (3 g, 21.7 mmol). The reaction mixture was stirred
at 80.degree. C. for 4 h. The solid was removed by filtration and
the filtrate was concentrated to give the title compound which was
used in the next step without further purification.
Step 2: 2-(p-tolyloxy)acetic acid
##STR00488##
[0316] To a solution of ethyl 2-(p-tolyloxy)acetate (200 mg, 1
mmol) in EtOH (10 ml) was added 10% NaOH solution(10 ml) at
26.degree. C. The mixture was stirred for 30 min, concentrated then
water (20 mL) added to it before washing with ethyl acetate
(2.times.20 mL). The aqueous layer was acidified with 2N HCL until
pH 3 and extracted with EA (2.times.20 ml). The organic layer was
washed with brine (30 mL), dried over Na.sub.2SO.sub.4 and
concentrated to give the title compound which was used in next step
without further purification.
Step 3:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(p-tolyl-
oxy)acetamide
##STR00489##
[0318] A mixture of compound 2-(p-tolyloxy)acetic acid (100 mg,
0.60 mmol), 1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol
(124 mg, 0.60 mmol), BOP-Cl (183 mg, 0.72 mmol) and DIPEA (1 mL) in
DCM (10 mL) was stirred at room temperature for 4 h. The solvent
was removed by concentration and the crude product was purified by
pre-HPLC to give the title compound (27.8 mg, yield 13.1%). .sup.1H
NMR (500 MHz, MeOD): .delta. 7.32-7.25 (m, 3H), 7.20 (d, J=7.2 Hz,
1H), 7.12 (d, J=8.4 Hz, 2H), 6.90 (d, J=8.8 Hz, 2H), 4.65-4.52
(br.s, 1H), 4.52 (s, 2H), 4.46-4.30 (br.s, 1H), 4.30-4.24 (m, 1H),
3.85-3.70 (br.s, 1H), 3.43 (d, J=5.6 Hz, 1H), 3.26-3.17 (m, 4H),
2.26 (s, 3H) ppm; ESI-MS (m/z): 355.2 [M+1].sup.+.
Compound 12
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(4-methoxyphenox-
y)acetamide
##STR00490##
[0319] Step 1: ethyl 2-(4-methoxyphenoxy)acetate
##STR00491##
[0321] To a solution of 4-methoxyphenol (500 mg, 4.03 mmol) in
CH.sub.3CN (10 mL) was added ethyl 2-bromo-2-methylpropanoate (807
mg, 4.84 mmol) and K.sub.2CO.sub.3 (3 g, 21.7 mmol) at 25.degree.
C. The mixture was refluxed for 16 h. The mixture was then diluted
with water (100 mL), extracted with ethyl acetate (2.times.50 mL)
and the combined organic layers washed with brine (30 mL), dried
over Na.sub.2SO.sub.4 and concentrated to give the title compound
which was used in next step without further purification.
Step 2: 2-(4-methoxyphenoxy)acetic acid
##STR00492##
[0323] To a solution of ethyl 2-methyl-2-phenoxypropanoate (210 mg,
1 mmol) in EtOH (10 ml) was added 10% NaOH aqueous solution (10 mL)
at 26.degree. C. The mixture was stirred for 30 min and then
concentrated before the addition of water (20 mL) and washing with
ethyl acetate (2.times.20 mL). The aqueous layer was acidified with
2N HCL until pH 3 and extracted with ethyl acetate (2.times.20 ml).
The organic layer was washed with brine (30 mL), dried over
Na.sub.2SO.sub.4 and concentrated to give the title compound which
was used in next step without further purification.
Step 3:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(4-metho-
xyphenoxy)acetamide
##STR00493##
[0325] A mixture of compound 2-(4-methoxyphenoxy)acetic acid (100
mg, 0.55 mmol),
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (113.7 mg,
0.55 mmol), BOP-Ci (177.2 mg, 0.696 mmol) and DIPEA (1 mL) in DCM
(10 mL) was stirred at room temperature for 4 h. The solvent was
removed by concentration and the crude product was purified by
pre-HPLC to give the title compound (13.3 mg, yield 6.5%). .sup.1H
NMR (500 MHz, MeOD): .delta. 7.11-7.06 (m, 3H), 7.02-7.00 (m, 1H),
6.90-6.60 (m, 4H), 4.45 (s, 2H), 4.01-3.98 (m, 1H), 3.73 (s, 3H),
3.66 (s, 1H), 3.41-3.39 (m, 2H), 2.90-2.87 (m, 2H), 2.81-2.76 (m,
2H), 2.55 (d, J=6.0 Hz, 2H) ppm; ESI-MS (m/z): 371.2
[M+1].sup.+.
Compound 15
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(o-tolyloxy)acet-
amide
##STR00494##
[0326] Step 1: 2-(o-tolyloxy)acetic acid
##STR00495##
[0328] To a stirred solution of o-cresol (200 mg, 1.85 mmol) in
MeCN (5 mL) was added ethyl bromoacetate (461 mg, 2.78 mmol) and
K.sub.2CO.sub.3 (766 mg, 5.55 mmol). The mixture was stirred at
80.degree. C. for 4 hours. The mixture was filtered and the
filtrate concentrated. NaOH (150 mg, 3.70 mmol) and H.sub.2O/EtOH
(1:1, 10 mL) was then added to the mixture and the mixture stirred
at 50.degree. C. for 4 hours. After cooling, the mixture was
acidified by adding 1M HCl then extracted with ethyl acetate
(2.times.30 mL). The combined organic extracts were washed with
brine (30 mL), dried over anhydrous Na.sub.2SO.sub.4 and
concentrated. The residue was used directly in the next step
without further purification.
Step 2:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(o-tolyl-
oxy)acetamide
##STR00496##
[0330] To a stirred mixture of 2-(o-tolyloxy)acetic acid (100 mg,
0.60 mmol) in DCM (5 mL) was added DIEA (116 mg, 0.90 mmol),
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (124 mg,
0.60 mmol) and Bop-Cl (183 mg, 0.72 mmol). The mixture was stirred
at 25.degree. C. for 16 hours then quenched by addition of water
(20 mL). The resulting mixture was extracted with DCM (3.times.20
mL). The combined extracts were washed with brine (20 mL), dried
over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was
purified by prep-HPLC to afford the product (79 mg, Yield 37%).
.sup.1H NMR (400 MHz, MeOD) .delta. 7.14-7.01 (m, 6H), 6.95-6.82
(m, 2H), 4.55 (s, 2H), 4.04-4.01 (m, 1H), 3.72-3.63 (m, 2H),
3.55-3.46 (m, 1H), 3.42-3.34 (m, 1H), 2.93-2.84 (m, 2H), 2.83-2.74
(m, 2H), 2.57 (d, J=6.3 Hz, 2H), 2.27 (s, 3H). LCMS (m/z): 355.1
(M+1).
Compound 19
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-phenoxypropanami-
de
##STR00497##
[0331] Step 1: ethyl 2-phenoxypropanoate
##STR00498##
[0333] To a solution of NaH (765 mg, 31.89 mmol) in DMF (10 mL) was
added phenol (1 g, 10.63 mmol) at 25.degree. C. The mixture was
heated at reflux temperature for 15 min, and then ethyl
2-bromopropanoate (2.3 g, 12.75 mmol) was added. The resulting
mixture was stirred at 25.degree. C. for another 16 h before
quenching with water (50 mL). The mixture was extracted with ethyl
acetate (3.times.20 mL). The combined organic layers were washed
with brine (30 mL), dried over Na.sub.2SO.sub.4 and concentrated to
give the title compound (1.8 g, 85.7%) as colorless oil which was
used in next step without further purification.
Step 2: 2-phenoxypropanoic acid
##STR00499##
[0335] To a solution of ethyl 2-phenoxypropanoate (1.8 g, 0.9 mmol)
in EtOH (16 ml) was added a solution of NaOH (0.44 g, 1.1 mmol) in
H.sub.2O (4 ml) at 25.degree. C. The mixture was stirred for 30 min
before being concentrated. The residue had water (20 mL) added and
washed with ethyl acetate (2.times.20 mL). The aqueous layer was
acidified with 2N HCL until pH 3 and extracted with ethyl acetate
(2.times.20 mL). The combined organic layers were washed with brine
(30 mL), dried over Na.sub.2SO.sub.4 and concentrated to give the
title compound (1.1 g, 73.3%) as a white solid which was used in
next step without further purification.
Step 3:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-phenoxyp-
ropanamide
##STR00500##
[0337] To a solution of 2-phenoxypropanoic acid (200 mg, 1.2 mmol)
in DMF (4 ml) was added TEA 364 mg, 3.6 mmol), HOBt (243 mg, 1.8
mmol), EDCI (346 mg, 1.8 mmol) and
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (297 mg,
1.44 mmol) at 24.degree. C. The reaction mixture was stirred for 16
h until TLC showed completion of the reaction. After evaporation of
the solvent, the residue was purified by prep-HPLC separation to
give the title compound as the formate salt (34 mg, 8%). .sup.1H
NMR (500 MHz, MeOD): .delta. 8.40 (s, 1H), 7.34-7.25 (m, 5H), 7.17
(d, J=7.2 Hz, 1H), 7.00-6.96 (m, 3H), 4.78-4.76 (m, 1H), 4.26-4.11
(m, 3H), 3.43-3.33 (m, 4H), 3.14-3.12 (m, 2H), 3.08-3.02 (m, 1H),
2.95-2.90 (m, 1H), 1.57 (d, J=6.4 Hz, 3H) ppm; ESI-MS (m/z): 354.2
[M+1].sup.+.
Compound 20
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-methoxyphenox-
y)acetamide
##STR00501##
[0338] Step 1: 2-(3-methoxyphenoxy)acetic acid
##STR00502##
[0340] To a stirred mixture of 3-methoxyphenol (200 mg, 1.61 mmol)
in MeCN (5 mL) was added ethyl bromoacetate (402 mg, 2.42 mmol) and
K.sub.2CO.sub.3 (672 mg, 4.83 mmol). The mixture was stirred at
80.degree. C. for 4 hours, filtered and the filtrate concentrated.
NaOH (129 mg, 3.22 mmol) and H2O/EtOH (1:1, 10 mL) was added to the
mixture. The reaction mixture was stirred at 50.degree. C. for 4
hours then acidified by 1M HCl and then extracted with ethyl
acetate (2.times.30 mL). The combined organic extracts were washed
with brine (30 mL), dried over anhydrous Na.sub.2SO.sub.4 and
concentrated with the residue used directly for the next step. LCMS
(m/z): 183.0 (M+1).
Step 2:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-metho-
xyphenoxy)acetamide
##STR00503##
[0342] To a stirred mixture of 2-(3-methoxyphenoxy)acetic acid (100
mg, 0.549 mmol) in DCM (5 mL) was added DIEA (106 mg, 0.824 mmol),
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (113 mg,
0.549 mmol) and Bop-Cl (168 mg, 0.66 mmol). The mixture was stirred
at 25.degree. C. for 16 hours then the reaction mixture was
quenched with water (20 mL), extracted with DCM (3.times.20 mL).
The combined extracts were washed with brine (20 mL), dried over
anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was
purified by prep-HPLC to afford the desired product (61 mg, Yield
32%). .sup.1H NMR (400 MHz, MeOD) .delta. 7.21 (dd,
J.sub.1=J.sub.2=8.0 Hz, 1H), 7.17-7.03 (m, 4H), 6.59-6.51 (m, 3H),
4.52 (s, 2H), 4.05-4.01 (m, 1H), 3.77 (s, 3H), 3.73 (d, J=2.8 Hz,
2H), 3.47-3.37 (m, 2H), 2.92 (d, J=5.2 Hz, 2H), 2.87 (d, J=5.2 Hz,
2H), 2.61 (d, J=6.0 Hz, 2H). LCMS (m/z): 371.1 (M+1).
Compound 21
2-(4-acetamidophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypro-
pyl)acetamide
##STR00504##
[0343] Step 1: ethyl 2-(4-acetamidophenoxy)acetate
##STR00505##
[0345] To a solution of N-(4-hydroxyphenyl)acetamide (500 mg, 3.31
mmol) in CH.sub.3CN (10 mL) was added ethyl
2-bromo-2-methylpropanoate (663 mg, 3.97 mmol) and K.sub.2CO.sub.3
(3 g, 21.7 mmol) at 25.degree. C. The mixture was heated at reflux
for 16 h. The mixture had water (100 mL) added and extracted with
ethyl acetate (2.times.50 mL). The combined organic layers were
washed with brine (30 mL), dried over Na.sub.2SO.sub.4 and
concentrated to give the title compound which was used in next step
without further purification.
Step 2: 2-(4-acetamidophenoxy)acetic acid
##STR00506##
[0347] To a solution of ethyl 2-(4-acetamidophenoxy)acetate (237
mg, 1 mmol) in EtOH (10 ml) was added 10% NaOH aqueous solution (10
mL) at 26.degree. C. The mixture was stirred for 30 min and then
concentrated. The residue was diluted with water (20 mL) and washed
ethyl acetate (2.times.20 mL). The aqueous layer was then acidified
with 2N HCL until pH 3 and extracted with ethyl acetate (2.times.20
ml). The combined organic layers were washed with brine (30 mL),
dried over Na.sub.2SO.sub.4 and concentrated to give the title
compound which was used in next step without further
purification.
Step 3:
2-(4-acetamidophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hy-
droxypropyl)acetamide
##STR00507##
[0349] A mixture of compound 2-(4-methoxyphenoxy)acetic acid (100
mg, 0.51 mmol),
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (132 mg,
0.51 mmol), BOP-Cl (156 mg, 0.61 mmol) and DIPEA (1 mL) in DCM (10
mL) was stirred at room temperature for 4 h. The solvent was
removed by concentration and the crude product was purified by
pre-HPLC to give the title compound (7.8 mg, yield: 3.8%). .sup.1H
NMR (500 MHz, MeOD): .delta. 7.45 (d, d, J=8.8 Hz, 2H), 7.11-7.05
(m, 3H), 7.02-7.00 (m, 1H), 6.88 (d, J=8.8 Hz, 2H), 4.50 (s, 2H),
4.01-3.98 (m, 1H), 3.66 (d, J=3.2 Hz, 2H), 3.41 (dd, J=0.8, 6.0 Hz,
2H), 2.89-2.87 (m, 2H), 2.80-2.76 (m, 2H), 2.55 (d, J=6.0 Hz, 2H),
2.08 (s, 3H) ppm; ESI-MS (m/z): 398.2 [M+1].sup.+.
Compound 23
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-methyl-1H-in-
dazol-6-yl)oxy)acetamide
##STR00508##
[0350] Step 1: 1-methyl-1H-indazol-6-amine
##STR00509##
[0352] To a solution of 1-methyl-6-nitro-1H-indazole (1.2 g, 0.7
mmol) in EtOH (50 ml) and THF (15 ml) was added PtO.sub.2 (125 mg)
at 26.degree. C. The mixture was stirred for 1.5 h at 26.degree. C.
under a H.sub.2 atmosphere at 30 Psi. Once the reaction was
complete by TLC analysis, the mixture was filtered and the filtrate
concentrated to give the target crude product as a white solid
which was used in the next step without further purification (1.0
g, Yield 90%). LCMS (m/z): 148.1 (M+1).
Step 2: 1-methyl-1H-indazol-6-ol
##STR00510##
[0354] To a solution of 1-methyl-1H-indazol-6-amine (300 mg, 2.04
mmol) in H.sub.2O/H.sub.2SO.sub.4=1:1 (5 ml) was added NaNO.sub.2
(141 mg, 2.04 mmol) at 0.degree. C. The mixture was then stirred
for 2 h at 25.degree. C. before being added to water (0.5 ml) and
stirred for a further 2 h at 120.degree. C. Once the reaction was
complete by TLC, the mixture was treated with NaHCO.sub.3 until
pH=7. The mixture was then extracted with ethyl acetate (2.times.10
ml) and the organic layer washed with brine (20 ml), dried over
Na.sub.2SO.sub.4 and concentrated to give 1-methyl-1H-indazol-6-ol
as a red solid which was used in the next step without further
purification (300 mg, 99.0%). LCMS (m/z): 149.1 (M+1).
Step 3: 2-((1-methyl-1H-indazol-6-yl)oxy)acetic acid
##STR00511##
[0356] To a solution of NaH (146 mg, 6.06 mmol) in DMF (3 mL) was
added 1-methyl-1H-indazol-6-ol (300 mg, 2.02 mmol) at 25.degree. C.
After stirring for 5 minutes, ethyl 2-bromoacetate (406 mg, 2.43
mmol) was added and stirred for 16 h at 25.degree. C. The mixture
was then diluted with water (50 mL) and washed with ethyl acetate
(2.times.20 mL). The water layer was then acidified by adding with
2N HCL until pH3 and then extracted with ethylacetate (2.times.20
ml). The combined organic layers were washed with brine (30 mL),
dried over Na.sub.2SO.sub.4 and concentrated to give the crude
product (300 mg, 63.4%) as colorless oil. It was used in next step
without further purification. LCMS (m/z): 207.1 (M+1).
Step 4:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-meth-
yl-1H-indazol-6-yl)oxy)acetamide
##STR00512##
[0358] To a solution of 2-((1-methyl-1H-indazol-6-yl)oxy)acetic
acid (200 mg, 0.97 mmol) in DMF (4 ml) was added TEA (294 mg, 2.91
mmol), HOBt (196 mg, 1.45 mmol), EDCI (278 mg, 1.45 mmol) and
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (240 mg, 1.2
mmol) at 27.degree. C. The reaction mixture was stirred for 16 h at
27.degree. C. Once the reaction was complete and evaporation of the
solvent, the mixture was purified by preparative HPLC to give the
formate salt of the title compound (26 mg, 11.9%) as a white solid.
.sup.1H NMR (400 MHz, MeOD) .delta.7.91 (s, 1H), 7.67 (d, J=8.8 Hz,
1H), 7.27-7.13 (m, 4H), 7.02 (s, 1H), 6.95 (dd, J=8.8 Hz,
J.sub.2=2.0 Hz, 1H), 4.68 (s, 2H), 4.26-4.25 (m, 3H), 4.02 (s, 3H),
3.47-3.33 (m, 4H), 3.14-3.06 (m, 4H). LCMS (m/z): 395.2 (M+1).
Compound 24
2-(cyclohexyloxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)a-
cetamide
##STR00513##
[0359] Step 1: 2-(cyclohexyloxy)acetic acid
##STR00514##
[0361] To a solution of compound NaH (719 mg, 29.94 mmol) in DMF
(10 mL) was added cyclohexanol (1 g, 9.98 mmol) at 0.degree. C.
After stirring for 5 minutes, ethyl 2-bromoacetate (2 g, 11.98
mmol) was added and the mixture stirred for another 16 h. Once
complete, the mixture was treated with water (50 mL) and washed
with ethyl acetate (2.times.20 mL). The water layer was treat with
2N HCL until pH 3. The water layer was extracted with ethyl acetate
(2.times.20 ml) and the combined organic layers washed with brine
(30 mL), dried over Na.sub.2SO.sub.4 and concentrated to give the
desired product (500 mg, 27.8%) as colorless oil which was used in
next step without further purification.
Step 2:
2-(cyclohexyloxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxy-
propyl)acetamide
##STR00515##
[0363] To a solution of 2-(cyclohexyloxy)acetic acid (100 mg, 0.632
mmol) in DMF (3 ml) was added TEA (191 mg, 1.896 mmol), HOBT (128
mg, 0.948 mmol), EDCI (182 mg, 0.948 mmol) and
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (156 mg,
0.758 mmol) at 27.degree. C. The mixture was stirred for 16 h until
the reaction was complete. After evaporation of the solvent, the
residue was purified by prep-HPLC to afford the title compound as
the formate salt (26 mg, Yield 11.9%). .sup.1H NMR (400 MHz, MeOD):
.delta. 8.48 (s, 1H), 7.27-7.22 (m, 3H), 7.16-7.15 (m, 1H),
4.23-4.18 (m, 3H), 4.00 (s, 2H), 3.44-3.33 (m, 3H), 3.13-3.02 (m,
4H), 1.95-1.13 (m, 2H), 1.78-1.76 (m, 2H), 1.63-1.61 (m, 1H),
1.37-1.27 (m, 5H) ppm; ESI-MS (m/z): 469.3 [M+1].sup.+.
Compound 25
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-methyl-2-phenoxy-
propanamide
##STR00516##
[0364] Step 1: ethyl 2-methyl-2-phenoxypropanoate
##STR00517##
[0366] To a solution of phenol (2 g, 21.25 mmol) in CH.sub.3CN (50
mL) was added ethyl 2-bromo-2-methylpropanoate (5 g, 25.5 mmol) and
Cs.sub.2CO.sub.3 (20 g, 63.75 mmol) at 25.degree. C. The mixture
was heated at reflux for 16 h then water (100 mL) added and
extracted with ethyl acetate (2.times.50 mL). The organic layer was
washed with brine (30 mL), dried over Na.sub.2SO.sub.4 and
concentrated to give the title compound (2.1 g, 47.7%) as colorless
oil which was used in next step without further purification.
Step 2: 2-methyl-2-phenoxypropanoic acid
##STR00518##
[0368] To a solution of ethyl 2-methyl-2-phenoxypropanoate (2.0 g,
9.6 mmol) in EtOH (16 ml) was added a solution of NaOH (0.46 g,
11.5 mmol) in H.sub.2O (4 ml) at 26.degree. C. The mixture was
stirred for 30 min then concentrated. Water was added (20 mL) and
washed with ethyl acetate (2.times.20 mL). The aqueous layer was
acidified with 2N HCL until pH 3 and extracted with ethyl acetate
(2.times.20 ml). The combined organic layers were washed with brine
(30 mL), dried over Na.sub.2SO.sub.4 and concentrated to give the
title compound (1.6 g, 94.1%) as a white solid which was used in
next step without further purification.
Step 3:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-methyl-2-
-phenoxypropanamide
##STR00519##
[0370] To a solution of compound 2-methyl-2-phenoxypropanoic acid
(200 mg, 1.11 mmol) in DMF (4 ml) was added TEA (336 mg, 3.33
mmol), HOBt (225 mg, 1.66 mmol), EDCI (320 mg, 1.66 mmol) and
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (320 mg,
1.33 mmol) at 24.degree. C. The reaction mixture was stirred for 16
h at 24.degree. C. After evaporation of the solvent, the residue
was purified by prep-HPLC separation to give the title compound as
the formate salt (33 mg, 8%). .sup.1H NMR (500 MHz, MeOD): .delta.
8.40 (s, 1H), 7.32-7.25 (m, 5H), 7.19 (d, J=6.8 Hz, 1H), 7.09 (t,
J=7.6 Hz, 1H), 6.98-6.96 (m, 2H), 4.33 (s, 2H), 4.29-4.22 (m, 1H),
3.49 (t, J=6.4 Hz, 2H), 3.42 (d, J=5.6 Hz, 2H), 3.16-3.07 (m, 4H),
1.51 (s, 6H) ppm; ESI-MS (m/z): 369.5 [M+1].sup.+.
Compound 28
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-(methylsulfon-
amido)phenoxy)acetamide
##STR00520##
[0372] MsCl (23 mg, 0.2 mmol) was added to a cooled 0.degree. C.
stirred solution of
2-(3-aminophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl-
)acetamide (71 mg, 0.2 mmol) in Et.sub.3N (0.1 mL) and DCM (10 mL).
After stirred for 2 h, the solvent was removed by concentration.
The residue was purified by prep-HPLC to afford the title compound.
.sup.1H NMR (400 MHz, MeOD): .delta. 7.26-7.22 (m, 1H), 7.11-7.01
(m, 4H), 6.91-6.85 (m, 2H), 6.71-6.68 (m, 1H), 4.53 (s, 2H),
4.03-4.00 (m, 1H), 3.69 (s, 2H), 3.43-3.88 (m, 1H), 3.13-3.12 (m,
1H), 2.96 (s, 3H), 2.89 (d, J=6 Hz, 1H), 2.82 (d, J=5.6 Hz, 1H),
2.57 (t, J=6 Hz, 1H) ppm; ESI-MS (m/z): 434.1 [M+1].sup.+.
Compound 30
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-methyl-1H-be-
nzo[d]imidazol-6-yl)oxy)acetamide
##STR00521##
[0373] Step 1: 3-(methylamino)-4-nitrophenol
##STR00522##
[0375] The solution of 3-fluoro-4-nitrophenol (1 g, 6.37 mmol) in
aqueous MeNH.sub.2 solution (5 mL) was stirred at 85.degree. C. for
5 h. After cooling to room temperature, the solution was diluted
with water (30 mL) and concentrated HCl added to adjust to pH 1.
The resulting precipitate was collected by filtration and the solid
dried under vacuum to give the crude product which was used without
further purification (1.1 g, 95% yield). LCMS (m/z): 169.1
(M+1).
Step 2: 1-methyl-1H-benzo[d]imidazol-6-ol
##STR00523##
[0377] Fe Powder (4.33 g, 77.4 mmol) was added to a solution of
3-(methylamino)-4-nitrophenol (1.3 g, 7.74 mmol) in HCOOH (30 mL)
and the mixture heated to 100.degree. C. for 16 h. After cooling to
room temperature, MeOH (250 mL) was added to mixture and filtered
over a pad of Celite. The filtrate was collected, concentrated and
the residue purified by column chromatography to give the crude
desired product (1.2 g) and was used directly in the next step.
LCMS (m/z): 149.06 (M+1).
Step 3: ethyl 2-((1-methyl-1H-benzo[d]imidazol-6-yl)oxy)acetate
##STR00524##
[0379] The mixture of 1-methyl-1H-benzo[d]imidazol-6-ol (600 mg,
4.03 mmol), BrCH.sub.2COOEt (372 mg, 4.03 mmol) and K.sub.2CO.sub.3
(1.1 g, 8.06 mmol) in DMF (8 mL) was stirred at room temperature
for 16 h. DCM (100 mL) and water (100 mL) was then added to the
reaction and the organic layer washed with water (50 mL), brine (50
mL) and dried over Na.sub.2SO.sub.4 before filtering and
concentration to give the crude desired product (560 mg, Yield
60%). LCMS (m/z): 235.1 (M+1).
Step 4:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-meth-
yl-1H-benzo[d]imidazol-6-yl)oxy)acetamide
##STR00525##
[0381] A neat solution of ethyl
2-((1-methyl-1H-benzo[d]imidazol-6-yl)oxy)acetate (100 mg, 0.427
mmol) and 1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol
(88 mg, 0.427 mmol) was stirred at room temperature for 2 days
until TLC showed the completion of the reaction. The solution
concentrated and the residue purified by prep-HPLC to give desired
product as the TFA salt (19.1 mg, Yield 11.3%). .sup.1H NMR (400
MHz, MeOD). .delta. 9.21 (s, 1H), 7.80 (d, J=8.8 Hz, 1H), 7.49 (s,
1H), 7.40-7.20 (m, 5H), 4.74 (s, 2H), 4.53 (br.s, 2H), 4.37-4.32
(m, 1H), 4.11 (s, 3H), 3.68 (br.s, 2H), 3.53-3.12 (m, 6H). LCMS
(m/z): 395.1 (M+1).
Compound 31
2-(3-acetamidophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypro-
pyl)acetamide
##STR00526##
[0382] Step 1: 2-(3-acetamidophenoxy) acetic acid
##STR00527##
[0384] To a stirred mixture of N-(3-hydroxyphenyl)acetamide (300
mg, 2.0 mmol) in MeCN (5 mL) was added ethyl bromoacetate (500 mg,
3 mmol) and K.sub.2CO.sub.3 (828 mg, 6 mmol). The mixture was
stirred at 80.degree. C. for 4 hours, filtered and the filtrate was
concentrated. NaOH (80 mg, 2 mmol) and water:EtOH (1:1, 10 mL) was
added to the mixture. This mixture was then stirred at 50.degree.
C. for 4 hours before being acidified by 1M HCl, extracted with
ethyl acetate (2.times.30 mL). The combined extracts were washed
with brine (30 mL), dried over anhydrous Na.sub.2SO.sub.4 and
concentrated with the residue used directly for the next step. LCMS
(m/z): 183.0 (M+1).
Step 2:
2-(3-acetamidophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hy-
droxypropyl)acetamide
##STR00528##
[0386] To a stirred mixture of 2-(3-acetamidophenoxy) acetic acid
(85 mg, 0.41 mmol) in DCM (5 mL) was added TEA (0.5 mL),
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (83.8 mg,
0.41 mmol) and HATU (171 mg, 0.451 mmol). The mixture was stirred
at 25.degree. C. for 3 hours. The reaction mixture was quenched by
addition of water (20 mL) and extracted with DCM (3.times.20 mL)
and the combined extracts washed with brine (20 mL), dried over
anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was
purified by prep-HPLC to afford the title product (55 mg, Yield
35%). .sup.1H NMR (400 MHz, MeOD) .delta.=7.36 (s, 1H), 7.27-7.17
(m, 1H), 7.19-7.01 (m, 6H), 6.68-6.66 (m, 1H), 4.53 (s, 2H),
4.03-4.00 (m, 1H), 3.71-3.62 (m, 2H), 3.50-3.35 (m, 2H), 2.95-2.85
(m, 2H), 2.83-2.74 (m, 2H), 2.56 (d, J=6.3 Hz, 2H), 2.12 (s, 3H).
LCMS (m/z): 398.1 (M+1).
Compound 34
3-(2-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)-2-oxoeth-
oxy)benzamide
##STR00529##
[0387] Step 1: ethyl 2-(3-carbamoylphenoxy)acetate
##STR00530##
[0389] To a stirred mixture of 3-hydroxybenzamide (300 mg, 2.19
mmol) in MeCN (5 mL) was added ethyl bromoacetate (545 mg, 3.29
mmol) and K.sub.2CO.sub.3 (907 mg, 6.57 mmol). The mixture was
stirred at 80.degree. C. for 4 hours. The mixture was filtered and
the filtrate concentrated. The residue was directly for the next
step. LCMS (m/z): 224.1 (M+1).
Step 2:
3-(2-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)--
2-oxoethoxy)benzamide
##STR00531##
[0391] To a stirred mixture of ethyl 2-(3-carbamoylphenoxy)acetate
(150 mg, 0.673 mmol) in EtOH (1 mL) was added
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (138.6 mg,
0.673 mmol). The mixture was stirred at 120.degree. C. for 0.5 hour
under mediated heating. After evaporation of the solvent, the
residue was purified by prep-HPLC to afford the desired title
product (64 mg, Yield 25%). .sup.1H NMR (400 MHz, MeOD) .delta.
7.53 (d, J=8.0 Hz, 1H), 7.48 (s, 1H), 7.39 (dd, J.sub.1=J.sub.2=7.9
Hz, 1H), 7.12-7.00 (m, 5H), 4.60 (s, 2H), 4.04-4.01 (m, 1H),
3.74-3.65 (m, 2H), 3.47-3.39 (m, 2H), 2.95-2.87 (m, 2H), 2.85-2.77
(m, 2H), 2.58 (d, J=6.0 Hz, 2H). LCMS (m/z): 384.1 (M+1).
Compound 46
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-methyl-1H-be-
nzo[d]imidazol-5-yl)oxy)acetamide
##STR00532##
[0392] Step 1: 5-methoxy-1-methyl-1H-benzo[d]imidazole
##STR00533##
[0394] To a solution of NaH (972 mg, 40.5 mmol) in DMF (20 mL) was
added 5-methoxy-1H-benzo[d]imidazole (2.0 g, 13.5 mmol) at
27.degree. C. After stirring for 5 minutes, MeI (2.3 g, 16.2 mmol)
was added and the resulting mixture was stirred for 16 h. The
mixture was then diluted with water (100 mL) and extracted with
ethyl acetate (2.times.50 mL). The combined organic layers were
washed with brine (30 mL), dried over Na.sub.2SO.sub.4 and
concentrated to give the crude product (1.2 g, 54.5%) as a grown
solid. This crude was used in next step without further
purification. LCMS (m/z): 163.1 (M+1).
Step 2: 1-methyl-1H-benzo[d]imidazol-5-ol
##STR00534##
[0396] To a solution of 5-methoxy-1-methyl-1H-benzo[d]imidazole
(500 mg, 3.08 mmol) in CH.sub.2Cl.sub.2 (6 ml) was added BBr.sub.3
(3.1 g, 12.33 mmol) dropwise at 0.degree. C. After addition, the
mixture was stirred for 2 h at 0.degree. C. The mixture was then
quenched by slow addition to ice water (50 mL). The resulting
mixture was extracted with CH.sub.2Cl.sub.2 (2.times.20 mL). The
combined organic layers were washed with brine (30 mL), dried over
Na.sub.2SO.sub.4 and concentrated to give the title compound (100
mg, 21.9%) as a white solid which was used in next step without
further purification. LCMS (m/z): 149.1 (M+1).
Step 3: 2-((1-methyl-1H-benzo[d]imidazol-5-yl)oxy)acetic acid
##STR00535##
[0398] To a solution of NaH (204 mg, 8.49 mmol) in DMF (5 mL) was
added 1-methyl-1H-benzo[d]imidazol-5-ol (420 mg, 2.83 mmol) at
28.degree. C. After being stirred for 5 minutes, ethyl
2-bromoacetate (568 mg, 3.4 mmol) was added and the resulting
mixture stirred for a further 16 h under the reaction was complete
by TLC. The mixture was treated with water (50 mL) and extracted
with ethyl acetate (2.times.20 mL). The water layer was treated
with 2N HCl until pH 3 and extracted with ethyl acetate (2.times.20
mL). The combined organic layers were washed with brine (30 mL),
dried over Na.sub.2SO.sub.4 and concentrated to give the product
(160 mg, 24.1%) as white solid which was used in next step without
further purification. LCMS (m/z): 207.1 (M+1).
Step 4:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-meth-
yl-1H-benzo[d]imidazol-5-yl)oxy)acetamide
##STR00536##
[0400] To a solution of
2-((1-methyl-1H-benzo[d]imidazol-5-yl)oxy)acetic acid (160 mg,
0.776 mmol) in DMF (4 ml) was added TEA (336 mg, 3.33 mmol), HOBt
(157 mg, 1.164 mmol), EDCI (223 mg, 1.164 mmol) and
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (192 mg,
0.931 mmol) at 29.degree. C. The reaction mixture was stirred for
16 h at 29.degree. C. until TLC showed the reaction to be complete.
After evaporation of the solvent, the mixture was purified by
preparative HPLC to give the title compound (13.1 mg, 4.2%) as
colorless oil. .sup.1H NMR (400 MHz, MeOD) .delta. 8.05 (s, 1H),
7.45 (d, J=8.8 Hz, 1H), 7.21 (d, J=2.0 Hz, 1H), 7.09-6.97 (m, 5H),
4.59 (s, 2H), 4.03-3.97 (m, 1H), 3.84 (s, 3H), 3.65 (dd,
J.sub.1=14.8 Hz, J.sub.2=30.4 Hz, 2H), 3.47-3.42 (m, 2H), 2.88 (t,
J=5.6 Hz, 2H), 2.72 (t, J=5.6 Hz, 2H), 2.53-2.51 (m, 2H). LCMS
(m/z): 395.2 (M+1).
Compound 37
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(2-(methylsulfon-
yl)phenoxy)acetamide
##STR00537##
[0401] Step 1: ethyl 2-(2-(methylsulfonyl)phenoxy)acetate
##STR00538##
[0403] To a solution of 2-(methylsulfonyl)phenol (200 mg, 1.16
mmol) in CH.sub.3CN (10 mL) was added ethyl
2-bromo-2-methylpropanoate (232 mg, 1.39 mmol) and K.sub.2CO.sub.3
(690 mg, 5 mmol) at 25.degree. C. The mixture was refluxed for 16 h
and then quenched by additions of water (100 mL). The resulting
mixture was extracted with ethyl acetate (2.times.50 mL). The
combined organic layers were washed with brine (30 mL), dried over
Na.sub.2SO.sub.4 and concentrated to give the title compound which
was used in next step without further purification.
Step 2: 2-(2-(methylsulfonyl)phenoxy)acetic acid
##STR00539##
[0405] To a solution of ethyl 2-(2-(methylsulfonyl)phenoxy)acetate
(750 mg, 2.9 mmol) in EtOH (15 ml) was added 10% NaOH aqueous
solution (15 mL) at 26.degree. C. The mixture was stirred for 30
min and then concentrated and the residue diluted with water (20
mL) and washed with ethyl acetate (2.times.20 mL). The aqueous
layer was then acidified with 2N HCL until pH 3 and then extracted
with ethyl acetate (2.times.20 ml). The combined organic layers
were washed with brine (30 mL), dried over Na.sub.2SO.sub.4 and
concentrated to give the title compound which was used in next step
without further purification.
Step 3:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(2-(meth-
ylsulfonyl)phenoxy)acetamide
##STR00540##
[0407] A mixture of compound 2-(4-methoxyphenoxy)acetic acid (100
mg, 0.43 mmol),
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (89 mg, 0.43
mmol), BOP-Cl (171 mg, 0.67 mmol) and DIPEA (1 mL) in DCM (10 mL)
was stirred at room temperature for 4 h. The solvent was removed by
concentration and the crude product was purified by pre-HPLC to
give the title compound (12 mg, 6.7%). .sup.1H NMR (500 MHz, MeOD):
.delta. 7.96 (dd, J=1.6, 8.0 Hz, 1H), 7.75 (t, J=7.2 Hz, 1H),
7.29-7.24 (m, 2H), 7.13-7.02 (m, 4H), 4.82 (s, 2H), 4.06-4.03 (m,
1H), 3.72 (d, J=2.4 Hz, 2H), 3.53 (dd, J=7.2, 13.2 Hz, 1H),
3.37-3.33 (m, 1H), 3.29 (s, 3H), 2.93-2.90 (m, 2H), 2.86-2.83 (m,
2H), 2.62-2.6 (m, 2H) ppm; ESI-MS (m/z): 419.1 [M+1].sup.+.
Compound 39
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-ylox-
y)propanamide
##STR00541##
[0408] Step 1: ethyl 2-(quinolin-8-yloxy)propanoate
##STR00542##
[0410] To a solution of compound NaH (100 mg, 4.14 mmol) in DMF (3
mL) was added quinolin-8-ol (200 mg, 1.38 mmol) at 26.degree. C.
After stirred for 5 minutes, ethyl 2-bromopropanoate (300 mg, 1.65
mmol) was added and the reaction mixture stirred for 16 h at
26.degree. C. The mixture was then diluted with water (20 mL) and
extracted with ethyl acetate (3.times.10 mL). The combined organic
layers were washed with brine (20 mL), dried over Na.sub.2SO.sub.4
and concentrated to give ethyl 2-(quinolin-8-yloxy)propanoate (200
mg, 59.2%) as colorless oil which was used in next step without
further purification. (304 mg, Yield 90%).
Step 2: 2-(quinolin-8-yloxy)propanoic acid
##STR00543##
[0412] To a solution of ethyl 2-(quinolin-8-yloxy)propanoate (100
mg, 0.4 mmol) in EtOH (1 ml) was added a solution of NaOH (24 mg,
0.6 mmol) in H.sub.2O (0.5 ml) at 27.degree. C. The mixture was
stirred for 30 min at 27.degree. C. The mixture was then
concentrated and the residue treated with water (5 mL) and
extracted with ethyl acetate (2.times.5 mL). The water layer was
then treated with 2N HCl until pH 3 before being extracted with
ethyl acetate (2.times.5 ml). The organic layer was washed with
brine (30 mL), dried over Na.sub.2SO.sub.4 and concentrated to give
the title product (70 mg, 80.5%) as a white solid which was used in
next step without further purification.
Step 3:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinoli-
n-8-yloxy)propanamide
##STR00544##
[0414] To a solution of 2-(quinolin-8-yloxy)propanoic acid (60 mg,
0.276 mmol) in DMF (4 ml) was added TEA (84 mg, 1.1 mmol), HOBt (60
mg, 0.41 mmol), EDCI (79.8 mg, 0.41 mmol) and
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (72 mg,
0.331 mmol) at 28.degree. C. The reaction mixture was stirred for
16 h until TLC showed the reaction was completed. After evaporation
of the solvent, the residue was purified by HPLC separation to give
the TFA salt of
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-ylo-
xy)propanamide (23 mg, 20.5%) as a white solid. .sup.1H NMR (400
MHz, MeOD) .delta. 9.15 (d, J=4.0 Hz, 1H), 9.04-9.00 (m, 1H),
8.06-8.02 (m, 1H), 7.89-7.80 (m, 2H), 7.57 (d, J=7.6 Hz, 1H),
7.33-7.19 (m, 4H), 5.29-5.27 (m, 1H), 4.50 (br.s, 2H), 4.30-4.27
(m, 1H), 3.69 (br.s, 2H), 3.45-3.42 (m, 2H), 3.26-3.20 (m, 4H),
2.76 (d, J=9.6 Hz, 3H). LCMS (m/z): 406.2 (M+1).
Compound 43
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-methyl-2-(quinol-
in-8-yloxy)propanamide
##STR00545##
[0415] Step 1: ethyl 2-methyl-2-(quinolin-8-yloxy)propanoate
##STR00546##
[0417] To a solution of compound NaH (248 mg, 10.2 mmol) in DMF (10
mL) was added quinolin-8-ol (500 mg, 3.44 mmol) at 28.degree. C.
After stirring for 5 minutes, ethyl 2-bromo-2-methylpropanoate (806
mg, 4.13 mmol) was added and the reaction mixture was stirred for
an additional 16 h at 28.degree. C. until the reaction was complete
by TLC. The mixture was then diluted with water (50 mL) and
extracted with ethyl acetate (3.times.20 mL). The combined organic
layers were washed with brine (30 mL), dried over Na.sub.2SO.sub.4
and concentrated to give the crude product (400 mg, 44.8%) as
colorless oil which was used in next step without further
purification.
Step 2:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-methyl-2-
-(quinolin-8-yloxy)propanamide
##STR00547##
[0419] To a solution of ethyl
2-methyl-2-(quinolin-8-yloxy)propanoate (120 mg, 0.46 mmol) in EtOH
(0.5 ml) was added
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (0.46 mmol)
at 29.degree. C. The mixture was stirred for 1 hour at 120.degree.
C. under microwave heating. The solvent was removed and the residue
purified by prep-HPLC to afford the title compound (19.5 mg, Yield
10.1%). .sup.1H NMR (400 MHz, MeOD): .delta. 8.95-8.93 (m, 1H),
8.34 (dd, J=8.4, 1.6 Hz, 1H), 7.69 (d, J=8 Hz, 1H), 7.56-7.52 (m,
2H), 7.39 (d, J=7.6 Hz, 1H), 7.11-7.02 (m, 3H), 6.94 (d, J=6.4 Hz,
1H), 4.13-4.10 (m, 1H), 3.70-3.53 (m, 2H), 3.48-3.43 (m, 2H),
2.91-2.81 (m, 4H), 2.79-2.64 (m, 2H), 1.57 (s, 6H) ppm; ESI-MS
(m/z): 420.3 [M+1].sup.+.
Compound 44
(S)--N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-
-yloxy)acetamide
##STR00548##
[0421] To a stirred mixture of ethyl 2-(quinolin-8-yloxy)acetate
(250 mg, 1.08 mmol) in EtOH (2 mL) was added
(S)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (222 mg,
1.08 mmol). The mixture was stirred at 120.degree. C. for 0.5 hour
under microwave heating. After evaporation of the solvent, the
residue was purified first by prep-TLC and then prep-SFC to afford
(140 mg, Yield 36%). .sup.1H NMR (400 MHz, MeOD) .delta.=8.91 (d,
J=4.3 Hz, 1H), 8.42 (d, J=8.3 Hz, 1H), 7.69-7.54 (m, 3H), 7.30 (d,
J=7.3 Hz, 1H), 7.12-6.96 (m, 4H), 4.78 (s, 2H), 4.18-4.07 (m, 1H),
3.71 (s, 2H), 3.60-3.49 (m, 1H), 3.48-3.40 (m, 1H), 2.89 (d, J=5.8
Hz, 2H), 2.84 (d, J=4.8 Hz, 2H), 2.69-2.55 (m, 2H). LCMS (m/z):
392.1 (M+1).
Compound 45
(R)--N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-
-yloxy)acetamide
##STR00549##
[0423] To a stirred mixture of ethyl 2-(quinolin-8-yloxy)acetate
(250 mg, 1.08 mmol) in EtOH (2 mL) was added
(R)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (222 mg,
1.08 mmol). The mixture was stirred at 120.degree. C. for 0.5 hour
under microwave heating. After evaporation of the solvent, the
residue was purified first by prep-TLC and then by prep-SFC to
afford (160 mg Yield 40%). .sup.1H NMR (400 MHz, MeOD)
.delta.=8.791 (d, J=4.3 Hz, 1H), 8.30 (d, J=8.3 Hz, 1H), 7.51-7.46
(m, 3H), 7.17 (d, J=7.3 Hz, 1H), 6.94-6.85 (m, 4H), 4.65 (s, 2H),
4.00-3.99 (m, 1H), 3.59 (s, 2H), 3.44-3.32 (m, 2H), 2.77-2.71 (m,
4H), 2.53-2.51 (m, 2H). LCMS (m/z): 392.1 (M+1).
Compound 48
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((2-morpholinoqu-
inolin-8-yl)oxy)acetamide
##STR00550##
[0424] Step 1: 2-chloroquinolin-8-ol
##STR00551##
[0426] To a stirred mixture of quinoline-2,8-diol (Ig, 6.21 mmol)
was added POCl.sub.3 (10 mL) and the mixture stirred at 100.degree.
C. for 1 hour before cooling. The mixture was then poured into
ice-water (100 mL) slowly and filtered. The collected solid was
dried and used in next step without further purification. (780 mg,
Yield 70%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.39 (d,
J=8.8 Hz, 1H), 7.57 (d, J=8.8 Hz, 1H), 7.48-7.43 (m, 2H), 7.17 (dd,
J=8.8 Hz, J=1.6 Hz, 1H).
Step 2: 2-morpholinoquinolin-8-ol
##STR00552##
[0428] To a stirred mixture of 2-chloroquinolin-8-ol (1.7 g crude,
9.5 mmol) was added morpholine (5 mL). The mixture was heated at
reflux for 16 hours. After cooling, the mixture was diluted with
water (40 mL) and extracted with ethyl acetate (3.times.30 mL). The
combined organic layers were washed with brine (20 mL), dried over
anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was used
directly for the next step. (1.6 g, Yield 80%). LCMS (m/z): 231.1
(M+1).
Step 3: ethyl 2-((2-morpholinoquinolin-8-yl)oxy)acetate
##STR00553##
[0430] To a stirred mixture of 2-morpholinoquinolin-8-ol (200 mg,
0.87 mmol) in MeCN (5 mL) was added ethyl bromoacetate (216 mg,
1.31 mmol) and K.sub.2CO.sub.3 (360 mg, 2.61 mmol). The mixture was
stirred at 80.degree. C. for 4 hours. After filtration, the
filtrate was concentrated to give crude product which was used
directly for the next step (250 mg, Yield 90%).
Step 4:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((2-morp-
holinoquinolin-8-yl)oxy)acetamide
##STR00554##
[0432] To a stirred mixture of ethyl 2-((2-morpholinoquinolin-8-yl)
oxy) acetate (100 mg, 0.316 mmol) in EtOH (2 mL) was added
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl) propan-2-ol (65 mg,
0.316 mmol). The mixture was stirred at 120.degree. C. for 0.5 hour
under microwave conditions then after evaporation of solvent, the
reaction mixture was purified by prep-HPLC to afford the title
product (14 mg, Yield 10%). .sup.1H NMR (400 MHz, MeOD)
.delta.=8.04 (d, J=9.3 Hz, 1H), 7.43-7.36 (m, 1H), 7.23-7.16 (m,
3H), 7.10-7.00 (m, 3H), 6.97 (d, J=4.8 Hz, 1H), 4.77 (s, 2H), 3.96
(t, J=6.3 Hz, 1H), 3.87-3.83 (m, 4H), 3.77-3.72 (m, 4H), 3.63-3.53
(m, 2H), 3.52-3.45 (m, 1H), 3.40 (d, J=6.3 Hz, 1H), 2.83 (d, J=5.8
Hz, 2H), 2.75-2.67 (m, 2H), 2.56-2.37 (m, 2H). LCMS (m/z): 477.2
(M+1).
Compound 49
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(2-morpholinophe-
noxy)acetamide
##STR00555##
[0433] Step 1: 4-(2-methoxyphenyl)morpholine
##STR00556##
[0435] To a solution of 1-iodo-2-methoxybenzene (1 g, 4.28 mmol) in
dioxane (10 mL) was added morpholine (446.8 mg, 5.12 mmol),
Pd2(dba).sub.3 (100 mg, 0.1 mmol), Xantphos (200 mg, 0.3 mmol) and
t-BuONa (671 mg, 6.0 mmol). Under a N.sub.2 atmosphere the reaction
mixture was heated at reflux temperature for 16 h. The solvent was
then removed and the residue dissolved in ethyl acetate and washed
with water. The separated organic layer was concentrated to give
the crude product which was used in next step without further
purification (578 mg Yield 70%). LCMS (m/z): 194.1 (M+1).
Step 2: 2-morpholinophenol
##STR00557##
[0437] To a solution of 4-(2-methoxyphenyl)morpholine (200 mg, 1.02
mmol) in CH.sub.2Cl.sub.2 (20 mL) was added BBr.sub.3 (1 mL) at
0.degree. C. The mixture was stirred for 2 h at 0.degree. C. The
mixture was then added drop wise to ice-water (50 mL) and the
mixture treated with CH.sub.2Cl.sub.2 (2.times.20 mL). The combined
organic layers were washed with brine (30 mL), dried over
Na.sub.2SO.sub.4 and concentrated to give the yellow solid which
was used in next step without further purification (165 mg Yield
80%). LCMS (m/z): 180.1 (M+1).
Step 3: ethyl 2-(2-morpholinophenoxy)acetate
##STR00558##
[0439] A mixture of 2-morpholinophenol (100 mg, 0.56 mmol) and
ethyl 2-bromoacetate (200 mg, 0.672 mmol) in CH.sub.3CN (10 mL) was
added K.sub.2CO.sub.3 (772.8 mg, 5.6 mmol). The reaction mixture
was stirred at 80.degree. C. for 4 h. The solid was removed by
filtration and the filtrate concentrated to give a crude material,
which was used in the next step without further purification. (130
mg, Yield 90%).
Step 4:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(2-morph-
olinophenoxy)acetamide
##STR00559##
[0441] A mixture of ethyl 2-(2-morpholinophenoxy)acetate (53 mg,
0.2 mmol) and
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (41 mg, 0.2
mmol) in EtOH (1 mL) was stirred at 120.degree. C. over microwave
for 30 min. The solvent was removed by concentration and the crude
product was purified by prep-HPLC separation to afford product the
desired title compound (8.0 mg, Yield 10%). .sup.1H NMR (400 MHz,
MeOD): 7.09-6.98 (m, 8H), 4.64 (s, 2H), 3.95 (br.s, 1H), 3.89-3.87
(m, 4H), 3.63-3.46 (m, 2H), 3.33-3.30 (m, 1H), 3.07-3.03 (m, 4H),
2.88 (dd, J=6.0 Hz, 2H), 2.77 (dd, J=6.0 Hz, 2H), 2.49 (d, J=6.0
Hz, 2H). LCMS (m/z): 426.2 (M+1).
Compound 50
2-(2-(1H-pyrazol-3-yl)phenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hy-
droxypropyl)acetamide
##STR00560##
[0442] Step 1: ethyl 2-(2-(1H-pyrazol-3-yl)phenoxy)acetate
##STR00561##
[0444] To a solution of 2-(1H-pyrazol-3-yl)phenol (500 mg, 3.125
mmol), K.sub.2CO.sub.3 (517.5 mg, 3.75 mmol) and ethyl
2-bromoacetate (417.5 mg, 2.5 mmol) in MeCN (20 mL). The mixture
was stirred at room temperature for 2 h, at which time TLC showed
the completion of the reaction. The mixture was diluted with water
and extracted with EtOAc. The combined organic layers were washed
with brine, dried over Na.sub.2SO.sub.4 and concentrated to give
the crude product which was used in next step without further
purification.
Step 2:
2-(2-(1H-pyrazol-3-yl)phenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)--
yl)-2-hydroxypropyl)acetamide
##STR00562##
[0446] To a solution of ethyl 2-(2-(1H-pyrazol-3-yl)phenoxy)acetate
(100 mg, 0.41 mmol) in EtOH (10 mL) was added
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (84 mg, 0.41
mmol). The mixture was stirred at 120.degree. C. under microwave
heating for 2 h. The reaction mixture was diluted with ethyl
acetate (30 mL) and washed with water (10 mL), dried over
Na.sub.2SO.sub.4 and concentrated to give the crude product. The
residue was purified by prep-HPLC to afford the desired title
compound (85 mg, 44%). .sup.1H NMR (400 MHz, MeOD): .delta.
7.71-7.69 (m, 2H), 7.34-7.33 (m, 1H), 7.10-7.01 (m, 6H), 6.74 (d,
J=2 Hz, 1H), 4.67 (s, 1H), 4.04-4.02 (m, 1H), 3.67 (s, 2H),
3.50-3.49 (m, 1H), 3.37-3.33 (m, 1H), 2.89-2.87 (m, 2H), 2.81-2.78
(m, 2H), 2.58-2.57 (m, 2H) ppm; ESI-MS (m/z): 469.3
[M+1].sup.+.
Compound 54
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-(1-methyl-1H--
pyrazol-5-yl)phenyl)acetamide
##STR00563##
[0447] Step 1: ethyl
2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetate
##STR00564##
[0449] A mixture of ethyl 2-(3-bromophenyl)acetate (1.0 g, 4.1
mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane)
(1.34 g, 5.3 mmol), KOAc (862 mg, 8.8 mmol) and Pd(pddf)Cl.sub.2
(50 mg) in dioxane (15 mL) was stirred at 120.degree. C. for 16 h
under N.sub.2. The reaction mixture was concentrated and the
residue dissolved in water then extracted with EtOAc. The organic
layer was concentrated, and the residue purified by column
chromatography to give the product which was used directly in the
next step.
Step 2: ethyl 2-(3-(1-methyl-1H-pyrazol-5-yl)phenyl)acetate
##STR00565##
[0451] A mixture of ethyl
2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetate
(500 mg, 1.72 mmol), 5-bromo-1-methyl-1H-pyrazole (252 mg, 1.57
mmol), K.sub.2CO.sub.3 (651 mg, 4.71 mmol) and Pd(dppf)Cl.sub.2 (20
mg) in a solution of dioxane (10 mL) and H.sub.2O (2.5 mL) was
stirred at 120.degree. C. for 30 min under microwave. The catalyst
was filtered through a pad of celite and the filtrate concentrated.
The residue was purified by column chromatography to give the
desired product (270 mg, Yield 70%) and used directly in the next
step. LCMS (m/z): 245.1 (M+1).
Step 3: 2-(3-(1-methyl-1H-pyrazol-5-yl)phenyl)acetic acid
##STR00566##
[0453] To a solution of ethyl
2-(3-(1-methyl-1H-pyrazol-5-yl)phenyl)acetate (300 mg, 1.2 mmol) in
MeOH (6 mL) was added aqueous NaOH (1.5 mL, 40 W %). The mixture
was stirred at room temperature for 2 h. The reaction mixture was
concentrated and the residue dissolved in water and adjusted pH to
5-6 with 2N of HCl. The solution was then extracted with EtOAc and
the combined organic layers were concentrated to give the crude
product which was used directly in the next step. LCMS (m/z): 231.1
(M+1).
Step 4:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-(1-me-
thyl-1H-pyrazol-5-yl)phenyl)acetamide
##STR00567##
[0455] To a solution of
2-(3-(1-methyl-1H-pyrazol-5-yl)phenyl)acetic acid (150 mg, 0.69
mmol) in DCM (6 mL) was added EDCI (265 mg, 1.38 mmol), HOBt (186
mg, 1.38 mmol), Et.sub.3N (209 mg, 2.07 mmol) and
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (142 mg,
0.69 mmol). The mixture was stirred at room temperature for 16 h
then diluted with water (10 mL) and extracted with DCM (10
mL.times.3). The combined organic layers were concentrated. The
residue was purified by prep-HPLC to give the product as a
colorless oil (60 mg, Yield 21%). .sup.1H NMR (400 MHz, MeOD): 7.47
(s, 1H), 7.43-7.33 (m, 4H), 7.08-7.04 (m, 3H), 6.96-6.94 (m, 1H),
6.35 (s, 1H), 3.96-3.91 (m, 1H), 3.83 (s, 3H), 3.60-3.59 (m, 4H),
3.38-3.20 (m, 2H), 2.84 (t, J=6.0 Hz, 2H), 2.72 (t, J=6.0 Hz, 2H),
2.49 (d, J=6.4 Hz, 2H). LCMS (m/z): 405.2 (M+1).
Compound 60
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yl)a-
cetamide
##STR00568##
[0457] A solution of 2-(quinolin-8-yl)acetic acid (187 mg, 1 mmol),
HATU (387.6 mg, 1.02 mmol) and TEA (196.1 mg, 1.94 mmol) in DCM (10
mL) was stirred at room temperature for 10 min.
1-Amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (210 mg, 1.0
mmol) was then added, and the solution stirred at for another 3 h,
at which point LCMS indicated completion of the reaction. The
reaction mixture was diluted with water and extracted with DCM (10
mL.times.3). The organic layers combined and dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by prep-HPLC to give the desired compound (50 mg, Yield
13%). .sup.1H NMR (400 MHz, MeOD): 8.92 (d, J=2.8 Hz, 1H), 8.33 (d,
J=8.0 Hz, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.73 (d, J=6.8 Hz, 1H),
7.55-7.50 (m, 2H), 7.10-6.97 (m, 4H), 4.25 (dd, JI=10.8 Hz,
J.sub.2=14.0 Hz, 2H), 3.90 (m, 1H), 3.54-3.51 (m 2H), 3.32-3.25 (m,
2H), 2.82-2.80 (m, 2H), 2.67-2.66 (m, 2H), 2.40-2.39 (m, 2H). LCMS
(m/z): 376.1 (M+1).
Compound 62
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((5,6,7,8-tetrah-
ydronaphthalen-1-yl)oxy)acetamide
##STR00569##
[0458] Step 1: ethyl
2-((5,6,7,8-tetrahydronaphthalen-1-yl)oxy)acetate
##STR00570##
[0460] To a stirred mixture of 5,6,7,8-tetrahydronaphthalen-1-ol
(200 mg, 1.35 mmol) in MeCN (5 mL) was added ethyl bromoacetate
(269 mg, 1.62 mmol) and K.sub.2CO.sub.3 (372 mg, 2.70 mmol). The
mixture was stirred at 80.degree. C. for 4 hours. The mixture was
filtered, the filtrate concentrated to yield the desired product
which used directly for the next step without further purification
(300 mg, Yield 95%).
Step 2:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((5,6,7,-
8-tetrahydronaphthalen-1-yl)oxy)acetamide
##STR00571##
[0462] To a stirred mixture of ethyl
2-((5,6,7,8-tetrahydronaphthalen-1-yl)oxy)acetate (150 mg, 0.641
mmol) in EtOH (2 mL) was added
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (132 mg,
0.641 mmol). The mixture was stirred at 120.degree. C. for 0.5
hours under microwave heating. After evaporation of the solvent,
the residue was purified by prep-HPLC to afford the desired target
product
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((5,6,7,8-tetra-
hydronaphthalen-1-yl)oxy)acetamide (14 mg, Yield 6%). .sup.1H NMR
(400 MHz, MeOD) 0.3=7.13-7.00 (m, 5H), 6.74 (d, J=7.8 Hz, 1H), 6.65
(d, J=8.3 Hz, 1H), 4.53 (s, 2H), 4.02 (quin, J=5.9 Hz, 1H),
3.72-3.63 (m, 2H), 3.53-3.45 (m, 1H), 3.39 (d, J=6.3 Hz, 1H),
2.96-2.86 (m, 2H), 2.83-2.69 (m, 6H), 2.56 (d, J=6.3 Hz, 2H),
1.84-1.74 (m, 4H). LCMS (m/z): 395.1 (M+1).
Compound 66
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(naphthalen-1-yl-
oxy)acetamide
##STR00572##
[0463] Step 1: ethyl 2-(naphthalen-1-yloxy)acetate
##STR00573##
[0465] To a stirred mixture of naphthalen-1-ol (196 mg, 1.35 mmol)
in MeCN (5 mL) was added ethyl bromoacetate (269 mg, 1.62 mmol) and
K.sub.2CO.sub.3 (372 mg, 2.70 mmol). The mixture was stirred at
80.degree. C. for 4 hours. The mixture was filtered and the
filtrate concentrated. The residue was used directly for the next
step without further purification (300 mg, Yield 95%).
Step 2:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(naphtha-
len-1-yloxy)acetamide
##STR00574##
[0467] To a stirred mixture of ethyl 2-(naphthalen-1-yloxy)acetate
(150 mg, 0.641 mmol) in EtOH (2 mL) was added
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (132 mg,
0.641 mmol). The mixture was stirred at 120.degree. C. for 30
minutes under microwave mediated heating. After evaporation of the
solvent, the reaction mixture was purified by prep-HPLC to afford
the desired product (64 mg, Yield 25%). .sup.1H NMR (400 MHz,
METHANOL-d.sub.4)=8.41-8.31 (m, 1H), 7.90-7.80 (m, 1H), 7.58-7.46
(m, 3H), 7.44-7.35 (m, 1H), 7.18-6.95 (m, 5H), 6.91 (d, J=7.8 Hz,
1H), 4.76 (s, 2H), 4.05 (quin, J=6.0 Hz, 1H), 3.72-3.59 (m, 2H),
3.56-3.48 (m, 1H), 3.41 (dd, J=6.5, 13.6 Hz, 1H), 2.93-2.83 (m,
2H), 2.80-2.72 (m, 2H), 2.59-2.52 (m, 2H). LCMS (m/z): 391.2
(M+1).
Compound 71
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-(1-methylpip-
eridin-4-yl)-1H-indazol-5-yl)oxy)acetamide
##STR00575##
[0468] Step 1: tert-butyl
4-((methylsulfonyl)oxy)piperidine-1-carboxylate
##STR00576##
[0470] To a solution of tert-butyl
4-hydroxypiperidine-1-carboxylate (3.0 g, 14.9 mmol) in DCM (30 mL)
was added triethylamine (4.5 g, 44.8 mmol). To this mixture
methanesulfonyl chloride (5.1 g, 44.8 mmol) was added dropwise.
After addition, the mixture was stirred at 25.degree. C. for 3 h
and then filtered. The filtrate was washed with aqueous HCl, dried
over Na.sub.2SO.sub.4, and concentrated under reduced pressure to
give tert-butyl 4-((methylsulfonyl) oxy) piperidine-1-carboxylate
(3.58 g, yield 86%) as a white solid. This material was used in the
next step without further purification. LCMS (m/z): 280.2
[M+H].sup.+
[0471] .sup.1H NMR (400 MHz, CDCl3) .delta. ppm 4.75-4.82 (m, 1H)
3.60-3.71 (m, 2H) 3.21-3.32 (m, 2H) 2.95 (s, 3H) 1.78-1.85 (m, 2H)
1.67-1.78 (m, 2H) 1.35 (s, 9H)
Step 2
5-((tert-butyldimethylsilyl)oxy)-1H-indazole
##STR00577##
[0473] To a solution of 1H-indazol-5-ol (400 mg, 2.98 mmol) in DMF
(10 mL) was added TBDMSCl (537 mg, 3.58 mmol) and imidazole (405
mg, 5.96 mmol) at 0.degree. C. and the resulting mixture was
stirred at 25.degree. C. for 16 h. The reaction was quenched by
addition of water and the product extracted with ethyl acetate. The
organic phase was washed with brine, then dried over
Na.sub.2SO.sub.4, and concentrated under reduced pressure to give
5-((tert-butyldimethylsilyl)oxy)-1H-indazole(500 mg, yield 68%) as
a brown solid, which was used in the next step without further
purification. LCMS (m/z): 249.1 [M+H].sup.+
Step 3
tert-butyl
4-(5-hydroxy-1H-indazol-1-yl)piperidine-1-carboxylate
##STR00578##
[0475] To a solution of NaH (60% in mineral oil) (43.5 mg, 1.81
mmol) in DMF at 0.degree. C. was added
5-((tert-butyldimethylsilyl)oxy)-1H-indazole (300 mg, 1.21 mmol),
and the mixture was stirred at 0.degree. C. for 15 min. Tert-butyl
4-((methylsulfonyl)oxy) piperidine-1-carboxylate (243 mg, 1.21
mmol) was then added to the mixture at 0.degree. C. After addition,
the mixture was stirred at 85.degree. C. for 12 h. The mixture was
poured into water, and the product extracted with ethyl acetate.
The organic phase was washed with water, dried over
Na.sub.2SO.sub.4, concentrated under reduced pressure, and purified
by TLC (Pet.Ether:EtOAc=2:1) to give tert-butyl
4-(5-hydroxy-1H-indazol-1-yl) piperidine-1-carboxylate(250 mg,
yield 65%) as a colorless oil.
[0476] LCMS (m/z): 318.2 [M+H].sup.+
Step 4
tert-butyl
4-(5-(2-ethoxy-2-oxoethoxy)-1H-indazol-1-yl)piperidine-1-carbox-
ylate
##STR00579##
[0478] To a solution of tert-butyl
4-(5-hydroxy-1H-indazol-1-yl)piperidine-1-carboxylate (150 mg,
0.458 mmol) in DMF (10 mL) at 0.degree. C. was added NaH (60% in
mineral oil) (16.8 mg, 0.706 mmol) and ethyl 2-bromoacetate (119
mg, 0.706 mmol). The mixture was stirred at 25.degree. C. for 3 h,
poured into water, and the product extracted with ethyl acetate.
The organic phase was washed with water, dried over
Na.sub.2SO.sub.4, concentrated under reduced pressure and purified
by TLC (Pet.Ether:EtOAc=1:1) to give tert-butyl
4-(5-(2-ethoxy-2-oxoethoxy)-1H-indazol-1-yl)piperidine-1-carboxylate
(120 mg, yield 65%) as a colorless oil. LCMS (m/z): 404.2
[M+H].sup.+
Step 5
tert-butyl
4-(5-(2-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)a-
mino)-2-oxoethoxy)-1H-indazol-1-yl)piperidine-1-carboxylate
##STR00580##
[0480] To a solution of tert-butyl
4-(5-(2-ethoxy-2-oxoethoxy)-1H-indazol-1-yl)piperidine-1-carboxylate(100
mg, 0.248 mmol) in EtOH (2 mL) was added
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol(102.2 mg,
0.496 mmol) and the mixture was stirred at 120.degree. C. for 2 h
in microwave under N.sub.2. The mixture was allowed to cool,
concentrated under reduced pressure, purified by TLC
(Pet.Ether:EtOAc=1:1) to give tert-butyl
4-(5-(2-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)-2-ox-
oethoxy)-1H-indazol-1-yl)piperidine-1-carboxylate (70 mg, yield
50%) as a colorless oil. LCMS (m/z): 564.3 [M+H].sup.+
Compound 70
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-(piperidin-4-
-yl)-1H-indazol-5-yl)oxy)acetamide
##STR00581##
[0482] To tert-butyl
4-(5-(2-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)-2-ox-
oethoxy)-1H-indazol-1-yl)piperidine-1-carboxylate (110 mg, 0.195
mmol) was added EtOAc.HCl (10 mL), the solution was stirred at
25.degree. C. for 2 h, concentrated under reduced pressure, and
purified by prep-HPLC to give
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-(piperidin--
4-yl)-1H-indazol-5-yl)oxy)acetamide (85 mg, yield 94%) as a
colorless oil. .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. ppm
8.47 (br. s., 2H), 8.00 (s, 1H), 7.70 (d, J=8.78 Hz, 1H), 7.29-7.19
(m, 3H), 7.18-7.11 (m, 2H), 6.93 (dd, J=8.85, 1.95 Hz, 1H), 5.12
(s, 2H), 4.86-4.81 (m, 1H), 4.17 (s, 3H), 3.49-3.35 (m, 4H),
3.32-3.20 (m, 4H), 3.13-2.92 (m, 4H), 2.28-2.04 (m, 4H). LCMS
(m/z): 464.2 [M+H].sup.+
Compound 71
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-(1-methylpip-
eridin-4-yl)-1H-indazol-5-yl)oxy)acetamide
##STR00582##
[0484] To a solution of
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-(piperidin--
4-yl)-1H-indazol-5-yl)oxy)acetamide (50 mg, 0.108 mmol) in MeOH (5
mL) was added triethylamine (1 mL), HCHO (30%) (0.3 mL), and HOAC
(0.4 mL). The mixture was stirred at 25.degree. C. for 30 min, then
NaBH.sub.3CN (0.4 mg) was added, and the mixture was stirred at
25.degree. C. for an additional 1 h, concentrated under reduced
pressure, and purified by prep-HPLC to give
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-(1-methylpi-
peridin-4-yl)-1H-indazol-5-yl)oxy)acetamide (51.1 mg, yield 99%) as
a colorless oil.
[0485] .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. ppm 8.50
(br. s., 2H), 8.00 (s, 1H), 7.70 (d, J=8.78 Hz, 1H), 7.28-7.19 (m,
3H), 7.17-7.11 (m, 2H), 6.94 (dd, J=8.85, 1.82 Hz, 1H), 5.12 (s,
2H), 4.82 (br. s., 2H), 4.24-4.11 (m, 3H), 3.47-3.35 (m, 3H), 3.32
(br. s., 1H), 3.13-2.94 (m, 4H), 2.87 (s, 3H), 2.32-2.07 (m, 4H).
LCMS (m/z): 478.3 [M+H].sup.+
Compound 73
(R)--N--((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quino-
lin-8-yloxy)propanamide
##STR00583##
[0486] Step 1: (R)-Methyl 2-(quinolin-8-yloxy)propanoate
##STR00584##
[0488] To a stirred mixture of quinolin-8-ol (300 mg, 2.07 mmol) in
THF (5 mL) was added (S)-methyl 2-hydroxypropanoate (215 mg, 2.07
mmol), PPh.sub.3 (647 mg, 2.47 mmol) and DEAD (430 mg, 2.47 mmol).
The mixture was stirred at 25.degree. C. for 16 hours.
Subsequently, 1M HCl was added (10 mL) and the solution was washed
with EtOAc (20 mL.times.3). The pH of the aqueous solution was
raised by addition of aqueous NaHCO.sub.3 (10 mL), and then this
solution was washed with EtOAc (10 mL.times.3). The combined
organic extracts were washed with brine (20 mL), dried over
anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was
purified by silica column chromatography to afford the product as a
colorless oil (300 mg, 62.5% yield). LCMS (m/z): 233.1
[M+H].sup.+
Step 2:
(R)--N--((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)--
2-(quinolin-8-yloxy)propanamide
##STR00585##
[0490] To a stirred mixture of (R)-methyl
2-(quinolin-8-yloxy)propanoate (100 mg, 0.433 mmol) in EtOH (1 mL)
was added (R)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)
propan-2-ol (89.2 mg, 0.433 mmol). The mixture was stirred in a
sealed tube in a microwave apparatus at 120.degree. C. for 0.5
hour. After cooling to room temperature the solvent was evaporated,
and the residue was first purified by prep-TLC and then prep-HPLC
to afford the title compound (90 mg, yield: 51%). .sup.1H NMR (400
MHz, METHANOL-d.sub.4) .delta. ppm 8.89 (br. S., 1H), 8.29-8.42 (m,
1H), 7.49-7.65 (m, 3H), 7.28 (d, J=5.52 Hz, 1H), 6.89-7.13 (m, 4H),
5.01-5.13 (m, 1H), 4.10-3.84 (m, 1H), 3.45-3.60 (m, 2H), 3.35-3.43
(m, 2H), 2.81 (d, J=3.01 Hz, 2H), 2.65 (br. S., 2H), 2.34-2.49 (m,
2H) 1.71 (d, J=6.78 Hz, 3H), LCMS (m/z): 406.2 [M+H].sup.+
Compound 76
(S)--N--((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quino-
lin-8-yloxy)propanamide
##STR00586##
[0491] Step 1: (S)-methyl 2-(quinolin-8-yloxy)propanoate
##STR00587##
[0493] To a stirred mixture of quinolin-8-ol (300 mg, 2.07 mmol) in
THF (5 mL) was added (R)-methyl 2-hydroxypropanoate (215 mg, 2.07
mmol), PPh.sub.3 (647 mg, 2.47 mmol) and DEAD (430 mg, 2.47 mmol).
The mixture was stirred at 25.degree. C. for 16 hours.
Subsequently, 1M HCl was added (10 mL) and the solution was washed
with EtOAc (20 mL.times.3). The pH of the aqueous solution was
raised by addition of aqueous NaHCO.sub.3 (10 mL), and then this
solution was washed with EtOAc (10 mL.times.3). The combined
organic extracts were washed with brine (20 mL), dried over
anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was
directly for the next step. LCMS (m/z): 232.1/233.1 [M+H].sup.+
Step 2:
(S)--N--((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)--
2-(quinolin-8-yloxy)propanamide
##STR00588##
[0495] To a stirred mixture of (S)-methyl
2-(quinolin-8-yloxy)propanoate (100 mg, 0.433 mmol) in EtOH (1 mL)
was added (S)-1-amino-3-(3,4-dihydroisoquinolin-2
(1H)-yl)propan-2-ol (89.2 mg, 0.433 mmol). The mixture was stirred
in a sealed tube in a microwave apparatus at 120.degree. C. for 0.5
hour. After cooling to room temperature the solvent was evaporated,
and the residue was first purified by prep-TLC and then prep-HPLC
to afford the title compound (49 mg, yield: 28%). .sup.1H NMR (400
MHz, METHANOL-d.sub.4) .delta. ppm 8.86-8.97 (m, 1H), 8.38 (d,
J=7.03 Hz, 1H), 7.51-7.67 (m, 3H), 7.30 (d, J=7.28 Hz, 1H),
7.06-7.13 (m, 3H), 6.98 (d, J=6.53 Hz, 1H), 5.07 (q, J=6.53 Hz,
1H), 4.10-3.88 (m, 1H), 3.65-3.76 (m, 2H), 3.43-3.51 (m, 1H), 3.35
(br. s., 1H), 2.86 (dd, J=16.06, 3.76 Hz, 4H), 2.62 (d, J=6.02 Hz,
2H), 1.71 (d, J=6.53 Hz, 3H). LCMS (m/z): 406.2 [M+H]
Compound 80
N-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(4-methyl-3-oxo--
3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy)acetamide
##STR00589##
[0496] Step 1
2-Aminobenzene-1,3-diol
##STR00590##
[0498] A solution of 2-nitrobenzene-1,3-diol (5.00 g, 32.2 mmol) in
MeOH (100 mL) was stirred under H.sub.2 atmosphere (balloon) in the
presence of 10% Pd/C (200 mg) for 16 h at room temperature. The
reaction mixture was filtered and the filtrate was concentrated to
render a residue characterized as 2-aminobenzene-1,3-diol (3.0 g,
95% yield), used as such for the next reaction step.
Step 2
5-Hydroxy-2H-benzo[b][1,4]oxazin-3(4H)-one
##STR00591##
[0500] A stirred solution of 2-aminobenzene-1,3-diol (2.0 g, 16.0
mmol) and TEA (1.94 g, 19.2 mmol) in anhydrous DMF (30 mL) was
treated with 2-chloroacetyl chloride (1.81 g, 16.0 mmol) and
stirring continued for 16 h at room temperature, then
K.sub.2CO.sub.3 (2.65 g, 19.2 mmol) was added and the mixture
further stirred for 16 h at the same temperature. The reaction
mixture was diluted with DCM (100 mL), washed twice with water and
then with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated. The resulting residue was purified by
chromatographic column of silicagel to give desired product (1.7 g,
64% yield) LCMS (m/z): 166.1 [M+H].sup.+.
Step 3
Ethyl
2-(4-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy)acetate
##STR00592##
[0502] A stirred mixture of
5-hydroxy-2H-benzo[b][1,4]oxazin-3(4H)-one (100 mg, 0.604 mmol) and
K.sub.2CO.sub.3 (167 mg, 1.21 mmol) in anhydrous DMF (5 mL) was
treated with ethyl 2-bromoacetate (121 mg, 0.727 mmol) and stirring
continued at room temperature for 16 h. To this solution of crude
ethyl 2-(3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy) acetate
was added K.sub.2CO.sub.3 (39.6 mg, 0.287 mmol) followed by MeI
(40.7 mg, 0.287 mmol). After being stirred at room temperature for
16 h, the reaction mixture was partitioned between water (50 mL)
and DCM (100 mL). The organic layer was washed by water followed by
and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated and the resulting residue was purified by preparative
TLC to give desired product (43 mg, 56% yield). .sup.1H NMR (400
MHz, CDCl.sub.3): 6.99 (t, J=8.3 Hz, 1H), 6.74 (d, J=7.5 Hz, 1H),
6.55 (d, J=8.3 Hz, 1H), 4.69 (s, 2H), 4.51 (s, 2H), 4.30 (q, J=7.2
Hz, 2H), 3.56 (s, 3H), 1.34-1.33 (m, 1H), 1.33 (t, J=7.2 Hz,
3H)
Step 4
N-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(4-methyl-3-oxo--
3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy)acetamide
##STR00593##
[0504] A reaction vessel containing a mixture of ethyl
2-(4-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy)
acetate (43.0 mg, 0.162 mmol),
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl) propan-2-ol (33.0 mg,
0.163 mmol) and EtOH (0.5 mL) was placed in a microwave reactor and
the mixture irradiated at external temperature of 120.degree. C.
for 1 h. The reaction mixture was purified in two steps by
preparative TLC followed by preparative HPLC to render the title
product (19.2 mg, 19% yield)
[0505] .sup.1H NMR (400 MHz, METHANOL) 6 ppm: 8.44 (br. s., 1H),
7.31-7.19 (m, 3H), 7.16 (d, J=6.5 Hz, 1H), 7.05 (t, J=1.0 Hz, 1H),
6.76 (dd, J=3.0, 8.3 Hz, 2H), 4.76-4.63 (m, 2H), 4.55-4.43 (m, 2H),
4.33-4.17 (m, 3H), 3.50 (s, 3H), 3.46-3.36 (m, 4H), 3.17-3.00 (m,
4H). LCMS (m/z): 426.2 [M+H].sup.+.
Compound 98
(R)--N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((2-methoxy-
quinolin-8-yl)oxy)acetamide
##STR00594##
[0506] Step 1: 8-(benzyloxy)quinolin-2-ol
##STR00595##
[0508] To a solution of quinoline-2,8-diol (5.0 g, 31.1 mmol) in
i-PrOH (50 mL) was added BnBr (5.31 g, 31.1 mmol) and DBU (2.02 g,
5.32 mmol). The mixture was stirred at 80.degree. C. for 16 h. The
mixture was evaporated and to the residue was added DCM (100 mL),
and this solution was washed with 0.5 N NaOH (50 mL), 10% HCl (50
mL), and H.sub.2O (50 mL). The organic layer was evaporated to give
the desired compound (6.6 g, yield 85%). .sup.1HNMR (CDCl.sub.3,
400 MHz) .delta.: 9.16 (br. s., 1H), 7.74 (d, J=9.8 Hz, 1H),
7.49-7.36 (m, 5H), 7.20-7.10 (m, 2H), 7.09-7.03 (m, 1H), 6.67 (d,
J=9.5 Hz, 1H), 5.19 (s, 2H).
Step 2: 8-(benzyloxy)-2-chloroquinoline
##STR00596##
[0510] 8-(benzyloxy)quinolin-2-ol (6.6 g, 26.3 mmol) was dissolved
in POCl.sub.3 (50 mL). The mixture was stirred at 90.degree. C. for
16 h. The POCl.sub.3 was evaporated and to the residue was added
EtOAc (100 mL) and the solution was washed with a.q. NaHCO.sub.3
(80 mL) and H.sub.2O (80 mL). The EtOAc was removed under vacuum to
give the desired compound (6.0 g, yield 85%). LCMS (m/z): 270.1
[M+H].sup.+
Step 3: 8-(benzyloxy)-2-methoxyquinoline
##STR00597##
[0512] To a solution of MeONa (400 mg, 7.43 mmol) in MeOH (20 mL)
was added 8-(benzyloxy)-2-chloroquinoline (2.0 g, 7.43 mmol). The
mixture was stirred at 70.degree. C. for 16 h. To the mixture was
added H.sub.2O (20 mL) and the product extracted with toluene (30
mL.times.3). The combined organic layers were dried with
Na.sub.2SO.sub.4 and evaporated to give the desired compound (1.5
g, yield 79%). LCMS (m/z): 266.1 [M+H].sup.+
Step 4: 2-methoxyquinolin-8-ol
##STR00598##
[0514] To a solution of 8-(benzyloxy)-2-methoxyquinoline (2.2 g,
8.3 mmol) in EtOH (40 mL) was added Pd/C (230 mg). The mixture was
stirred at 25.degree. C. for 16 h under an atmosphere of H.sub.2.
The mixture was filtered, and the filtrate was evaporated to give
the desired compound (1.2 g, 83%). .sup.1HNMR (CDCl.sub.3, 400 MHz)
.delta.: 7.91 (d, J=8.8 Hz, 1H), 7.52 (br. s., 1H), 7.23-7.15 (m,
2H), 7.07 (dd, J=1.4, 7.2 Hz, 1H), 6.85 (d, J=8.8 Hz, 1H), 3.99 (s,
3H).
Step 5: ethyl 2-((2-methoxyquinolin-8-yl)oxy)acetate
##STR00599##
[0516] To a solution of 2-methoxyquinolin-8-ol (500 mg, 2.86 mmol)
in MeCN (10 mL) was added ethyl 2-bromoacetate (501 mg, 3.0 mmol)
and K.sub.2CO.sub.3 (789 mg, 5.72 mmol). The mixture was stirred at
80.degree. C. for 5 h. The mixture was filtered and the filtrate
evaporated to give the desired compound (700 mg, yield 94%). LCMS
(m/z): 262.1 [M+H].sup.+
Step 6:
(R)--N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((2-
-methoxyquinolin-8-yl)oxy)acetamide
##STR00600##
[0518] Ethyl 2-((2-methoxyquinolin-8-yl)oxy)acetate (100 mg, 0.383
mmol), (R)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol
(79.0 mg, 0.383 mmol) and EtOH (1 mL) were combined in a sealed
tube. The mixture was stirred at 120.degree. C. for 30 min in a
microwave. The EtOH was evaporated and the residue was purified by
pre-HPLC to give the desired product (77 mg, yield 48%). .sup.1HNMR
(MeOD-d.sub.4, 400 MHz) .delta.: 8.36 (br. s., 1H), 8.16 (d, J=8.8
Hz, 1H), 7.51 (d, J=7.5 Hz, 1H), 7.36 (t, J=7.9 Hz, 1H), 7.28-7.15
(m, 4H), 7.12 (d, J=6.8 Hz, 1H), 7.03 (d, J=8.8 Hz, 1H), 4.82 (s,
2H), 4.23-4.16 (m, 3H), 4.10 (s, 3H), 3.54-3.48 (m, 1H), 3.47-3.40
(m, 1H), 3.38-3.33 (m, 2H), 3.13-2.98 (m, 4H). LCMS (m/z): 422.2
[M+H].sup.+
Compound 102
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((2-(methylsulfo-
nyl)-1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetamide
##STR00601##
[0519] Step 1: 1,2,3,4-tetrahydroisoquinolin-5-ol
##STR00602##
[0521] A mixture of isoquinolin-5-ol (4 g, 27.6 mmol) and PtO.sub.2
(1.3 g) in HOAc (50 mL) was stirred under H.sub.2 (45 Psi) at room
temperature overnight. The mixture was filtered and the filtrate
was concentrated under vacuum to give the crude product (3.2 g,
80%) which was used in the next step without purification. LCMS
(m/z): 150.1 [M+H].sup.+.
Step 2: tert-butyl
3,4-dihydro-5-hydroxyisoquinoline-2(1H)-carboxylate
##STR00603##
[0523] A mixture of 1,2,3,4-tetrahydroisoquinolin-5-ol (2.06 g,
13.8 mmol) and Na.sub.2CO.sub.3 (2.93 g, 27.6 mmol) in DMF was
cooled with an ice-water bath. Then (Boc).sub.20 (3.61 g, 16.6
mmol) was added in three portions. The solution was then stirred at
room temperature overnight. The mixture was then filtered and the
filtrate was concentrated under vacuum to give the crude product
(3.1 g, 91%) which was used directly in the next step. LCMS (m/z):
250.2 [M+H].sup.+.
Step 3: tert-butyl
5-((ethoxycarbonyl)methoxy)-3,4-dihydroisoquinoline-2(1H)-carboxylate
##STR00604##
[0525] To a solution of tert-butyl
3,4-dihydro-5-hydroxyisoquinoline-2(1H)-carboxylate (750 mg, 3.01
mmol) and K.sub.2CO.sub.3 (498 mg, 3.61 mmol) in MeCN was added
ethyl 2-bromoacetate (603 mg, 3.61 mmol). The mixture was stirred
at room temperature overnight, the mixture was then filtered and
the filtrate was concentrated under vacuum to give the desired
product (900 mg, 90%). LCMS (m/z): 336.2 [M+H].sup.+.
Step 4: ethyl
2-((1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetate
##STR00605##
[0527] To a solution of tert-butyl
5-(2-ethoxy-2-oxoethoxy)-3,4-dihydroisoquinoline-2(1H)-carboxylate
(400 mg, 1.19 mmol) in ethyl acetate (10 mL), cooled in an
ice-water bath, was added (10 mL, 1N) drop wise. The mixture was
stirred at 25.degree. C. for 16 h and then concentrated under
vacuum to give the crude product (275 mg, 98%) which was used to
the next step without further purification.
Step 5: ethyl
2-((2-(methylsulfonyl)-1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetate
##STR00606##
[0529] To a solution of ethyl
2-((1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetate (300 mg, 1.28
mmol) and Et.sub.3N (387 mg, 3.83 mmol) in DCM (25 mL) cooled in an
ice-water bath was added MsCl (176.6 mg, 1.54 mmol) drop wise. The
mixture was stirred at 25.degree. C. for 16 h and then quenched
with aq.NH.sub.4Cl. The mixture was extracted with ethyl acetate
and the combined organic layers were concentrated under vacuum to
give the crude product (312 mg, 78%) which was used to the next
step without purification.
Step 6:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((2-(met-
hylsulfonyl)-1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetamide
##STR00607##
[0531] A mixture of ethyl
2-((2-(methylsulfonyl)-1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetate
(140 mg, 0.447 mmol) and
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (184 mg,
0.89 mmol) in EtOH (0.2 mL) was stirred at 120.degree. C. for 30
min under microwave conditions. The mixture was diluted with MeOH
and purified by prep-HPLC to afford
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((2-(methylsulf-
onyl)-1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetamide (53.5 mg,
25%). .sup.1H NMR (400 MHz, MeOD) .delta. 0.84 (s, 1H), 7.29-7.15
(m, 5H), 6.80 (d, J=8.2 Hz, 1H), 6.83 (d, J=7.7 Hz, 1H), 4.61 (s,
2H), 4.41 (s, 2H), 4.35-4.17 (m, 3H), 3.54 (t, J=6.1 Hz, 2H),
3.48-3.38 (m, 4H), 3.17-3.04 (m, 4H), 2.98 (t, J=6.0 Hz, 2H), 2.92
(s, 3H). LCMS (m/z): 474.2 [M+H].sup.+.
Compound 152
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(pyridin-3-ylmet-
hoxy)acetamide
##STR00608##
[0532] Step 1: Ethyl 2-(pyridin-3-ylmethoxy)acetate
##STR00609##
[0534] To a solution of NaH (330 mg, 13.7 mmol) in DMF (10 mL) was
added pyridin-3-ylmethanol (500 mg, 4.6 mmol) and the solution was
stirred at 27.degree. C. for 20 minutes. Ethyl 2-bromoacetate
(921.8 mg, 5.52 mmol) was then added and the reaction mixture
stirred at 27.degree. C. for further 16 h. Once the reaction was
complete by TLC analysis, the mixture was quenched by addition of
water (50 mL) and extracted with ethyl acetate (3.times.20 mL). The
combined organic layers were washed with brine (30 mL), dried over
Na.sub.2SO.sub.4 and concentrated to give the Ethyl
2-(pyridin-3-ylmethoxy)acetate (600 mg, 66.8%) as colorless oil
which was used in next step without further purification. LCMS:
196.1 [M+H].sup.+.
Step 2:
N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(pyridin-
-3-ylmethoxy)acetamide
##STR00610##
[0536] To a solution of Ethyl 2-(pyridin-3-ylmethoxy)acetate (100
mg, 0.51 mmol) in EtOH (0.5 ml) was added
1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (105 mg,
0.51 mmol) at 28.degree. C. The mixture was stirred at 120.degree.
C. under microwave heating for 1 hour until the reaction was
completed by TLC. After evaporation of the solvent, the residue was
purified by HPLC separation to give the title compound (30 mg,
yield: 16.5%) as a white solid. LCMS: 356.2 [M+H].sup.+. .sup.1H
NMR (MeOD, 400 MHz) .delta. 8.53 (s, 1H), 8.50 (d, J=4.8 Hz, 1H),
7.85 (d, J=8.0 Hz, 1H), 7.462 (dd, J=4.8 Hz, J.sub.2=8.0 Hz, 1H),
7.19-7.03 (m, 4H), 4.58 (s, 2H), 4.02 (br. s, 3H), 3.72 (s, 2H),
3.42-3.36 (m, 2H), 2.93-2.83 (m, 4H), 2.62-2.61 (m, 2H).
Compound 175
Step 1:
(S)-3-(1H-benzo[d]imidazol-2-yl)-N-(3-(3,4-dihydroisoquinolin-2(1H-
)-yl)-2-hydroxypropyl)propanamide
##STR00611##
[0538] To a solution of 3-(1H-benzo[d]imidazol-2-yl)propanoic acid
(300 mg, 1.579 mmol) in DCM (10 mL) was added HATU (722 mg, 1.895
mmol) and TEA (478 mg, 4.737 mmol). After stirring for 30 min at
room temperature, (S)-1-amino-3-(3,4-dihydro
isoquinolin-2(1H)-yl)propan-2-ol (488 mg, 2.368 mmol) was added and
the resulting mixture then stirred at room temperature for 16 h.
After completion of the reaction the solvent was evaporated at
reduced pressure and the residue purified by preparative HPLC to
give the title compound (159.1 mg, 26%). .sup.1H NMR (400 MHz,
METHANOL-d.sub.4) 6=7.51 (br. s., 2H), 7.22-7.16 (m, 2H), 7.13-7.06
(m, 3H), 7.02-6.97 (m, 1H), 3.98-3.90 (m, 1H), 3.66-3.55 (m, 2H),
3.37 (dd, J=4.9, 13.7 Hz, 1H), 3.25-3.17 (m, 3H), 2.89-2.83 (m,
2H), 2.79 (t, J=7.5 Hz, 2H), 2.74-2.69 (m, 2H), 2.50-2.45 (m, 2H).
LCMS (m/z): 379.1 [M+H].sup.+.
Compound 192
Step 1:
(R)-3-(1H-benzo[d]imidazol-2-yl)-N-(3-(3,4-dihydroisoquinolin-2(1H-
)-yl)-2-hydroxypropyl)propanamide
##STR00612##
[0540] To a solution of 3-(1H-benzo[d]imidazol-2-yl)propanoic acid
(300 mg, 1.579 mmol) in DCM (10 mL) was added HATU (722 mg, 1.895
mmol) and TEA (478 mg, 4.737 mmol). After stirring for 30 min at
room temperature, (R)-1-amino-3-(3,4-dihydro
isoquinolin-2(1H)-yl)propan-2-ol (488 mg, 2.368 mmol) was added and
the resulting mixture then stirred at room temperature for 16 h.
Once TLC analysis showed the reaction to be complete, the solvent
was evaporated at reduced pressure and the residue purified by
preparative HPLC to give the title compound (184.6 mg, 30.9%) as
white solid. .sup.1H NMR (400 MHz, METHANOL-d.sub.4)
.delta.=7.56-7.46 (m, 2H), 7.22-7.17 (m, 2H), 7.13-7.07 (m, 3H),
7.02-6.97 (m, 1H), 3.94 (t, J=5.8 Hz, 1H), 3.61 (d, J=3.3 Hz, 2H),
3.40-3.35 (m, 1H), 3.25-3.18 (m, 3H), 2.89-2.84 (m, 2H), 2.79 (t,
J=7.4 Hz, 2H), 2.74-2.69 (m, 2H), 2.47 (d, J=6.5 Hz, 2H). LCMS
(m/z): 379.1 [M+H].sup.+.
LC-MS Conditions
[0541] Method A (LCMS-B (0-60AB_ELSD_2MIN))
[0542] Experiments performed on an Agilent 1200 HPLC (with a PDA
detector and a ELSD detector) with Agilent 6100 MSD mass
spectrometer using ESI as ionization source using an
Xtimate.TM.-C18 30*2.1 mm column and a 0.8 ml/minute flow rate.
Acquire Time: 2 min, Wavelength: UV220, Oven Temp.: 50.degree. C.
The solvent system was a gradient starting with 100% water
containing 0.038% TFA (solvent A) and 0% acetonitrile containing
0.02% TFA (solvent B), followed by a gradient up to 40% solvent A
and 60% solvent B over the next 0.9 minutes. This was maintained
for 0.6 minutes before returning to 100% solvent A over the next
0.5 minute. Total run time was 2 min.
Method B (LCMS-C(10-80_AB))
[0543] Experiments performed on an SHIMADZU 20 A HPLC (with a PDA
detector) with SHIMADZU 2010EV MSD mass spectrometer using ESI as
ionization source using an Xtimate.TM.-C18 30*2.1 mm column and a
1.2 ml/minute flow rate. The solvent system was a gradient starting
with 90% water containing 0.038% TFA (solvent A) and 10%
acetonitrile containing 0.02% TFA (solvent B), followed by a
gradient up to 20% solvent A and 80% solvent B over the next 0.9
minutes. This was maintained for 0.6 minutes before returning to
90% solvent A and 10% solvent B over the next 0.5 minute. Total run
time was 2 min.
Method C (LCMS-E(5-95AB_220&254 nm))
[0544] Experiments performed on an SHIMADZU 20 A HPLC (with a PDA
detector) with SHIMADZU 2010EV MSD mass spectrometer using ESI as
ionization source using an Merk RP-18e 2*25 mm column and a 1.5
ml/minute flow rate. The solvent system was a gradient starting
with 95% water containing 0.038% TFA (solvent A) and 5%
acetonitrile containing 0.02% TFA (solvent B), followed by a
gradient up to 5% solvent A and 95% solvent B over the next 0.7
minutes. This was maintained for 0.4 minutes before returning to
95% solvent A and 5% solvent B over the next 0.4 minute. Total run
time was 1.5 min.
Method D (LCMS-A(0-30_AB))
[0545] Experiments performed on an SHIMADZU 20 A HPLC (with a PDA
detector) with SHIMADZU 2010EV MSD mass spectrometer using ESI as
ionization source using an Xtimate.TM.-C18 30*2.1 mm column and a
1.2 ml/minute flow rate. The solvent system was a gradient starting
with 100% water containing 0.038% TFA (solvent A) and 0%
acetonitrile containing 0.02% TFA (solvent B), followed by a
gradient up to 70% solvent A and 30% solvent B over the next 0.9
minutes. This was maintained for 0.6 minutes before returning to
100% solvent A over the next 0.5 minute. Total run time was 2
min.
General HPLC Conditions (Acidic)
[0546] Mobile phase A: 4 L H.sub.2O\1.5 ml TFA; Mobile phase B: 4 L
ACN\0.75 ml TFA
[0547] Column: HPLC-D: Innovation C18 UPLC Column 2.1.times.30 mm,
2.6 um
[0548] HPLC-E: Xtimate C18 2.1*30 mm*3 um
[0549] HPLC-H: Innovation C18 UPLC Column 2.1.times.30 mm, 2.6
um
[0550] Column temperature: 50.degree. C.; Wavelength: 220 nm &
254 nm & 215 nm
General HPLC Conditions (Basic)
[0551] Mobile phase A: 4 L H.sub.2O\2 ml NH.sub.4OH; Mobile phase
B: Acetonitrile
[0552] Column: HPLC-B: XBridge C18 2.1*50 mm, 5 um
[0553] HPLC-C: Xbridge shield RP18 2.1*50 mm, 5 u
[0554] Column temperature: 30.degree. C.; Wavelength: 220 nm &
254 nm & 215 nm
General HPLC Conditions (Neutral)
[0555] Mobile phase A: H.sub.2O; Mobile phase B: Acetonitrile
[0556] Column: HPLC-B: XBridge C18 2.1*50 mm, 5 um
[0557] HPLC-C: Xbridge shield RP18 2.1*50 mm, 5 um
[0558] Column temperature: 30.degree. C.; Wavelength: 220 nm &
254 nm & 215 nm
Method A (0-30AB_6MIN)
[0559] Flow Rate: 0.8 ml/min
[0560] Gradient: 0% B to 30% B in 4.2 min, holding 30% B for 1 min,
30% B to 0% B in 0.01 min, holding 0% B for 1.09 min and then
end.
Method B (0-60AB_6MIN)
[0561] Flow Rate: 0.8 ml/min
[0562] Gradient: 0% B to 60% B in 4.2 min, holding 60% B for 1 min,
60% B to 0% B in 0.01 min, holding 0% B for 1.09 min and then
end.
Method C (10-80AB_6MIN)
[0563] Flow Rate: 0.8 ml/min
[0564] Gradient: 10% B to 80% B in 4.2 min, holding 80% B for 1
min, 80% B to 10% B in 0.01 min, holding 10% B for 1.09 min and
then end.
Chiral HPLC Conditions:
Method A (OJ-H):
[0565] Column: Chiralcel OJ-H 250.times.4.6 mm I.D., 5 um
[0566] Mobile phase: A/B=90/10, A: Hexane with 0.1% DEA, B:
Ethanol
[0567] Flow rate: 0.5 mL/min
[0568] Wavelength: 220 nm
Method B (OD-H):
[0569] Column: Chiralcel OD-H 250.times.4.6 mm I.D., 5 um
[0570] Mobile phase: A/B=90/10, A: Hexane with 0.1% DEA, B:
Ethanol
[0571] Flow rate: 0.5 mL/min
[0572] Wavelength: 220 nm
Method C (AD-H):
[0573] Column: Chiralpak AD-H 250.times.4.6 mm I.D., 5 um
[0574] Mobile phase: A/B=90/10, A: Hexane with 0.1% DEA, B:
Ethanol
[0575] Flow rate: 0.5 mL/min
[0576] Wavelength: 220 nm
Method D (AS-H):
[0577] Column: Chiralpak OJ-H 250.times.4.6 mm I.D., 5 um
[0578] Mobile phase: A/B=90/10, A: Hexane with 0.1% DEA, B:
Ethanol
[0579] Flow rate: 0.5 mL/min
[0580] Wavelength: 220 nm
Biological Assays
PRMT5 Biochemical Assay
[0581] General Materials.
[0582] S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH),
bicine, KCl, Tween20, dimethylsulfoxide (DMSO), bovine skin gelatin
(BSG), and Tris(2-carboxyethyl)phosphine hydrochloride solution
(TCEP) were purchased from Sigma-Aldrich at the highest level of
purity possible. .sup.3H-SAM was purchase from American
Radiolabeled Chemicals with a specific activity of 80 Ci/mmol.
384-well streptavidin Flashplates were purchased from
PerkinElmer.
[0583] Substrates.
[0584] Peptide representative of human histone H4 residues 1-15 was
synthesized with a C-terminal linker-affinity tag motif and a
C-terminal amide cap by 21.sup.st Century Biochemicals. The peptide
was high high-perfomance liquid chromatography (HPLC) purified to
greater than 95% purity and confirmed by liquid chromatography mass
spectrometry (LC-MS). The sequence was
Ac-SGRGKGGKGLGKGGA[K-Biot]-amide (SEQ ID NO.:3).
[0585] Molecular Biology:
[0586] Full-length human PRMT5 (NM_006109.3) transcript variant 1
clone was amplified from a fetal brain cDNA library, incorporating
flanking 5' sequence encoding a FLAG tag (MDYKDDDDK) (SEQ ID NO.:4)
fused directly to Ala 2 of PRMT5. Full-length human MEP50
(NM_024102) clone was amplified from a human testis cDNA library
incorporating a 5' sequence encoding a 6-histidine tag (MHHHHHH)
(SEQ ID NO.:5) fused directly to Arg 2 of MEP50. The amplified
genes were sublconed into pENTR/D/TEV (Life Technologies) and
subsequently transferred by Gateway.TM. attL.times.attR
recombination to pDEST8 baculvirus expression vector (Life
Technologies).
[0587] Protein Expression.
[0588] Recombinant baculovirus and Baculovirus-Infected Insect
Cells (BIIC) were generated according to Bac-to-Bac kit
instructions (Life Technologies) and Wasilko, 2006, respectively.
Protein over-expression was accomplished by infecting exponentially
growing Spodoptera frugiperda (SF9) cell culture at
1.2.times.10.sup.6 cell/ml with a 5000 fold dilution of BIIC stock.
Infections were carried out at 27.degree. C. for 72 hours,
harvested by centrifugation, and stored at -80.degree. C. for
purification.
[0589] Protein Purification.
[0590] Expressed full-length human Flag-PRMT5/6His-MeP50 protein
complex was purified from cell paste by NiNTA agarose affinity
chromatography after a five hour equilibration of the resin with
buffer containing 50 mM Tris-HCL, pH 8.0, 25 mM NaCl, and 1 mM TCEP
at 4.degree. C., to minimize the adsorption of tubulin impurity by
the resin. Flag-PRMT5/6His-MeP50 was eluted with 300 mM Imidazole
in the same buffer. The purity of recovered protein was 87%.
Reference: Wasilko, D. J. and S. E. Lee: "TIPS: titerless
infected-cells preservation and scale-up" Bioprocess J., 5 (2006),
pp. 29-32.
[0591] Predicted Translations:
TABLE-US-00006 Flag-PRMT5 (SEQ ID NO.: 6) MDYKDDDDKA AMAVGGAGGS
RVSSGRDLNC VPEIADTLGA VAKQGFDFLC MPVFHPRFKR EFIQEPAKNR PGPQTRSDLL
LSGRDWNTLI VGKLSPWIRP DSKVEKIRRN SEAAMLQELN FGAYLGLPAF LLPLNQEDNT
NLARVLTNHI HTGHHSSMFW MRVPLVAPED LRDDIIENAP TTHTEEYSGE EKTWMWWHNF
RTLCDYSKRI AVALEIGADL PSNHVIDRWL GEPIKAAILP TSIFLTNKKG FPVLSKMHQR
LIFRLLKLEV QFIITGTNHH SEKEFCSYLQ YLEYLSQNRP PPNAYELFAK GYEDYLQSPL
QPLMDNLESQ TYEVFEKDPI KYSQYQQAIY KCLLDRVPEE EKDTNVQVLM VLGAGRGPLV
NASLRAAKQA DRRIKLYAVE KNPNAVVTLE NWQFEEWGSQ VTVVSSDMRE WVAPEKADII
VSELLGSFAD NELSPECLDG AQHFLKDDGV SIPGEYTSFL APISSSKLYN EVRACREKDR
DPEAQFEMPY VVRLHNFHQL SAPQPCFTFS HPNRDPMIDN NRYCTLEFPV EVNTVLHGFA
GYFETVLYQD ITLSIRPETH SPGMFSWFPI LFPIKQPITV REGQTICVRF WRCSNSKKVW
YEWAVTAPVC SAIHNPTGRS YTIG L 6His-MEP50 (SEQ ID NO.: 7) MHHHHHHRKE
TPPPLVPPAA REWNLPPNAP ACMERQLEAA RYRSDGALLL GASSLSGRCW AGSLWLFKDP
CAAPNEGFCS AGVQTEAGVA DLTWVGERGI LVASDSGAVE LWELDENETL IVSKFCKYEH
DDIVSTVSVL SSGTQAVSGS KDICIKVWDL AQQVVLSSYR AHAAQVTCVA ASPHKDSVFL
SCSEDNRILL WDTRCPKPAS QIGCSAPGYL PTSLAWHPQQ SEVFVFGDEN GTVSLVDTKS
TSCVLSSAVH SQCVTGLVFS PHSVPFLASL SEDCSLAVLD SSLSELFRSQ AHRDFVRDAT
WSPLNHSLLT TVGWDHQVVH HVVPTEPLPA PGPASVTE
[0592] General Procedure for PRMT5/MEP50 Enzyme Assays on Peptide
Substrates.
[0593] The assays were all performed in a buffer consisting of 20
mM Bicine (pH=7.6), 1 mM TCEP, 0.005% BSG, and 0.002% Tween20,
prepared on the day of use. Compounds in 100% DMSO (1 ul) were
spotted into a polypropylene 384-well V-bottom plates (Greiner)
using a Platemate Plus outfitted with a 384-channel head (Thermo
Scientific). DMSO (lul) was added to Columns 11, 12, 23, 24, rows
A-H for the maximum signal control and lul of SAH, a known product
and inhibitor of PRMT5/MEP50, was added to columns 11, 12, 23, 24,
rows I-P for the minimum signal control. A cocktail (40 ul)
containing the PRMT5/MEP50 enzyme and the peptide was added by
Multidrop Combi (Thermo-Fisher). The compounds were allowed to
incubate with PRMT5/MEP50 for 30 min at 25 degrees Celsius, then a
cocktail (10 ul) containing .sup.3H-SAM was added to initiate the
reaction (final volume=51 ul). The final concentrations of the
components were as follows: PRMT5/MEP50 was 4 nM, .sup.3H-SAM was
75 nM, peptide was 40 nM, SAH in the minimum signal control wells
was 100 uM, and the DMSO concentration was 1%. The assays were
stopped by the addition of non-radioactive SAM (10 ul) to a final
concentration of 600 uM, which dilutes the .sup.3H-SAM to a level
where its incorporation into the peptide substrate is no longer
detectable. 50 ul of the reaction in the 384-well polypropylene
plate was then transferred to a 384-well Flashplate and the
biotinylated peptides were allowed to bind to the streptavidin
surface for at least 1 hour before being washed three times with
0.1% Tween20 in a Biotek ELx405 plate washer. The plates were then
read in a PerkinElmer TopCount plate reader to measure the quantity
of .sup.3H-labeled peptide bound to the Flashplate surface,
measured as disintegrations per minute (dpm) or alternatively,
referred to as counts per minute (cpm).
% inhibition calculation ##EQU00001## % inh = 100 - ( dpm cmpd -
dpm min dpm max - dpm min ) .times. 100 ##EQU00001.2##
Where dpm=disintegrations per minute, cmpd=signal in assay well,
and min and max are the respective minimum and maximum signal
controls.
Four - parameter IC 50 fit ##EQU00002## Y = Bottom + ( Top - Bottom
) ( 1 + ( X IC 50 ) Hill Coefficient ##EQU00002.2##
Where top and bottom are the normally allowed to float, but may be
fixed at 100 or 0 respectively in a 3-parameter fit. The Hill
Coefficient normally allowed to float but may also be fixed at 1 in
a 3-parameter fit. Y is the % inhibition and X is the compound
concentration.
Z-138 Methylation Assay
[0594] Z-138 suspension cells were purchased from ATCC (American
Type Culture Collection, Manassas, Va.). RPMI/Glutamax medium,
penicillin-streptomycin, heat inactivated fetal bovine serum, and
D-PBS were purchased from Life Technologies, Grand Island, N.Y.,
USA. Odyssey blocking buffer, 800 CW goat anti-rabbit IgG (H+L)
antibody, and Licor Odyssey infrared scanner were purchased from
Licor Biosciences, Lincoln, Nebr., USA. Symmetric di-methyl
arginine antibody was purchased from EMD Millipore, Billerica,
Mass., USA. 16% Paraformaldehyde was purchased from Electron
Microscopy Sciences, Hatfield, Pa., USA.
[0595] Z-138 suspension cells were maintained in growth medium
(RPMI 1640 supplemented with 10% v/v heat inactivated fetal bovine
serum and 100 units/mL penicillin-streptomycin) and cultured at
37.degree. C. under 5% CO.sub.2.
[0596] Cell Treatment, in Cell Western (ICW) for Detection of
Symmetric Di-Methyl Arginine and DNA content.
[0597] Z-138 cells were seeded in assay medium at a concentration
of 50,000 cells per mL to a 384-well cell culture plate with 50
.mu.L per well. Compound (100 nL) from 384 well source plates was
added directly to 384 well cell plate. Plates were incubated at
37.degree. C., 5% CO.sub.2 for 96 hours. After four days of
incubation, 40 .mu.L of cells from incubated plates were added to
poly-D-lysine coated 384 well culture plates (BD Biosciences
356697). Plates were incubated at room temperature for 30 minutes
then incubated at 37.degree. C., 5% CO.sub.2 for 5 hours. After the
incubation, 40 .mu.L per well of 8% paraformaldehyde in PBS (16%
paraformaldahyde was diluted to 8% in PBS) was added to each plate
and incubated for 30 minutes. Plates were transferred to a Biotek
405 plate washer and washed 5 times with 100 L per well of wash
buffer (IX PBS with 0.1% Triton X-100 (v/v)). Next 30 .mu.L per
well of Odyssey blocking buffer were added to each plate and
incubated 1 hour at room temperature. Blocking buffer was removed
and 20 .mu.L per well of primary antibody was added (symmetric
di-methyl arginine diluted 1:100 in Odyssey buffer with 0.1% Tween
20 (v/v)) and plates were incubated overnight (16 hours) at
4.degree. C. Plates were washed 5 times with 100 .mu.L per well of
wash buffer. Next 20 .mu.L per well of secondary antibody was added
(1:200 800 CW goat anti-rabbit IgG (H+L) antibody, 1:1000 DRAQ5
(Biostatus limited) in Odyssey buffer with 0.1% Tween 20 (v/v)) and
incubated for 1 hour at room temperature. The plates were washed 5
times with 100 .mu.L per well wash buffer then 1 time with 100
.mu.L per well of water. Plates were allowed to dry at room
temperature then imaged on the Licor Odyssey machine which measures
integrated intensity at 700 nm and 800 nm wavelengths. Both 700 and
800 channels were scanned.
[0598] Calculations:
[0599] First, the ratio for each well was determined by:
( symmetric di - methyl Arginine 800 nm value DRAQS 700 nm value )
##EQU00003##
[0600] Each plate included fourteen control wells of DMSO only
treatment (minimum inhibition) as well as fourteen control wells
for maximum inhibition treated with 3 .mu.M of a reference compound
(Background wells). The average of the ratio values for each
control type was calculated and used to determine the percent
inhibition for each test well in the plate. Reference compound was
serially diluted three-fold in DMSO for a total of nine test
concentrations, beginning at 3 .mu.M. Percent inhibition was
determined and IC.sub.50 curves were generated using triplicate
wells per concentration of compound.
Percent Inhibition = 100 - ( ( ( Individual Test Sample Ratio ) - (
Background Avg Ratio ) ( Minimum Inhibition Ratio ) - ( Background
Average Ratio ) ) * 100 ) ##EQU00004##
Z-138 Proliferation Assay
[0601] Z-138 suspension cells were purchased from ATCC (American
Type Culture Collection, Manassas, Va.). RPMI/Glutamax medium,
penicillin-streptomycin, heat inactivated fetal bovine serum were
purchased from Life Technologies, Grand Island, N.Y., USA. V-bottom
polypropylene 384-well plates were purchased from Greiner Bio-One,
Monroe, N.C., USA. Cell culture 384-well white opaque plates were
purchased from Perkin Elmer, Waltham, Mass., USA. Cell-Titer
Glo.RTM. was purchased from Promega Corporation, Madison, Wis.,
USA. SpectraMax M5 plate reader was purchased from Molecular
Devices LLC, Sunnyvale, Calif., USA.
[0602] Z-138 suspension cells were maintained in growth medium
(RPMI 1640 supplemented with 10% v/v heat inactivated fetal bovine
serum and cultured at 37.degree. C. under 5% CO.sub.2. Under assay
conditions, cells were incubated in assay medium (RPMI 1640
supplemented with 10% v/v heat inactivated fetal bovine serum and
100 units/mL penicillin-streptomycin) at 37.degree. C. under 5%
CO.sub.2.
[0603] For the assessment of the effect of compounds on the
proliferation of the Z-138 cell line, exponentially growing cells
were plated in 384-well white opaque plates at a density of 10,000
cells/ml in a final volume of 50 .mu.l of assay medium. A compound
source plate was prepared by performing triplicate nine-point
3-fold serial dilutions in DMSO, beginning at 10 mM (final top
concentration of compound in the assay was 20 .mu.M and the DMSO
was 0.2%). A 100 nL aliquot from the compound stock plate was added
to its respective well in the cell plate. The 100% inhibition
control consisted of cells treated with 200 nM final concentration
of staurosporine and the 0% inhibition control consisted of DMSO
treated cells. After addition of compounds, assay plates were
incubated for 5 days at 37.degree. C., 5% CO.sub.2, relative
humidity >90%.
[0604] Cell viability was measured by quantitation of ATP present
in the cell cultures, adding 35 .mu.l of Cell Titer Glo.RTM.
reagent to the cell plates. Luminescence was read in the SpectraMax
M5 microplate reader. The concentration of compound inhibiting cell
viability by 50% was determined using a 4-parametric fit of the
normalized dose response curves.
[0605] Results for certain compounds described herein are shown in
Table 2.
TABLE-US-00007 TABLE 2 Biological Assay Results Cmpd No Biochemical
IC.sub.50 ICW EC.sub.50 Proliferation EC.sub.50 1 B B -- 2 C -- --
3 C -- -- 4 A -- -- 5 D -- -- 6 A A B 7 B B D 8 B B D 9 B D D 10 C
-- -- 11 B B D 12 B B D 13 C -- -- 14 C -- -- 15 B B C 16 B B D 17
B B D 18 C C ** 19 A B C 20 A B C 21 B B ** 22 A B C 23 A B C 24 B
-- -- 25 B -- -- 26 A B C 27 A B C 28 A B C 29 B B ** 30 B B D 31 C
B D 32 B B D 33 C -- -- 34 B B D 35 B B D 36 B B ** 37 A A C 38 A A
C 39 A A B 40 A B C 41 C -- -- 42 B B ** 43 A B C 44 B B D 45 A A B
46 B B D 47 A B C 48 A B D 49 A B C 50 A A C 51 D -- -- 52 C -- --
53 A B C 54 B B -- 55 B -- -- 56 C -- -- 57 D -- -- 58 D -- -- 59 C
-- -- 60 B C -- 61 C -- -- 62 C -- -- 63 D -- -- 64 A B C 65 A B C
66 A B C 67 A A C 68 A B ** 69 B C -- 70 A B ** 71 A B ** 72 C --
-- 73 A A B 74 A B C 75 A A C 76 A B C 77 A A C 78 B B -- 79 A B C
80 A B D 81 A A B 82 A A C 83 B B D 84 A B C 85 C C -- 86 A B D 87
C -- -- 88 A B D 89 B C -- 90 A B D 91 A B C 92 A A C 93 A A C 94 A
B D 95 A B D 96 A B D 97 B B C 98 A A C 99 A B C 100 A A C 101 A A
C 102 A A C 103 A B ** 104 B C ** 105 A B C 106 B B ** 107 A A C
108 A B D 109 A A B 110 A A B 111 A A B 112 B B ** 113 B B D 114 B
C ** 115 D -- -- 116 C C ** 117 B B C 118 B C ** 119 A B D 120 B C
** 121 C -- -- 122 B C ** 123 A B C 124 C C ** 125 C -- -- 126 E --
-- 127 B C ** 128 E -- -- 129 B C ** 130 A B C 131 C -- -- 132 C --
-- 133 * -- -- 134 B C ** 135 C -- -- 136 C -- -- 137 B C ** 138 B
C -- 139 * -- -- 140 C -- -- 141 C -- -- 142 B B ** 143 C -- -- 144
* -- -- 145 C -- -- 146 A B -- 147 * -- -- 148 * -- -- 149 A B --
150 B -- -- 151 B -- -- 152 C -- -- 153 A B D 154 C -- -- 155 A A C
156 C -- -- 157 C -- -- 158 A B C 159 A A C 160 B C ** 161 B C **
162 C C ** 163 A B C 164 A A B 165 B B ** 166 C -- -- 167 C -- --
168 A A C 169 A A C 170 A B D 171 C -- -- 172 B B D 173 C -- -- 174
B B ** 175 A A B 176 C -- -- 177 B B D 178 B B C 179 B B D 180 C --
-- 181 A A -- 182 B B -- 183 B B -- 184 C -- -- 185 B -- -- 186 C
-- -- 187 C -- -- 188 C -- -- 189 B -- -- 190 A -- -- 191 A -- --
192 B -- -- 193 C -- -- 194 B F ** 195 C F ** 196 B F ** 197 A B D
198 A F ** 199 C F ** 200 C F ** 201 A A C 202 A B D 203 B B ** 204
B F D 205 C F D 206 C F ** 207 B B D 208 B B D 209 B B D 210 B B D
211 C F ** 212 C -- -- 213 B B ** 214 A A B 215 A F ** 216 B B **
217 B B ** 218 B F ** 219 A A B 220 A B D 221 A B D 222 B B C 223 B
B C 224 A A C 225 A A B 226 A A B 227 A A B 228 A A B 229 B B D 230
C -- -- 231 B F ** 232 A A C 233 C -- -- 234 B F ** 235 C -- -- 236
B F D 237 B F ** 238 A A B 239 A A C 240 B B D 241 A A B 242 A A C
243 A A C 244 A F ** 245 B F **
246 A B ** 247 B F ** 248 C -- -- 249 A A C 250 A A C 251 B -- --
252 -- A A 253 A B D 254 B F D 255 B B ** 256 B B ** 257 A A B 258
B B C 259 B B ** 260 A A C 261 A B C 262 A B D 263 B B D 264 B F D
265 B F ** 266 B F ** 267 B F ** 268 C -- -- 269 B B G 270 C -- --
271 A A C 272 A -- -- 273 A B D 274 A A B 275 A A A 276 A A A 277 A
A A 278 A A A For Table 2, "A" indicates an IC.sub.50 or EC.sub.50
<0.100 .mu.M, "B" indicates an IC.sub.50 or EC.sub.50 of
0.101-1.000 .mu.M, "C" indicates an IC.sub.50 or EC.sub.50 of
1.001-10.000 .mu.M, "D" indicates an IC.sub.50 or EC.sub.50 of
10.001-50 .mu.M, and "E" indicates an IC.sub.50 or EC.sub.50 >50
.mu.M, "--" indicates no data, "F" indicates an IC.sub.50 or
EC.sub.50 >1 .mu.M, "G" indicates an IC.sub.50 or EC.sub.50
>5 .mu.M, "*" indicates an IC.sub.50 or EC.sub.50 >10 .mu.M,
"**" indicates an IC.sub.50 or EC.sub.50 >20 .mu.M.
Other Embodiments
[0606] The foregoing has been a description of certain non-limiting
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
71637PRTHomo sapiens 1Met Ala Ala Met Ala Val Gly Gly Ala Gly Gly
Ser Arg Val Ser Ser 1 5 10 15 Gly Arg Asp Leu Asn Cys Val Pro Glu
Ile Ala Asp Thr Leu Gly Ala 20 25 30 Val Ala Lys Gln Gly Phe Asp
Phe Leu Cys Met Pro Val Phe His Pro 35 40 45 Arg Phe Lys Arg Glu
Phe Ile Gln Glu Pro Ala Lys Asn Arg Pro Gly 50 55 60 Pro Gln Thr
Arg Ser Asp Leu Leu Leu Ser Gly Arg Asp Trp Asn Thr 65 70 75 80 Leu
Ile Val Gly Lys Leu Ser Pro Trp Ile Arg Pro Asp Ser Lys Val 85 90
95 Glu Lys Ile Arg Arg Asn Ser Glu Ala Ala Met Leu Gln Glu Leu Asn
100 105 110 Phe Gly Ala Tyr Leu Gly Leu Pro Ala Phe Leu Leu Pro Leu
Asn Gln 115 120 125 Glu Asp Asn Thr Asn Leu Ala Arg Val Leu Thr Asn
His Ile His Thr 130 135 140 Gly His His Ser Ser Met Phe Trp Met Arg
Val Pro Leu Val Ala Pro 145 150 155 160 Glu Asp Leu Arg Asp Asp Ile
Ile Glu Asn Ala Pro Thr Thr His Thr 165 170 175 Glu Glu Tyr Ser Gly
Glu Glu Lys Thr Trp Met Trp Trp His Asn Phe 180 185 190 Arg Thr Leu
Cys Asp Tyr Ser Lys Arg Ile Ala Val Ala Leu Glu Ile 195 200 205 Gly
Ala Asp Leu Pro Ser Asn His Val Ile Asp Arg Trp Leu Gly Glu 210 215
220 Pro Ile Lys Ala Ala Ile Leu Pro Thr Ser Ile Phe Leu Thr Asn Lys
225 230 235 240 Lys Gly Phe Pro Val Leu Ser Lys Met His Gln Arg Leu
Ile Phe Arg 245 250 255 Leu Leu Lys Leu Glu Val Gln Phe Ile Ile Thr
Gly Thr Asn His His 260 265 270 Ser Glu Lys Glu Phe Cys Ser Tyr Leu
Gln Tyr Leu Glu Tyr Leu Ser 275 280 285 Gln Asn Arg Pro Pro Pro Asn
Ala Tyr Glu Leu Phe Ala Lys Gly Tyr 290 295 300 Glu Asp Tyr Leu Gln
Ser Pro Leu Gln Pro Leu Met Asp Asn Leu Glu 305 310 315 320 Ser Gln
Thr Tyr Glu Val Phe Glu Lys Asp Pro Ile Lys Tyr Ser Gln 325 330 335
Tyr Gln Gln Ala Ile Tyr Lys Cys Leu Leu Asp Arg Val Pro Glu Glu 340
345 350 Glu Lys Asp Thr Asn Val Gln Val Leu Met Val Leu Gly Ala Gly
Arg 355 360 365 Gly Pro Leu Val Asn Ala Ser Leu Arg Ala Ala Lys Gln
Ala Asp Arg 370 375 380 Arg Ile Lys Leu Tyr Ala Val Glu Lys Asn Pro
Asn Ala Val Val Thr 385 390 395 400 Leu Glu Asn Trp Gln Phe Glu Glu
Trp Gly Ser Gln Val Thr Val Val 405 410 415 Ser Ser Asp Met Arg Glu
Trp Val Ala Pro Glu Lys Ala Asp Ile Ile 420 425 430 Val Ser Glu Leu
Leu Gly Ser Phe Ala Asp Asn Glu Leu Ser Pro Glu 435 440 445 Cys Leu
Asp Gly Ala Gln His Phe Leu Lys Asp Asp Gly Val Ser Ile 450 455 460
Pro Gly Glu Tyr Thr Ser Phe Leu Ala Pro Ile Ser Ser Ser Lys Leu 465
470 475 480 Tyr Asn Glu Val Arg Ala Cys Arg Glu Lys Asp Arg Asp Pro
Glu Ala 485 490 495 Gln Phe Glu Met Pro Tyr Val Val Arg Leu His Asn
Phe His Gln Leu 500 505 510 Ser Ala Pro Gln Pro Cys Phe Thr Phe Ser
His Pro Asn Arg Asp Pro 515 520 525 Met Ile Asp Asn Asn Arg Tyr Cys
Thr Leu Glu Phe Pro Val Glu Val 530 535 540 Asn Thr Val Leu His Gly
Phe Ala Gly Tyr Phe Glu Thr Val Leu Tyr 545 550 555 560 Gln Asp Ile
Thr Leu Ser Ile Arg Pro Glu Thr His Ser Pro Gly Met 565 570 575 Phe
Ser Trp Phe Pro Ile Leu Phe Pro Ile Lys Gln Pro Ile Thr Val 580 585
590 Arg Glu Gly Gln Thr Ile Cys Val Arg Phe Trp Arg Cys Ser Asn Ser
595 600 605 Lys Lys Val Trp Tyr Glu Trp Ala Val Thr Ala Pro Val Cys
Ser Ala 610 615 620 Ile His Asn Pro Thr Gly Arg Ser Tyr Thr Ile Gly
Leu 625 630 635 2620PRTHomo sapiens 2Met Arg Gly Pro Asn Ser Gly
Thr Glu Lys Gly Arg Leu Val Ile Pro 1 5 10 15 Glu Lys Gln Gly Phe
Asp Phe Leu Cys Met Pro Val Phe His Pro Arg 20 25 30 Phe Lys Arg
Glu Phe Ile Gln Glu Pro Ala Lys Asn Arg Pro Gly Pro 35 40 45 Gln
Thr Arg Ser Asp Leu Leu Leu Ser Gly Arg Asp Trp Asn Thr Leu 50 55
60 Ile Val Gly Lys Leu Ser Pro Trp Ile Arg Pro Asp Ser Lys Val Glu
65 70 75 80 Lys Ile Arg Arg Asn Ser Glu Ala Ala Met Leu Gln Glu Leu
Asn Phe 85 90 95 Gly Ala Tyr Leu Gly Leu Pro Ala Phe Leu Leu Pro
Leu Asn Gln Glu 100 105 110 Asp Asn Thr Asn Leu Ala Arg Val Leu Thr
Asn His Ile His Thr Gly 115 120 125 His His Ser Ser Met Phe Trp Met
Arg Val Pro Leu Val Ala Pro Glu 130 135 140 Asp Leu Arg Asp Asp Ile
Ile Glu Asn Ala Pro Thr Thr His Thr Glu 145 150 155 160 Glu Tyr Ser
Gly Glu Glu Lys Thr Trp Met Trp Trp His Asn Phe Arg 165 170 175 Thr
Leu Cys Asp Tyr Ser Lys Arg Ile Ala Val Ala Leu Glu Ile Gly 180 185
190 Ala Asp Leu Pro Ser Asn His Val Ile Asp Arg Trp Leu Gly Glu Pro
195 200 205 Ile Lys Ala Ala Ile Leu Pro Thr Ser Ile Phe Leu Thr Asn
Lys Lys 210 215 220 Gly Phe Pro Val Leu Ser Lys Met His Gln Arg Leu
Ile Phe Arg Leu 225 230 235 240 Leu Lys Leu Glu Val Gln Phe Ile Ile
Thr Gly Thr Asn His His Ser 245 250 255 Glu Lys Glu Phe Cys Ser Tyr
Leu Gln Tyr Leu Glu Tyr Leu Ser Gln 260 265 270 Asn Arg Pro Pro Pro
Asn Ala Tyr Glu Leu Phe Ala Lys Gly Tyr Glu 275 280 285 Asp Tyr Leu
Gln Ser Pro Leu Gln Pro Leu Met Asp Asn Leu Glu Ser 290 295 300 Gln
Thr Tyr Glu Val Phe Glu Lys Asp Pro Ile Lys Tyr Ser Gln Tyr 305 310
315 320 Gln Gln Ala Ile Tyr Lys Cys Leu Leu Asp Arg Val Pro Glu Glu
Glu 325 330 335 Lys Asp Thr Asn Val Gln Val Leu Met Val Leu Gly Ala
Gly Arg Gly 340 345 350 Pro Leu Val Asn Ala Ser Leu Arg Ala Ala Lys
Gln Ala Asp Arg Arg 355 360 365 Ile Lys Leu Tyr Ala Val Glu Lys Asn
Pro Asn Ala Val Val Thr Leu 370 375 380 Glu Asn Trp Gln Phe Glu Glu
Trp Gly Ser Gln Val Thr Val Val Ser 385 390 395 400 Ser Asp Met Arg
Glu Trp Val Ala Pro Glu Lys Ala Asp Ile Ile Val 405 410 415 Ser Glu
Leu Leu Gly Ser Phe Ala Asp Asn Glu Leu Ser Pro Glu Cys 420 425 430
Leu Asp Gly Ala Gln His Phe Leu Lys Asp Asp Gly Val Ser Ile Pro 435
440 445 Gly Glu Tyr Thr Ser Phe Leu Ala Pro Ile Ser Ser Ser Lys Leu
Tyr 450 455 460 Asn Glu Val Arg Ala Cys Arg Glu Lys Asp Arg Asp Pro
Glu Ala Gln 465 470 475 480 Phe Glu Met Pro Tyr Val Val Arg Leu His
Asn Phe His Gln Leu Ser 485 490 495 Ala Pro Gln Pro Cys Phe Thr Phe
Ser His Pro Asn Arg Asp Pro Met 500 505 510 Ile Asp Asn Asn Arg Tyr
Cys Thr Leu Glu Phe Pro Val Glu Val Asn 515 520 525 Thr Val Leu His
Gly Phe Ala Gly Tyr Phe Glu Thr Val Leu Tyr Gln 530 535 540 Asp Ile
Thr Leu Ser Ile Arg Pro Glu Thr His Ser Pro Gly Met Phe 545 550 555
560 Ser Trp Phe Pro Ile Leu Phe Pro Ile Lys Gln Pro Ile Thr Val Arg
565 570 575 Glu Gly Gln Thr Ile Cys Val Arg Phe Trp Arg Cys Ser Asn
Ser Lys 580 585 590 Lys Val Trp Tyr Glu Trp Ala Val Thr Ala Pro Val
Cys Ser Ala Ile 595 600 605 His Asn Pro Thr Gly Arg Ser Tyr Thr Ile
Gly Leu 610 615 620 316PRTArtificial SequenceSynthetic Polypeptide
3Ser Gly Arg Gly Lys Gly Gly Lys Gly Leu Gly Lys Gly Gly Ala Lys 1
5 10 15 49PRTArtificial SequenceSynthetic Polypeptide 4Met Asp Tyr
Lys Asp Asp Asp Asp Lys 1 5 57PRTArtificial SequenceSynthetic
Polypeptide 5Met His His His His His His 1 5 6645PRTArtificial
SequenceSynthetic Polypeptide 6Met Asp Tyr Lys Asp Asp Asp Asp Lys
Ala Ala Met Ala Val Gly Gly 1 5 10 15 Ala Gly Gly Ser Arg Val Ser
Ser Gly Arg Asp Leu Asn Cys Val Pro 20 25 30 Glu Ile Ala Asp Thr
Leu Gly Ala Val Ala Lys Gln Gly Phe Asp Phe 35 40 45 Leu Cys Met
Pro Val Phe His Pro Arg Phe Lys Arg Glu Phe Ile Gln 50 55 60 Glu
Pro Ala Lys Asn Arg Pro Gly Pro Gln Thr Arg Ser Asp Leu Leu 65 70
75 80 Leu Ser Gly Arg Asp Trp Asn Thr Leu Ile Val Gly Lys Leu Ser
Pro 85 90 95 Trp Ile Arg Pro Asp Ser Lys Val Glu Lys Ile Arg Arg
Asn Ser Glu 100 105 110 Ala Ala Met Leu Gln Glu Leu Asn Phe Gly Ala
Tyr Leu Gly Leu Pro 115 120 125 Ala Phe Leu Leu Pro Leu Asn Gln Glu
Asp Asn Thr Asn Leu Ala Arg 130 135 140 Val Leu Thr Asn His Ile His
Thr Gly His His Ser Ser Met Phe Trp 145 150 155 160 Met Arg Val Pro
Leu Val Ala Pro Glu Asp Leu Arg Asp Asp Ile Ile 165 170 175 Glu Asn
Ala Pro Thr Thr His Thr Glu Glu Tyr Ser Gly Glu Glu Lys 180 185 190
Thr Trp Met Trp Trp His Asn Phe Arg Thr Leu Cys Asp Tyr Ser Lys 195
200 205 Arg Ile Ala Val Ala Leu Glu Ile Gly Ala Asp Leu Pro Ser Asn
His 210 215 220 Val Ile Asp Arg Trp Leu Gly Glu Pro Ile Lys Ala Ala
Ile Leu Pro 225 230 235 240 Thr Ser Ile Phe Leu Thr Asn Lys Lys Gly
Phe Pro Val Leu Ser Lys 245 250 255 Met His Gln Arg Leu Ile Phe Arg
Leu Leu Lys Leu Glu Val Gln Phe 260 265 270 Ile Ile Thr Gly Thr Asn
His His Ser Glu Lys Glu Phe Cys Ser Tyr 275 280 285 Leu Gln Tyr Leu
Glu Tyr Leu Ser Gln Asn Arg Pro Pro Pro Asn Ala 290 295 300 Tyr Glu
Leu Phe Ala Lys Gly Tyr Glu Asp Tyr Leu Gln Ser Pro Leu 305 310 315
320 Gln Pro Leu Met Asp Asn Leu Glu Ser Gln Thr Tyr Glu Val Phe Glu
325 330 335 Lys Asp Pro Ile Lys Tyr Ser Gln Tyr Gln Gln Ala Ile Tyr
Lys Cys 340 345 350 Leu Leu Asp Arg Val Pro Glu Glu Glu Lys Asp Thr
Asn Val Gln Val 355 360 365 Leu Met Val Leu Gly Ala Gly Arg Gly Pro
Leu Val Asn Ala Ser Leu 370 375 380 Arg Ala Ala Lys Gln Ala Asp Arg
Arg Ile Lys Leu Tyr Ala Val Glu 385 390 395 400 Lys Asn Pro Asn Ala
Val Val Thr Leu Glu Asn Trp Gln Phe Glu Glu 405 410 415 Trp Gly Ser
Gln Val Thr Val Val Ser Ser Asp Met Arg Glu Trp Val 420 425 430 Ala
Pro Glu Lys Ala Asp Ile Ile Val Ser Glu Leu Leu Gly Ser Phe 435 440
445 Ala Asp Asn Glu Leu Ser Pro Glu Cys Leu Asp Gly Ala Gln His Phe
450 455 460 Leu Lys Asp Asp Gly Val Ser Ile Pro Gly Glu Tyr Thr Ser
Phe Leu 465 470 475 480 Ala Pro Ile Ser Ser Ser Lys Leu Tyr Asn Glu
Val Arg Ala Cys Arg 485 490 495 Glu Lys Asp Arg Asp Pro Glu Ala Gln
Phe Glu Met Pro Tyr Val Val 500 505 510 Arg Leu His Asn Phe His Gln
Leu Ser Ala Pro Gln Pro Cys Phe Thr 515 520 525 Phe Ser His Pro Asn
Arg Asp Pro Met Ile Asp Asn Asn Arg Tyr Cys 530 535 540 Thr Leu Glu
Phe Pro Val Glu Val Asn Thr Val Leu His Gly Phe Ala 545 550 555 560
Gly Tyr Phe Glu Thr Val Leu Tyr Gln Asp Ile Thr Leu Ser Ile Arg 565
570 575 Pro Glu Thr His Ser Pro Gly Met Phe Ser Trp Phe Pro Ile Leu
Phe 580 585 590 Pro Ile Lys Gln Pro Ile Thr Val Arg Glu Gly Gln Thr
Ile Cys Val 595 600 605 Arg Phe Trp Arg Cys Ser Asn Ser Lys Lys Val
Trp Tyr Glu Trp Ala 610 615 620 Val Thr Ala Pro Val Cys Ser Ala Ile
His Asn Pro Thr Gly Arg Ser 625 630 635 640 Tyr Thr Ile Gly Leu 645
7348PRTArtificial SequenceSynthetic Polypeptide 7Met His His His
His His His Arg Lys Glu Thr Pro Pro Pro Leu Val 1 5 10 15 Pro Pro
Ala Ala Arg Glu Trp Asn Leu Pro Pro Asn Ala Pro Ala Cys 20 25 30
Met Glu Arg Gln Leu Glu Ala Ala Arg Tyr Arg Ser Asp Gly Ala Leu 35
40 45 Leu Leu Gly Ala Ser Ser Leu Ser Gly Arg Cys Trp Ala Gly Ser
Leu 50 55 60 Trp Leu Phe Lys Asp Pro Cys Ala Ala Pro Asn Glu Gly
Phe Cys Ser 65 70 75 80 Ala Gly Val Gln Thr Glu Ala Gly Val Ala Asp
Leu Thr Trp Val Gly 85 90 95 Glu Arg Gly Ile Leu Val Ala Ser Asp
Ser Gly Ala Val Glu Leu Trp 100 105 110 Glu Leu Asp Glu Asn Glu Thr
Leu Ile Val Ser Lys Phe Cys Lys Tyr 115 120 125 Glu His Asp Asp Ile
Val Ser Thr Val Ser Val Leu Ser Ser Gly Thr 130 135 140 Gln Ala Val
Ser Gly Ser Lys Asp Ile Cys Ile Lys Val Trp Asp Leu 145 150 155 160
Ala Gln Gln Val Val Leu Ser Ser Tyr Arg Ala His Ala Ala Gln Val 165
170 175 Thr Cys Val Ala Ala Ser Pro His Lys Asp Ser Val Phe Leu Ser
Cys 180 185 190 Ser Glu Asp Asn Arg Ile Leu Leu Trp Asp Thr Arg Cys
Pro Lys Pro 195 200 205 Ala Ser Gln Ile Gly Cys Ser Ala Pro Gly Tyr
Leu Pro Thr Ser Leu 210 215 220 Ala Trp His Pro Gln Gln Ser Glu Val
Phe Val Phe Gly Asp Glu Asn 225 230 235 240 Gly Thr Val Ser Leu Val
Asp Thr Lys Ser Thr Ser Cys Val Leu Ser 245 250 255 Ser Ala Val His
Ser Gln Cys Val Thr Gly Leu Val Phe Ser Pro His 260 265 270 Ser Val
Pro Phe Leu Ala Ser Leu Ser Glu Asp Cys Ser Leu Ala Val 275 280 285
Leu Asp Ser Ser Leu Ser Glu Leu Phe Arg Ser Gln Ala His Arg Asp 290
295 300 Phe Val Arg Asp Ala Thr Trp Ser Pro Leu Asn His Ser Leu Leu
Thr 305 310 315 320 Thr Val Gly Trp Asp His Gln Val Val His His Val
Val Pro Thr Glu 325 330
335 Pro Leu Pro Ala Pro Gly Pro Ala Ser Val Thr Glu 340 345
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