U.S. patent application number 16/897690 was filed with the patent office on 2021-08-19 for substituted purine and 7-deazapurine compounds.
The applicant listed for this patent is Epizyme, Inc.. Invention is credited to Richard CHESWORTH, Scott R. DAIGLE, Kevin W. KUNTZ, Edward J. OLHAVA, Roy M. POLLOCK, Victoria M. RICHON.
Application Number | 20210252035 16/897690 |
Document ID | / |
Family ID | 1000005078766 |
Filed Date | 2021-08-19 |
United States Patent
Application |
20210252035 |
Kind Code |
A1 |
OLHAVA; Edward J. ; et
al. |
August 19, 2021 |
SUBSTITUTED PURINE AND 7-DEAZAPURINE COMPOUNDS
Abstract
The present invention relates to substituted purine and
7-deazapurine compounds. The present invention also relates to
pharmaceutical compositions containing these compounds and methods
of treating disorders in which DOT1-mediated protein methylation
plays a part, such as cancer and neurological disorders, by
administering these compounds and pharmaceutical compositions to
subjects in need thereof.
Inventors: |
OLHAVA; Edward J.; (Newton,
MA) ; CHESWORTH; Richard; (Concord, MA) ;
KUNTZ; Kevin W.; (Woburn, MA) ; RICHON; Victoria
M.; (Wellesley, MA) ; POLLOCK; Roy M.;
(Medford, MA) ; DAIGLE; Scott R.; (Newburyport,
MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Epizyme, Inc. |
Cambridge |
MA |
US |
|
|
Family ID: |
1000005078766 |
Appl. No.: |
16/897690 |
Filed: |
June 10, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15802979 |
Nov 3, 2017 |
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16897690 |
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14752023 |
Jun 26, 2015 |
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15802979 |
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14055510 |
Oct 16, 2013 |
9096634 |
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14752023 |
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13310157 |
Dec 2, 2011 |
8580762 |
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14055510 |
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61419661 |
Dec 3, 2010 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07H 19/14 20130101;
A61K 31/519 20130101; A61K 31/7076 20130101; A61K 31/52 20130101;
C07H 19/16 20130101; C07D 487/04 20130101; C07D 473/34 20130101;
A61K 31/7064 20130101 |
International
Class: |
A61K 31/7076 20060101
A61K031/7076; C07D 487/04 20060101 C07D487/04; C07D 473/34 20060101
C07D473/34; C07H 19/14 20060101 C07H019/14; C07H 19/16 20060101
C07H019/16; A61K 31/519 20060101 A61K031/519; A61K 31/52 20060101
A61K031/52; A61K 31/7064 20060101 A61K031/7064 |
Claims
1. (canceled)
2. A method of treating cancer comprising administering to a
subject in need thereof a therapeutically effective amount of a
DOT1L inhibitor.
3. A method of treating hematological cancer comprising
administering to a subject in need thereof a therapeutically
effective amount of a DOT1L inhibitor.
4. A method of treating leukemia comprising administering to a
subject in need thereof a therapeutically effective amount of a
DOT1L inhibitor.
5. The method of claim 4, wherein the leukemia is acute myeloid
leukemia, acute lymphocytic leukemia or mixed lineage leukemia.
6. The method of claim 4, wherein the leukemia is a MLL-rearranged
leukemia.
7. The method of claim 4, wherein the leukemia is a leukemia
characterized by a partial tandem duplication of the MLL gene
(MLL-PTD).
8. The method of claim 2, wherein the DOT1L inhibitor is a compound
having the structure of Formula (II): ##STR00433## or a
pharmaceutically acceptable salt thereof, wherein A is O or
CH.sub.2; Q is H, NH.sub.2, NHR.sub.b, NR.sub.bR.sub.c, R.sub.b, or
OR.sub.b, in which each of R.sub.b and R, independently is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
7-membered heterocycloalkyl, 5 to 10-membered heteroaryl, or
-M.sub.1-T.sub.1 in which M.sub.1 is a bond or C.sub.1-C.sub.6
alkyl linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxyl and T.sub.1 is C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, or R.sub.b and R.sub.c, together with the N
atom to which they attach, form 4 to 7-membered heterocycloalkyl
having 0 or 1 additional heteroatoms to the N atom optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and each of R.sub.b, R.sub.c, and T.sub.1 is
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, and 5 to
6-membered heteroaryl; X is N or CR.sub.x, in which R.sub.x is H,
halo, hydroxyl, carboxyl, cyano, or R.sub.S1, R.sub.S1 being amino,
C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and R.sub.S1 being optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl; L.sub.1 is N(Y),
S, SO, or SO.sub.2; L.sub.2 is CO or absent when L.sub.1 is N(Y) or
L.sub.2 is absent when L.sub.1 is S, SO, or SO.sub.2, in which Y is
H, R.sub.d, SO.sub.2R.sub.d, or COR.sub.d when L.sub.2 is absent,
or Y is H or R.sub.d when L.sub.2 is CO, R.sub.d being
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl, and
R.sub.d being optionally substituted with one or more substituents
selected from the group consisting of C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6
alkylsulfonyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, and 5 to
6-membered heteroaryl and with C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl further optionally substituted with
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, halo, hydroxyl, carboxyl, C(O)OH, C(O)O--C.sub.1-C.sub.6
alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl; each of
R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, and R.sub.7,
independently, is H, halo, hydroxyl, carboxyl, cyano, R.sub.S2,
R.sub.S2 being amino, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl, and
each R.sub.S2 being optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl; R.sub.8 is H,
halo or R.sub.S3, R.sub.S3 being C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl, and R.sub.S3
being optionally substituted with one or more substituents selected
from the group consisting of halo, hydroxyl, carboxyl, cyano amino,
C.sub.1-C.sub.6 alkoxyl, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, and C.sub.3-C.sub.8 cycloalkyl;
R.sub.9 is ##STR00434## in which each of R.sub.e, R.sub.f, R.sub.g,
and R.sub.h, independently is -M.sub.2-T.sub.2, in which M.sub.2 is
a bond, SO.sub.2, SO, S, CO, CO.sub.2, O, O--C.sub.1-C.sub.4 alkyl
linker, C.sub.1-C.sub.4 alkyl linker, NH, or N(R.sub.t), R.sub.t
being C.sub.1-C.sub.6 alkyl, and T.sub.2 is H, halo, or R.sub.S4,
R.sub.S4 being C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 8-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, and each of O--C.sub.1-C.sub.4 alkyl
linker, C.sub.1-C.sub.4 alkyl linker, R.sub.t, and R.sub.S4 being
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, R.sub.i is H or
C.sub.1-C.sub.6 alkyl optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, D is O, NR.sub.j,
or CR.sub.jR.sub.k, each of R.sub.j and R.sub.k independently being
H or C.sub.1-C.sub.6 alkyl, or R.sub.j and R.sub.k taken together,
with the carbon atom to which they are attached, form a
C.sub.3-C.sub.10 cycloalkyl ring, and E is-M.sub.3-T.sub.3, M.sub.3
being a bond or C.sub.1-C.sub.6 alkyl linker optionally substituted
with halo or cyano, T.sub.3 being C.sub.3-C.sub.10 cycloalkyl,
C.sub.6-C.sub.10 aryl, 5 to 10-membered heteroaryl, or 4 to
10-membered heterocycloalkyl, and T.sub.3 being optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, thiol, carboxyl, cyano, nitro,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 haloalkoxyl, C.sub.1-C.sub.6 alkylthio,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.1-C.sub.6 haloalkylsulfonyl,
C.sub.1-C.sub.6 alkylcarbonyl, C.sub.1-C.sub.6 alkoxycarbonyl, oxo,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.4-C.sub.12
alkylcycloalkyl, C.sub.6-C.sub.10 aryl, C.sub.6-C.sub.10 aryloxyl,
C.sub.7-C.sub.14 alkylaryl, C.sub.6-C.sub.10 aminoaryloxyl,
C.sub.6-C.sub.10 arylthio, 4 to 6-membered heterocycloalkyl
optionally substituted with halo, C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 haloalkyl, 5 to 6-membered heteroaryl optionally
substituted with halo, C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.6
alkyl that is substituted with hydroxy, halo, C.sub.1-C.sub.6
alkoxycarbonyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl,
4 to 6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl
optionally further substituted with halo, hydroxyl, or
C.sub.1-C.sub.6 alkoxyl; q is 0, 1, 2, 3, or 4; m is 0, 1, or 2;
and n is 0, 1, or 2.
9. The method of claim 3, wherein the DOT1L inhibitor is a compound
having the structure of Formula (II): ##STR00435## or a
pharmaceutically acceptable salt thereof, wherein A is O or
CH.sub.2; Q is H, NH.sub.2, NHR.sub.b, NR.sub.bR.sub.c, R.sub.b, or
OR.sub.b, in which each of R.sub.b and R, independently is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
7-membered heterocycloalkyl, 5 to 10-membered heteroaryl, or
-M.sub.1-T.sub.1 in which M.sub.1 is a bond or C.sub.1-C.sub.6
alkyl linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxyl and T.sub.1 is C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, or R.sub.b and R.sub.c, together with the N
atom to which they attach, form 4 to 7-membered heterocycloalkyl
having 0 or 1 additional heteroatoms to the N atom optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and each of R.sub.b, R.sub.c, and T.sub.1 is
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, and 5 to
6-membered heteroaryl; X is N or CR.sub.x, in which R.sub.x is H,
halo, hydroxyl, carboxyl, cyano, or R.sub.S1, R.sub.S1 being amino,
C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and R.sub.S1 being optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl; L.sub.1 is N(Y),
S, SO, or SO.sub.2; L.sub.2 is CO or absent when L.sub.1 is N(Y) or
L.sub.2 is absent when L.sub.1 is S, SO, or SO.sub.2, in which Y is
H, R.sub.d, SO.sub.2R.sub.d, or COR.sub.d when L.sub.2 is absent,
or Y is H or R.sub.d when L.sub.2 is CO, R.sub.d being
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl, and
R.sub.d being optionally substituted with one or more substituents
selected from the group consisting of C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6
alkylsulfonyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, and 5 to
6-membered heteroaryl and with C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl further optionally substituted with
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, halo, hydroxyl, carboxyl, C(O)OH, C(O)O--C.sub.1-C.sub.6
alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl; each of
R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, and R.sub.7,
independently, is H, halo, hydroxyl, carboxyl, cyano, R.sub.S2,
R.sub.S2 being amino, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl, and
each R.sub.S2 being optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl; R.sub.8 is H,
halo or R.sub.S3, R.sub.S3 being C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl, and R.sub.S3
being optionally substituted with one or more substituents selected
from the group consisting of halo, hydroxyl, carboxyl, cyano amino,
C.sub.1-C.sub.6 alkoxyl, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, and C.sub.3-C.sub.8 cycloalkyl;
R.sub.9 is ##STR00436## in which each of R.sub.e, R.sub.f, R.sub.g,
and R.sub.h, independently is -M.sub.2-T.sub.2, in which M.sub.2 is
a bond, SO.sub.2, SO, S, CO, CO.sub.2, O, O--C.sub.1-C.sub.4 alkyl
linker, C.sub.1-C.sub.4 alkyl linker, NH, or N(R.sub.t), R.sub.t
being C.sub.1-C.sub.6 alkyl, and T.sub.2 is H, halo, or R.sub.S4,
R.sub.S4 being C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 8-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, and each of O--C.sub.1-C.sub.4 alkyl
linker, C.sub.1-C.sub.4 alkyl linker, R.sub.t, and R.sub.S4 being
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, R.sub.i is H or
C.sub.1-C.sub.6 alkyl optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, D is O, NR.sub.j,
or CR.sub.jR.sub.k, each of R.sub.j and R.sub.k independently being
H or C.sub.1-C.sub.6 alkyl, or R.sub.j and R.sub.k taken together,
with the carbon atom to which they are attached, form a
C.sub.3-C.sub.10 cycloalkyl ring, and E is-M.sub.3-T.sub.3, M.sub.3
being a bond or C.sub.1-C.sub.6 alkyl linker optionally substituted
with halo or cyano, T.sub.3 being C.sub.3-C.sub.10 cycloalkyl,
C.sub.6-C.sub.10 aryl, 5 to 10-membered heteroaryl, or 4 to
10-membered heterocycloalkyl, and T.sub.3 being optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, thiol, carboxyl, cyano, nitro,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 haloalkoxyl, C.sub.1-C.sub.6 alkylthio,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.1-C.sub.6 haloalkylsulfonyl,
C.sub.1-C.sub.6 alkylcarbonyl, C.sub.1-C.sub.6 alkoxycarbonyl, oxo,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.4-C.sub.12
alkylcycloalkyl, C.sub.6-C.sub.10 aryl, C.sub.6-C.sub.10 aryloxyl,
C.sub.7-C.sub.14 alkylaryl, C.sub.6-C.sub.10 aminoaryloxyl,
C.sub.6-C.sub.10 arylthio, 4 to 6-membered heterocycloalkyl
optionally substituted with halo, C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 haloalkyl, 5 to 6-membered heteroaryl optionally
substituted with halo, C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.6
alkyl that is substituted with hydroxy, halo, C.sub.1-C.sub.6
alkoxycarbonyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl,
4 to 6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl
optionally further substituted with halo, hydroxyl, or
C.sub.1-C.sub.6 alkoxyl; q is 0, 1, 2, 3, or 4; m is 0, 1, or 2;
and n is 0, 1, or 2.
10. The method of claim 4, wherein the DOT1L inhibitor is a
compound having the structure of Formula (II): ##STR00437## or a
pharmaceutically acceptable salt thereof, wherein A is O or
CH.sub.2; Q is H, NH.sub.2, NHR.sub.b, NR.sub.bR.sub.c, R.sub.b, or
OR.sub.b, in which each of R.sub.b and R.sub.c, independently is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
7-membered heterocycloalkyl, 5 to 10-membered heteroaryl, or
-M.sub.1-T.sub.1 in which M.sub.1 is a bond or C.sub.1-C.sub.6
alkyl linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxyl and T.sub.1 is C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, or R.sub.b and R.sub.c, together with the N
atom to which they attach, form 4 to 7-membered heterocycloalkyl
having 0 or 1 additional heteroatoms to the N atom optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and each of R.sub.b, R.sub.c, and T.sub.1 is
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, and 5 to
6-membered heteroaryl; X is N or CR.sub.x, in which R.sub.x is H,
halo, hydroxyl, carboxyl, cyano, or R.sub.S1, R.sub.S1 being amino,
C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and R.sub.S1 being optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl; L.sub.1 is N(Y),
S, SO, or SO.sub.2; L.sub.2 is CO or absent when L.sub.1 is N(Y) or
L.sub.2 is absent when L.sub.1 is S, SO, or SO.sub.2, in which Y is
H, R.sub.d, SO.sub.2R.sub.d, or COR.sub.d when L.sub.2 is absent,
or Y is H or R.sub.d when L.sub.2 is CO, R.sub.d being
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl, and
R.sub.d being optionally substituted with one or more substituents
selected from the group consisting of C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6
alkylsulfonyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, and 5 to
6-membered heteroaryl and with C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl further optionally substituted with
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, halo, hydroxyl, carboxyl, C(O)OH, C(O)O--C.sub.1-C.sub.6
alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl; each of
R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, and R.sub.7,
independently, is H, halo, hydroxyl, carboxyl, cyano, R.sub.S2,
R.sub.S2 being amino, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl, and
each R.sub.S2 being optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl; R.sub.8 is H,
halo or R.sub.S3, R.sub.S3 being C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl, and R.sub.S3
being optionally substituted with one or more substituents selected
from the group consisting of halo, hydroxyl, carboxyl, cyano amino,
C.sub.1-C.sub.6 alkoxyl, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, and C.sub.3-C.sub.8 cycloalkyl;
R.sub.9 is ##STR00438## in which each of R.sub.e, R.sub.f, R.sub.g,
and R.sub.h, independently is -M.sub.2-T.sub.2, in which M.sub.2 is
a bond, SO.sub.2, SO, S, CO, CO.sub.2, O, O--C.sub.1-C.sub.4 alkyl
linker, C.sub.1-C.sub.4 alkyl linker, NH, or N(R.sub.t), R.sub.t
being C.sub.1-C.sub.6 alkyl, and T.sub.2 is H, halo, or R.sub.S4,
R.sub.S4 being C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 8-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, and each of O--C.sub.1-C.sub.4 alkyl
linker, C.sub.1-C.sub.4 alkyl linker, R.sub.t, and R.sub.S4 being
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, R.sub.i is H or
C.sub.1-C.sub.6 alkyl optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, D is O, NR.sub.j,
or CR.sub.jR.sub.k, each of R.sub.j and R.sub.k independently being
H or C.sub.1-C.sub.6 alkyl, or R.sub.j and R.sub.k taken together,
with the carbon atom to which they are attached, form a
C.sub.3-C.sub.10 cycloalkyl ring, and E is-M.sub.3-T.sub.3, M.sub.3
being a bond or C.sub.1-C.sub.6 alkyl linker optionally substituted
with halo or cyano, T.sub.3 being C.sub.3-C.sub.10 cycloalkyl,
C.sub.6-C.sub.10 aryl, 5 to 10-membered heteroaryl, or 4 to
10-membered heterocycloalkyl, and T.sub.3 being optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, thiol, carboxyl, cyano, nitro,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 haloalkoxyl, C.sub.1-C.sub.6 alkylthio,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.1-C.sub.6 haloalkylsulfonyl,
C.sub.1-C.sub.6 alkylcarbonyl, C.sub.1-C.sub.6 alkoxycarbonyl, oxo,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.4-C.sub.12
alkylcycloalkyl, C.sub.6-C.sub.10 aryl, C.sub.6-C.sub.10 aryloxyl,
C.sub.7-C.sub.14 alkylaryl, C.sub.6-C.sub.10 aminoaryloxyl,
C.sub.6-C.sub.10 arylthio, 4 to 6-membered heterocycloalkyl
optionally substituted with halo, C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 haloalkyl, 5 to 6-membered heteroaryl optionally
substituted with halo, C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.6
alkyl that is substituted with hydroxy, halo, C.sub.1-C.sub.6
alkoxycarbonyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl,
4 to 6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl
optionally further substituted with halo, hydroxyl, or
C.sub.1-C.sub.6 alkoxyl; q is 0, 1, 2, 3, or 4; m is 0, 1, or 2;
and n is 0, 1, or 2.
11. The method of claim 8, wherein A is O.
12. The method of claim 9, wherein A is O.
13. The method of claim 10, wherein A is O.
14. The method of claim 8, wherein Q is NH.sub.2 or NHR.sub.b, in
which R.sub.b is -M.sub.1-T.sub.1, M.sub.1 being a bond or
C.sub.1-C.sub.6 alkyl linker and T.sub.1 being C.sub.3-C.sub.8
cycloalkyl.
15. The method of claim 9, wherein Q is NH.sub.2 or NHR.sub.b, in
which R.sub.b is -M.sub.1-T.sub.1, M.sub.1 being a bond or
C.sub.1-C.sub.6 alkyl linker and T.sub.1 being C.sub.3-C.sub.8
cycloalkyl.
16. The method of claim 10, wherein Q is NH.sub.2 or NHR.sub.b, in
which R.sub.b is -M.sub.1-T.sub.1, M.sub.1 being a bond or
C.sub.1-C.sub.6 alkyl linker and T.sub.1 being C.sub.3-C.sub.8
cycloalkyl.
17. The method of claim 2, wherein the DOT1L inhibitor is a
compound selected from: ##STR00439## ##STR00440## ##STR00441##
##STR00442## ##STR00443## ##STR00444## ##STR00445## ##STR00446##
##STR00447## ##STR00448## ##STR00449## ##STR00450## ##STR00451##
##STR00452## ##STR00453## ##STR00454## ##STR00455## ##STR00456##
##STR00457## ##STR00458## ##STR00459## ##STR00460## ##STR00461##
##STR00462## ##STR00463## ##STR00464## ##STR00465## ##STR00466##
##STR00467## ##STR00468## ##STR00469## ##STR00470## ##STR00471##
##STR00472## ##STR00473## ##STR00474## ##STR00475## ##STR00476##
##STR00477## ##STR00478## ##STR00479## and pharmaceutically
acceptable salts thereof.
18. The method of claim 3, wherein the DOT1L inhibitor is a
compound selected from: ##STR00480## ##STR00481## ##STR00482##
##STR00483## ##STR00484## ##STR00485## ##STR00486## ##STR00487##
##STR00488## ##STR00489## ##STR00490## ##STR00491## ##STR00492##
##STR00493## ##STR00494## ##STR00495## ##STR00496## ##STR00497##
##STR00498## ##STR00499## ##STR00500## ##STR00501## ##STR00502##
##STR00503## ##STR00504## ##STR00505## ##STR00506## ##STR00507##
##STR00508## ##STR00509## and pharmaceutically acceptable salts
thereof.
19. The method of claim 4, wherein the DOT1L inhibitor is a
compound selected from: ##STR00510## ##STR00511## ##STR00512##
##STR00513## ##STR00514## ##STR00515## ##STR00516## ##STR00517##
##STR00518## ##STR00519## ##STR00520## ##STR00521## ##STR00522##
##STR00523## ##STR00524## ##STR00525## ##STR00526## ##STR00527##
##STR00528## ##STR00529## ##STR00530## ##STR00531## ##STR00532##
##STR00533## ##STR00534## ##STR00535## ##STR00536## ##STR00537##
##STR00538## ##STR00539## ##STR00540## ##STR00541## ##STR00542##
##STR00543## ##STR00544## ##STR00545## ##STR00546## ##STR00547##
##STR00548## ##STR00549## ##STR00550## ##STR00551## ##STR00552##
##STR00553## and pharmaceutically acceptable salts thereof.
20. The method of claim 3, wherein the DOT1L inhibitor is Compound
2: ##STR00554## or a pharmaceutically acceptable salt thereof.
21. The method of claim 4, wherein the DOT1L inhibitor is Compound
2: ##STR00555## or a pharmaceutically acceptable salt thereof.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation of U.S. application Ser.
No. 15/802,979, filed Nov. 3, 2017, which is a continuation of U.S.
application Ser. No. 14/752,023, filed Jun. 26, 2015 (now
abandoned), which is a continuation of U.S. application Ser. No.
14/055,510, filed Oct. 16, 2013, now U.S. Pat. No. 9,096,634, which
is a continuation of U.S. application Ser. No. 13/310,157, filed
Dec. 2, 2011, now U.S. Pat. No. 8,580,762, which claims priority
to, and the benefit of, U.S. provisional application No.
61/419,661, filed Dec. 3, 2010, the contents of each of which are
incorporated herein by reference in their entireties.
BACKGROUND OF THE INVENTION
[0002] In eukaryotic cells, DNA is packaged with histones to form
chromatin. Approximately 150 base pairs of DNA are wrapped twice
around an octamer of histones (two each of histones 2A, 2B, 3, and
4) to form a nucleosome, the basic unit of chromatin. Changes in
the ordered structure of chromatin can lead to alterations in
transcription of associated genes. This process is highly
controlled because changes in gene expression patterns can
profoundly affect fundamental cellular processes such as
differentiation, proliferation, and apoptosis. Control of changes
in chromatin structure (and hence of transcription) is mediated by
covalent modifications to histones, most notably of their
N-terminal tails. These modifications are often referred to as
epigenetic because they can lead to heritable changes in gene
expression, but do not affect the sequence of the DNA itself.
Covalent modifications (for example, methylation, acetylation,
phosphorylation, and ubiquitination) of the side chains of amino
acids are enzymatically mediated.
[0003] The selective addition of methyl groups to specific amino
acid sites on histones is controlled by the action of a unique
family of enzymes known as histone methyltransferases (HMTs). The
level of expression of a particular gene is influenced by the
presence or absence of a methyl group at a relevant histone site.
The specific effect of a methyl group at a particular histone site
persists until the methyl group is removed by a histone
demethylase, or until the modified histone is replaced through
nucleosome turnover. In a like manner, other enzyme classes can
decorate DNA and histones with other chemical species and still
other enzymes can remove these species to provide temporal control
of gene expression.
[0004] The orchestrated collection of biochemical systems behind
transcriptional regulation must be tightly controlled in order for
cell growth and differentiation to proceed optimally. Disease
states result when these controls are disrupted by aberrant
expression and/or activity of the enzymes responsible for DNA and
histone modification. In human cancers, for example, there is a
growing body of evidence to suggest that dysregulated epigenetic
enzyme activity contributes to the uncontrolled cell proliferation
associated with cancer as well as other cancer-relevant phenotypes
such as enhanced cell migration and invasion. Beyond cancer, there
is growing evidence for a role of epigenetic enzymes in a number of
other human diseases, including metabolic diseases (such as
diabetes), inflammatory diseases (such as Crohn's disease),
neurodegenerative diseases (such as Alzheimer's disease) and
cardiovascular diseases. Therefore, selectively modulating the
aberrant action of epigenetic enzymes holds great promise for the
treatment of a range of diseases.
[0005] There is an ongoing need for new agents which modulate the
aberrant action of epigenetic enzymes. The present invention
provides compounds that meet this need.
SUMMARY OF THE INVENTION
[0006] The invention provides compounds useful for modulating the
aberrant action of epigenetic enzymes. The present invention also
provides pharmaceutically acceptable salts, esters, and/or
N-oxides, of these compounds.
[0007] In one aspect, the present invention features a substituted
purine or 7-deazapurine compound of Formula (I) below or a
pharmaceutically acceptable salt or ester thereof.
##STR00001##
In this formula,
[0008] A is O or CH.sub.2;
[0009] each of G and J, independently, is H, halo, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl or OR.sub.a, R.sub.a being H,
C.sub.1-C.sub.6 alkyl or C(O)--C.sub.1-C.sub.6 alkyl, wherein
C(O)O--C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkyl or
C(O)--C.sub.1-C.sub.6 alkyl is optionally substituted with one or
more substituents selected from the group consisting of halo, cyano
hydroxyl, carboxyl, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, and
C.sub.3-C.sub.8 cycloalkyl;
[0010] Q is H, NH.sub.2, NHR.sub.b, NR.sub.bR.sub.c, R.sub.b, or
OR.sub.b, in which each of R.sub.b and R.sub.c independently is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
7-membered heterocycloalkyl, 5 to 10-membered heteroaryl, or
-M.sub.1-T.sub.1 in which M.sub.1 is a bond or C.sub.1-C.sub.6
alkyl linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxyl and T.sub.1 is C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, or R.sub.b and R.sub.c, together with the N
atom to which they attach, form 4 to 7-membered heterocycloalkyl
having 0 or 1 additional heteroatoms to the N atom optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and each of R.sub.b, R.sub.c, and T.sub.1 is
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, and 5 to
6-membered heteroaryl;
[0011] X is N or CR.sub.x, in which R.sub.x is H, halo, hydroxyl,
carboxyl, cyano, or R.sub.S1, R.sub.S1 being amino, C.sub.1-C.sub.6
alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and R.sub.S1 being optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl;
[0012] L.sub.1 is N(Y), S, SO, or SO.sub.2;
[0013] L.sub.2 is CO or absent when L.sub.1 is N(Y) or L.sub.2 is
absent when L.sub.1 is S, SO, or SO.sub.2, in which Y is H, Rd,
SO.sub.2R.sub.a, or COR.sub.a when L.sub.2 is absent, or Y is H or
Rd when L.sub.2 is CO, R.sub.a being C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl, and Rd being
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 alkylsulfonyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl and with
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl further optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl;
[0014] each of R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6, and R.sub.7, independently, is H, halo, hydroxyl,
carboxyl, cyano, R.sub.S2, R.sub.S2 being amino, C.sub.1-C.sub.6
alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl, and each R.sub.S2 being optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, and 5 to 6-membered heteroaryl;
[0015] R.sub.8 is H, halo or R.sub.S3, R.sub.S3 being
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl, and R.sub.S3 being optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano amino, C.sub.1-C.sub.6 alkoxyl,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, and
C.sub.3-C.sub.8 cycloalkyl;
[0016] R.sub.9 is
##STR00002##
in which each of R.sub.e, R.sub.f, R.sub.g, and R.sub.h,
independently is -M.sub.2-T.sub.2, in which M.sub.2 is a bond,
SO.sub.2, SO, S, CO, CO.sub.2, O, O--C.sub.1-C.sub.4 alkyl linker,
C.sub.1-C.sub.4 alkyl linker, NH, or N(R.sub.t), R.sub.t being
C.sub.1-C.sub.6 alkyl, and T.sub.2 is H, halo, or R.sub.S4,
R.sub.S4 being C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 8-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, and each of O--C.sub.1-C.sub.4 alkyl
linker, C.sub.1-C.sub.4 alkyl linker, R.sub.t, and R.sub.S4 being
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, R.sub.i is H or
C.sub.1-C.sub.6 alkyl optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, D is O, NR.sub.j,
or CR.sub.jR.sub.k, each of R.sub.j and R.sub.k independently being
H or C.sub.1-C.sub.6 alkyl, or R.sub.j and R.sub.k taken together,
with the carbon atom to which they are attached, form a
C.sub.3-C.sub.10 cycloalkyl ring, and E is-M.sub.3-T.sub.3, M.sub.3
being a bond or C.sub.1-C.sub.6 alkyl linker optionally substituted
with halo or cyano, T.sub.3 being C.sub.3-C.sub.10 cycloalkyl,
C.sub.6-C.sub.10 aryl, 5 to 10-membered heteroaryl, or 4 to
10-membered heterocycloalkyl, and T.sub.3 being optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, thiol, carboxyl, cyano, nitro,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 haloalkoxyl, C.sub.1-C.sub.6 alkylthio,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.1-C.sub.6 haloalkylsulfonyl,
C.sub.1-C.sub.6 alkylcarbonyl, C.sub.1-C.sub.6 alkoxycarbonyl, oxo,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.4-C.sub.12
alkylcycloalkyl, C.sub.6-C.sub.10 aryl, C.sub.6-C.sub.10 aryloxyl,
C.sub.7-C.sub.14 alkylaryl, C.sub.6-C.sub.10 aminoaryloxyl,
C.sub.6-C.sub.10 arylthio, 4 to 6-membered heterocycloalkyl
optionally substituted with halo, C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 haloalkyl, 5 to 6-membered heteroaryl optionally
substituted with halo, C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.6
alkyl that is substituted with hydroxy, halo, C.sub.1-C.sub.6
alkoxycarbonyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl,
4 to 6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl
optionally further substituted with halo, hydroxyl, or
C.sub.1-C.sub.6 alkoxyl;
[0017] q is 0, 1, 2, 3, or 4;
[0018] m is 0, 1, or 2; and
[0019] n is 0, 1, or 2.
[0020] One subset of the compounds of Formula (I) includes those of
Formula (II):
##STR00003##
[0021] Another subset of the compounds of Formula (I) includes
those of Formula (IIIa), (IIIb) or (IIIc):
##STR00004##
[0022] The compounds of Formulae (I), (II), (IIIa), (IIIb), (IIIc)
and (IV) can include one or more of the following features.
[0023] The sum of m and n is at least 1.
[0024] m is 1 or 2 and n is 0.
[0025] m is 2 and n is 0.
[0026] A is CH.sub.2.
[0027] A is O.
[0028] L.sub.1 is N(Y).
[0029] L.sub.1 is SO or SO.sub.2.
[0030] Y is R.sub.d.
[0031] R.sub.d is C.sub.1-C.sub.6 alkyl.
[0032] L.sub.2 is absent.
[0033] At least one of R.sub.e, R.sub.f, R.sub.g, and R.sub.h is
halo (such as F, Cl, and Br), C.sub.1-C.sub.6 alkoxyl optionally
substituted with one or more halo (such as OCH.sub.3,
OCH.sub.2CH.sub.3, O-iPr, and OCF.sub.3), C.sub.1-C.sub.6
alkylsulfonyl optionally substituted with one or more halo (such as
SO.sub.2ClF.sub.3), or C.sub.1-C.sub.6 alkyl optionally substituted
with one or more halo (such as CH.sub.3, i-Pr, t-Bu, and
CF.sub.3).
[0034] R.sub.i is H or C.sub.1-C.sub.6 alkyl.
[0035] R.sub.9 is
##STR00005##
[0036] D is O.
[0037] D is NR.sub.j, e.g., NH.
[0038] D is CR.sub.jR.sub.k, e.g., CH.sub.2, CHCH.sub.3, or
C(CH.sub.3).sub.2.
[0039] E is -M.sub.3-T.sub.3, in which M.sub.3 is a bond or
C.sub.1-C.sub.3 alkyl linker, T.sub.3 is phenyl, naphthyl, thienyl,
cyclopropyl, or cyclohexyl, and T.sub.3 is optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, thiol, carboxyl, cyano, nitro, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
haloalkoxyl, C.sub.1-C.sub.6 alkylthio, C.sub.1-C.sub.6
alkylsulfonyl, C.sub.1-C.sub.6 alkylcarbonyl, C.sub.1-C.sub.6
alkoxycarbonyl, oxo, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.4-C.sub.12 alkylcycloalkyl, C.sub.6-C.sub.10 aryl,
C.sub.6-C.sub.10 aryloxyl, C.sub.7-C.sub.14 alkylaryl,
C.sub.6-C.sub.10 aminoaryloxyl, C.sub.6-C.sub.10 arylthio, 4 to
6-membered heterocycloalkyl optionally substituted with
C.sub.1-C.sub.4 alkyl, 5 to 6-membered heteroaryl optionally
substituted with C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.6 alkyl
that is substituted with hydroxy, C.sub.1-C.sub.6 alkoxycarbonyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl.
[0040] T.sub.3 is phenyl optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano, nitro, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkoxyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 haloalkoxyl,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.6-C.sub.10 aryl, and
C.sub.6-C.sub.10 aryloxyl, and C.sub.7-C.sub.14 alkylaryl.
[0041] X is N.
[0042] X is CR.sub.x, e.g., CH.
[0043] Q is NH.sub.2 or NHR.sub.b, in which R.sub.b is
-M.sub.1-T.sub.1, M.sub.1 being a bond or C.sub.1-C.sub.6 alkyl
linker and T.sub.1 being C.sub.3-C.sub.8 cycloalkyl.
[0044] Q is H.
[0045] R.sub.9 is
##STR00006##
[0046] At least one of R.sub.e, R.sub.f, R.sub.g, and R.sub.h is
selected from the group consisting of F, Cl, CF.sub.3, OCF.sub.3,
SO.sub.2ClF.sub.3, C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.4
alkoxyl.
[0047] R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6,
R.sub.7, and R.sub.8 are each H.
[0048] The invention also relates to a compound of Formula (IV) or
its N-oxide or a pharmaceutically acceptable salt thereof:
##STR00007##
wherein, A is O or CH.sub.2; Q is H, NH.sub.2, NHR.sub.b,
NR.sub.bR.sub.c, OH, R.sub.b, or OR.sub.b, in which each of R.sub.b
and R.sub.c independently is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 7-membered heterocycloalkyl, 5 to
10-membered heteroaryl, or -M.sub.1-T.sub.1 in which M.sub.1 is a
bond or C.sub.1-C.sub.6 alkyl linker optionally substituted with
halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxyl and T.sub.1 is
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 10-membered heteroaryl, or R.sub.b and
R.sub.c, together with the N atom to which they attach, form 4 to
7-membered heterocycloalkyl having 0 or 1 additional heteroatoms to
the N atom optionally substituted with C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl,
carboxyl, C(O)OH, C(O)O--C.sub.1-C.sub.6 alkyl,
OC(O)--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl, and
each of R.sub.b, R.sub.c, and T.sub.1 is optionally substituted
with one or more substituents selected from C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl; X is N or
CR.sub.x, in which R.sub.x is H, halo, hydroxyl, carboxyl, cyano,
or R.sub.S1, R.sub.S1 being amino, C.sub.1-C.sub.6 alkoxyl,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl, and
R.sub.S1 being optionally substituted with one or more substituents
selected from halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, and 5 to 6-membered heteroaryl; Y is
H, R.sub.d, SO.sub.2R.sub.d, or COR.sub.d, R.sub.d being
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl, and
R.sub.d being optionally substituted with one or more substituents
selected from C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 alkylsulfonyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl and with
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl further optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl; each of R.sub.1 and R.sub.2 independently,
is H, halo, hydroxyl, carboxyl, cyano, R.sub.S2, R.sub.S2 being
amino, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl, and each
R.sub.S2 being optionally substituted with one or more substituents
selected from halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, and 5 to 6-membered heteroaryl; each
of R.sub.e, R.sub.f, R.sub.g, and R.sub.h, independently is
-M.sub.2-T.sub.2, in which M.sub.2 is a bond, SO.sub.2, SO, S, CO,
CO.sub.2, O, O--C.sub.1-C.sub.4 alkyl linker, C.sub.1-C.sub.4 alkyl
linker, NH, or N(R.sub.t), R.sub.t being C.sub.1-C.sub.6 alkyl, and
T.sub.2 is H, halo, or R.sub.S4, R.sub.S4 being C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 8-membered
heterocycloalkyl, or 5 to 10-membered heteroaryl, and each of
O--C.sub.1-C.sub.4 alkyl linker, C.sub.1-C.sub.4 alkyl linker,
R.sub.t, and R.sub.S4 being optionally substituted with one or more
substituents selected from halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, and m is 0, 1, or
2.
[0049] For example, A is O. In certain compounds of Formula (IV), A
is O and m is 2.
[0050] In certain compounds of Formula (IV), X is N.
[0051] For example, in certain compounds, Q is NH.sub.2 or
NHR.sub.b, in which R.sub.b is -M.sub.1-T.sub.1, M.sub.1 being a
bond or C.sub.1-C.sub.6 alkyl linker and T.sub.1 being
C.sub.3-C.sub.8 cycloalkyl
[0052] For example, in certain compounds of Formula (IV), R.sub.1
and R.sub.2 are each H.
[0053] In certain compounds of Formula (IV), Y is R.sub.d. For
example, R.sub.d is C.sub.1-C.sub.6 alkyl optionally substituted
with C.sub.3-C.sub.8 cycloalkyl or halo. For example, R is
C.sub.3-C.sub.8 cycloalkyl optionally substituted with
C.sub.1-C.sub.6 alkyl or halo.
[0054] The invention also relates to a compound of Formula (IV),
wherein at least one of R.sub.e, R.sub.f, R.sub.g, and R.sub.h is
halo, C.sub.1-C.sub.6 alkoxyl optionally substituted with one or
more halo; C.sub.1-C.sub.6 alkylsulfonyl optionally substituted
with one or more halo; C.sub.1-C.sub.6 alkyl optionally substituted
with one or more substituents selected from CN, halo,
C.sub.3-C.sub.8 cycloalkyl, hydroxy, and C.sub.1-C.sub.6 alkoxyl;
C.sub.3-C.sub.8 cycloalkyl optionally substituted with one or more
C.sub.1-C.sub.6 alkyl or CN; or 4 to 8-membered heterocycloalkyl
optionally substituted with one or more substituents selected from
CN, halo, hydroxy, C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6
alkoxyl. For example, the compound of Formula (IV) has at least one
of R.sub.e, R.sub.f, R.sub.g, and R.sub.h selected from F; C.sub.1;
Br; CF.sub.3; OCF.sub.3; SO.sub.2ClF.sub.3; oxetanyl optionally
substituted with one or more substituents selected from CN, halo,
hydroxy, C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxyl;
C.sub.3-C.sub.8 cycloalkyl optionally substituted with one or more
substituents selected from C.sub.1-C.sub.4 alkyl; and
C.sub.1-C.sub.4 alkyl optionally substituted with one or more
substituents selected from halo, C.sub.3-C.sub.8 cycloalkyl,
hydroxy and C.sub.1-C.sub.6 alkoxyl.
[0055] For example, the invention relates to compounds of Formula
(IV) where at least one of R.sub.f and R.sub.g is alkyl, optionally
substituted with hydroxyl. For example, the invention relates to
compounds where at least one of R.sub.f and R.sub.g is t-butyl
substituted with hydroxyl.
[0056] The invention relates to a compound selected from Compounds
1-140. The invention also relates to a salt of a compound selected
from Compounds 1-140. The invention also relates to an N-oxide of
compound selected from Compounds 1-140. The invention also relates
to a salt of an N-oxide of compound selected from Compounds 1-140.
For example, the invention relates to a compound selected from
Compounds 1-7, 9-109, and 111-140.
[0057] The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of a compound of Formula (IV)
and a pharmaceutically acceptable carrier.
[0058] The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of a salt of a compound of
Formula (IV) and a pharmaceutically acceptable carrier.
[0059] The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of a hydrate of a compound of
Formula (IV) and a pharmaceutically acceptable carrier.
[0060] The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of a compound selected from
Compounds 1-140 and a pharmaceutically acceptable carrier. The
invention also relates to a pharmaceutical composition of a
therapeutically effective amount of a salt of a compound selected
from Compounds 1-140 and a pharmaceutically acceptable carrier. The
invention also relates to a pharmaceutical composition of a
therapeutically effective amount of an N-oxide of a compound
selected from Compounds 1-140 and a pharmaceutically acceptable
carrier. The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of an N-oxide of salt of a
compound selected from Compounds 1-140 and a pharmaceutically
acceptable carrier. The invention also relates to a pharmaceutical
composition of a therapeutically effective amount of a hydrate of a
compound selected from Compounds 1-140 and a pharmaceutically
acceptable carrier.
[0061] The present invention provides pharmaceutical compositions
comprising one or more compounds of Formula (I), (II), (IIIa),
(IIIb), (IIIc) or (IV), and one or more pharmaceutically acceptable
carriers.
[0062] The present invention provides methods of treating or
preventing cancer. The present invention provides methods of
treating cancer. The present invention also provides methods of
preventing cancer. The method includes administering to a subject
in need thereof a therapeutically effective amount of the compound
of Formula (I), (II), (IIIa), (IIIb), or (IIIc). The cancer can be
a hematological cancer. Preferably, the cancer is leukemia. More
preferably, the cancer is acute myeloid leukemia, acute lymphocytic
leukemia or mixed lineage leukemia.
[0063] The present invention provides methods of treating or
preventing a disease or disorder mediated by translocation of a
gene on chromosome 11q23. The present invention provides methods of
treating a disease or disorder mediated by translocation of a gene
on chromosome 11q23. The present invention also provides methods of
preventing a disease or disorder mediated by translocation of a
gene on chromosome 11q23. The method includes administering to a
subject in need thereof a therapeutically effective amount of the
compound of Formula (I), (II), (IIIa), (IIIb), (IIIc) or (IV).
[0064] The present invention provides methods of treating or
preventing a disease or disorder in which DOT1-mediated protein
methylation plays a part or a disease or disorder mediated by
DOT1-mediated protein methylation. The present invention provides
methods of treating a disease or disorder in which DOT1-mediated
protein methylation plays a part or a disease or disorder mediated
by DOT1-mediated protein methylation. The present invention also
provides methods of preventing a disease or disorder in which
DOT1-mediated protein methylation plays a part or a disease or
disorder mediated by DOT1-mediated protein methylation. The method
includes administering to a subject in need thereof a
therapeutically effective amount of the compound of Formula (I),
(II), (IIIa), (IIIb), (IIIc) or (IV).
[0065] The present invention provides methods of inhibiting DOT1L
activity in a cell. The method includes contacting the cell with an
effective amount of one or more of the compound of Formula (I),
(II), (IIIa), (IIIb), (IIIc) or (IV).
[0066] Still another aspect of the invention relates to a method of
reducing the level of Histone H3 Lysine residue 79 (H3-K79)
methylation in a cell. The method includes contacting a cell with a
compound of the present invention. Such method can be used to
ameliorate any condition which is caused by or potentiated by the
activity of DOT1 through H3-K79 methylation.
[0067] The present invention relates to use of the compounds
disclosed herein in preparation of a medicament for treating or
preventing cancer. The use includes a compound of Formula (I),
(II), (IIIa), (IIIb), (IIIc) or (IV) for administration to a
subject in need thereof in a therapeutically effective amount. The
cancer can be a hematological cancer. Preferably, the cancer is
leukemia. More preferably, the cancer is acute myeloid leukemia,
acute lymphocytic leukemia or mixed lineage leukemia.
[0068] The present invention provides use of the compounds
disclosed herein in preparation of a medicament for treating or
preventing a disease or disorder mediated by translocation of a
gene on chromosome 11q23. The use includes a compound of Formula
(I), (II), (IIIa), (IIIb), (IIIc) or (IV) for administration to a
subject in need thereof in a therapeutically effective amount.
[0069] The present invention provides use of the compounds
disclosed herein in preparation of a medicament for treating or
preventing a disease or disorder in which DOT1-mediated protein
methylation plays a part or a disease or disorder mediated by
DOT1-mediated protein methylation. The use includes a compound of
Formula (I), (II), (IIIa), (IIIb), (IIIc) or (IV) for
administration to a subject in need thereof in a therapeutically
effective amount.
[0070] The present invention provides use of the compounds
disclosed herein for inhibiting DOT1L activity in a cell. The use
includes contacting the cell with an effective amount of one or
more of the compound of Formula (I), (II), (IIIa), (IIIb), (IIIc)
or (IV).
[0071] Still another aspect of the invention relates to a use of
the compounds disclosed herein for reducing the level of Histone H3
Lysine residue 79 (H3-K79) methylation in a cell. The use includes
contacting a cell with a compound of the present invention. Such
use can ameliorate any condition which is caused by or potentiated
by the activity of DOT1 through H3-K79 methylation.
[0072] In the formulae presented herein, the variables can be
selected from the respective groups of chemical moieties later
defined in the detailed description.
[0073] In addition, the invention provides methods of synthesizing
the foregoing compounds. Following synthesis, a therapeutically
effective amount of one or more of the compounds can be formulated
with a pharmaceutically acceptable carrier for administration to a
mammal, particularly humans, for use in modulating an epigenetic
enzyme. In certain embodiments, the compounds of the present
invention are useful for treating, preventing, or reducing the risk
of cancer or for the manufacture of a medicament for treating,
preventing, or reducing the risk of cancer. Accordingly, the
compounds or the formulations can be administered, for example, via
oral, parenteral, otic, ophthalmic, nasal, or topical routes, to
provide an effective amount of the compound to the mammal.
[0074] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. In the
specification, the singular forms also include the plural unless
the context clearly dictates otherwise. Although methods and
materials similar or equivalent to those described herein can be
used in the practice or testing of the present invention, suitable
methods and materials are described below. All publications, patent
applications, patents and other references mentioned herein are
incorporated by reference. The references cited herein are not
admitted to be prior art to the claimed invention. In the case of
conflict, the present specification, including definitions, will
control. In addition, the materials, methods and examples are
illustrative only and are not intended to be limiting.
[0075] Other features and advantages of the invention will be
apparent from the following detailed description and claims.
DESCRIPTION OF THE FIGURES
[0076] FIGS. 1A and 1B are respectively a table and a plot
demonstrating the potency and selectivity of the anti-proliferative
activity of Compound 2 using a panel of MLL-rearranged and
non-MLL-rearranged human leukemia cell lines. The cell lines used
in the study are listed in FIG. 1A
[0077] FIG. 2 is a plot showing the tumor growth over 21 days of
dosing.
[0078] FIG. 3A is a plot showing the estimated steady state plasma
concentrations of Compound 2 in Groups 4 and 5 as determined by the
averaged blood samples taken on days 7, 14 and 21.
[0079] FIG. 3B is a plot showing the Compound 2 plasma
concentrations plotted against time after ip injection.
DETAILED DESCRIPTION OF THE INVENTION
[0080] The present invention provides a family of compounds that
can be used to selectively modulate the aberrant action of an
epigenetic enzyme. Further, the compounds can be used to treat or
prevent a disease state in a mammal caused or mediated by aberrant
action of an epigenetic enzyme. The present invention includes
pharmaceutically acceptable salts, esters, tautomers, and N-oxides
of these compounds.
[0081] The present invention provides novel substituted purine and
7-deazapurine compounds, synthetic methods for making the
compounds, pharmaceutical compositions containing them and various
uses of the compounds.
1. SUBSTITUTED PURINE COMPOUNDS AND SUBSTITUTED 7-DEAZAPURINE
COMPOUNDS
[0082] The present invention provides the compounds of Formula
(I):
##STR00008##
or a pharmaceutically acceptable salt or ester thereof,
wherein:
[0083] A is O or CH.sub.2;
[0084] each of G and J, independently, is H, halo, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl or OR.sub.a, R.sub.a being H,
C.sub.1-C.sub.6 alkyl or C(O)--C.sub.1-C.sub.6 alkyl, wherein
C(O)O--C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkyl or
C(O)--C.sub.1-C.sub.6 alkyl is optionally substituted with one or
more substituents selected from the group consisting of halo, cyano
hydroxyl, carboxyl, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, and
C.sub.3-C.sub.8 cycloalkyl;
[0085] Q is H, NH.sub.2, NHR.sub.b, NR.sub.bR.sub.c, R.sub.b, or
OR.sub.b, in which each of R.sub.b and R.sub.c independently is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
7-membered heterocycloalkyl, 5 to 10-membered heteroaryl, or
-M.sub.1-T.sub.1 in which M.sub.1 is a bond or C.sub.1-C.sub.6
alkyl linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxyl and T.sub.1 is C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, or R.sub.b and R.sub.c, together with the N
atom to which they attach, form 4 to 7-membered heterocycloalkyl
having 0 or 1 additional heteroatoms to the N atom optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and each of R.sub.b, R.sub.c, and T.sub.1 is
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, and 5 to
6-membered heteroaryl;
[0086] X is N or CR.sub.x, in which R.sub.x is H, halo, hydroxyl,
carboxyl, cyano, or R.sub.S1, R.sub.S1 being amino, C.sub.1-C.sub.6
alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and R.sub.S1 being optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl;
[0087] L.sub.1 is N(Y), S, SO, or SO.sub.2;
[0088] L.sub.2 is CO or absent when L.sub.1 is N(Y) or L.sub.2 is
absent when L.sub.1 is S, SO, or SO.sub.2, in which Y is H,
R.sub.d, SO.sub.2R.sub.a, or COR.sub.a when L.sub.2 is absent, or Y
is H or R.sub.d when L.sub.2 is CO, R.sub.d being C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl, and R.sub.d being
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 alkylsulfonyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl and with
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl further optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl;
[0089] each of R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6, and R.sub.7, independently, is H, halo, hydroxyl,
carboxyl, cyano, R.sub.S2, R.sub.S2 being amino, C.sub.1-C.sub.6
alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl, and each R.sub.S2 being optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, and 5 to 6-membered heteroaryl;
[0090] R.sub.8 is H, halo or R.sub.S3, R.sub.S3 being
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl, and R.sub.S3 being optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano amino, C.sub.1-C.sub.6 alkoxyl,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, and
C.sub.3-C.sub.8 cycloalkyl;
[0091] R.sub.9 is
##STR00009##
in which each of R.sub.e, R.sub.f, R.sub.g, and R.sub.h,
independently is -M.sub.2-T.sub.2, in which M.sub.2 is a bond,
SO.sub.2, SO, S, CO, CO.sub.2, O, O--C.sub.1-C.sub.4 alkyl linker,
C.sub.1-C.sub.4 alkyl linker, NH, or N(R.sub.t), R.sub.t being
C.sub.1-C.sub.6 alkyl, and T.sub.2 is H, halo, or R.sub.S4,
R.sub.S4 being C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 8-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, and each of O--C.sub.1-C.sub.4 alkyl
linker, C.sub.1-C.sub.4 alkyl linker, R.sub.t, and R.sub.S4 being
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, R.sub.i is H or
C.sub.1-C.sub.6 alkyl optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, D is O, NR.sub.j,
or CR.sub.jR.sub.k, each of R.sub.j and R.sub.k independently being
H or C.sub.1-C.sub.6 alkyl, or R.sub.j and R.sub.k taken together,
with the carbon atom to which they are attached, form a
C.sub.3-C.sub.10 cycloalkyl ring, and E is-M.sub.3-T.sub.3, M.sub.3
being a bond or C.sub.1-C.sub.6 alkyl linker optionally substituted
with halo or cyano, T.sub.3 being C.sub.3-C.sub.10 cycloalkyl,
C.sub.6-C.sub.10 aryl, 5 to 10-membered heteroaryl, or 4 to
10-membered heterocycloalkyl, and T.sub.3 being optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, thiol, carboxyl, cyano, nitro,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 haloalkoxyl, C.sub.1-C.sub.6 alkylthio,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.1-C.sub.6 haloalkylsulfonyl,
C.sub.1-C.sub.6 alkylcarbonyl, C.sub.1-C.sub.6 alkoxycarbonyl, oxo,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.4-C.sub.12
alkylcycloalkyl, C.sub.6-C.sub.10 aryl, C.sub.6-C.sub.10 aryloxyl,
C.sub.7-C.sub.14 alkylaryl, C.sub.6-C.sub.10 aminoaryloxyl,
C.sub.6-C.sub.10 arylthio, 4 to 6-membered heterocycloalkyl
optionally substituted with halo, C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 haloalkyl, 5 to 6-membered heteroaryl optionally
substituted with halo, C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.6
alkyl that is substituted with hydroxy, halo, C.sub.1-C.sub.6
alkoxycarbonyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl,
4 to 6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl
optionally further substituted with halo, hydroxyl, or
C.sub.1-C.sub.6 alkoxyl;
[0092] q is 0, 1, 2, 3, or 4;
[0093] m is 0, 1, or 2; and
[0094] n is 0, 1, or 2.
[0095] For example, the sum of m and n is at least 1.
[0096] For example, m is 1 or 2 and n is 0.
[0097] For example, m is 2 and n is 0
[0098] For example, A is CH.sub.2.
[0099] For example, A is O.
[0100] For example, L.sub.1 is N(Y).
[0101] For example, L.sub.1 is SO or SO.sub.2.
[0102] For example, Y is R.sub.d.
[0103] For example, R.sub.d is C.sub.1-C.sub.6 alkyl.
[0104] For example, L.sub.2 is absent.
[0105] For example, each of G and J independently is OR.sub.a.
[0106] For example, R.sub.a is H.
[0107] For example, R.sub.9 is
##STR00010##
For example, R.sub.9 is
##STR00011##
[0108] For example, at least one of R.sub.e, R.sub.f, R.sub.g, and
R.sub.h is halo (such as F, Cl, and Br), C.sub.1-C.sub.6 alkoxyl
optionally substituted with one or more halo (such as OCH.sub.3,
OCH.sub.2CH.sub.3, O-iPr, and OCF.sub.3), C.sub.1-C.sub.6
alkylsulfonyl optionally substituted with one or more halo (such as
SO.sub.2ClF.sub.3), or C.sub.1-C.sub.6 alkyl optionally substituted
with one or more halo (such as CH.sub.3, i-propyl, n-butyl, and
CF.sub.3).
[0109] For example, R.sub.i is H or C.sub.1-C.sub.6 alkyl (e.g.,
methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, t-butyl,
n-pentyl, s-pentyl and n-hexyl).
[0110] For example,
##STR00012##
is unsubstituted benzimidazolyl or one of the following groups:
##STR00013## ##STR00014## ##STR00015##
[0111] For example, R.sub.9 is
##STR00016##
[0112] For example, D is O.
[0113] For example, D is NR.sub.j.
[0114] For example, R.sub.j is H.
[0115] For example, D is CR.sub.jR.sub.k.
[0116] For example, each of R.sub.j and R.sub.k is H.
[0117] For example, E is -M.sub.3-T.sub.3, in which M.sub.3 is a
bond or C.sub.1-C.sub.3 alkyl linker, T.sub.3 is phenyl, naphthyl,
thienyl, cyclopropyl, or cyclohexyl, and T.sub.3 is optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, thiol, carboxyl, cyano, nitro,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 haloalkoxyl, C.sub.1-C.sub.6 alkylthio,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.1-C.sub.6 alkylcarbonyl,
C.sub.1-C.sub.6 alkoxycarbonyl, oxo, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.4-C.sub.12 alkylcycloalkyl, C.sub.6-C.sub.10
aryl, C.sub.6-C.sub.10 aryloxyl, C.sub.7-C.sub.14 alkylaryl,
C.sub.6-C.sub.10 aminoaryloxyl, C.sub.6-C.sub.10 arylthio, 4 to
6-membered heterocycloalkyl optionally substituted with
C.sub.1-C.sub.4 alkyl, 5 to 6-membered heteroaryl optionally
substituted with C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.6 alkyl
that is substituted with hydroxy, C.sub.1-C.sub.6 alkoxycarbonyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl.
[0118] For example, T.sub.3 is phenyl optionally substituted with
one or more substituents selected from the group consisting of
halo, hydroxyl, carboxyl, cyano, nitro, C.sub.1-C.sub.6 alkyl
(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl,
t-butyl, n-pentyl, s-pentyl and n-hexyl), C.sub.1-C.sub.6 alkoxyl,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 haloalkoxyl,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.6-C.sub.10 aryl (e.g., phenyl
or naphthyl), and C.sub.6-C.sub.10 aryloxyl, and C.sub.7-C.sub.14
alkylaryl.
[0119] For example, E is
##STR00017## ##STR00018## ##STR00019## ##STR00020##
[0120] For example, X is N.
[0121] For example, X is CR.sub.x.
[0122] For example, X is CH.
[0123] For example, Q is NH.sub.2 or NHR.sub.b, in which R.sub.b is
-M.sub.1-T.sub.1, M.sub.1 being a bond or C.sub.1-C.sub.6 alkyl
linker and T.sub.1 being C.sub.3-C.sub.8 cycloalkyl.
[0124] For example, Q is H.
[0125] For example, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6, R.sub.7, and R.sub.8 are each H.
[0126] For example, when R.sub.8 is halo and is attached to the
same carbon atom as J, then J is not hydroxyl.
[0127] For example, when R.sub.8 is halo and is attached to the
same carbon atom as G, then G is not hydroxyl.
[0128] For example, T.sub.2 is not halo when M.sub.2 is SO.sub.2,
SO, S, CO or O.
[0129] For example, T.sub.2 is a 4-8 membered heterocycloalkyl
which is bound to M.sub.2 via a heteroatom.
[0130] For example, T.sub.2 is a 4-8 membered heterocycloalkyl
which is bound to M.sub.2 via a N atom.
[0131] For example, T.sub.2 is a 4-8 membered heterocycloalkyl
which is bound to M.sub.2 via a C atom.
[0132] The present invention provides the compounds of Formula
(II):
##STR00021##
or a pharmaceutically acceptable salt or ester thereof,
wherein.
[0133] A is O or CH.sub.2;
[0134] Q is H, NH.sub.2, NHR.sub.b, NR.sub.bR.sub.c, R.sub.b, or
OR.sub.b, in which each of R.sub.b and R.sub.c independently is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
7-membered heterocycloalkyl, 5 to 10-membered heteroaryl, or
-M.sub.1-T.sub.1 in which M.sub.1 is a bond or C.sub.1-C.sub.6
alkyl linker optionally substituted with halo, cyano, hydroxyl, or
C.sub.1-C.sub.6 alkoxyl and T.sub.1 is C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, or R.sub.b and R.sub.c, together with the N
atom to which they attach, form 4 to 7-membered heterocycloalkyl
having 0 or 1 additional heteroatoms to the N atom optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and each of R.sub.b, R.sub.c, and T.sub.1 is
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, and 5 to
6-membered heteroaryl;
[0135] X is N or CR.sub.x, in which R.sub.x is H, halo, hydroxyl,
carboxyl, cyano, or R.sub.S1, R.sub.S1 being amino, C.sub.1-C.sub.6
alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and R.sub.S1 being optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl;
[0136] L.sub.1 is N(Y), S, SO, or SO.sub.2;
[0137] L.sub.2 is CO or absent when L.sub.1 is N(Y) or L.sub.2 is
absent when L.sub.1 is S, SO, or SO.sub.2, in which Y is H,
R.sub.d, SO.sub.2R.sub.a, or COR.sub.a when L.sub.2 is absent, or Y
is H or R.sub.d when L.sub.2 is CO, R.sub.d being C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl, and R.sub.d being
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 alkylsulfonyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl and with
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl further optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl;
[0138] each of R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6, and R.sub.7, independently, is H, halo, hydroxyl,
carboxyl, cyano, R.sub.S2, R.sub.S2 being amino, C.sub.1-C.sub.6
alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl, and each R.sub.S2 being optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, and 5 to 6-membered heteroaryl;
[0139] R.sub.8 is H, halo or R.sub.S3, R.sub.S3 being
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl, and R.sub.S3 being optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano amino, C.sub.1-C.sub.6 alkoxyl,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, and
C.sub.3-C.sub.8 cycloalkyl;
[0140] R.sub.9 is
##STR00022##
in which each of R.sub.e, R.sub.f, R.sub.g, and R.sub.h,
independently is -M.sub.2-T.sub.2, in which M.sub.2 is a bond,
SO.sub.2, SO, S, CO, CO.sub.2, O, O--C.sub.1-C.sub.4 alkyl linker,
C.sub.1-C.sub.4 alkyl linker, NH, or N(R.sub.t), R.sub.t being
C.sub.1-C.sub.6 alkyl, and T.sub.2 is H, halo, or R.sub.S4,
R.sub.S4 being C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 8-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, and each of O--C.sub.1-C.sub.4 alkyl
linker, C.sub.1-C.sub.4 alkyl linker, R.sub.t, and R.sub.S4 being
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, R.sub.i is H or
C.sub.1-C.sub.6 alkyl optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, D is O, NR.sub.j,
or CR.sub.jR.sub.k, each of R.sub.j and R.sub.k independently being
H or C.sub.1-C.sub.6 alkyl, or R.sub.j and R.sub.k taken together,
with the carbon atom to which they are attached, form a
C.sub.3-C.sub.10 cycloalkyl ring, and E is -M.sub.3-T.sub.3,
M.sub.3 being a bond or C.sub.1-C.sub.6 alkyl linker optionally
substituted with halo or cyano, T.sub.3 being C.sub.3-C.sub.10
cycloalkyl, C.sub.6-C.sub.10 aryl, 5 to 10-membered heteroaryl, or
4 to 10-membered heterocycloalkyl, and T.sub.3 being optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, thiol, carboxyl, cyano, nitro,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 haloalkoxyl, C.sub.1-C.sub.6 alkylthio,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.1-C.sub.6 haloalkylsulfonyl,
C.sub.1-C.sub.6 alkylcarbonyl, C.sub.1-C.sub.6 alkoxycarbonyl, oxo,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.4-C.sub.12
alkylcycloalkyl, C.sub.6-C.sub.10 aryl, C.sub.6-C.sub.10 aryloxyl,
C.sub.7-C.sub.14 alkylaryl, C.sub.6-C.sub.10 aminoaryloxyl,
C.sub.6-C.sub.10 arylthio, 4 to 6-membered heterocycloalkyl
optionally substituted with halo, C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 haloalkyl, 5 to 6-membered heteroaryl optionally
substituted with halo, C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.6
alkyl that is substituted with hydroxy, halo, C.sub.1-C.sub.6
alkoxycarbonyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl,
4 to 6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl
optionally further substituted with halo, hydroxyl, or
C.sub.1-C.sub.6 alkoxyl; [0141] q is 0, 1, 2, 3, or 4; [0142] m is
0, 1, or 2; and [0143] n is 0, 1, or 2.
[0144] For example, the sum of m and n is at least 1.
[0145] For example, m is 1 or 2 and n is 0.
[0146] For example, m is 2 and n is 0
[0147] For example, A is CH.sub.2.
[0148] For example, A is O.
[0149] For example, L.sub.1 is N(Y).
[0150] For example, L.sub.1 is SO or SO.sub.2.
[0151] For example, Y is R.sub.d.
[0152] For example, R.sub.d is C.sub.1-C.sub.6 alkyl.
[0153] For example, L.sub.2 is absent.
[0154] For example, R.sub.9 is
##STR00023##
For example, R.sub.9 is
##STR00024##
[0155] For example, at least one of R.sub.e, R.sub.f, R.sub.g, and
R.sub.h is halo (such as F, Cl, and Br), C.sub.1-C.sub.6 alkoxyl
optionally substituted with one or more halo (such as OCH.sub.3,
OCH.sub.2CH.sub.3, O-iPr, and OCF.sub.3), C.sub.1-C.sub.6
alkylsulfonyl optionally substituted with one or more halo (such as
SO.sub.2ClF.sub.3), or C.sub.1-C.sub.6 alkyl optionally substituted
with one or more halo (such as CH.sub.3, i-propyl, n-butyl, and
CF.sub.3).
[0156] For example, R.sub.i is H or C.sub.1-C.sub.6 alkyl (e.g.,
methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, t-butyl,
n-pentyl, s-pentyl or n-hexyl).
[0157] For example,
##STR00025##
is unsubstituted benzimidazolyl or one of the following groups:
##STR00026## ##STR00027## ##STR00028##
[0158] For example, R.sub.9 is
##STR00029##
[0159] For example, D is O.
[0160] For example, D is NR.sub.j
[0161] For example, R.sub.j is H.
[0162] For example, D is CR.sub.jR.sub.k.
[0163] For example, each of R.sub.j and R.sub.k is H.
[0164] For example, E is -M.sub.3-T.sub.3, in which M.sub.3 is a
bond or C.sub.1-C.sub.3 alkyl linker, T.sub.3 is phenyl, naphthyl,
thienyl, cyclopropyl, or cyclohexyl, and T.sub.3 is optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, thiol, carboxyl, cyano, nitro,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 haloalkoxyl, C.sub.1-C.sub.6 alkylthio,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.1-C.sub.6 alkylcarbonyl,
C.sub.1-C.sub.6 alkoxycarbonyl, oxo, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.4-C.sub.12 alkylcycloalkyl, C.sub.6-C.sub.10
aryl, C.sub.6-C.sub.10 aryloxyl, C.sub.7-C.sub.14 alkylaryl,
C.sub.6-C.sub.10 aminoaryloxyl, C.sub.6-C.sub.10 arylthio, 4 to
6-membered heterocycloalkyl optionally substituted with
C.sub.1-C.sub.4 alkyl, 5 to 6-membered heteroaryl optionally
substituted with C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.6 alkyl
that is substituted with hydroxy, C.sub.1-C.sub.6 alkoxycarbonyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl.
[0165] For example, T.sub.3 is phenyl optionally substituted with
one or more substituents selected from the group consisting of
halo, hydroxyl, carboxyl, cyano, nitro, C.sub.1-C.sub.6 alkyl
(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl,
t-butyl, n-pentyl, s-pentyl and n-hexyl), C.sub.1-C.sub.6 alkoxyl,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 haloalkoxyl,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.6-C.sub.10 aryl (e.g., phenyl
or naphthyl), and C.sub.6-C.sub.10 aryloxyl, and C.sub.7-C.sub.14
alkylaryl.
[0166] For example, E is
##STR00030## ##STR00031## ##STR00032## ##STR00033##
[0167] For example, X is N.
[0168] For example, X is CR.
[0169] For example, X is CH.
[0170] For example, Q is NH.sub.2 or NHR.sub.b, in which R.sub.b is
-M.sub.1-T.sub.1, M.sub.1 being a bond or C.sub.1-C.sub.6 alkyl
linker and T.sub.1 being C.sub.3-C.sub.8 cycloalkyl.
[0171] For example, Q is H.
[0172] For example, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6, R.sub.7, and R.sub.8 are each H.
[0173] For example, when R.sub.8 is halo and is attached to the
same carbon atom as J, then J is not hydroxyl.
[0174] For example, when R.sub.8 is halo and is attached to the
same carbon atom as G, then G is not hydroxyl.
[0175] For example, T.sub.2 is not halo when M.sub.2 is SO.sub.2,
SO, S, CO or O.
[0176] For example, T.sub.2 is a 4-8 membered heterocycloalkyl
which is bound to M.sub.2 via a heteroatom.
[0177] For example, T.sub.2 is a 4-8 membered heterocycloalkyl
which is bound to M.sub.2 via a N atom.
[0178] For example, T.sub.2 is a 4-8 membered heterocycloalkyl
which is bound to M.sub.2 via a C atom.
[0179] The present invention provides the compounds of Formula
(IIIa) or (IIIb):
##STR00034##
or a pharmaceutically acceptable salt or ester thereof,
wherein:
[0180] A is O or CH.sub.2;
[0181] each of G and J, independently, is H, halo, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl or OR.sub.a, R.sub.a being H,
C.sub.1-C.sub.6 alkyl or C(O)--C.sub.1-C.sub.6 alkyl, wherein
C(O)O--C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkyl or
C(O)--C.sub.1-C.sub.6 alkyl is optionally substituted with one or
more substituents selected from the group consisting of halo, cyano
hydroxyl, carboxyl, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, and
C.sub.3-C.sub.8 cycloalkyl;
[0182] Q is H, NH.sub.2, NHR.sub.b, NR.sub.bR.sub.c, R.sub.b, or
OR.sub.b, in which each of R.sub.b and R.sub.c independently is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
7-membered heterocycloalkyl, 5 to 10-membered heteroaryl, or
-M.sub.1-T.sub.1 in which M.sub.1 is a bond or C.sub.1-C.sub.6
alkyl linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxyl and T.sub.1 is C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, or R.sub.b and R.sub.c, together with the N
atom to which they attach, form 4 to 7-membered heterocycloalkyl
having 0 or 1 additional heteroatoms to the N atom optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and each of R.sub.b, R.sub.c, and T.sub.1 is
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, and 5 to
6-membered heteroaryl;
[0183] X is N or CR.sub.x, in which R.sub.x is H, halo, hydroxyl,
carboxyl, cyano, or R.sub.S1, R.sub.S1 being amino, C.sub.1-C.sub.6
alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and R.sub.S1 being optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl;
[0184] L.sub.1 is N(Y), S, SO, or SO.sub.2;
[0185] L.sub.2 is CO or absent when L.sub.1 is N(Y) or L.sub.2 is
absent when L.sub.1 is S, SO, or SO.sub.2, in which Y is H,
R.sub.d, SO.sub.2R.sub.a, or COR.sub.a when L.sub.2 is absent, or Y
is H or R.sub.d when L.sub.2 is CO, R.sub.d being C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl, and R.sub.d being
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 alkylsulfonyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl and with
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl further optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl;
[0186] each of R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6, and R.sub.7, independently, is H, halo, hydroxyl,
carboxyl, cyano, R.sub.S2, R.sub.S2 being amino, C.sub.1-C.sub.6
alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl, and each R.sub.S2 being optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, and 5 to 6-membered heteroaryl;
[0187] R.sub.8 is H, halo or R.sub.S3, R.sub.S3 being
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl, and R.sub.S3 being optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano amino, C.sub.1-C.sub.6 alkoxyl,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, and
C.sub.3-C.sub.8 cycloalkyl;
[0188] each of R.sub.e, R.sub.f, R.sub.g, and R.sub.h,
independently is -M.sub.2-T.sub.2, in which M.sub.2 is a bond,
SO.sub.2, SO, S, CO, CO.sub.2, O, O--C.sub.1-C.sub.4 alkyl linker,
C.sub.1-C.sub.4 alkyl linker, NH, or N(R.sub.t), R.sub.t being
C.sub.1-C.sub.6 alkyl, and T.sub.2 is H, halo, or R.sub.S4,
R.sub.S4 being C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 8-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, and each of O--C.sub.1-C.sub.4 alkyl
linker, C.sub.1-C.sub.4 alkyl linker, R.sub.t, and R.sub.S4 being
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl,
[0189] R.sub.i is H or C.sub.1-C.sub.6 alkyl optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl;
[0190] q is 0, 1, 2, 3, or 4;
[0191] m is 0, 1, or 2; and
[0192] n is 0, 1, or 2.
[0193] For example, the sum of m and n is at least 1.
[0194] For example, m is 1 or 2 and n is 0.
[0195] For example, m is 2 and n is 0
[0196] For example, A is CH.sub.2.
[0197] For example, A is O.
[0198] For example, L.sub.1 is N(Y).
[0199] For example, L.sub.1 is SO or SO.sub.2.
[0200] For example, Y is R.sub.d.
[0201] For example, R.sub.d is C.sub.1-C.sub.6 alkyl.
[0202] For example, L.sub.2 is absent.
[0203] For example, each of G and J independently is OR.sub.a.
[0204] For example, R.sub.a is H.
[0205] For example, at least one of R.sub.e, R.sub.f, R.sub.g, and
R.sub.h is halo (such as F, Cl, and Br), C.sub.1-C.sub.6 alkoxyl
optionally substituted with one or more halo (such as OCH.sub.3,
OCH.sub.2CH.sub.3, O-iPr, and OCF.sub.3), C.sub.1-C.sub.6
alkylsulfonyl optionally substituted with one or more halo (such as
SO.sub.2ClF.sub.3), or C.sub.1-C.sub.6 alkyl optionally substituted
with one or more halo (such as CH.sub.3, i-propyl, n-butyl, and
CF.sub.3).
[0206] For example, R.sub.i is H or C.sub.1-C.sub.6 alkyl (e.g.,
methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, t-butyl,
n-pentyl, s-pentyl and n-hexyl).
[0207] For example,
##STR00035##
is unsubstituted benzimidazolyl or one of the following groups:
##STR00036## ##STR00037##
[0208] For example, X is N.
[0209] For example, X is CR.sub.x.
[0210] For example, X is CH.
[0211] For example, Q is NH.sub.2 or NHR.sub.b, in which R.sub.b is
-M.sub.1-T.sub.1, M.sub.1 being a bond or C.sub.1-C.sub.6 alkyl
linker and T.sub.1 being C.sub.3-C.sub.8 cycloalkyl.
[0212] For example, Q is H.
[0213] For example, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6, R.sub.7, and R.sub.8 are each H.
[0214] For example, when R.sub.8 is halo and is attached to the
same carbon atom as J, then J is not hydroxyl.
[0215] For example, when R.sub.8 is halo and is attached to the
same carbon atom as G, then G is not hydroxyl.
[0216] For example, T.sub.2 is not halo when M.sub.2 is SO.sub.2,
SO, S, CO or O.
[0217] For example, T.sub.2 is a 4-8 membered heterocycloalkyl
which is bound to M.sub.2 via a heteroatom.
[0218] For example, T.sub.2 is a 4-8 membered heterocycloalkyl
which is bound to M.sub.2 via a N atom.
[0219] For example, T.sub.2 is a 4-8 membered heterocycloalkyl
which is bound to M.sub.2 via a C atom.
[0220] The present invention provides the compounds of Formula
(IIIc):
##STR00038##
or a pharmaceutically acceptable salt or ester thereof,
wherein:
[0221] A is O or CH.sub.2;
[0222] each of G and J, independently, is H, halo, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl or OR.sub.a, R.sub.a being H,
C.sub.1-C.sub.6 alkyl or C(O)--C.sub.1-C.sub.6 alkyl, wherein
C(O)O--C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkyl or
C(O)--C.sub.1-C.sub.6 alkyl is optionally substituted with one or
more substituents selected from the group consisting of halo, cyano
hydroxyl, carboxyl, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, and
C.sub.3-C.sub.8 cycloalkyl;
[0223] Q is H, NH.sub.2, NHR.sub.b, NR.sub.bR.sub.c, R.sub.b, or
OR.sub.b, in which each of R.sub.b and R.sub.c independently is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
7-membered heterocycloalkyl, 5 to 10-membered heteroaryl, or
-M.sub.1-T.sub.1 in which M.sub.1 is a bond or C.sub.1-C.sub.6
alkyl linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxyl and T.sub.1 is C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, or R.sub.b and R.sub.c, together with the N
atom to which they attach, form 4 to 7-membered heterocycloalkyl
having 0 or 1 additional heteroatoms to the N atom optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and each of R.sub.b, R.sub.c, and T.sub.1 is
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, and 5 to
6-membered heteroaryl;
[0224] X is N or CR.sub.x, in which R.sub.x is H, halo, hydroxyl,
carboxyl, cyano, or R.sub.S1, R.sub.S1 being amino, C.sub.1-C.sub.6
alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and R.sub.S1 being optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl;
[0225] L.sub.1 is N(Y), S, SO, or SO.sub.2;
[0226] L.sub.2 is CO or absent when L.sub.1 is N(Y) or L.sub.2 is
absent when L.sub.1 is S, SO, or SO.sub.2, in which Y is H,
R.sub.d, SO.sub.2R.sub.a, or COR.sub.a when L.sub.2 is absent, or Y
is H or R.sub.d when L.sub.2 is CO, R.sub.d being C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl, and R.sub.d being
optionally substituted with one or more substituents selected from
the group consisting of C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 alkylsulfonyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl and with
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl further optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl;
[0227] each of R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6, and R.sub.7, independently, is H, halo, hydroxyl,
carboxyl, cyano, R.sub.S2, R.sub.S2 being amino, C.sub.1-C.sub.6
alkoxyl, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl, and each R.sub.S2 being optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, and 5 to 6-membered heteroaryl;
[0228] R.sub.8 is H, halo or R.sub.S3, R.sub.S3 being
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl, and R.sub.S3 being optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
carboxyl, cyano amino, C.sub.1-C.sub.6 alkoxyl,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, and
C.sub.3-C.sub.8 cycloalkyl;
[0229] D is O, NR.sub.j, or CR.sub.jR.sub.k, each of R.sub.j and
R.sub.k independently being H or C.sub.1-C.sub.6 alkyl, or R.sub.j
and R.sub.k taken together, with the carbon atom to which they are
attached, form a C.sub.3-C.sub.10 cycloalkyl ring;
[0230] E is -M.sub.3-T.sub.3, M.sub.3 being a bond or
C.sub.1-C.sub.6 alkyl linker optionally substituted with halo or
cyano, T.sub.3 being C.sub.3-C.sub.10 cycloalkyl, C.sub.6-C.sub.10
aryl, 5 to 10-membered heteroaryl, or 4 to 10-membered
heterocycloalkyl, and T.sub.3 being optionally substituted with one
or more substituents selected from the group consisting of halo,
hydroxyl, thiol, carboxyl, cyano, nitro, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6
alkoxyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 haloalkoxyl,
C.sub.1-C.sub.6 alkylthio, C.sub.1-C.sub.6 alkylsulfonyl,
C.sub.1-C.sub.6 haloalkylsulfonyl, C.sub.1-C.sub.6 alkylcarbonyl,
C.sub.1-C.sub.6 alkoxycarbonyl, oxo, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.4-C.sub.12 alkylcycloalkyl, C.sub.6-C.sub.10
aryl, C.sub.6-C.sub.10 aryloxyl, C.sub.7-C.sub.14 alkylaryl,
C.sub.6-C.sub.10 aminoaryloxyl, C.sub.6-C.sub.10 arylthio, 4 to
6-membered heterocycloalkyl optionally substituted with halo,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 haloalkyl, 5 to 6-membered
heteroaryl optionally substituted with halo, C.sub.1-C.sub.4 alkyl,
and C.sub.1-C.sub.6 alkyl that is substituted with hydroxy, halo,
C.sub.1-C.sub.6 alkoxycarbonyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl optionally further substituted with halo,
hydroxyl, or C.sub.1-C.sub.6 alkoxyl;
[0231] q is 0, 1, 2, 3, or 4;
[0232] m is 0, 1, or 2; and
[0233] n is 0, 1, or 2.
[0234] For example, the sum of m and n is at least 1.
[0235] For example, m is 1 or 2 and n is 0.
[0236] For example, m is 2 and n is 0
[0237] For example, A is CH.sub.2.
[0238] For example, A is O.
[0239] For example, L.sub.1 is N(Y).
[0240] For example, L.sub.1 is SO or SO.sub.2.
[0241] For example, Y is R.sub.d.
[0242] For example, R.sub.d is C.sub.1-C.sub.6 alkyl.
[0243] For example, L.sub.2 is absent.
[0244] For example, each of G and J independently is OR.sub.a.
[0245] For example, R.sub.a is H.
[0246] For example, D is O.
[0247] For example, D is NR.sub.j.
[0248] For example, R.sub.j is H.
[0249] For example, D is CR.sub.jR.sub.k.
[0250] For example, each of R.sub.j and R.sub.k is H.
[0251] For example, E is -M.sub.3-T.sub.3, in which M.sub.3 is a
bond or C.sub.1-C.sub.3 alkyl linker, T.sub.3 is phenyl, naphthyl,
thienyl, cyclopropyl, or cyclohexyl, and T.sub.3 is optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, thiol, carboxyl, cyano, nitro,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 haloalkoxyl, C.sub.1-C.sub.6 alkylthio,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.1-C.sub.6 alkylcarbonyl,
C.sub.1-C.sub.6 alkoxycarbonyl, oxo, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.4-C.sub.12 alkylcycloalkyl, C.sub.6-C.sub.10
aryl, C.sub.6-C.sub.10 aryloxyl, C.sub.7-C.sub.14 alkylaryl,
C.sub.6-C.sub.10 aminoaryloxyl, C.sub.6-C.sub.10 arylthio, 4 to
6-membered heterocycloalkyl optionally substituted with
C.sub.1-C.sub.4 alkyl, 5 to 6-membered heteroaryl optionally
substituted with C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.6 alkyl
that is substituted with hydroxy, C.sub.1-C.sub.6 alkoxycarbonyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl.
[0252] For example, T.sub.3 is phenyl optionally substituted with
one or more substituents selected from the group consisting of
halo, hydroxyl, carboxyl, cyano, nitro, C.sub.1-C.sub.6 alkyl
(e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl,
t-butyl, n-pentyl, s-pentyl and n-hexyl), C.sub.1-C.sub.6 alkoxyl,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 haloalkoxyl,
C.sub.1-C.sub.6 alkylsulfonyl, C.sub.6-C.sub.10 aryl (e.g., phenyl
or naphthyl), and C.sub.6-C.sub.10 aryloxyl, and C.sub.7-C.sub.14
alkylaryl.
[0253] For example, E is
##STR00039## ##STR00040## ##STR00041##
[0254] For example, X is N.
[0255] For example, X is CR,
[0256] For example, X is CH.
[0257] For example, Q is NH.sub.2 or NHR.sub.b, in which R.sub.b is
-M.sub.1-T.sub.1, M.sub.1 being a bond or C.sub.1-C.sub.6 alkyl
linker and T.sub.1 being C.sub.3-C.sub.8 cycloalkyl.
[0258] For example, Q is H.
[0259] For example, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6, R.sub.7, and R.sub.8 are each H.
[0260] For example, when R.sub.8 is halo and is attached to the
same carbon atom as J, then J is not hydroxyl.
[0261] For example, when R.sub.8 is halo and is attached to the
same carbon atom as G, then G is not hydroxyl.
[0262] For example, T.sub.2 is not halo when M.sub.2 is SO.sub.2,
SO, S, CO or O.
[0263] For example, T.sub.2 is a 4-8 membered heterocycloalkyl
which is bound to M.sub.2 via a heteroatom.
[0264] For example, T.sub.2 is a 4-8 membered heterocycloalkyl
which is bound to M.sub.2 via a N atom.
[0265] For example, T.sub.2 is a 4-8 membered heterocycloalkyl
which is bound to M.sub.2 via a C atom.
[0266] The invention also relates to a compound of Formula (IV) or
its N-oxide or a pharmaceutically acceptable salt thereof:
##STR00042##
Q is H, NH.sub.2, NHR.sub.b, NR.sub.bR.sub.c, OH, R.sub.b, or
OR.sub.b, in which each of R.sub.b and R.sub.c independently is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
7-membered heterocycloalkyl, 5 to 10-membered heteroaryl, or
-M.sub.1-T.sub.1 in which M.sub.1 is a bond or C.sub.1-C.sub.6
alkyl linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxyl and T.sub.1 is C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
10-membered heteroaryl, or R.sub.b and R.sub.c, together with the N
atom to which they attach, form 4 to 7-membered heterocycloalkyl
having 0 or 1 additional heteroatoms to the N atom optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl, and each of R.sub.b, R.sub.c, and T.sub.1 is
optionally substituted with one or more substituents selected from
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, and 5 to 6-membered heteroaryl; X is N
or CR.sub.x, in which R.sub.x is H, halo, hydroxyl, carboxyl,
cyano, or R.sub.S1, R.sub.S1 being amino, C.sub.1-C.sub.6 alkoxyl,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl, and
R.sub.S1 being optionally substituted with one or more substituents
selected from halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, and 5 to 6-membered heteroaryl; Y is
H, R.sub.d, SO.sub.2R.sub.d, or COR.sub.d, R.sub.d being
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, or 5 to 6-membered heteroaryl, and
R.sub.d being optionally substituted with one or more substituents
selected from C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 alkylsulfonyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl and with
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, or 5 to 6-membered heteroaryl further optionally
substituted with C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, halo, hydroxyl, carboxyl, C(O)OH,
C(O)O--C.sub.1-C.sub.6 alkyl, OC(O)--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 6-membered heterocycloalkyl, or 5 to
6-membered heteroaryl; each of R.sub.1 and R.sub.2 independently,
is H, halo, hydroxyl, carboxyl, cyano, R.sub.S2, R.sub.S2 being
amino, C.sub.1-C.sub.6 alkoxyl, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl, and each
R.sub.S2 being optionally substituted with one or more substituents
selected from halo, hydroxyl, carboxyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
6-membered heterocycloalkyl, and 5 to 6-membered heteroaryl; each
of R.sub.e, R.sub.f, R.sub.g, and R.sub.h, independently is
-M.sub.2-T.sub.2, in which M.sub.2 is a bond, SO.sub.2, SO, S, CO,
CO.sub.2, O, O--C.sub.1-C.sub.4 alkyl linker, C.sub.1-C.sub.4 alkyl
linker, NH, or N(R.sub.t), R.sub.t being C.sub.1-C.sub.6 alkyl, and
T.sub.2 is H, halo, or R.sub.S4, R.sub.S4 being C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 8-membered
heterocycloalkyl, or 5 to 10-membered heteroaryl, and each of
O--C.sub.1-C.sub.4 alkyl linker, C.sub.1-C.sub.4 alkyl linker,
R.sub.t, and R.sub.S4 being optionally substituted with one or more
substituents selected from halo, hydroxyl, carboxyl, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 6-membered
heterocycloalkyl, and 5 to 6-membered heteroaryl, and m is 0, 1, or
2.
[0267] For example, A is O. In certain compounds of Formula (IV), A
is O and m is 2.
[0268] In certain compounds of Formula (IV), X is N.
[0269] For example, in certain compounds, Q is NH.sub.2 or
NHR.sub.b, in which R.sub.b is -M.sub.1-T.sub.1, M.sub.1 being a
bond or C.sub.1-C.sub.6 alkyl linker and T.sub.1 being
C.sub.3-C.sub.8 cycloalkyl
[0270] For example, in certain compounds of Formula (IV), R.sub.1
and R.sub.2 are each H.
[0271] In certain compounds of Formula (IV), Y is R.sub.d. For
example, R.sub.d is C.sub.1-C.sub.6 alkyl optionally substituted
with C.sub.3-C.sub.8 cycloalkyl or halo. For example, R.sub.d is
C.sub.3-C.sub.8 cycloalkyl optionally substituted with
C.sub.1-C.sub.6 alkyl or halo.
[0272] The invention also relates to a compound of Formula (IV),
wherein at least one of R.sub.e, R.sub.f, R.sub.g, and R.sub.h is
halo, C.sub.1-C.sub.6 alkoxyl optionally substituted with one or
more halo; C.sub.1-C.sub.6 alkylsulfonyl optionally substituted
with one or more halo; C.sub.1-C.sub.6 alkyl optionally substituted
with one or more substituents selected from CN, halo,
C.sub.3-C.sub.8 cycloalkyl, hydroxy, and C.sub.1-C.sub.6 alkoxyl;
C.sub.3-C.sub.8 cycloalkyl optionally substituted with one or more
C.sub.1-C.sub.6 alkyl or CN; or 4 to 8-membered heterocycloalkyl
optionally substituted with one or more substituents selected from
CN, halo, hydroxy, C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6
alkoxyl. For example, the compound of Formula (IV) has at least one
of R.sub.e, R.sub.f, R.sub.g, and R.sub.h selected from F; Cl; Br;
CF.sub.3; OCF.sub.3; SO.sub.2ClF.sub.3; oxetanyl optionally
substituted with one or more substituents selected from CN, halo,
hydroxy, C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxyl;
C.sub.3-C.sub.8 cycloalkyl optionally substituted with one or more
substituents selected from C.sub.1-C.sub.4 alkyl; and
C.sub.1-C.sub.4 alkyl optionally substituted with one or more
substituents selected from halo, C.sub.3-C.sub.8 cycloalkyl,
hydroxy and C.sub.1-C.sub.6 alkoxyl.
[0273] For example, the invention relates to compounds of Formula
(IV) where at least one of R.sub.f and R.sub.g is alkyl, optionally
substituted with hydroxyl. For example, the invention relates to
compounds where at least one of R.sub.f and R.sub.g is t-butyl
substituted with hydroxyl.
[0274] The invention relates to a compound selected from Compounds
1-140. The invention also relates to a salt of a compound selected
from Compounds 1-140. The invention also relates to an N-oxide of
compound selected from Compounds 1-140. The invention also relates
to a salt of an N-oxide of compound selected from Compounds 1-140.
For example, the invention relates to a compound selected from
Compounds 1-7, 9-109, and 111-140.
[0275] The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of a compound of Formula (IV)
and a pharmaceutically acceptable carrier.
[0276] The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of a salt of a compound of
Formula (IV) and a pharmaceutically acceptable carrier.
[0277] The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of a hydrate of a compound of
Formula (IV) and a pharmaceutically acceptable carrier.
[0278] The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of a compound selected from
Compounds 1-140 and a pharmaceutically acceptable carrier.
[0279] The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of a salt of a compound
selected from Compounds 1-140 and a pharmaceutically acceptable
carrier. The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of an N-oxide of a compound
selected from Compounds 1-140 and a pharmaceutically acceptable
carrier. The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of an N-oxide of salt of a
compound selected from Compounds 1-140 and a pharmaceutically
acceptable carrier. The invention also relates to a pharmaceutical
composition of a therapeutically effective amount of a hydrate of a
compound selected from Compounds 1-140 and a pharmaceutically
acceptable carrier.
[0280] The present invention provides pharmaceutical a composition
comprising one or more compounds of Formula (I), (II), (IIIa),
(IIIb), or (IIIc), and one or more pharmaceutically acceptable
carriers.
[0281] The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of a salt of a compound of
Formula (I), (II), (IIIa), (IIIb), or (IIIc) and a pharmaceutically
acceptable carrier.
[0282] The invention also relates to a pharmaceutical composition
of a therapeutically effective amount of a hydrate of a compound of
Formula (I), (II), (IIIa), (IIIb), or (IIIc) and a pharmaceutically
acceptable carrier.
[0283] The present invention provides methods of treating or
preventing cancer. The present invention provides methods of
treating cancer. The present invention also provides methods of
preventing cancer. The method includes administering to a subject
in need thereof a therapeutically effective amount of the compound
of Formula (I), (II), (IIIa), (IIIb), or (IIIc). The cancer can be
a hematological cancer. Preferably, the cancer is leukemia. More
preferably, the cancer is acute myeloid leukemia, acute lymphocytic
leukemia or mixed lineage leukemia.
[0284] The present invention provides methods of treating or
preventing a disease or disorder mediated by translocation of a
gene on chromosome 11q23. The present invention provides methods of
treating a disease or disorder mediated by translocation of a gene
on chromosome 11q23. The present invention also provides methods of
preventing a disease or disorder mediated by translocation of a
gene on chromosome 11q23. The method includes administering to a
subject in need thereof a therapeutically effective amount of the
compound of Formula (I), (II), (IIIa), (IIIb), (IIIc) or (IV).
[0285] The present invention provides methods of treating or
preventing a disease or disorder in which DOT1-mediated protein
methylation plays a part or a disease or disorder mediated by
DOT1-mediated protein methylation. The present invention provides
methods of treating a disease or disorder in which DOT1-mediated
protein methylation plays a part or a disease or disorder mediated
by DOT1-mediated protein methylation. The present invention also
provides methods of preventing a disease or disorder in which
DOT1-mediated protein methylation plays a part or a disease or
disorder mediated by DOT1-mediated protein methylation. The method
includes administering to a subject in need thereof a
therapeutically effective amount of the compound of Formula (I),
(II), (IIIa), (IIIb), (IIIc) or (IV).
[0286] The present invention provides methods of inhibiting DOT1L
activity in a cell. The method includes contacting the cell with an
effective amount of one or more of the compound of Formula (I),
(II), (IIIa), (IIIb), (IIIc) or (IV).
[0287] Still another aspect of the invention relates to a method of
reducing the level of Histone H3 Lysine residue 79 (H3-K79)
methylation in a cell. The method includes contacting a cell with a
compound of the present invention. Such method can be used to
ameliorate any condition which is caused by or potentiated by the
activity of DOT1 through H3-K79 methylation.
[0288] The present invention relates to use of the compounds
disclosed herein in preparation of a medicament for treating or
preventing cancer. The use includes a compound of Formula (I),
(II), (IIIa), (IIIb), (IIIc) or (IV) for administration to a
subject in need thereof in a therapeutically effective amount. The
cancer can be a hematological cancer. Preferably, the cancer is
leukemia. More preferably, the cancer is acute myeloid leukemia,
acute lymphocytic leukemia or mixed lineage leukemia.
[0289] The present invention provides use of the compounds
disclosed herein in preparation of a medicament for treating or
preventing a disease or disorder mediated by translocation of a
gene on chromosome 11q23. The use includes a compound of Formula
(I), (II), (IIIa), (IIIb), (IIIc) or (IV) for administration to a
subject in need thereof in a therapeutically effective amount.
[0290] The present invention provides use of the compounds
disclosed herein in preparation of a medicament for treating or
preventing a disease or disorder in which DOT1-mediated protein
methylation plays a part or a disease or disorder mediated by
DOT1-mediated protein methylation. The use includes a compound of
Formula (I), (II), (IIIa), (IIIb), (IIIc) or (IV) for
administration to a subject in need thereof in a therapeutically
effective amount.
[0291] The present invention provides use of the compounds
disclosed herein for inhibiting DOT1L activity in a cell. The use
includes contacting the cell with an effective amount of one or
more of the compound of Formula (I), (II), (IIIa), (IIIb), (IIIc)
or (IV).
[0292] Still another aspect of the invention relates to a use of
the compounds disclosed herein for reducing the level of Histone H3
Lysine residue 79 (H3-K79) methylation in a cell. The use includes
contacting a cell with a compound of the present invention. Such
use can ameliorate any condition which is caused by or potentiated
by the activity of DOT1 through H3-K79 methylation.
[0293] In the formulae presented herein, the variables can be
selected from the respective groups of chemical moieties later
defined in the detailed description.
[0294] In addition, the invention provides methods of synthesizing
the foregoing compounds. Following synthesis, a therapeutically
effective amount of one or more of the compounds can be formulated
with a pharmaceutically acceptable carrier for administration to a
mammal, particularly humans, for use in modulating an epigenetic
enzyme. In certain embodiments, the compounds of the present
invention are useful for treating, preventing, or reducing the risk
of cancer or for the manufacture of a medicament for treating,
preventing, or reducing the risk of cancer. Accordingly, the
compounds or the formulations can be administered, for example, via
oral, parenteral, otic, ophthalmic, nasal, or topical routes, to
provide an effective amount of the compound to the mammal.
[0295] Representative compounds of the present invention include
compounds listed in Table 1.
TABLE-US-00001 TABLE 1 Data (MS Cmpd or No. Structure Chemical Name
NMR) 1 ##STR00043## (2R,3S,4R,5R)-2- (((3-(2-(1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)-5-(6-
amino-9H-purin-9- yl)tetrahydrofuran- 3,4-diol 2 ##STR00044##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-((((1r,3S)-3-(2-
(5-(tert-butyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(isopropyl)amino) methyl) tetrahydrofuran- 3,4-diol 563.4 (M +
H).sup.+ 3 ##STR00045## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-((((1s,3R)-3-(2- (5-(tert-butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isopropyl)amino) methyl) tetrahydrofuran-
3,4-diol 563.5 (M + H).sup.+ 4 ##STR00046## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9- yl)-5-((((1s,3R)-3-(2- ((trifluoromethyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (isopropyl)amino) methyl)
tetrahydrofuran- 3,4-diol 609.2 (M + H).sup.+ 5 ##STR00047##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-((((1r,3S)-3-(2-
(5-chloro-6- (trifluoromethyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isopropyl)amino) methyl) tetrahydrofuran-
3,4-diol 609.2 (M + H).sup.+ 6 ##STR00048## (2R,3R,4S,5R)-2-(4-
amino-7H-pyrrolo [2,3-d]pyrimidin- 7-yl)-5-(((3-((5-(tert-
butyl)-1H-benzo[d] imidazol-2-yl) methyl)cyclobutyl)
(methyl)amino)methyl) tetrahydrofuran- 3,4-diol 520.4 (M + H).sup.+
7 ##STR00049## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- (((3-(2-(6-chloro-5- (trifluoromethyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (methyl)amino)methyl)
cyclopentane-1,2- diol 579.7 (M + H).sup.+ 8 ##STR00050## 1-(3-
((((2R,3S,4R,5R)-5- (6-amino-9H-purin-9- yl)-3,4-
dihydroxytetrahydrofuran- 2- yl)methyl)(methyl)
amino)cyclobutyl)-3- (4-tert- butylphenyl)urea 525.5 (M + H).sup.+
9 ##STR00051## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(6- chloro-5-
(trifluoromethyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(methyl)amino)methyl) cyclopentane-1,2- diol 578.3 (M + H).sup.+ 10
##STR00052## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((methyl((1r,3S)-3- (2-(5-
(trifluoromethyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
amino)methyl) cyclopentane-1,2-diol 544.3 (M + H).sup.+ 11
##STR00053## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((methyl((1s,3R)-3- 2-(5-
(trifluoromethyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
amino)methyl) cyclopentane-1,2-diol 544.3 (M + H).sup.+ 12
##STR00054## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((((1s,3R)-3-(2-(6- chloro-5-
(trifluoromethyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(methyl)amino)methyl) cyclopentane-1,2- diol 578.3 (M + H).sup.+ 13
##STR00055## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((methyl(3-(2-(5- (trifluoromethyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
cyclopentane-1,2-diol 544.5 (M + H).sup.+ 14 ##STR00056##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
(((3-(2-(5-(tert- butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (methyl)amino)methyl) cyclopentane-1,2- diol
532.3 (M + H).sup.+ 15 ##STR00057## (1R,2S,3R,5R)-3-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-5- ((isopropyl((3-((5-
(trifluoromethyl)-1H- benzo[d]imidazol-2- yl)methyl)cyclobutyl)
methyl)amino)methyl) cyclopentane-1,2- diol 572.4 (M + H).sup.+ 16
##STR00058## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- (((3-(5-chloro-6- (trifluoromethyl)-1H-
benzo[d]imidazol-2- yl)cyclobutyl)(methyl) amino)methyl)
cyclopentane-1,2-diol 550.3 (M + H).sup.+ 17 ##STR00059##
(1R,2R,4S)-2-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-4-
(((3-(2-(5-chloro-6- (trifluoromethyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (methyl)amino)methyl) cyclopentanol 562.3 (M +
H).sup.+ 18 ##STR00060## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((methyl((3-((5- (trifluoromethyl)-1H-
benzo[d]imidazol-2- yl)methyl)cyclobutyl) methyl)amino)methyl)
cyclopentane-1,2- diol 544 (M + H).sup.+ 19 ##STR00061##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
(((3-(2-(5-(tert- butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isopropyl)amino)methyl) cyclopentane-1,2-
diol NMR data 20 ##STR00062## (1R,2S,3R,5R)-3-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-5- (((3-(2-(6-chloro-5-
(trifluoromethyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(isopropyl)amino)methyl) cyclopentane- 1,2-diol NMR data 21
##STR00063## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((((3-((6-chloro-5- (trifluoromethyl)-1H-
benzo[d]imidazol-2- yl)methyl)cyclobutyl) methyl)(isopropyl)
amino)methyl) cyclopentane-1,2-diol 606.3 (M + H).sup.+ 22
##STR00064## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- (((3-((5-(tert-butyl)- 1H- benzo[d]imidazol-2-
yl)methyl)cyclobutyl) methyl)(isopropyl) amino)methyl)
cyclopentane-1,2-diol 560.4 (M + H).sup.+ 23 ##STR00065##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
(((3-(2-(5,6-dichloro- 1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(methyl)amino)methyl) cyclopentane-1,2- diol NMR data 24
##STR00066## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((methyl(3-(2-(5- (trifluoromethoxy)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
cyclopentane-1,2-diol 558.2 (M - H).sup.+ 25 ##STR00067##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
(((3-(2-(5-(tert- butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (ethyl)amino)methyl) cyclopentane-1,2-diol
546.3 (M + H).sup.+ 26 ##STR00068## (1R,2S,3R,5R)-3-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-5- (((3-(2-(5-bromo-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (methyl)amino)methyl)
cyclopentane-1,2- diol 554.1 (M + H).sup.+ 27 ##STR00069##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-((isopropyl(3-
(2-(5-(1- methylcyclobutyl)- 1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) amino)methyl) tetrahydrofuran- 3,4-diol 575.5
(M + H).sup.+ 28 ##STR00070## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5- ((isopropyl((1r,3S)-3- (2-(5-(1- methylcyclobutyl)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran- 3,4-diol 575.5 (M + H).sup.+ 29 ##STR00071##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((methyl(3-(2-(5-(1- methylcyclobutyl)- 1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) amino)methyl) cyclopentane-1,2-diol 544.4 (M +
H).sup.+ 30 ##STR00072## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5- ((methyl((1r,3S)-3- (2-(5-(1- methylcyclobutyl)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran- 3,4-diol 547.6 (M + H).sup.+ 31 ##STR00073##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5- ((isopropyl((1s,3R)-
3-(2-(5-(1- methylcyclobutyl)- 1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) amino)methyl) tetrahydrofuran- 3,4-diol 575.6
(M + H).sup.+ 32 ##STR00074## (1R,2S,3R,5R)-3-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(5-
(tert-butyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(methyl)amino)methyl) cyclopentane-1,2- diol 532.4 (M + H).sup.+ 33
##STR00075## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-((methyl(3-(2- (5-(1- methylcyclobutyl)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran- 3,4-diol 547.3 (M + H).sup.+ 34 ##STR00076##
2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5- ((methyl((1s,3R)-3-
(2-(5-(1- methylcyclobutyl)- 1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) amino)methyl) tetrahydrofuran- 3,4-diol 547.5
(M + H).sup.+ 35 ##STR00077## (1R,2S,3R,5R)-3-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-5- ((((1s,3R)-3-(2-(5-
(tert-butyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(methyl)amino)methyl) cyclopentane-1,2- diol NMR data 36
##STR00078## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((methyl((1r,3S)-3- (2-(5-(1-
methylcyclobutyl)- 1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
amino)methyl) cyclopentane-1,2-diol 544.4 (M + H).sup.+ 37
##STR00079## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((methyl((1r,3S)-3- (2-(5- (trifluoromethoxy)-
1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
cyclopentane-1,2-diol 558.3 (M - H).sup.- 38 ##STR00080##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((((1r,3S)-3-(2-(5- (tert-butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (ethyl)amino)methyl) cyclopentane-1,2-diol
546.3 (M + H).sup.+ 39 ##STR00081## (1R,2S,3R,5R)-3-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-5- ((methyl((1s,3R)-3- (2-(5-(1-
methylcyclobutyl)- 1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
amino)methyl) cyclopentane-1,2-diol 544.3 (M + H).sup.+ 40
##STR00082## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(5- (tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (cyclopropylmethyl)
amino)methyl) cyclopentane-1,2-diol NMR data 41 ##STR00083##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((((1r,3S)-3-(2-(5- (tert-butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isopropyl)amino) methyl)cyclopentane-1,2-
diol NMR data 42 ##STR00084## (1R,2S,3R,5R)-3-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(5-
(tert-butyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(cyclobutylmethyl) amino)methyl) cyclopentane-1,2-diol 586.3 (M +
H).sup.+ 43 ##STR00085## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- (((3-(2-(5-(tert- butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (cyclobutyl)amino)
methyl)cyclopentane-1,2- diol 572.2 (M + H).sup.+ 44 ##STR00086##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
(((3-(2-(5-(tert- butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (cyclopropylmethyl) amino)methyl)
cyclopentane-1,2-diol 572.6 (M + H).sup.+ 45 ##STR00087##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
(((3-(2-(5-(tert- butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isobutyl)amino)methyl) cyclopentane-1,2- diol
574.6 (M + H).sup.+ 46 ##STR00088## (1R,2S,3R,5R)-3-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(5-
(tert-butyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(cyclobutyl)amino) methyl)cyclopentane-1,2- diol 572.6 (M +
H).sup.+ 47 ##STR00089## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(5- bromo-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (methyl)amino)methyl)
cyclopentane-1,2- diol 556.0 (M + H).sup.+ 48 ##STR00090##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((((1s,3R)-3-(2-(5- (tert-butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isobutyl)amino)methyl) cyclopentane-1,2- diol
572.3 (M - H).sup.- 49 ##STR00091## (1R,2S,3R,5R)-3-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-5- ((((1s,3R)-3-(2-(5-
(tert-butyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(ethyl)amino)methyl) cyclopentane-1,2-diol 546.3 (M + H).sup.+
50 ##STR00092## (1R,2S,3R,5R)-3-(6- amino-9H-purin-9-
yl)-5-(((3-(2-(5-(tert- butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isopropyl)amino) methyl)cyclopentane-1,2-
diol 561.4 (M + H).sup.+ 51 ##STR00093## (1R,2S,3R,5R)-3-(4-
amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5- ((((1s,3R)-3-(2-(5-
(tert-butyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(cyclobutyl)amino) methyl)cyclopentane-1,2- diol 572.7 (M +
H).sup.+ 52 ##STR00094## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(5,6- dichloro-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (methyl)amino)methyl)
cyclopentane-1,2- diol NMR data 53 ##STR00095## (1R,2S,3R,5R)-3-(4-
amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(5-
(tert-butyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(isobutyl)amino) methyl)cyclopentane-1,2- diol 572.3 (M - H).sup.-
54 ##STR00096## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((((1s,3R)-3-(2-(5- (tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (cyclopropylmethyl)
amino)methyl) cyclopentane- 1,2-diol NMR data 55 ##STR00097##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((((1s,3R)-3-(2-(5- bromo-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (methyl)amino)methyl) cyclopentane-1,2- diol
NMR data 56 ##STR00098## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((isopropyl(3-(2-(5- (trifluoromethoxy)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
cyclopentane-1,2-diol 588.2 (M + H).sup.+ 57 ##STR00099##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((methyl((1s,3R)-3- (2-(5- (trifluoromethoxy)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
cyclopentane-1,2-diol 560.1 (M + H).sup.+ 58 ##STR00100##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((((1s,3R)-3-(2-(5,6- dichloro-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (methyl)amino)methyl) cyclopentane-1,2- diol
NMR data 59 ##STR00101## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((((1s,3R)-3-(2-(5- (tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (cyclobutylmethyl)
amino)methyl) cyclopentane-1,2-diol 586.4 (M + H).sup.+ 60
##STR00102## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((isopropyl((1r,3S)-3- (2-(5-
(trifluoromethoxy)- 1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
amino)methyl) cyclopentane-1,2-diol 588.2 (M + H).sup.+ 61
##STR00103## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((isopropyl((1s,3R)- 3-(2-(5-
(trifluoromethoxy)- 1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
amino)methyl) cyclopentane-1,2-diol 588.7 (M + H).sup.+ 62
##STR00104## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((methyl(3-(2-(5- (oxetan-3-yl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
cyclopentane-1,2-diol NMR data 63 ##STR00105## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9- yl)-5- ((methyl((1r,3S)-3- (2-(5-(oxetan-3-yl)-
1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran-3,4-diol 535.4 (M + H).sup.+ 64 ##STR00106##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-((methyl(3-(2-
(5-(oxetan-3-yl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
amino)methyl) tetrahydrofuran-3,4-diol 535.3 (M + H).sup.+ 65
##STR00107## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-
((methyl((1s,3R)-3- (2-(5-(oxetan-3-yl)- 1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) amino)methyl) tetrahydrofuran-3,4-diol 535.4
(M + H).sup.+ 66 ##STR00108## (1R,2S,3R,5R)-3-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-5- (((3-(2-(5-(tert- butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (2,2,2-
trifluoroethyl)amino) methyl)cyclopentane- 1,2-diol 600.2 (M +
H).sup.+ 67 ##STR00109## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-(((3-(2-(5- cyclobutyl-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isopropyl)amino) methyl)tetrahydrofuran-
3,4-diol 561.5 (M + H).sup.+ 68 ##STR00110## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9- yl)-5-(((3-(2-(5-(1- methoxy-2-
methylpropan-2-yl)- 1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(methyl)amino)methyl) tetrahydrofuran-3,4- diol 565.4 (M + H).sup.+
69 ##STR00111## (1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(5- (tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (methyl)amino)methyl)
cyclopentane-1,2- diol 532.3 (M + H).sup.+ 70 ##STR00112##
(1R,2S,3R,5R)-3-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((((1s,3R)-3-(2-(6- (tert-butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (methyl)amino)methyl) cyclopentane-1,2- diol
532.3 (M + H).sup.+ 71 ##STR00113## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9- yl)-5-((((1r,3S)-3-(2- (5-(tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (methyl)amino)methyl)
tetrahydrofuran-3,4- diol 535.3 (M + H).sup.+ 72 ##STR00114##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-((((1s,3R)-3-(2-
(5-(tert-butyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(methyl)amino)methyl) tetrahydrofuran-3,4- diol 535.3 (M + H).sup.+
73 ##STR00115## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-((((1r,3S)-3-(2- (5-(tert-butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (ethyl)amino)methyl) tetrahydrofuran-3,4-diol
549.3 (M + H).sup.+ 74 ##STR00116## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9- yl)-5-((((1s,3R)-3-(2- (5-(tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (ethyl)amino)methyl)
tetrahydrofuran-3,4-diol 549.3 (M + H).sup.+ 75 ##STR00117##
(2R,3R,4S,5R)-2-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((((1s,3R)-3-(2-(5- (tert-butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isopropyl)amino) methyl)tetrahydrofuran-
3,4-diol 562.5 (M + H).sup.+ 76 ##STR00118## (2R,3R,4S,5R)-2-(4-
amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5- ((isopropyl((1s,3R)-
3-(2-(5- (trifluoromethoxy)- 1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) amino)methyl) tetrahydrofuran-3,4-diol 590.3
(M + H).sup.+ 77 ##STR00119## (2R,3R,4S,5R)-2-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-5- ((((1s,3R)-3-(2-(5-
(tert-butyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (2,2,2-
trifluoroethyl)amino) methyl)tetrahydrofuran- 3,4-diol 602.3 (M +
H).sup.+ 78 ##STR00120## (2R,3R,4S,5R)-2-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(5- (tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (2,2,2-
trifluoroethyl)amino) methyl)tetrahydrofuran- 3,4-diol 602.3 (M +
H).sup.+ 79 ##STR00121## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-(((3-(2-(5-(tert- butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (ethyl)amino)methyl) tetrahydrofuran-3,4-diol
549.3 (M + H).sup.+ 80 ##STR00122## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9- yl)-5-((((1s,3R)-3-(2- (5-(tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (2,2,2-
trifluoroethyl)amino) methyl)tetrahydrofuran- 3,4-diol 603.3 (M +
H).sup.+ 81 ##STR00123## (1R,2R,4S)-2-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-4- ((((1r,3R)-3-(2-(5- chloro-6-
(trifluoromethyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(methyl)amino)methyl) cyclopentanol 562.3 (M + H).sup.+ 82
##STR00124## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-(((3-(2-(5-(tert- butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (2,2,2- trifluoroethyl)amino)
methyl)tetrahydrofuran- 3,4-diol 603.3 (M + H).sup.+ 83
##STR00125## (1R,2R,4S)-2-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-4- (((3-(2-(5-(tert- butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (isopropyl)amino)
methyl)cyclopentanol 544.5 (M + H).sup.+ 84 ##STR00126##
(1R,2R,4S)-2-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-4-
(((3-(2-(5-chloro-6- (trifluoromethyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isopropyl)amino) methyl)cyclopentanol 590.3
(M + H).sup.+ 85 ##STR00127## (1R,2R,4S)-2-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-4- ((((1s,3S)-3-(2-(5- chloro-6-
(trifluoromethyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(methyl)amino)methyl) cyclopentanol 562.3 (M + H).sup.+ 86
##STR00128## (2R,3R,4S,5R)-2-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(5- (tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (methyl)amino)methyl)
tetrahydrofuran-3,4- diol 534.3 (M + H).sup.+ 87 ##STR00129##
(2R,3R,4S,5R)-2-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((isopropyl((1r,3S)-3- (2-(5- (trifluoromethoxy)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran-3,4-diol 590.3 (M + H).sup.+ 88 ##STR00130##
(2R,3R,4S,5R)-2-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((((1s,3R)-3-(2-(5- (tert-butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (ethyl)amino)methyl) tetrahydrofuran-3,4-diol
548.3 (M + H).sup.+ 89 ##STR00131## (2R,3R,4S,5R)-2-(4- amino-7H-
pyrrolo[2,3- d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(5-
(tert-butyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(ethyl)amino)methyl) tetrahydrofuran-3,4-diol 548.3 (M + H).sup.+
90 ##STR00132## (2R,3R,4S,5R)-2-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((((1r,3S)-3-(2-(5- (tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (isopropyl)amino)
methyl)tetrahydrofuran- 3,4-diol 562.5 (M + H).sup.+ 91
##STR00133## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-
((isopropyl((1r,3S)-3- (2-(5- (trifluoromethoxy)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran-3,4-diol 591.2 (M + H).sup.+ 92 ##STR00134##
(2R,3R,4S,5R)-2-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((((1s,3R)-3-(2-(5- (tert-butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (methyl)amino)methyl) tetrahydrofuran-3,4-
diol 534.3 (M + H).sup.+ 93 ##STR00135## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9- yl)-5- ((isopropyl((1s,3R)- 3-(2-(5-
(trifluoromethoxy)- 1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
amino)methyl) tetrahydrofuran-3,4-diol 591.3 (M + H).sup.+ 94
##STR00136## (2R,3R,4S,5R)-2-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-5- ((methyl((1r,3S)-3- (2-(5- (trifluoromethoxy)-
1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran-3,4-diol 562.2 (M + H).sup.+ 95 ##STR00137##
(2R,3R,4S,5R)-2-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-5-
((methyl((1s,3R)-3- (2-(5- (trifluoromethoxy)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran-3,4-diol 562.3 (M + H).sup.+ 96 ##STR00138##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5- ((methyl((1r,3S)-3-
(2-(5- (trifluoromethoxy)- 1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) amino)methyl) tetrahydrofuran-3,4-diol 563.3
(M + H).sup.+ 97 ##STR00139## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5- ((methyl((1s,3R)-3- (2-(5- (trifluoromethoxy)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran-3,4-diol 563.3 (M + H).sup.+ 98 ##STR00140##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-((((1r,3S)-3-(2-
(5-(tert-butyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
amino)methyl) tetrahydrofuran-3,4-diol 521.3 (M + H).sup.+ 99
##STR00141## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-((((1s,3R)-3-(2- (5-(tert-butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) amino)methyl) tetrahydrofuran-3,4-diol 521.3
(M + H).sup.+ 100 ##STR00142## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9-
yl)-5-((isopropyl(3- (2-(5- (trifluoromethoxy)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran-3,4-diol 591.3 (M + H).sup.+ 101 ##STR00143##
(1R,2R,4S)-2-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-4-
((((1r,3R)-3-(2-(5- (tert-butyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isopropyl)amino) methyl)cyclopentanol 544.1
(M + H).sup.+ 102 ##STR00144## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9- yl)-5-((((1r,3S)-3-(2- (5-(tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (2,2,2-
trifluoroethyl)amino) methyl)tetrahydrofuran- 3,4-diol 603.3 (M +
H).sup.+ 103 ##STR00145## (1R,2R,4S)-2-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-4- ((((1s,3S)-3-(2-(5- chloro-6-
(trifluoromethyl)-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(isopropyl)amino) methyl)cyclopentanol 589.9 (M + H).sup.+ 104
##STR00146## (1R,2R,4S)-2-(4- amino-7H- pyrrolo[2,3-
d]pyrimidin-7-yl)-4- ((((1s,3S)-3-(2-(5- (tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (isopropyl)amino)
methyl)cyclopentanol 544.1 (M + H).sup.+ 105 ##STR00147##
(1R,2R,4S)-2-(4- amino-7H- pyrrolo[2,3- d]pyrimidin-7-yl)-4-
((((1r,3R)-3-(2-(5- chloro-6- (trifluoromethyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (isopropyl)amino)
methyl)cyclopentanol 589.9 (M + H).sup.+ 106 ##STR00148##
(1r,3S)-N- (((2R,3S,4R,5R)-5- (6-amino-9H-purin-9- yl)-3,4-
dihydroxytetrahydrofuran- 2-yl)methyl)-3- (2-(5-(tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)-N- isopropylcyclobutanamine oxide
579.4 (M + H).sup.+ 107 ##STR00149## (R,1s,3R)-N-
(((2R,3S,4R,5R)-5- (6-amino-9H-purin-9- yl)-3,4-
dihydroxytetrahydrofuran- 2-yl)methyl)-3- (2-(5-(tert-butyl)-1H-
benzo[d]imidazol-2- yl)ethyl)-N- isopropylcyclobutanamine oxide
579.4 (M + H).sup.+ 108 ##STR00150## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9- yl)-5-((((1r,3S)-3-(2- (5-(1-hydroxy-2-
methylpropan-2-yl)- 1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(isopropyl)amino) methyl)tetrahydrofuran- 3,4-diol 579.4 (M +
H).sup.+ 109 ##STR00151## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-((((1s,3R)-3-(2- (5-(1-hydroxy-2- methylpropan-2-yl)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (isopropyl)amino)
methyl)tetrahydrofuran- 3,4-diol 579.4 (M + H).sup.+ 110
##STR00152## 1-((3- ((((2R,3S,4R,5R)-5- (6-amino-9H-purin-9-
yl)-3,4- dihydroxytetrahydrofuran- 2- yl)methyl)(methyl)
amino)cyclobutyl) methyl)-3-(4-(tert- butyl)phenyl)urea 539.3 (M +
H).sup.+ 111 ##STR00153## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-(((3-(2-(5- cyclobutyl-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isopropyl)amino) methyl)tetrahydrofuran-
3,4-diol 561 (M + H).sup.+ 112 ##STR00154## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9- yl)-5-(((3-(2-(5- cyclopropyl-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (isopropyl)amino)methyl)
tetrahydrofuran- 3,4-diol 547 (M + H).sup.+ 113 ##STR00155##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-((isopropyl(3-
(2-(5-(2,2,2- trifluoroethyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) amino)methyl) tetrahydrofuran-3,4-diol 589 (M
+ H).sup.+ 114 ##STR00156## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-((((1s,3R)-3-(2- (5-cyclobutyl-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (isopropyl)amino) methyl)tetrahydrofuran-
3,4-diol 561 (M + H).sup.+ 115 ##STR00157## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9- yl)-5-((((1r,3S)-3-(2- (5-cyclobutyl-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (isopropyl)amino)
methyl)tetrahydrofuran- 3,4-diol 561 (M + H).sup.+ 116 ##STR00158##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-((((1r,3S)-3-(2-
(5-cyclopropyl-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(isopropyl)amino) methyl)tetrahydrofuran- 3,4-diol 547 (M +
H).sup.+ 117 ##STR00159## 1-(2-(2-(3- ((((2R,3S,4R,5R)-5-
(6-amino-9H-purin-9- yl)-3,4- dihydroxytetrahydrofuran-
2-yl)methyl)(isopropyl) amino)cyclobutyl) ethyl)-1H-
benzo[d]imidazol-5-yl) cyclobutanecarbonitrile 586 (M + H).sup.+
118 ##STR00160## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-((isopropyl(3- (2-(5-(1-methoxy-2- methylpropan-2-yl)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran-3,4-diol 593 (M + H).sup.+ 119 ##STR00161##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-((((1s,3R)-3-(2-
(5-cyclopropyl-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(isopropyl)amino) methyl)tetrahydrofuran- 3,4-diol 547 (M +
H).sup.+ 120 ##STR00162## 2-(2-(2-(3- ((((2R,3S,4R,5R)-5-
(6-amino-9H-purin-9- yl)-3,4- dihydroxytetrahydrofuran-
2-yl)methyl)(isopropyl) amino)cyclobutyl) ethyl)-1H-
benzo[d]imidazol-5- yl)-2- methylpropanenitrile 574 (M + H).sup.+
121 ##STR00163## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-
((isopropyl((1s,3R)- 3-(2-(5-(1-methoxy- 2-methylpropan-2- yl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran-3,4-diol 593 (M + H).sup.+ 122 ##STR00164##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5- ((isopropyl((1r,3S)-3-
(2-(5-(1-methoxy-2- methylpropan-2-yl)- 1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) amino)methyl) tetrahydrofuran-3,4-diol 593 (M
+ H).sup.+ 123 ##STR00165## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5- ((isopropyl((1s,3R)- 3-(2-(5-(2,2,2- trifluoroethyl)-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran-3,4-diol 589 (M + H).sup.+ 124 ##STR00166##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5- ((isopropyl((1r,3S)-3-
(2-(5-(2,2,2- trifluoroethyl)-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) amino)methyl) tetrahydrofuran-3,4-diol 589 (M
+ H).sup.+ 125 ##STR00167## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-(((3-(2-(5- cyclobutyl-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (methyl)amino)methyl) tetrahydrofuran-3,4-
diol 533 (M + H).sup.+ 126 ##STR00168## 1-(2-(2-(3-
((((2R,3S,4R,5R)-5- (6-amino-9H-purin-9- yl)-3,4-
dihydroxytetrahydrofuran- 2-yl)methyl)(isopropyl) amino)cyclobutyl)
ethyl)-1H- benzo[d]imidazol-5-yl) cyclopropanecarbonitrile 572 (M +
H).sup.+ 127 ##STR00169## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-(((3-(2-(5- cyclopropyl-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (methyl)amino)methyl) tetrahydrofuran-3,4-
diol 519 (M + H).sup.+ 128 ##STR00170## 2-(2-(2-((1S,3r)-3-
((((2R,3S,4R,5R)-5- (6-amino-9H-purin-9- yl)-3,4-
dihydroxytetrahydrofuran- 2-yl)methyl)(isopropyl) amino)cyclobutyl)
ethyl)-1H- benzo[d]imidazol-5- yl)-2- methylpropanenitrile 574 (M +
H).sup.+ 129 ##STR00171## 2-(2-(2-((1R,3s)-3- ((((2R,3S,4R,5R)-5-
(6-amino-9H-purin-9- yl)-3,4- dihydroxytetrahydrofuran-
2-yl)methyl)(isopropyl) amino)cyclobutyl) ethyl)-1H-
benzo[d]imidazol-5- yl)-2- methylpropanenitrile 574 (M + H).sup.+
130 ##STR00172## 1-(2-(2-(3- ((((2R,3S,4R,5R)-5-
(6-amino-9H-purin-9- yl)-3,4- dihydroxytetrahydrofuran-
2-yl)methyl)(methyl) amino)cyclobutyl) ethyl)-1H-
benzo[d]imidazol-5-yl) cyclopropanecarbonitrile 544 (M + H).sup.+
131 ##STR00173## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-((((1r,3S)-3-(2- (5-cyclobutyl-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (methyl)amino)methyl) tetrahydrofuran-3,4-
diol 533 (M + H).sup.+ 132 ##STR00174## (2R,3R,4S,5R)-2-(6-
amino-9H-purin-9- yl)-5-((((1s,3R)-3-(2- (5-cyclobutyl-1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) (methyl)amino)methyl)
tetrahydrofuran-3,4- diol 533 (M + H).sup.+ 133 ##STR00175##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-((((1r,3S)-3-(2-
(5-cyclopropyl-1H- benzo[d]imidazol-2- yl)ethyl)cyclobutyl)
(methyl)amino)methyl) tetrahydrofuran-3,4- diol 519 (M + H).sup.+
134 ##STR00176## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-((((1s,3R)-3-(2- (5-cyclopropyl-1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (methyl)amino)methyl) tetrahydrofuran-3,4-
diol 519 (M + H).sup.+ 135 ##STR00177## 1-(2-(2-((1S,3r)-3-
((((2R,3S,4R,5R)-5- (6-amino-9H-purin-9- yl)-3,4-
dihydroxytetrahydrofuran- 2-yl)methyl)(isopropyl) amino)cyclobutyl)
ethyl)-1H- benzo[d]imidazol-5-yl) cyclopropanecarbonitrile 572 (M +
H).sup.+ 136 ##STR00178## 1-(2-(2-((1R,3s)-3- ((((2R,3S,4R,5R)-5-
(6-amino-9H-purin-9- yl)-3,4- dihydroxytetrahydrofuran-
2-yl)methyl)(isopropyl) amino)cyclobutyl) ethyl)-1H-
benzo[d]imidazol-5-yl) cyclopropanecarbonitrile 572 (M + H).sup.+
137 ##STR00179## 1-(2-(2-((1S,3r)-3- ((((2R,3S,4R,5R)-5-
(6-amino-9H-purin-9- yl)-3,4- dihydroxytetrahydrofuran-
2-yl)methyl)(methyl) amino)cyclobutyl) ethyl)-1H-
benzo[d]imidazol-5-yl) cyclopropanecarbonitrile 544 (M + H).sup.+
138 ##STR00180## 1-(2-(2-((1R,3s)-3- ((((2R,3S,4R,5R)-5-
(6-amino-9H-purin-9- yl)-3,4- dihydroxytetrahydrofuran-
2-yl)methyl)(methyl) amino)cyclobutyl)ethyl)- 1H-
benzo[d]imidazol-5-yl) cyclopropanecarbonitrile 544 (M + H).sup.+
139 ##STR00181## (2R,3R,4S,5R)-2-(6- amino-9H-purin-9-
yl)-5-((isopropyl(3- (2-(5-(1- methylcyclopropyl)- 1H-
benzo[d]imidazol-2- yl)ethyl)cyclobutyl) amino)methyl)
tetrahydrofuran-3,4-diol 561 (M + H).sup.+ 140 ##STR00182##
(2R,3R,4S,5R)-2-(6- amino-9H-purin-9- yl)-5-(((3-(2-(5-(1-
methoxy-2- methylpropan-2-yl)- 1H- benzo[d]imidazol-2-
yl)ethyl)cyclobutyl) (methyl)amino)methyl) tetrahydrofuran-3,4-
diol 565 (M + H).sup.+
[0296] As used herein, "alkyl", "C.sub.1, C.sub.2, C.sub.3,
C.sub.4, C.sub.5 or C.sub.6 alkyl" or "C.sub.1-C.sub.6 alkyl" is
intended to include C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5 or
C.sub.6 straight chain (linear) saturated aliphatic hydrocarbon
groups and C.sub.3, C.sub.4, C.sub.5 or C.sub.6 branched saturated
aliphatic hydrocarbon groups. For example, C.sub.1-C.sub.6 alkyl is
intended to include C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5 and
C.sub.6 alkyl groups. Examples of alkyl include, moieties having
from one to six carbon atoms, such as, but not limited to, methyl,
ethyl, n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-pentyl,
s-pentyl or n-hexyl.
[0297] In certain embodiments, a straight chain or branched alkyl
has six or fewer carbon atoms (e.g., C.sub.1-C.sub.6 for straight
chain, C.sub.3-C.sub.6 for branched chain), and in another
embodiment, a straight chain or branched alkyl has four or fewer
carbon atoms.
[0298] As used herein, the term "cycloalkyl" refers to a saturated
or unsaturated nonaromatic hydrocarbon mono- or multi-ring system
having 3 to 30 carbon atoms (e.g., C.sub.3-C.sub.10). Examples of
cycloalkyl include, but are not limited to, cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl,
cyclopentenyl, cyclohexenyl, cycloheptenyl, and adamantyl. The term
"heterocycloalkyl" refers to a saturated or unsaturated nonaromatic
5-8 membered monocyclic, 8-12 membered bicyclic, or 11-14 membered
tricyclic ring system having one or more heteroatoms (such as O, N,
S, or Se). Examples of heterocycloalkyl groups include, but are not
limited to, piperazinyl, pyrrolidinyl, dioxanyl, morpholinyl, and
tetrahydrofuranyl.
[0299] The term "optionally substituted alkyl" refers to
unsubstituted alkyl or alkyl having designated substituents
replacing one or more hydrogen atoms on one or more carbons of the
hydrocarbon backbone. Such substituents can include, for example,
alkyl, alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,
alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato,
amino (including alkylamino, dialkylamino, arylamino, diarylamino
and alkylarylamino), acylamino (including alkylcarbonylamino,
arylcarbonylamino, carbamoyl and ureido), amidino, imino,
sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,
alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an
aromatic or heteroaromatic moiety.
[0300] An "arylalkyl" or an "aralkyl" moiety is an alkyl
substituted with an aryl (e.g., phenylmethyl (benzyl)). An
"alkylaryl" moiety is an aryl substituted with an alkyl (e.g.,
methylphenyl).
[0301] As used herein, "alkyl linker" is intended to include
C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5 or C.sub.6 straight
chain (linear) saturated divalent aliphatic hydrocarbon groups and
C.sub.3, C.sub.4, C.sub.5 or C.sub.6 branched saturated aliphatic
hydrocarbon groups. For example, C.sub.1-C.sub.6 alkyl linker is
intended to include C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5 and
C.sub.6 alkyl linker groups. Examples of alkyl linker include,
moieties having from one to six carbon atoms, such as, but not
limited to, methyl (--CH.sub.2--), ethyl (--CH.sub.2CH.sub.2--),
n-propyl (--CH.sub.2CH.sub.2CH.sub.2--), i-propyl
(--CHCH.sub.3CH.sub.2--), n-butyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--), s-butyl
(--CHCH.sub.3CH.sub.2CH.sub.2--), i-butyl
(--C(CH.sub.3).sub.2CH.sub.2--), n-pentyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2--), s-pentyl
(--CHCH.sub.3CH.sub.2CH.sub.2CH.sub.2--) or n-hexyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2--).
[0302] "Alkenyl" includes unsaturated aliphatic groups analogous in
length and possible substitution to the alkyls described above, but
that contain at least one double bond. For example, the term
"alkenyl" includes straight chain alkenyl groups (e.g., ethenyl,
propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl,
decenyl), and branched alkenyl groups. In certain embodiments, a
straight chain or branched alkenyl group has six or fewer carbon
atoms in its backbone (e.g., C.sub.2-C.sub.6 for straight chain,
C.sub.3-C.sub.6 for branched chain). The term "C.sub.2-C.sub.6"
includes alkenyl groups containing two to six carbon atoms. The
term "C.sub.3-C.sub.6" includes alkenyl groups containing three to
six carbon atoms.
[0303] The term "optionally substituted alkenyl" refers to
unsubstituted alkenyl or alkenyl having designated substituents
replacing one or more hydrogen atoms on one or more hydrocarbon
backbone carbon atoms. Such substituents can include, for example,
alkyl, alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,
alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato,
amino (including alkylamino, dialkylamino, arylamino, diarylamino
and alkylarylamino), acylamino (including alkylcarbonylamino,
arylcarbonylamino, carbamoyl and ureido), amidino, imino,
sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,
alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, heterocyclyl, alkylaryl, or an aromatic or
heteroaromatic moiety.
[0304] "Alkynyl" includes unsaturated aliphatic groups analogous in
length and possible substitution to the alkyls described above, but
which contain at least one triple bond. For example, "alkynyl"
includes straight chain alkynyl groups (e.g., ethynyl, propynyl,
butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl),
and branched alkynyl groups. In certain embodiments, a straight
chain or branched alkynyl group has six or fewer carbon atoms in
its backbone (e.g., C.sub.2-C.sub.6 for straight chain,
C.sub.3-C.sub.6 for branched chain). The term "C.sub.2-C.sub.6"
includes alkynyl groups containing two to six carbon atoms. The
term "C.sub.3-C.sub.6" includes alkynyl groups containing three to
six carbon atoms.
[0305] The term "optionally substituted alkynyl" refers to
unsubstituted alkynyl or alkynyl having designated substituents
replacing one or more hydrogen atoms on one or more hydrocarbon
backbone carbon atoms. Such substituents can include, for example,
alkyl, alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,
alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato,
amino (including alkylamino, dialkylamino, arylamino, diarylamino
and alkylarylamino), acylamino (including alkylcarbonylamino,
arylcarbonylamino, carbamoyl and ureido), amidino, imino,
sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,
alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an
aromatic or heteroaromatic moiety.
[0306] Other optionally substituted moieties (such as optionally
substituted cycloalkyl, heterocycloalkyl, aryl, or heteroaryl)
include both the unsubstituted moieties and the moieties having one
or more of the designated substituents.
[0307] "Aryl" includes groups with aromaticity, including
"conjugated," or multicyclic systems with at least one aromatic
ring and do not contain any heteroatom in the ring structure.
Examples include phenyl, benzyl, 1,2,3,4-tetrahydronaphthalenyl,
etc.
[0308] "Heteroaryl" groups are aryl groups, as defined above,
except having from one to four heteroatoms in the ring structure,
and may also be referred to as "aryl heterocycles" or
"heteroaromatics." As used herein, the term "heteroaryl" is
intended to include a stable 5- or 6-membered monocyclic or 7-, 8-,
9-, 10-, 11- or 12-membered bicyclic aromatic heterocyclic ring
which consists of carbon atoms and one or more heteroatoms, e.g., 1
or 1-2 or 1-3 or 1-4 or 1-5 or 1-6 heteroatoms, or e.g., 1, 2, 3,
4, 5, or 6 heteroatoms, independently selected from the group
consisting of nitrogen, oxygen and sulfur. The nitrogen atom may be
substituted or unsubstituted (i.e., N or NR wherein R is H or other
substituents, as defined). The nitrogen and sulfur heteroatoms may
optionally be oxidized (i.e., N.fwdarw.O and S(O).sub.p, where p=1
or 2). It is to be noted that total number of S and O atoms in the
aromatic heterocycle is not more than 1.
[0309] Examples of heteroaryl groups include pyrrole, furan,
thiophene, thiazole, isothiazole, imidazole, triazole, tetrazole,
pyrazole, oxazole, isoxazole, pyridine, pyrazine, pyridazine,
pyrimidine, and the like.
[0310] Furthermore, the terms "aryl" and "heteroaryl" include
multicyclic aryl and heteroaryl groups, e.g., tricyclic, bicyclic,
e.g., naphthalene, benzoxazole, benzodioxazole, benzothiazole,
benzoimidazole, benzothiophene, methylenedioxyphenyl, quinoline,
isoquinoline, naphthrydine, indole, benzofuran, purine, benzofuran,
deazapurine, indolizine.
[0311] In the case of multicyclic aromatic rings, only one of the
rings needs to be aromatic (e.g., 2,3-dihydroindole), although all
of the rings may be aromatic (e.g., quinoline). The second ring can
also be fused or bridged.
[0312] The aryl or heteroaryl aromatic ring can be substituted at
one or more ring positions with such substituents as described
above, for example, alkyl, alkenyl, alkynyl, halogen, hydroxyl,
alkoxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkylaminocarbonyl,
aralkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl,
arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylthiocarbonyl, phosphate, phosphonato,
phosphinato, amino (including alkylamino, dialkylamino, arylamino,
diarylamino and alkylarylamino), acylamino (including
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),
amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,
sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an
aromatic or heteroaromatic moiety. Aryl groups can also be fused or
bridged with alicyclic or heterocyclic rings, which are not
aromatic so as to form a multicyclic system (e.g., tetralin,
methylenedioxyphenyl).
[0313] As used herein, "carbocycle" or "carbocyclic ring" is
intended to include any stable monocyclic, bicyclic or tricyclic
ring having the specified number of carbons, any of which may be
saturated, unsaturated, or aromatic. For example, a
C.sub.3-C.sub.14 carbocycle is intended to include a monocyclic,
bicyclic or tricyclic ring having 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13 or 14 carbon atoms. Examples of carbocycles include, but are not
limited to, cyclopropyl, cyclobutyl, cyclobutenyl, cyclopentyl,
cyclopentenyl, cyclohexyl, cycloheptenyl, cycloheptyl,
cycloheptenyl, adamantyl, cyclooctyl, cyclooctenyl,
cyclooctadienyl, fluorenyl, phenyl, naphthyl, indanyl, adamantyl
and tetrahydronaphthyl. Bridged rings are also included in the
definition of carbocycle, including, for example,
[3.3.0]bicyclooctane, [4.3.0]bicyclononane, [4.4.0]bicyclodecane
and [2.2.2]bicyclooctane. A bridged ring occurs when one or more
carbon atoms link two non-adjacent carbon atoms. In one embodiment,
bridge rings are one or two carbon atoms. It is noted that a bridge
always converts a monocyclic ring into a tricyclic ring. When a
ring is bridged, the substituents recited for the ring may also be
present on the bridge. Fused (e.g., naphthyl, tetrahydronaphthyl)
and spiro rings are also included.
[0314] As used herein, "heterocycle" includes any ring structure
(saturated or partially unsaturated) which contains at least one
ring heteroatom (e.g., N, O or S). Examples of heterocycles
include, but are not limited to, morpholine, pyrrolidine,
tetrahydrothiophene, piperidine, piperazine and
tetrahydrofuran.
[0315] Examples of heterocyclic groups include, but are not limited
to, acridinyl, azocinyl, benzimidazolyl, benzofuranyl,
benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl,
benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl,
benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl,
carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl,
2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran,
furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl,
1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl,
3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl,
isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl,
methylenedioxyphenyl, morpholinyl, naphthyridinyl,
octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl,
1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl,
1,2,4-oxadiazol5(4H)-one, oxazolidinyl, oxazolyl, oxindolyl,
pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl,
phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl,
piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl,
pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl,
pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole,
pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl,
pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl,
quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,
tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,
tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl,
1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl,
thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl,
thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl,
1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and
xanthenyl.
[0316] The term "substituted," as used herein, means that any one
or more hydrogen atoms on the designated atom is replaced with a
selection from the indicated groups, provided that the designated
atom's normal valency is not exceeded, and that the substitution
results in a stable compound. When a substituent is oxo or keto
(i.e., .dbd.O), then 2 hydrogen atoms on the atom are replaced.
Keto substituents are not present on aromatic moieties. Ring double
bonds, as used herein, are double bonds that are formed between two
adjacent ring atoms (e.g., C.dbd.C, C.dbd.N or N.beta.N). "Stable
compound" and "stable structure" are meant to indicate a compound
that is sufficiently robust to survive isolation to a useful degree
of purity from a reaction mixture, and formulation into an
efficacious therapeutic agent.
[0317] When a bond to a substituent is shown to cross a bond
connecting two atoms in a ring, then such substituent may be bonded
to any atom in the ring. When a substituent is listed without
indicating the atom via which such substituent is bonded to the
rest of the compound of a given formula, then such substituent may
be bonded via any atom in such formula. Combinations of
substituents and/or variables are permissible, but only if such
combinations result in stable compounds.
[0318] When any variable (e.g., R.sub.1) occurs more than one time
in any constituent or formula for a compound, its definition at
each occurrence is independent of its definition at every other
occurrence. Thus, for example, if a group is shown to be
substituted with 0-2 R.sub.1 moieties, then the group may
optionally be substituted with up to two R.sub.1 moieties and
R.sub.1 at each occurrence is selected independently from the
definition of R.sub.1. Also, combinations of substituents and/or
variables are permissible, but only if such combinations result in
stable compounds.
[0319] The term "hydroxy" or "hydroxyl" includes groups with an
--OH or --O.sup.-.
[0320] As used herein, "halo" or "halogen" refers to fluoro,
chloro, bromo and iodo. The term "perhalogenated" generally refers
to a moiety wherein all hydrogen atoms are replaced by halogen
atoms. The term "haloalkyl" or "haloalkoxyl" refers to an alkyl or
alkoxyl substituted with one or more halogen atoms.
[0321] The term "carbonyl" includes compounds and moieties which
contain a carbon connected with a double bond to an oxygen atom.
Examples of moieties containing a carbonyl include, but are not
limited to, aldehydes, ketones, carboxylic acids, amides, esters,
anhydrides, etc.
[0322] The term "carboxyl" refers to --COOH or its C.sub.1-C.sub.6
alkyl ester.
[0323] "Acyl" includes moieties that contain the acyl radical
(R--C(O)--) or a carbonyl group. "Substituted acyl" includes acyl
groups where one or more of the hydrogen atoms are replaced by, for
example, alkyl groups, alkynyl groups, halogen, hydroxyl,
alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, amino (including alkylamino,
dialkylamino, arylamino, diarylamino and alkylarylamino), acylamino
(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and
ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,
thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,
sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,
alkylaryl, or an aromatic or heteroaromatic moiety.
[0324] "Aroyl" includes moieties with an aryl or heteroaromatic
moiety bound to a carbonyl group. Examples of aroyl groups include
phenylcarboxy, naphthyl carboxy, etc.
[0325] "Alkoxyalkyl," "alkylaminoalkyl," and "thioalkoxyalkyl"
include alkyl groups, as described above, wherein oxygen, nitrogen,
or sulfur atoms replace one or more hydrocarbon backbone carbon
atoms.
[0326] The term "alkoxy" or "alkoxyl" includes substituted and
unsubstituted alkyl, alkenyl and alkynyl groups covalently linked
to an oxygen atom. Examples of alkoxy groups or alkoxyl radicals
include, but are not limited to, methoxy, ethoxy, isopropyloxy,
propoxy, butoxy and pentoxy groups. Examples of substituted alkoxy
groups include halogenated alkoxy groups. The alkoxy groups can be
substituted with groups such as alkenyl, alkynyl, halogen,
hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, amino (including alkylamino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moieties.
Examples of halogen substituted alkoxy groups include, but are not
limited to, fluoromethoxy, difluoromethoxy, trifluoromethoxy,
chloromethoxy, dichloromethoxy and trichloromethoxy.
[0327] The term "ether" or "alkoxy" includes compounds or moieties
which contain an oxygen bonded to two carbon atoms or heteroatoms.
For example, the term includes "alkoxyalkyl," which refers to an
alkyl, alkenyl, or alkynyl group covalently bonded to an oxygen
atom which is covalently bonded to an alkyl group.
[0328] The term "ester" includes compounds or moieties which
contain a carbon or a heteroatom bound to an oxygen atom which is
bonded to the carbon of a carbonyl group. The term "ester" includes
alkoxycarboxy groups such as methoxycarbonyl, ethoxycarbonyl,
propoxycarbonyl, butoxycarbonyl, pentoxycarbonyl, etc.
[0329] The term "thioalkyl" includes compounds or moieties which
contain an alkyl group connected with a sulfur atom. The thioalkyl
groups can be substituted with groups such as alkyl, alkenyl,
alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, carboxyacid,
alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,
alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl,
alkoxyl, amino (including alkylamino, dialkylamino, arylamino,
diarylamino and alkylarylamino), acylamino (including
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),
amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,
sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an
aromatic or heteroaromatic moieties.
[0330] The term "thiocarbonyl" or "thiocarboxy" includes compounds
and moieties which contain a carbon connected with a double bond to
a sulfur atom.
[0331] The term "thioether" includes moieties which contain a
sulfur atom bonded to two carbon atoms or heteroatoms. Examples of
thioethers include, but are not limited to alkthioalkyls,
alkthioalkenyls, and alkthioalkynyls. The term "alkthioalkyls"
include moieties with an alkyl, alkenyl, or alkynyl group bonded to
a sulfur atom which is bonded to an alkyl group. Similarly, the
term "alkthioalkenyls" refers to moieties wherein an alkyl, alkenyl
or alkynyl group is bonded to a sulfur atom which is covalently
bonded to an alkenyl group; and alkthioalkynyls" refers to moieties
wherein an alkyl, alkenyl or alkynyl group is bonded to a sulfur
atom which is covalently bonded to an alkynyl group.
[0332] As used herein, "amine" or "amino" refers to unsubstituted
or substituted --NH.sub.2. "Alkylamino" includes groups of
compounds wherein nitrogen of --NH.sub.2 is bound to at least one
alkyl group. Examples of alkylamino groups include benzylamino,
methylamino, ethylamino, phenethylamino, etc. "Dialkylamino"
includes groups wherein the nitrogen of --NH.sub.2 is bound to at
least two additional alkyl groups. Examples of dialkylamino groups
include, but are not limited to, dimethylamino and diethylamino.
"Arylamino" and "diarylamino" include groups wherein the nitrogen
is bound to at least one or two aryl groups, respectively.
"Aminoaryl" and "aminoaryloxy" refer to aryl and aryloxy
substituted with amino. "Alkylarylamino," "alkylaminoaryl" or
"arylaminoalkyl" refers to an amino group which is bound to at
least one alkyl group and at least one aryl group. "Alkaminoalkyl"
refers to an alkyl, alkenyl, or alkynyl group bound to a nitrogen
atom which is also bound to an alkyl group. "Acylamino" includes
groups wherein nitrogen is bound to an acyl group. Examples of
acylamino include, but are not limited to, alkylcarbonylamino,
arylcarbonylamino, carbamoyl and ureido groups.
[0333] The term "amide" or "aminocarboxy" includes compounds or
moieties that contain a nitrogen atom that is bound to the carbon
of a carbonyl or a thiocarbonyl group. The term includes
"alkaminocarboxy" groups that include alkyl, alkenyl or alkynyl
groups bound to an amino group which is bound to the carbon of a
carbonyl or thiocarbonyl group. It also includes "arylaminocarboxy"
groups that include aryl or heteroaryl moieties bound to an amino
group that is bound to the carbon of a carbonyl or thiocarbonyl
group. The terms "alkylaminocarboxy", "alkenylaminocarboxy",
"alkynylaminocarboxy" and "arylaminocarboxy" include moieties
wherein alkyl, alkenyl, alkynyl and aryl moieties, respectively,
are bound to a nitrogen atom which is in turn bound to the carbon
of a carbonyl group. Amides can be substituted with substituents
such as straight chain alkyl, branched alkyl, cycloalkyl, aryl,
heteroaryl or heterocycle. Substituents on amide groups may be
further substituted.
[0334] Compounds of the present invention that contain nitrogens
can be converted to N-oxides by treatment with an oxidizing agent
(e.g., 3-chloroperoxybenzoic acid (mCPBA) and/or hydrogen
peroxides) to afford other compounds of the present invention.
Thus, all shown and claimed nitrogen-containing compounds are
considered, when allowed by valency and structure, to include both
the compound as shown and its N-oxide derivative (which can be
designated as N.fwdarw.O or N+-O.sup.-). Furthermore, in other
instances, the nitrogens in the compounds of the present invention
can be converted to N-hydroxy or N-alkoxy compounds. For example,
N-hydroxy compounds can be prepared by oxidation of the parent
amine by an oxidizing agent such as m-CPBA. All shown and claimed
nitrogen-containing compounds are also considered, when allowed by
valency and structure, to cover both the compound as shown and its
N-hydroxy (i.e., N--OH) and N-alkoxy (i.e., N--OR, wherein R is
substituted or unsubstituted C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkenyl, C.sub.1-C.sub.6 alkynyl, 3-14-membered carbocycle or
3-14-membered heterocycle) derivatives.
[0335] In the present specification, the structural formula of the
compound represents a certain isomer for convenience in some cases,
but the present invention includes all isomers, such as geometrical
isomers, optical isomers based on an asymmetrical carbon,
stereoisomers, tautomers, and the like. In addition, a crystal
polymorphism may be present for the compounds represented by the
formula. It is noted that any crystal form, crystal form mixture,
or anhydride or hydrate thereof is included in the scope of the
present invention. Furthermore, so-called metabolite which is
produced by degradation of the present compound in vivo is included
in the scope of the present invention.
[0336] "Isomerism" means compounds that have identical molecular
formulae but differ in the sequence of bonding of their atoms or in
the arrangement of their atoms in space. Isomers that differ in the
arrangement of their atoms in space are termed "stereoisomers."
Stereoisomers that are not mirror images of one another are termed
"diastereoisomers," and stereoisomers that are non-superimposable
mirror images of each other are termed "enantiomers" or sometimes
optical isomers. A mixture containing equal amounts of individual
enantiomeric forms of opposite chirality is termed a "racemic
mixture."
[0337] A carbon atom bonded to four nonidentical substituents is
termed a "chiral center."
[0338] "Chiral isomer" means a compound with at least one chiral
center. Compounds with more than one chiral center may exist either
as an individual diastereomer or as a mixture of diastereomers,
termed "diastereomeric mixture." When one chiral center is present,
a stereoisomer may be characterized by the absolute configuration
(R or S) of that chiral center. Absolute configuration refers to
the arrangement in space of the substituents attached to the chiral
center. The substituents attached to the chiral center under
consideration are ranked in accordance with the Sequence Rule of
Cahn, Ingold and Prelog. (Cahn et al., Angew. Chem. Inter. Edit.
1966, 5, 385; errata 511; Cahn et al., Angew. Chem. 1966, 78, 413;
Cahn and Ingold, J. Chem. Soc. 1951 (London), 612; Cahn et al.,
Experientia 1956, 12, 81; Cahn, J. Chem. Educ. 1964, 41, 116).
[0339] "Geometric isomer" means the diastereomers that owe their
existence to hindered rotation about double bonds or a cycloalkyl
linker (e.g., 1,3-cylcobutyl). These configurations are
differentiated in their names by the prefixes cis and trans, or Z
and E, which indicate that the groups are on the same or opposite
side of the double bond in the molecule according to the
Cahn-Ingold-Prelog rules.
[0340] It is to be understood that the compounds of the present
invention may be depicted as different chiral isomers or geometric
isomers. It should also be understood that when compounds have
chiral isomeric or geometric isomeric forms, all isomeric forms are
intended to be included in the scope of the present invention, and
the naming of the compounds does not exclude any isomeric
forms.
[0341] For example, compounds of Formula (I) includes those of the
following chiral isomers and geometric isomers.
##STR00183##
[0342] Furthermore, the structures and other compounds discussed in
this invention include all atropic isomers thereof. "Atropic
isomers" are a type of stereoisomer in which the atoms of two
isomers are arranged differently in space. Atropic isomers owe
their existence to a restricted rotation caused by hindrance of
rotation of large groups about a central bond. Such atropic isomers
typically exist as a mixture, however as a result of recent
advances in chromatography techniques, it has been possible to
separate mixtures of two atropic isomers in select cases.
[0343] "Tautomer" is one of two or more structural isomers that
exist in equilibrium and is readily converted from one isomeric
form to another. This conversion results in the formal migration of
a hydrogen atom accompanied by a switch of adjacent conjugated
double bonds. Tautomers exist as a mixture of a tautomeric set in
solution. In solutions where tautomerization is possible, a
chemical equilibrium of the tautomers will be reached. The exact
ratio of the tautomers depends on several factors, including
temperature, solvent and pH. The concept of tautomers that are
interconvertable by tautomerizations is called tautomerism.
[0344] Of the various types of tautomerism that are possible, two
are commonly observed. In keto-enol tautomerism a simultaneous
shift of electrons and a hydrogen atom occurs. Ring-chain
tautomerism arises as a result of the aldehyde group (--CHO) in a
sugar chain molecule reacting with one of the hydroxy groups (--OH)
in the same molecule to give it a cyclic (ring-shaped) form as
exhibited by glucose.
[0345] Common tautomeric pairs are: ketone-enol, amide-nitrile,
lactam-lactim, amide-imidic acid tautomerism in heterocyclic rings
(e.g., in nucleobases such as guanine, thymine and cytosine),
amine-enamine and enamine-enamine. Benzimidazoles also exhibit
tautomerism, when the benzimidazole contains one or more
substituents in the 4, 5, 6 or 7 positions, the possibility of
different isomers arises. For example,
2,5-dimethyl-1H-benzo[d]imidazole can exist in equilibrium with its
isomer 2,6-dimethyl-1H-benzo[d]imidazole via tautomerization.
##STR00184##
[0346] Another example of tautomerism is shown below.
##STR00185##
[0347] 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 the compounds does not exclude
any tautomer form.
[0348] The term "crystal polymorphs", "polymorphs" or "crystal
forms" means crystal structures in which a compound (or a salt or
solvate thereof) can crystallize in different crystal packing
arrangements, all of which have the same elemental composition.
Different crystal forms usually have different X-ray diffraction
patterns, infrared spectral, melting points, density hardness,
crystal shape, optical and electrical properties, stability and
solubility. Recrystallization solvent, rate of crystallization,
storage temperature, and other factors may cause one crystal form
to dominate. Crystal polymorphs of the compounds can be prepared by
crystallization under different conditions.
[0349] Compounds of the invention may be crystalline,
semi-crystalline, non-crystalline, amorphous, mesomorphous,
etc.
[0350] The compounds of Formula (I), (II), (IIIa), (IIIb), (IIIc)
or (IV) include the compounds themselves, as well as their
N-oxides, salts, their solvates, and their prodrugs, if applicable.
A salt, for example, can be formed between an anion and a
positively charged group (e.g., amino) on a substituted purine or
7-deazapurine compound. Suitable anions include chloride, bromide,
iodide, sulfate, bisulfate, sulfamate, nitrate, phosphate, citrate,
methanesulfonate, trifluoroacetate, glutamate, glucuronate,
glutarate, malate, maleate, succinate, fumarate, tartrate,
tosylate, salicylate, lactate, naphthalenesulfonate, and acetate.
Likewise, a salt can also be formed between a cation and a
negatively charged group (e.g., carboxylate) on a substituted
purine or 7-deazapurine compound. Suitable cations include sodium
ion, potassium ion, magnesium ion, calcium ion, and an ammonium
cation such as tetramethylammonium ion. The substituted purine or
7-deazapurine compounds also include those salts containing
quaternary nitrogen atoms. Examples of prodrugs include esters and
other pharmaceutically acceptable derivatives, which, upon
administration to a subject, are capable of providing active
substituted purine or 7-deazapurine compounds.
[0351] Additionally, the compounds of the present invention, for
example, the salts of the compounds, can exist in either hydrated
or unhydrated (the anhydrous) form or as solvates with other
solvent molecules. Nonlimiting examples of hydrates include
hemihydrates, monohydrates, dihydrates, trihydrates, etc.
Nonlimiting examples of solvates include ethanol solvates, acetone
solvates, etc.
[0352] "Solvate" means solvent addition forms that contain either
stoichiometric or non stoichiometric amounts of solvent. Some
compounds have a tendency to trap a fixed molar ratio of solvent
molecules in the crystalline solid state, thus forming a solvate.
If the solvent is water the solvate formed is a hydrate; and if the
solvent is alcohol, the solvate formed is an alcoholate. Hydrates
are formed by the combination of one or more molecules of water
with one molecule of the substance in which the water retains its
molecular state as H.sub.2O. A hemihydrate is formed by the
combination of one molecule of water with more than one molecule of
the substance in which the water retains its molecular state as
H.sub.2O.
[0353] As used herein, the term "analog" refers to a chemical
compound that is structurally similar to another but differs
slightly in composition (as in the replacement of one atom by an
atom of a different element or in the presence of a particular
functional group, or the replacement of one functional group by
another functional group). Thus, an analog is a compound that is
similar or comparable in function and appearance, but not in
structure or origin to the reference compound.
[0354] As defined herein, the term "derivative" refers to compounds
that have a common core structure, and are substituted with various
groups as described herein. For example, all of the compounds
represented by Formula (I) are substituted purine compounds or
substituted 7-deazapurine compounds, and have Formula (I) as a
common core.
[0355] The term "bioisostere" refers to a compound resulting from
the exchange of an atom or of a group of atoms with another,
broadly similar, atom or group of atoms. The objective of a
bioisosteric replacement is to create a new compound with similar
biological properties to the parent compound. The bioisosteric
replacement may be physicochemically or topologically based.
Examples of carboxylic acid bioisosteres include, but are not
limited to, acyl sulfonimides, tetrazoles, sulfonates and
phosphonates. See, e.g., Patani and LaVoie, Chem. Rev. 96,
3147-3176, 1996.
[0356] The present invention is intended to include all isotopes of
atoms occurring in the present compounds. Isotopes include those
atoms having the same atomic number but different mass numbers. By
way of general example and without limitation, isotopes of hydrogen
include tritium and deuterium, and isotopes of carbon include C-13
and C-14.
2. SYNTHESIS OF SUBSTITUTED PURINE COMPOUNDS AND SUBSTITUTED
7-DEAZAPURINE COMPOUNDS
[0357] The present invention provides methods for the synthesis of
the compounds of Formulae (I), (II), (IIIa), (IIIb), (IIIc) and
(IV). The present invention also provides detailed methods for the
synthesis of various disclosed compounds of the present invention
according to the following schemes as shown in the Examples.
[0358] Throughout the description, where compositions are described
as having, including, or comprising specific components, it is
contemplated that compositions also consist essentially of, or
consist of, the recited components. Similarly, where methods or
processes are described as having, including, or comprising
specific process steps, the processes also consist essentially of,
or consist of, the recited processing steps. Further, it should be
understood that the order of steps or order for performing certain
actions is immaterial so long as the invention remains operable.
Moreover, two or more steps or actions can be conducted
simultaneously.
[0359] The synthetic processes of the invention can tolerate a wide
variety of functional groups, therefore various substituted
starting materials can be used. The processes generally provide the
desired final compound at or near the end of the overall process,
although it may be desirable in certain instances to further
convert the compound to a pharmaceutically acceptable salt, ester,
or prodrug thereof.
[0360] Compounds of the present invention can be prepared in a
variety of ways using commercially available starting materials,
compounds known in the literature, or from readily prepared
intermediates, by employing standard synthetic methods and
procedures either known to those skilled in the art, or which will
be apparent to the skilled artisan in light of the teachings
herein. Standard synthetic methods and procedures for the
preparation of organic molecules and functional group
transformations and manipulations can be obtained from the relevant
scientific literature or from standard textbooks in the field.
Although not limited to any one or several sources, classic texts
such as Smith, M. B., March, J., March's Advanced Organic
Chemistry: Reactions, Mechanisms, and Structure, 5.sup.th edition,
John Wiley & Sons: New York, 2001; Greene, T. W., Wuts, P. G.
M., Protective Groups in Organic Synthesis, 3.sup.rd edition, John
Wiley & Sons: New York, 1999; R. Larock, Comprehensive Organic
Transformations, VCH Publishers (1989); L. Fieser and M. Fieser,
Fieser and Fieser's Reagentsfor Organic Synthesis, John Wiley and
Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for
Organic Synthesis, John Wiley and Sons (1995), incorporated by
reference herein, are useful and recognized reference textbooks of
organic synthesis known to those in the art. The following
descriptions of synthetic methods are designed to illustrate, but
not to limit, general procedures for the preparation of compounds
of the present invention.
[0361] Compounds of the present invention can be conveniently
prepared by a variety of methods familiar to those skilled in the
art. The compounds of this invention with Formulae (I), (II),
(IIIa), (IIIb), (IIIc) and (IV) may be prepared according to the
procedures illustrated in Schemes A-W below, from commercially
available starting materials or starting materials which can be
prepared using literature procedures. The R groups (such as R, R',
and R.sub.a) in Schemes A-P may correspond to variables (i.e.,
R.sub.1, R.sub.2, R.sub.b, and R.sub.c) as defined in Formula (I),
(II), (IIIa), (IIIb), (IIIc) or (IV), unless otherwise specified.
"PG" in the schemes refers to a protecting group.
[0362] One of ordinary skill in the art will note that, during the
reaction sequences and synthetic schemes described herein, the
order of certain steps may be changed, such as the introduction and
removal of protecting groups.
[0363] One of ordinary skill in the art will recognize that certain
groups may require protection from the reaction conditions via the
use of protecting groups. Protecting groups may also be used to
differentiate similar functional groups in molecules. A list of
protecting groups and how to introduce and remove these groups can
be found in Greene, T. W., Wuts, P. G. M., Protective Groups in
Organic Synthesis, 3.sup.rd edition, John Wiley & Sons: New
York, 1999.
[0364] Preferred protecting groups include, but are not limited
to:
[0365] For the hydroxyl moiety: TBS, benzyl, THP, Ac
[0366] For carboxylic acids: benzyl ester, methyl ester, ethyl
ester, allyl ester
[0367] For amines: Cbz, BOC, DMB
[0368] For diols: Ac (.times.2) TBS (.times.2), or when taken
together acetonides
[0369] For thiols: Ac
[0370] For benzimidazoles: SEM, benzyl, PMB, DMB
[0371] For aldehydes: di-alkyl acetals such as dimethoxy acetal or
diethyl acetyl.
[0372] In the reaction schemes described herein, multiple
stereoisomers may be produced. When no particular stereoisomer is
indicated, it is understood to mean all possible stereoisomers that
could be produced from the reaction. A person of ordinary skill in
the art will recognize that the reactions can be optimized to give
one isomer preferentially, or new schemes may be devised to produce
a single isomer. If mixtures are produced, techniques such as
preparative thin layer chromatography, preparative HPLC,
preparative chiral HPLC, or preparative SFC may be used to separate
the isomers.
[0373] The following abbreviations are used throughout the
specification and are defined below:
[0374] AA ammonium acetate
[0375] Ac acetyl
[0376] ACN acetonitrile
[0377] AcOH acetic acid
[0378] atm atmosphere
[0379] Bn benzyl
[0380] BOC tert-butoxy carbonyl
[0381] BOP (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium
hexafluorophosphate
[0382] Cbz benzyloxycarbonyl
[0383] COMU
(1-cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbeni-
um hexafluorophosphate
[0384] d days
[0385] DBU 1,8-diazabicyclo[5.4.0]undec-7-ene
[0386] DCE 1,2 dichloroethane
[0387] DCM dichloromethane
[0388] DEA diethylamine
[0389] DEAD diethyl azodicarboxylate
[0390] DIAD diisopropyl azodicarboxylate
[0391] DiBAL-H diisobutylalumininium hydride
[0392] DIPEA N,N-diisopropylethylamine (Hunig's base)
[0393] DMAP N,N-dimethyl-4-aminopyridine
[0394] DMB 2,4 dimethoxybenzyl
[0395] DMF dimethylformamide
[0396] DMSO dimethylsulfoxide
[0397] DPPA diphenylphosphoryl azide
[0398] EA or EtOAc ethylacetate
[0399] EDC or EDCl
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide
[0400] ELS Evaporative Light Scattering
[0401] ESI- Electrospray negative mode
[0402] ESI+ Electrospray positive mode
[0403] Et.sub.2O diethyl ether
[0404] Et.sub.3N or TEA triethylamine
[0405] EtOH ethanol
[0406] FA formic acid
[0407] FC flash chromatography
[0408] h hours
[0409] H.sub.2O water
[0410] HATU O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate
[0411] HCl hydrochloric acid
[0412] HOAT 1-hydroxy-7-azabenzotriazole
[0413] HOBt 1-hydroxybenzotriazole
[0414] HOSu N-hydroxysuccinimide
[0415] HPLC high performance liquid chromatography
[0416] Inj. Vol. injection volume
[0417] I.V. or IV intravenous
[0418] KHMDs potassium hexamethyldisilazide
[0419] LC/MS or LC-MS liquid chromatography mass spectrum
[0420] LDA lithium diisopropylamide
[0421] LG leaving group
[0422] LiHMs lithium hexamethyldisilazide
[0423] M Molar
[0424] m/z mass/charge ratio
[0425] m-CPBA meta-chloroperbenzoic acid
[0426] MeCN acetonitrile
[0427] MeOD d.sub.4-methanol
[0428] MeOH methanol
[0429] MgSO.sub.4 magnesium sulfate
[0430] min minutes
[0431] MS mass spectrometry or mass spectrum
[0432] Ms mesyl
[0433] MsCl methanesulfonyl chloride
[0434] MsO mesylate
[0435] MWI microwave irradiation
[0436] Na.sub.2CO.sub.3 sodium carbonate
[0437] NaHCO.sub.3 sodium bicarbonate
[0438] NaHMDs sodium hexamethyldisilazide
[0439] NaOH sodium hydroxide
[0440] NIS N-iodosuccinimide
[0441] NMR Nuclear Magnetic Resonance
[0442] o/n or O/N overnight
[0443] PE petroleum ether
[0444] PG protecting group
[0445] PMB para-methoxybenzyl
[0446] PPAA 1-propanephosphonic acid cyclic anhydride
[0447] ppm parts per million
[0448] prep HPLC preparative high performance liquid
chromatography
[0449] prep TLC preparative thin layer chromatography
[0450] p-TsOH para-toluenesulfonic acid
[0451] rt or RT room temperature
[0452] SEM 2-(trimethylsilyl)ethoxymethyl
[0453] SEMCI (trimethylsilyl)ethoxymethyl chloride
[0454] SFC supercritical chromatography
[0455] SGC silica gel chromatography
[0456] STAB sodium triacetoxyborohydride
[0457] TBAF tetra-n-butylammonium fluoride
[0458] TFA trifluoroacetic acid
[0459] TfO triflate
[0460] THE tetrahydrofuran
[0461] THP tetrahydropyran
[0462] TLC thin layer chromatography
[0463] Ts tosyl
[0464] TsOH tosic acid
[0465] UV ultraviolet
[0466] The invention provides methods for making the compounds of
the invention. The following schemes depict exemplary chemistries
available for synthesizing the compounds of the invention.
##STR00186##
[0467] 5'-Amino purine-ribose intermediates (A-V) can be
synthesized as depicted in Scheme A above. A suitable protected
6-C.sub.1 adenosine derivative (A-I) is converted into a 6-amino
derivative (A-II) by treatment with the appropriate amine
(including ammonia) in the presence of a base such as Et.sub.3N,
K.sub.2CO.sub.3 or Hunig's base in solvent such as MeCN or DMF,
THF, iPrOH or a mixture thereof. If required, the reaction may be
heated to 100.degree. C. (if the temperature required is greater
than the boiling point of one or more of the components in the
mixture, the reaction may be performed in a sealed tube). The R
groups in the scheme may represent alkyl protecting groups (e.g.,
2, 4 dimethoxybenzyl). The 6-amino product (A-II) may be
transformed into the 5'-azido intermediate (A-III) by converting
the 5'-hydroxyl group into a leaving group such as MsO (i.e.,
CH.sub.3S(O).sub.2O) by treatment with methanesulfonyl chloride
(MsCl) in the presence of a base such as Et.sub.3N, pyridine or
K.sub.2CO.sub.3 in an inert solvent such as CH.sub.2Cl.sub.2, THF,
MeCN, DMF or a mixture thereof. The 5'-leaving group is then
displaced with azide anion from NaN.sub.3 in an inert solvent such
as DMF. Alternatively (A-II) may be directly transformed into
(A-III) by treatment with DPPA, Ph.sub.3P, and DIAD in a solvent
such as THF. The azido group of (A-III) may be reduced to the
primary amine (A-IV) by reduction with H.sub.2 in the presence of a
metal catalyst (e.g. Pd/C, PtO.sub.2) or by a Staudinger reaction
with a phosphine such as Ph.sub.3P or PMe.sub.3. The primary amine
(A-IV) may be converted into the secondary amine (A-V) by treatment
with the appropriate ketone or aldehyde in the presence of a
suitable reducing agent such as NaBH(OAc).sub.3 or NaCNBH.sub.3.
Additional reagents such as Ti(OiPr)4 may be added.
[0468] Alternatively the 5'-hydroxy intermediate (A-II) may be
treated with the sulfonamide (A-VI), DEAD and Ph.sub.3P in an inert
solvent such as THF. The resultant sulfonamide product may then be
treated with benzenethiol in the presence of a base such as
K.sub.2CO.sub.3, Cs.sub.2CO.sub.3 to give the secondary amine
(A-V).
[0469] These reaction sequences above may also be applied to lyxose
derivatives starting from (A-VII) to give the diastereomer with
opposite configuration at the 5' position.
##STR00187##
[0470] A similar set of reaction sequences may be employed for
2'-deoxy, or 3'-deoxy, or substituted ribose or lyxose (A-VIII)
above to obtain 5'-amino purine-ribose/lyxose intermediates.
[0471] An alternative method for introduction of a 6-NH.sub.2
group, as shown below, is via treating (A-IX) derivatives with
NaN.sub.3 to produce a 6-azido intermediate followed by reduction
to the NH.sub.2 moiety A-X with a trialkyl phosphine such as
PMe.sub.3 or PPh.sub.3.
##STR00188##
##STR00189##
[0472] 5'-Amino-7-deazapurine-ribose intermediates (B-V) can be
synthesized as depicted in Scheme B above. A suitable protected
7-deazapurine-ribose intermediate containing a 6-chloro substituent
(B-I) may be converted into the corresponding 6-amino derivative
(B-II) via treatment with the appropriate amine (including ammonia)
in the presence of a base such as Et.sub.3N, K.sub.2CO.sub.3 or
Hunig's base in solvent such as MeCN or DMF, THF, iPrOH or a
mixture thereof. If required, the reaction may be heated to
100.degree. C. (if the temperature required is greater than the
boiling point of one or more of the components in the mixture, the
reaction may be performed in a sealed tube). The R groups in the
scheme may represent alkyl protecting groups (e.g. 2, 4
dimethoxybenzyl). The 6-amino product (B-II) may be transformed
into the 5'-azido intermediate (B-III) by converting the
5'-hydroxyl group into a leaving group such as MsO by treatment
with MsCl in the presence of a base such as Et.sub.3N, pyridine or
K.sub.2CO.sub.3 in an inert solvent such as CH.sub.2Cl.sub.2, THF,
MeCN, DMF or a mixture thereof. The 5'-leaving group is then
displaced with azide anion from an azide source such as NaN.sub.3
in an inert solvent such as DMF. Alternatively (B-II) may be
directly transformed into (B-III) by treatment with DPPA, Ph.sub.3P
and DIAD in a solvent such as THF. The azido group of (B-III) may
be reduced to the primary amine (B-IV) by reduction with H2 in the
presence of a metal catalyst (e.g. Pd/C, PtO.sub.2) or by a
Staudinger reaction with a phosphine such as Ph.sub.3P or
PMe.sub.3. The primary amine (B-IV) may be converted into the
secondary amine (B-V) by treatment with the appropriate ketone or
aldehyde in the presence of a suitable reducing agent such as
NaBH(OAc).sub.3 or NaCNBH.sub.3. Additional reagents such as
Ti(OiPr).sub.4 may be added. Alternatively the 5'-hydroxy
intermediate (B-II) may be treated with the sulfonamide (B-VI),
DEAD and Ph.sub.3P in an inert solvent such as THF. The resultant
sulfonamide product may then be treated with benzenethiol in the
presence of a base such as K.sub.2CO.sub.3, Cs.sub.2CO.sub.3 to
give the secondary amine (B-V). These reaction sequences may also
be applied to lyxose derivatives starting from (B-VII) to give the
diastereoisomer with opposite configuration at the 5'-position
##STR00190##
[0473] A similar set of reaction sequences may be employed for
2'-deoxy, or 3'-deoxy, or substituted ribose or lyxose (B-VIII)
above to obtain 5'-amino 7-deazapurine-ribose/lyxose
intermediates.
[0474] An alternative method for introduction of a 6-NH.sub.2
group, as shown below, is via treating (B-IX) derivatives with
NaN.sub.3 to produce a 6-azido intermediate followed by reduction
to the NH.sub.2 moiety (B-X) with a trialkyl phosphine such as
PMe.sub.3 or PPh.sub.3.
##STR00191##
##STR00192##
[0475] The 5'-amino purine carbocyclic intermediates (C-X) may be
prepared as depicted in Scheme C. The cyclopentane (C-I) is
optionally protected by methods known to those of ordinary skill in
the art to give (C-II). (C-II) is treated with the appropriate
4,6-dichloropyrimidine-5-amine in the presence of a base such as
Et.sub.3N in a protic solvent such as n-butanol. The reaction is
heated or submitted to microwave conditions to give the
intermediate (C-III). The purine intermediate (C-V) is produced by
treating (C-III) with the orthoester (C-IV) in the presence of an
acid, such as AcOH. The reaction is usually heated. The 6-amino
substituent may be introduced by treatment with the appropriate
amine (including ammonia) in the presence of a base such as
Et.sub.3N, K.sub.2CO.sub.3 or Hunig's base in solvent such as MeCN
or DMF, THF, iPrOH, or a mixture thereof. If required, the reaction
may be heated to 100.degree. C. (if the temperature required is
greater than the boiling point of one or more of the components in
the mixture, the reaction may be performed in a sealed tube). The R
groups in the scheme may represent alkyl protecting groups (e.g.,
2, 4 dimethoxybenzyl). The 6-amino product (C-VI) may be
transformed into the 5'-azido intermediate (C-VII) by converting
the 5'-hydroxyl group into a leaving group such as MsO by treatment
with MsCl in the presence of a base such as Et.sub.3N, pyridine or
K.sub.2CO.sub.3 in an inert solvent such as CH.sub.2Cl.sub.2, THF,
MeCN, DMF or a mixture thereof. The 5'-leaving group is then
displaced with azide anion from NaN.sub.3 in an inert solvent such
as DMF. Alternatively (C-VI) may be directly transformed into
(C-VII) by treatment with DPPA, Ph.sub.3P and DIAD in a solvent
such as THF. The azido group of (C-VII) may be reduced to the
primary amine (C-VIII) by reduction with H2 in the presence of a
metal catalyst (e.g. Pd/C, PtO.sub.2) or by a Staudinger reaction
with a phosphine such as Ph.sub.3P or PMe.sub.3. The primary amine
(C-VIII) may be converted into the secondary amine (C-X) by
treatment with the appropriate ketone or aldehyde in the presence
of a suitable reducing agent such as NaBH(OAc).sub.3 or
NaCNBH.sub.3. Additional reagents such as Ti(OiPr).sub.4 may be
added. Alternatively the 5'-hydroxy intermediate (C-VI) may be
treated with the sulfonamide (C-IX), DEAD and Ph.sub.3P in an inert
solvent such as THF. The resultant sulfonamide product may then be
treated with benzenethiol in the presence of a base such as
K.sub.2CO.sub.3, Cs.sub.2CO.sub.3 to give the secondary amine
(C-X).
##STR00193##
[0476] The 5'-amino 7-deazapurine carbocyclic intermediates (D-X)
may be prepared as depicted in Scheme D. The cyclopentane (D-I) is
optionally protected by methods known to those of ordinary skill in
the art to give (D-II). (D-II) is treated with the appropriate
4,6-dichloropyrimidine (D-III) in the presence of a base such as
Et.sub.3N in a protic solvent such as EtOH, n-butanol. The reaction
is heated to give the intermediate (D-IV). The intermediate (D-V)
is produced by treating (D-IV) with an acid, such as HCl or
AcOH.
[0477] The 5'-hydroxyl of (D-V) may be transformed into the
5'-azido intermediate (D-VI) by initially converting the
5'-hydroxyl group into a leaving group such as MsO by treatment
with MsCl in the presence of a base such as Et.sub.3N, pyridine or
K.sub.2CO.sub.3 in an inert solvent such as CH.sub.2Cl.sub.2, THF,
MeCN, DMF or a mixture thereof and then displacing the leaving
group with azide anion from NaN.sub.3 in an inert solvent such as
DMF. Alternatively (D-V) may be directly transformed into (D-VI) by
treatment with DPPA, Ph.sub.3P and DIAD in a solvent such as
THF.
[0478] The 6-amino substituent may be introduced by treatment of
(D-VI) with the appropriate amine (including ammonia) in the
presence of a base such as Et.sub.3N, K.sub.2CO.sub.3 or Hunig's
base in solvent such as MeCN or DMF, THF, iPrOH or a mixture
thereof. If required, the reaction may be heated to 100.degree. C.
(if the temperature required is greater than the boiling point of
one or more of the components in the mixture, the reaction may be
performed in a sealed tube). The R groups in the scheme may
represent alkyl protecting groups (e.g. 2, 4 dimethoxybenzyl). The
azido group of (D-VII) may be reduced to the primary amine (D-VIII)
by reduction with H2 in the presence of a metal catalyst (e.g.
Pd/C, PtO.sub.2) or by a Staudinger reaction with a phosphine such
as Ph.sub.3P or PMe.sub.3. The primary amine (D-VIII) may be
converted into the secondary amine (D-X) by treatment with the
appropriate ketone or aldehyde in the presence of a suitable
reducing agent such as NaBH(OAc).sub.3 or NaCNBH.sub.3. Additional
reagents such as Ti(OiPr).sub.4 may be added. Alternatively the
5'-hydroxy intermediate (D-V) may be treated with the sulfonamide
(D-IX), DEAD and Ph.sub.3P in an inert solvent such as THF. 6-Amino
group may then be introduced using conditions similar to those used
for converting (D-VI) into (D-VII). The resultant sulfonamide
product may then be treated with benzenethiol in the presence of a
base such as K.sub.2CO.sub.3, Cs.sub.2CO.sub.3 to give the
secondary amine (D-X).
[0479] A person of ordinary skill will recognize that having an
appropriately substituted dichloropyrimidine (D-III) will allow for
substitution on the 7-deazapurine moiety.
##STR00194##
[0480] To generate the appropriate intermediates of opposite
stereochemistry at 5'-position, a reaction sequence as depicted in
Scheme E above may be followed. The cyclopentane (E-I) is
optionally protected, upon which the 5'-hydroxyl is converted into
a leaving group via treatment with MsCl in the presence of
Et.sub.3N in a solvent such a CH.sub.2Cl.sub.2. The 5'-leaving
group is displaced with Me.sub.2NH, by treatment with Me.sub.2NH as
a 2.0M solution in THE in a sealed tube. The reaction is heated to
40-80.degree. C. The resultant tertiary amine is oxidized with an
oxidant such as mCPBA in a solvent such a CH.sub.2Cl.sub.2 to give
the corresponding N-oxide. The N-oxide is then subjected to heat,
50-120.degree. C. in an inert solvent such N,N-dimethylacetamide to
give the alkene (E-III). The alkene is subjected to
hydroboration/oxidative work up to produce the inverted 5'
stereoisomer (E-IV). Suitable hydroboration reagents include
BH.sub.3-THF and suitable oxidative work-up conditions include
H.sub.2O.sub.2/NaOH. The intermediate (E-IV) may then be subjected
to the reaction sequences depicted in schemes C and D to furnish
the intermediates (E-V) and (E-VI).
##STR00195##
[0481] The use of intermediates (F-I) and (F-III) in the procedures
outlined in Schemes C and D allows for the synthesis of purine and
7-deazapurine deoxy carbocyclic derivatives. The ribose based
intermediates (F-V), (F-VI), (F-VIII) and (F-IX) may be used in
reaction procedures similar to those described in Schemes A and B
above.
##STR00196##
[0482] Cyclobutanes of formulae (G-VIII), (G-XV) and (G-XXI) may be
synthesized as depicted in Scheme G. The alkenyl esters (G-I) may
be subjected to a [2+2] cycloaddition with trichloroacetyl chloride
in the presence of Zn/Cu couple in an inert solvent such as
Et.sub.2O, DME, THE or a mixture thereof. Alternatively the [2+2]
cycloaddition reaction may be performed using Zn dust under
sonication conditions. The dichlorides (G-II) are reduced via
treatment with Zn powder in the presence of a proton donor such as
NH.sub.4C.sub.1 in a solvent such as MeOH. The cyclobutanones
(G-IV) (which include (G-III)) may be further elaborated by
treatment with a phosphonate (G-V) to give the .alpha., .beta.
unsaturated esters (G-VI). The acid (VI) is converted to the
Weinreb amide (G-VII) under standard conditions (e.g. iso-butyl
chloroformate, Hunig's base, N,O-dimethyl hydroxylamine). The
double bond may then be reduced via hydrogenation using H.sub.2 in
the presence of a metal catalyst such as Pd/C, PtO.sub.2 or
Pd(OH).sub.2 to give the cyclobutane intermediates (G-VIII).
[0483] The cyclobutanones (G-IX) may be treated with the Wittig
reagent (G-X) to give the cyclobutane enol ether (G-XI) which upon
deprotection gives the corresponding acid (G-XII).
[0484] The cyclobutanones (G-IX) may also be treated with the
stabilized phosphonate (G-V) in the presence of a base such as
KOtBu, LDA, NaHMDS, KHMDS or LiHMDS or with Et.sub.3N in the
presence of LiCl in an inert solvent to give the .alpha., .beta.
unsaturated ester (G-XIII) which can be reduced to the (G-XIV) by
treatment with H.sub.2 in the presence of a metal catalyst such as
Pd/C, Pd(OH).sub.2 or PtO.sub.2 in an inert solvent. The acid
functionality of (G-XIV) may be converted into the corresponding
Weinreb amide by treatment with N,O-dimethylhydroxylamine in
presence of a suitable coupling agent such as
iso-butylchloroformate and a base such as Hunig's base to give
(G-XV).
[0485] The cyclobutanones (G-XVI) may also be treated with
N,O-dimethylhydroxylamine in presence of a suitable coupling agent
such as iso-butylchloroformate and a base such as Hunig's base to
give the corresponding Weinreb amide (G-XVII) which upon reductive
amination with an ammonia equivalent followed by deprotection as
needed gives the amine (G-XVIII). Suitable ammonia equivalents
include benzhydryl amine, NH.sub.3, NH.sub.4Cl, BnNH.sub.2,
PMB-NH.sub.2, 2,4 DMB-NH.sub.2 which may be treated with the ketone
(G-XVII) and a suitable reducing agent such as NaCN(BH.sub.3) or
Na(OAc).sub.3BH in the presence of an acid if required such as HCl
or AcOH. Protecting groups on the reductive amination products may
be removed by methods known to those of ordinary skill in the art.
Alternatively the ketone (G-XVII) can be treated with hydroxyl
amine to form the corresponding oxime which then can be reduced
with H2 in the presence of a metal catalyst such as Pd/C, PtO.sub.2
or Pd(OH).sub.2 to give the intermediate (G-XVIII).
[0486] The cyclobutane (G-IV) may converted into the amine (G-XXI)
via a multi-step sequence involving treating (G-IV) with the
phosphorane (G-V) to produce the enol ether (G-XIX). Treatment of
(G-XIX) with which is then N,O-dimethylhydroxylamine in presence of
a suitable coupling agent such as iso-butyl chloroformate and a
base such as Hunig's base to give the corresponding Weinreb amide
(G-XX) which after aqueous hydrolysis of the enol ether (e.g.
TsOH/H.sub.2O, HCl/H.sub.2O) and reductive amination with an
ammonia equivalent followed by deprotection as needed gives the
amine (G-XXI). Suitable ammonia equivalents include benzhydryl
amine, NH.sub.3, NH.sub.4Cl, BnNH.sub.2, PMB-NH.sub.2, 2,4
DMB-NH.sub.2. Suitable reducing agents for the reductive amination
include NaCN(BH.sub.3) or Na(OAc).sub.3BH used in the presence of
an acid if required such as HCl or AcOH. Protecting groups on the
reductive amination products may be removed by methods known to
those of ordinary skill in the art.
##STR00197##
[0487] The cyclobutanones (H-I) can be converted into the
benzimidazoles (H-III), ureas (H-IV) and amides (H-V) via the
reaction sequences depicted in Scheme H. The benzimidazoles (H-III)
may be formed by treating the acids (H-I) with the appropriate
benzene diamine (H-II) in the presence of a suitable coupling agent
(e.g., HATU, PPAA, COMU, EDC, EDCI), in the presence of a base
(e.g., Et.sub.3N, Hunig's base, K.sub.2CO.sub.3). Additional
reagents such as HOAT, HOBt or HOSu may be added if necessary. The
resultant amino-amides are then cyclized to the benzimidazoles in
the presence of acid, e.g. AcOH which may also serve as the
solvent. The reaction is normally carried at a temperatures ranging
from RT to 80.degree. C. The ureas (H-IV) may be prepared by
converting the acids (H-I) as follows. The acid is reduced to the
corresponding primary alcohol using a reagent such as LiAlH.sub.4
or BH.sub.3.THF. If needed the ketone functionality may first be
protected (e.g., as a ketal) prior to reduction and subsequently
deprotected. The primary alcohol is then converted into a leaving
group such as a mesylate. The resultant leaving group is displaced
with azide from a source such as NaN.sub.3. The azido product
compound is reduced to the corresponding amino compound using
H.sub.2 in the presence of a metal catalyst such as Pd/C or via a
Staudinger reaction with a phosphine such as PMe.sub.3 or
PPh.sub.3. The urea (H-IV) is then formed by treatment of the
primary amine with the appropriate isocyanate, R-C.dbd.N.dbd.O in
the presence of a base such as Et.sub.3N or K.sub.2CO.sub.3 in an
inert solvent such as CH.sub.2Cl.sub.2. The amides (H-V) are formed
by treating the acids (H-I) with the appropriate amine R--NH.sub.2
is the presence of a suitable coupling agent (e.g. HATU, PPAA,
COMU, EDC, EDCI), in the presence of a base (e.g. Et.sub.3N,
Hunig's base, K.sub.2CO.sub.3). Additional reagents such as HOAT,
HOBt or HOSu may be added if necessary.
##STR00198##
[0488] As shown in Scheme I above, the Weinreb amides (I-I) and
(I-V) may be transformed into the benzimidazole aldehydes (I-III),
urea aldehydes (I-IV) and (I-VI) and amide aldehydes (I-V) using
similar procedures as described in Scheme H followed by Weinreb
amide reduction. The reduction of the Weinreb amide can be
performed using DiBAL-H in an inert solvent such as
CH.sub.2Cl.sub.2, THE at a temperature of 10 to -78.degree. C. The
amino Weinreb amide (I-V) may also be converted into the urea
(I-VI) via treatment with the appropriate isocyanate followed by
reduction with DiBAL-H.
##STR00199##
[0489] The benzimidazole (J-V), urea (J-VI) and amide (J-VII)
intermediates may be synthesized as depicted in Scheme J. The
cyclobutanones (J-I) may be subjected to a Wittig reaction with the
phosphorane (J-II) to produce the enol ether (J-III). The enol
ether acid is then subjected to reactions conditions similar to
those as described in scheme H to produce the corresponding enol
ether benzimidazoles, ureas and amides, which upon treatment with
aqueous acid, such as TsOH/H.sub.2O, HCl generates the
corresponding aldehyde intermediates (J-V), (J-VI) and (J-VII)
respectively.
##STR00200##
[0490] The formula (K-V)
##STR00201##
in Scheme K above represents the intermediates (K-I through K-IV)
below and their corresponding 2'- or 3-deoxy intermediates, whose
syntheses are described in Schemes A-F.
##STR00202##
[0491] As shown in Scheme K, the ketones (K-VI), (K-VII) and
(K-VIII) and the aldehydes (K-XII), (K-XIII) and (K-XIV) are
converted into the corresponding benzimidazoles (K-IX) and (K-XV),
ureas (K-X) and (K-XVI) and amides (K-XI) and (K-XVII) via
reductive amination with (K-V). The reductive amination can be
performed with a suitable reducing agent such as NaCN(BH.sub.3) or
Na(OAc).sub.3BH in the presence of an acid if required such as HCl
or AcOH or a Lewis acid/dehydrating agent such as Ti(OiPr).sub.4 or
MgSO.sub.4.
##STR00203##
[0492] In an alternative reaction sequence, the target
benzimidazoles (L-V), ureas (L-VI) and amides (L-VII) may be
prepared by the reaction sequence depicted in Scheme L. The amines
(L-I), where Q and R.sub.c have the same definitions as in Scheme
K, are treated with the cyclobutanes (L-II) under reductive
amination conditions using a reducing agent such as NaCNBH.sub.3 or
Na(OAc).sub.3BH in the presence of an acid such as HCl or AcOH or a
Lewis acid such as Ti(OiPr).sub.4, to give the acid (L-III). The
acid may be converted into the corresponding target benzimidazoles
(L-V), ureas (L-VI) and amides (L-VII) using reaction conditions
similar to those in scheme H.
##STR00204##
[0493] The enol ethers (M-1) may be hydrolyzed under acidic
conditions (e.g. TsOH/H.sub.2O, HCl/H.sub.2O) to give the aldehydes
(M-II). Reductive amination of (M-II) is conducted with amines
(M-III) where Q and R.sub.c have the same meaning as in scheme K,
using a reducing agent such as NaCNBH.sub.3 or Na(OAc).sub.3BH in
the presence of an acid such as HCl or AcOH or a Lewis
acid/dehydrating agent such as Ti(OiPr).sub.4 or MgSO.sub.4.
Subsequent ester protecting group removal is then carried out to
give the acid (M-IV). The acids (M-V) (which include the acids
(M-IV)) may be transformed into the corresponding target
benzimidazoles (M-VII), ureas (M-VIII) and amides (M-IX) using
reaction conditions similar to those described in scheme H.
##STR00205##
[0494] The benzimidazoles, ureas and amides of formula (N-XIII) may
be synthesized as depicted in Scheme N. The aldehyde (N-I) where X
represents a benzimidazole, urea or amide functionality, may be
converted in the corresponding acid via oxidation with a reagent
such as NaClO.sub.2. The cyclobutanones of formula (N-III) may be
treated with phosphonates of formula (N-IV) and a suitable base
such as LiHMDS, NaHMDS, KHMDS, KOtBu, LDA or Et.sub.3N/LiCl in an
inert solvent to give the cyclobutanes (N-V), which upon reduction
via treatment with H.sub.2 in the presence of suitable metal
catalyst such as Pd/C, PtO.sub.2 or Pd(OH).sub.2 gives the acid
(N-VI).
[0495] The acids of formula (N-VII) which include the acids of
formula (N-VI) may be converted into the corresponding
benzimidazoles (N-IX), ureas (N-X) and amides (N-XI) via a series
of reactions similar to those depicted in scheme H. The
benzimidazoles, ureas and amides of formula (N-II) may then be
converted to the benzimidazoles, ureas and amides of formula
(N-XIII) via an amide coupling reaction with the amine (N-XIII)
where R.sub.c and Q have the same definitions as in Scheme K, using
a suitable coupling agent (e.g. HATU, PPAA, COMU, EDC, EDCI), in
the presence of a base (e.g. Et.sub.3N, Hunig's base,
K.sub.2CO.sub.3). Additional reagents such as HOAT, HOBt or HOSu
may be added if necessary.
##STR00206## ##STR00207##
[0496] The thioethers (O-XIII), sulfoxides (O-XIV) and sulfones
(O-XV) may be synthesized as depicted in Scheme O. The
cyclobutanones (O-I), where X represents benzimidazole, urea or
amide functionality, may be reduced with a reducing reagent such as
NaBH.sub.4 to give the corresponding alcohol, which in turn may
converted to a leaving group such a MsO, by treatment with MsCl
with a base such as K.sub.2CO.sub.3 in an inert solvent. The
cyclobutane (O-II) is then converted into the corresponding thiol
(O-III) by first treating with a sulfur based nucleophile such as
KSAc followed by hydrolysis of the thioester under basic
conditions, e.g. LiOH/H.sub.2O/MeOH.
[0497] The aldehydes (O-IV) may be converted into the corresponding
thiols via a reaction sequence similar to that as described for
converting (O-I) into (O-III).
[0498] The cyclobutanones (O-VII) may be converted into acids
(O-IX) by treatment with a phosphonate of formula (O-VIII) in the
presence of a suitable base (Cs.sub.2CO.sub.3, K.sub.2CO.sub.3,
LiHMDS, NaHMDS, KHMDS, KOtBu, LDA), followed by reduction using
H.sub.2 in the presence of a suitable metal catalyst such as Pd/C,
PtO.sub.2 or Pd(OH).sub.2. The acid is the selectively reduced to
the primary alcohol using a reagent such as BH.sub.3:THF or by
reaction with carbonyldiimidazole, Et.sub.3N followed by
NaBH.sub.4. The primary alcohol is converted to the corresponding
thiol using reactions similar to those used for the synthesis of
(O-III) and (O-VI) from their secondary alcohol intermediates.
[0499] The thiols (O-XI) may then be treated with the intermediates
(O-XII) where Q' represents the intermediates (O-XVI), (O-XVII),
(O-XVIII) and (O-XIX) depicted below (and also the corresponding
2'- or 3'-deoxy intermediates) and LG represents a leaving group
such as Cl, TfO or MsO. The reaction may carried out in the
presence of a suitable base such as K.sub.2CO.sub.3,
Cs.sub.2CO.sub.3, Et.sub.3N or Hunig's base to give the thioethers
(O-XIII) which may be converted into the corresponding sulfoxides
(O-XIV) or sulfones (O-XV) via treatment with suitable oxidizing
agents such as H.sub.2O.sub.2 or mCPBA.
##STR00208##
##STR00209##
[0500] The amine, amide and sulfonamide target molecules may be
synthesized from the amines (P-I), (P-II) and (P-III) using
standard reaction conditions. The amide target molecules may be
produced by treating the amines with the appropriate carboxylic
acid in the presence of a suitable coupling agent(e.g. HATU, PPAA,
COMU, EDC, EDCI), in the presence of a base (e.g. Et.sub.3N,
Hunig's base, K.sub.2CO.sub.3). Additional reagents such as HOAt,
HOBt or HOSu may be added if necessary. The sulfonamide target
molecules may be produced may treating the amines with the
appropriate sulfonyl chloride in the presence of a base such as
K.sub.2CO.sub.3, or Et.sub.3N. The amine target molecules may be
formed via a reductive amination reaction with the appropriate
aldehyde or ketone in the presence of a suitable reducing agent
such as NaBH(OAc).sub.3 or NaCNBH.sub.3. Additional reagents such
as acids AcOH or HCl or the Lewis acid/dehydrating agents
Ti(OiPr).sub.4 or MgSO.sub.4 may be added.
##STR00210##
##STR00211##
##STR00212##
##STR00213##
##STR00214##
##STR00215##
##STR00216##
[0501] Throughout the description, where compositions are described
as having, including, or comprising specific components, or where
processes are described as having, including, or comprising
specific process steps, it is contemplated that compositions of the
present invention also consist essentially of, or consist of, the
recited components, and that the processes of the present invention
also consist essentially of, or consist of, the recited processing
steps. Further, it should be understood that the order of steps or
order for performing certain actions are immaterial so long as the
invention remains operable. Moreover, two or more steps or actions
can be conducted simultaneously.
[0502] Compounds designed, selected and/or optimized by methods
described above, once produced, can be characterized using a
variety of assays known to those skilled in the art to determine
whether the compounds have biological activity. For example, the
molecules can be characterized by conventional assays, including
but not limited to those assays described below, to determine
whether they have a predicted activity, binding activity and/or
binding specificity.
[0503] Furthermore, high-throughput screening can be used to speed
up analysis using such assays. As a result, it can be possible to
rapidly screen the molecules described herein for activity, using
techniques known in the art. General methodologies for performing
high-throughput screening are described, for example, in Devlin
(1998) High Throughput Screening, Marcel Dekker; and U.S. Pat. No.
5,763,263. High-throughput assays can use one or more different
assay techniques including, but not limited to, those described
herein.
[0504] To further assess a compound's drug-like properties,
measurements of inhibition of cytochrome P450 enzymes and phase II
metabolizing enzyme activity can also be measured either using
recombinant human enzyme systems or more complex systems like human
liver microsomes. Further, compounds can be assessed as substrates
of these metabolic enzyme activities as well. These activities are
useful in determining the potential of a compound to cause
drug-drug interactions or generate metabolites that retain or have
no useful antimicrobial activity.
[0505] To get an estimate of the potential of the compound to be
orally bioavailable, one can also perform solubility and Caco-2
assays. The latter is a cell line from human epithelium that allows
measurement of drug uptake and passage through a Caco-2 cell
monolayer often growing within wells of a 24-well microtiter plate
equipped with a 1 micron membrane. Free drug concentrations can be
measured on the basolateral side of the monolayer, assessing the
amount of drug that can pass through the intestinal monolayer.
Appropriate controls to ensure monolayer integrity and tightness of
gap junctions are needed. Using this same system one can get an
estimate of P-glycoprotein mediated efflux. P-glycoprotein is a
pump that localizes to the apical membrane of cells, forming
polarized monolayers. This pump can abrogate the active or passive
uptake across the Caco-2 cell membrane, resulting in less drug
passing through the intestinal epithelial layer. These results are
often done in conjunction with solubility measurements and both of
these factors are known to contribute to oral bioavailability in
mammals. Measurements of oral bioavailability in animals and
ultimately in man using traditional pharmacokinetic experiments
will determine the absolute oral bioavailability.
[0506] Experimental results can also be used to build models that
help predict physical-chemical parameters that contribute to
drug-like properties. When such a model is verified, experimental
methodology can be reduced, with increased reliance on the model
predictability.
3. METHODS OF TREATMENT
[0507] Mixed lineage leukemia (MLL) is a genetically distinct form
of acute leukemia that constitutes over 70% of infant leukemias and
approximately 10% of adult acute myeloid leukemias (AML) (Hess, J.
L. (2004), Trends Mol Med 10, 500-507; Krivtsov, A. V., and
Armstrong, S. A. (2007), Nat Rev Cancer 7, 823-833). MLL represents
a particularly aggressive form of leukemia and patients with this
disease generally have poor prognoses; these patients often suffer
from early relapse after treatment with current chemotherapies.
There is thus a great and present need for new treatment modalities
for patients suffering with MLL.
[0508] A universal hallmark of MLL disease is a chromosomal
translocation affecting the MLL gene on chromosome 11q23 (Hess,
2004; Krivtsov and Armstrong, 2007). Normally, the MLL gene encodes
for a SET-domain histone methyltransferase that catalyzes the
methylation of lysine 4 of histone H3 (H3K4) at specific gene loci
(Milne et al. (2002) Mol Cell 10, 1107-1117; Nakamura et al.
(2002), Mol Cell 10, 1119-1128). Gene localization is conferred by
specific interactions with recognition elements within MLL,
external to the SET-domain (Ayton et al. (2004) Mol Cell Biol 24,
10470-10478; Slany et al., (1998) Mol Cell Biol 18, 122-129;
Zeleznik-Le et al. (1994) Proc Natl Acad Sci USA 91, 10610-10614).
In the disease-linked translocations, the catalytic SET-domain is
lost and the remaining MLL protein is fused to a variety of
partners, including members of the AF and ENL family of proteins
such as AF4, AF9, AF10 and ENL (Hess, 2004; Krivtsov and Armstrong,
2007; Slany (2009) Haematologica 94, 984-993). These fusion
partners are capable of interacting directly, or indirectly, with
another histone methyltransferase, DOT1L (Bitoun et al. (2007) Hum
Mol Genet 16, 92-106; Mohan et al. (2010) Genes Dev. 24, 574-589;
Mueller et al. (2007) Blood 110, 4445-4454; Mueller et al. (2009)
PLoS Biol 7, e1000249; Okada et al. (2005) Cell 121, 167-178; Park
et al. (2010) Protein J 29, 213-223; Yokoyama et al. (2010) Cancer
Cell 17, 198-212; Zhang et al. (2006) J Biol Chem 281,
18059-18068). As a result, translocation products retain
gene-specific recognition elements within the remainder of the MLL
protein, but also gain the ability to recruit DOT1L, to these
locations (Monroe et al. (2010) Exp Hematol. 2010 Sep. 18. [Epub
ahead of print] Pubmed PMID: 20854876; Mueller et al., 2007;
Mueller et al., 2009; Okada et al., 2005). DOT1L catalyzes the
methylation of H3K79, a chromatin modification associated with
actively transcribed genes (Feng et al. (2002) Curr Biol 12,
1052-1058; Steger et al. (2008) Mol Cell Biol 28, 2825-2839). The
ectopic H3K79 methylation that results from MLL fusion protein
recruitment of DOT1L leads to enhanced expression of leukemogenic
genes, including HOXA9 and MEIS1 (Guenther et al. (2008) Genes
& Development 22, 3403-3408; Krivtsov et al. (2008) Nat Rev
Cancer 7, 823-833; Milne et al. (2005) Cancer Res 65, 11367-11374;
Monroe et al., 2010; Mueller et al., 2009; Okada et al., 2005;
Thiel et al. (2010) Cancer Cell 17, 148-159). Hence, while DOT1L is
not genetically altered in the disease per se, its mislocated
enzymatic activity is a direct consequence of the chromosomal
translocation affecting MLL patients; thus, DOT1L has been proposed
to be a catalytic driver of leukemogenesis in this disease
(Krivtsov et al., 2008; Monroe et al., 2010; Okada et al., 2005;
Yokoyama et al. (2010) Cancer Cell 17, 198-212). Further support
for a pathogenic role of DOT1L in MLL comes from studies in model
systems that demonstrate a requirement for DOT1L in propagating the
transforming activity of MLL fusion proteins (Mueller et al., 2007;
Okada et al., 2005).
[0509] Evidence indicates that the enzymatic activity of DOT1L is
critical to pathogenesis in MLL and inhibition of DOT1L may provide
a pharmacologic basis for therapeutic intervention in this disease.
Compound treatment results in selective, concentration-dependent
killing of leukemia cells bearing the MLL-translocation without
effect on non-MLL transformed cells. Gene expression analysis of
inhibitor treated cells shows downregulation of genes aberrantly
over expressed in MLL-rearranged leukemias and similarities with
gene expression changes caused by genetic knockout of the Dot1L
gene in a mouse model of MLL-AF9 leukemia.
[0510] The present invention provides methods for the treatment of
a cell proliferative disorder in a subject in need thereof by
administering to a subject in need of such treatment, a
therapeutically effective amount of a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof. The cell proliferative
disorder can be cancer or a precancerous condition. The present
invention further provides the use of a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, for the preparation of a
medicament useful for the treatment of a cell proliferative
disorder.
[0511] The present invention provides methods for the treatment of
hematological cancer or hematologic tumors in a subject in need
thereof by administering to a subject in need of such treatment, a
therapeutically effective amount of a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof. The present invention
further provides the use of a compound of the present invention, or
a pharmaceutically acceptable salt, prodrug, metabolite, polymorph
or solvate thereof, for the preparation of a medicament useful for
the treatment of hematological cancer or hematologic tumors.
[0512] The present invention provides methods for the treatment of
leukemia in a subject in need thereof by administering to a subject
in need of such treatment, a therapeutically effective amount of a
compound of the present invention, or a pharmaceutically acceptable
salt, prodrug, metabolite, polymorph or solvate thereof. The
leukemia can be acute or chronic leukemia. Preferably, the leukemia
is acute myeloid leukemia, acute lymphocytic leukemia or mixed
lineage leukemia. The present invention further provides the use of
a compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
for the preparation of a medicament useful for the treatment of
leukemia.
[0513] The present invention provides methods for the treatment of
a disease or disorder mediated by translocation of a gene on
chromosome 11q23 in a subject in need thereof by administering to a
subject in need of such treatment, a therapeutically effective
amount of a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof. The gene can be the MLL gene. The present
invention further provides the use of a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, for the preparation of a
medicament useful for the treatment of a disease or disorder
mediated by translocation of a gene on chromosome 11q23.
[0514] The present invention provides methods for the treatment of
a disease or disorder mediated by DOT1 (e.g., DOT1L)-mediated
protein methylation in a subject in need thereof by administering
to a subject in need of such treatment, a therapeutically effective
amount of a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof. The present invention further provides the use of
a compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
for the preparation of a medicament useful for the treatment of a
disease or disorder mediated by DOT1L-mediated protein
methylation.
[0515] The present invention provides methods for the treatment of
a disorder the course of which is influenced by modulating the
methylation status of histones or other proteins, wherein said
methylation status is mediated at least in part by the activity of
DOT1L. Modulation of the methylation status of histones can in turn
influence the level of expression of target genes activated by
methylation, and/or target genes suppressed by methylation. The
method includes administering to a subject in need of such
treatment, a therapeutically effective amount of a compound of the
present invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph, solvate, or stereoisomeror thereof.
[0516] The disorder in which DOT1L-mediated protein methylation
plays a part can be cancer or a precancerous condition or a
neurological disease. The present invention further provides the
use of a compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
for the preparation of a medicament useful for the treatment of
cancer or a neurological disease.
[0517] The present invention also provides methods of protecting
against a disorder in which DOT1L-mediated protein methylation
plays a part in a subject in need thereof by administering a
therapeutically effective amount of compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, to a subject in need of
such treatment. The disorder can be cancer or a neurological
disease. The present invention also provides the use of compound of
the present invention, or a pharmaceutically acceptable salt,
prodrug, metabolite, polymorph, solvate, or stereoisomeror thereof,
for the preparation of a medicament useful for the prevention of a
cell proliferative disorder.
[0518] The compounds of this invention can be used to modulate
protein (e.g., histone) methylation, e.g., to modulate histone
methyltransferase or histone demethylase enzyme activity. Histone
methylation has been reported to be involved in aberrant expression
of certain genes in cancers, and in silencing of neuronal genes in
non-neuronal cells. The compounds described herein can be used to
treat these diseases, i.e., to decreases methylation or restores
methylation to roughly its level in counterpart normal cells.
[0519] In general, compounds that are methylation modulators can be
used for modulating cell proliferation, generally. For example, in
some cases excessive proliferation may be reduced with agents that
decrease methylation, whereas insufficient proliferation may be
stimulated with agents that increase methylation. Accordingly,
diseases that may be treated by the compounds of the invention
include hyperproliferative diseases, such as benign cell growth and
malignant cell growth.
[0520] As used herein, a "subject in need thereof" is a subject
having a cell proliferative disorder, or a subject having an
increased risk of developing a cell proliferative disorder relative
to the population at large. The subject can have cancer or
pre-cancer. Preferably, a subject in need thereof has cancer. More
preferably, a hematologic cancer or leukemia. A "subject" includes
a mammal. The mammal can be e.g., any mammal, e.g., a human,
primate, bird, mouse, rat, fowl, dog, cat, cow, horse, goat, camel,
sheep or a pig. Preferably, the mammal is a human.
[0521] As used herein, the term "cell proliferative disorder"
refers to conditions in which unregulated or abnormal growth, or
both, of cells can lead to the development of an unwanted condition
or disease, which may or may not be cancerous. Exemplary cell
proliferative disorders of the invention encompass a variety of
conditions wherein cell division is deregulated. Exemplary cell
proliferative disorder include, but are not limited to, neoplasms,
benign tumors, malignant tumors, pre-cancerous conditions, in situ
tumors, encapsulated tumors, metastatic tumors, liquid tumors,
solid tumors, immunological tumors, hematological tumors, cancers,
carcinomas, leukemias, lymphomas, sarcomas, and rapidly dividing
cells. The term "rapidly dividing cell" as used herein is defined
as any cell that divides at a rate that exceeds or is greater than
what is expected or observed among neighboring or juxtaposed cells
within the same tissue. A cell proliferative disorder includes a
precancer or a precancerous condition. A cell proliferative
disorder includes cancer. Preferably, the methods provided herein
are used to treat or alleviate a symptom of cancer. The term
"cancer" includes solid tumors, as well as, hematologic tumors
and/or malignancies. A "precancer cell" or "precancerous cell" is a
cell manifesting a cell proliferative disorder that is a precancer
or a precancerous condition. A "cancer cell" or "cancerous cell" is
a cell manifesting a cell proliferative disorder that is a cancer.
Any reproducible means of measurement may be used to identify
cancer cells or precancerous cells. Cancer cells or precancerous
cells can be identified by histological typing or grading of a
tissue sample (e.g., a biopsy sample). Cancer cells or precancerous
cells can be identified through the use of appropriate molecular
markers.
[0522] Exemplary non-cancerous conditions or disorders include, but
are not limited to, rheumatoid arthritis; inflammation; autoimmune
disease; lymphoproliferative conditions; acromegaly; rheumatoid
spondylitis; osteoarthritis; gout, other arthritic conditions;
sepsis; septic shock; endotoxic shock; gram-negative sepsis; toxic
shock syndrome; asthma; adult respiratory distress syndrome;
chronic obstructive pulmonary disease; chronic pulmonary
inflammation; inflammatory bowel disease; Crohn's disease;
psoriasis; eczema; ulcerative colitis; pancreatic fibrosis; hepatic
fibrosis; acute and chronic renal disease; irritable bowel
syndrome; pyresis; restenosis; cerebral malaria; stroke and
ischemic injury; neural trauma; Alzheimer's disease; Huntington's
disease; Parkinson's disease; acute and chronic pain; allergic
rhinitis; allergic conjunctivitis; chronic heart failure; acute
coronary syndrome; cachexia; malaria; leprosy; leishmaniasis; Lyme
disease; Reiter's syndrome; acute synovitis; muscle degeneration,
bursitis; tendonitis; tenosynovitis; herniated, ruptures, or
prolapsed intervertebral disk syndrome; osteopetrosis; thrombosis;
restenosis; silicosis; pulmonary sarcosis; bone resorption
diseases, such as osteoporosis; graft-versus-host reaction;
Multiple Sclerosis; lupus; fibromyalgia; AIDS and other viral
diseases such as Herpes Zoster, Herpes Simplex I or II, influenza
virus and cytomegalovirus; and diabetes mellitus.
[0523] Exemplary cancers include, but are not limited to,
adrenocortical carcinoma, AIDS-related cancers, AIDS-related
lymphoma, anal cancer, anorectal cancer, cancer of the anal canal,
appendix cancer, childhood cerebellar astrocytoma, childhood
cerebral astrocytoma, basal cell carcinoma, skin cancer
(non-melanoma), biliary cancer, extrahepatic bile duct cancer,
intrahepatic bile duct cancer, bladder cancer, uringary bladder
cancer, bone and joint cancer, osteosarcoma and malignant fibrous
histiocytoma, brain cancer, brain tumor, brain stem glioma,
cerebellar astrocytoma, cerebral astrocytoma/malignant glioma,
ependymoma, medulloblastoma, supratentorial primitive
neuroectodeimal tumors, visual pathway and hypothalamic glioma,
breast cancer, bronchial adenomas/carcinoids, carcinoid tumor,
gastrointestinal, nervous system cancer, nervous system lymphoma,
central nervous system cancer, central nervous system lymphoma,
cervical cancer, childhood cancers, chronic lymphocytic leukemia,
chronic myelogenous leukemia, chronic myeloproliferative disorders,
colon cancer, colorectal cancer, cutaneous T-cell lymphoma,
lymphoid neoplasm, mycosis fungoides, Seziary Syndrome, endometrial
cancer, esophageal cancer, extracranial germ cell tumor,
extragonadal germ cell tumor, extrahepatic bile duct cancer, eye
cancer, intraocular melanoma, retinoblastoma, gallbladder cancer,
gastric (stomach) cancer, gastrointestinal carcinoid tumor,
gastrointestinal stromal tumor (GIST), germ cell tumor, ovarian
germ cell tumor, gestational trophoblastic tumor glioma, head and
neck cancer, hepatocellular (liver) cancer, Hodgkin lymphoma,
hypopharyngeal cancer, intraocular melanoma, ocular cancer, islet
cell tumors (endocrine pancreas), Kaposi Sarcoma, kidney cancer,
renal cancer, kidney cancer, laryngeal cancer, acute lymphoblastic
leukemia, acute lymphocytic leukemia, acute myeloid leukemia,
chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy
cell leukemia, lip and oral cavity cancer, liver cancer, lung
cancer, non-small cell lung cancer, small cell lung cancer,
AIDS-related lymphoma, non-Hodgkin lymphoma, primary central
nervous system lymphoma, Waldenstram macroglobulinemia,
medulloblastoma, melanoma, intraocular (eye) melanoma, merkel cell
carcinoma, mesothelioma malignant, mesothelioma, metastatic
squamous neck cancer, mouth cancer, cancer of the tongue, multiple
endocrine neoplasia syndrome, mycosis fungoides, myelodysplastic
syndromes, myelodysplastic/myeloproliferative diseases, chronic
myelogenous leukemia, acute myeloid leukemia, multiple myeloma,
chronic myeloproliferative disorders, nasopharyngeal cancer,
neuroblastoma, oral cancer, oral cavity cancer, oropharyngeal
cancer, ovarian cancer, ovarian epithelial cancer, ovarian low
malignant potential tumor, pancreatic cancer, islet cell pancreatic
cancer, paranasal sinus and nasal cavity cancer, parathyroid
cancer, penile cancer, pharyngeal cancer, pheochromocytoma,
pineoblastoma and supratentorial primitive neuroectodermal tumors,
pituitary tumor, plasma cell neoplasm/multiple myeloma,
pleuropulmonary blastoma, prostate cancer, rectal cancer, renal
pelvis and ureter, transitional cell cancer, retinoblastoma,
rhabdomyosarcoma, salivary gland cancer, ewing family of sarcoma
tumors, Kaposi Sarcoma, soft tissue sarcoma, uterine cancer,
uterine sarcoma, skin cancer (non-melanoma), skin cancer
(melanoma), merkel cell skin carcinoma, small intestine cancer,
soft tissue sarcoma, squamous cell carcinoma, stomach (gastric)
cancer, supratentorial primitive neuroectodermal tumors, testicular
cancer, throat cancer, thymoma, thymoma and thymic carcinoma,
thyroid cancer, transitional cell cancer of the renal pelvis and
ureter and other urinary organs, gestational trophoblastic tumor,
urethral cancer, endometrial uterine cancer, uterine sarcoma,
uterine corpus cancer, vaginal cancer, vulvar cancer, and Wilm's
Tumor.
[0524] A "cell proliferative disorder of the hematologic system" is
a cell proliferative disorder involving cells of the hematologic
system. A cell proliferative disorder of the hematologic system can
include lymphoma, leukemia, myeloid neoplasms, mast cell neoplasms,
myelodysplasia, benign monoclonal gammopathy, lymphomatoid
granulomatosis, lymphomatoid papulosis, polycythemia vera, chronic
myelocytic leukemia, agnogenic myeloid metaplasia, and essential
thrombocythemia. A cell proliferative disorder of the hematologic
system can include hyperplasia, dysplasia, and metaplasia of cells
of the hematologic system. Preferably, compositions of the present
invention may be used to treat a cancer selected from the group
consisting of a hematologic cancer of the present invention or a
hematologic cell proliferative disorder of the present invention. A
hematologic cancer of the present invention can include multiple
myeloma, lymphoma (including Hodgkin's lymphoma, non-Hodgkin's
lymphoma, childhood lymphomas, and lymphomas of lymphocytic and
cutaneous origin), leukemia (including childhood leukemia,
hairy-cell leukemia, acute lymphocytic leukemia, acute myelocytic
leukemia, chronic lymphocytic leukemia, chronic myelocytic
leukemia, chronic myelogenous leukemia, and mast cell leukemia),
myeloid neoplasms and mast cell neoplasms.
[0525] A "cell proliferative disorder of the lung" is a cell
proliferative disorder involving cells of the lung. Cell
proliferative disorders of the lung can include all forms of cell
proliferative disorders affecting lung cells. Cell proliferative
disorders of the lung can include lung cancer, a precancer or
precancerous condition of the lung, benign growths or lesions of
the lung, and malignant growths or lesions of the lung, and
metastatic lesions in tissue and organs in the body other than the
lung. Preferably, compositions of the present invention may be used
to treat lung cancer or cell proliferative disorders of the lung.
Lung cancer can include all forms of cancer of the lung. Lung
cancer can include malignant lung neoplasms, carcinoma in situ,
typical carcinoid tumors, and atypical carcinoid tumors. Lung
cancer can include small cell lung cancer ("SCLC"), non-small cell
lung cancer ("NSCLC"), squamous cell carcinoma, adenocarcinoma,
small cell carcinoma, large cell carcinoma, adenosquamous cell
carcinoma, and mesothelioma. Lung cancer can include "scar
carcinoma," bronchioalveolar carcinoma, giant cell carcinoma,
spindle cell carcinoma, and large cell neuroendocrine carcinoma.
Lung cancer can include lung neoplasms having histologic and
ultrastructual heterogeneity (e.g., mixed cell types).
[0526] Cell proliferative disorders of the lung can include all
forms of cell proliferative disorders affecting lung cells. Cell
proliferative disorders of the lung can include lung cancer,
precancerous conditions of the lung. Cell proliferative disorders
of the lung can include hyperplasia, metaplasia, and dysplasia of
the lung. Cell proliferative disorders of the lung can include
asbestos-induced hyperplasia, squamous metaplasia, and benign
reactive mesothelial metaplasia. Cell proliferative disorders of
the lung can include replacement of columnar epithelium with
stratified squamous epithelium, and mucosal dysplasia. Individuals
exposed to inhaled injurious environmental agents such as cigarette
smoke and asbestos may be at increased risk for developing cell
proliferative disorders of the lung. Prior lung diseases that may
predispose individuals to development of cell proliferative
disorders of the lung can include chronic interstitial lung
disease, necrotizing pulmonary disease, scleroderma, rheumatoid
disease, sarcoidosis, interstitial pneumonitis, tuberculosis,
repeated pneumonias, idiopathic pulmonary fibrosis, granulomata,
asbestosis, fibrosing alveolitis, and Hodgkin's disease.
[0527] A "cell proliferative disorder of the colon" is a cell
proliferative disorder involving cells of the colon. Preferably,
the cell proliferative disorder of the colon is colon cancer.
Preferably, compositions of the present invention may be used to
treat colon cancer or cell proliferative disorders of the colon.
Colon cancer can include all forms of cancer of the colon. Colon
cancer can include sporadic and hereditary colon cancers. Colon
cancer can include malignant colon neoplasms, carcinoma in situ,
typical carcinoid tumors, and atypical carcinoid tumors. Colon
cancer can include adenocarcinoma, squamous cell carcinoma, and
adenosquamous cell carcinoma. Colon cancer can be associated with a
hereditary syndrome selected from the group consisting of
hereditary nonpolyposis colorectal cancer, familial adenomatous
polyposis, Gardner's syndrome, Peutz-Jeghers syndrome, Turcot's
syndrome and juvenile polyposis. Colon cancer can be caused by a
hereditary syndrome selected from the group consisting of
hereditary nonpolyposis colorectal cancer, familial adenomatous
polyposis, Gardner's syndrome, Peutz-Jeghers syndrome, Turcot's
syndrome and juvenile polyposis.
[0528] Cell proliferative disorders of the colon can include all
forms of cell proliferative disorders affecting colon cells. Cell
proliferative disorders of the colon can include colon cancer,
precancerous conditions of the colon, adenomatous polyps of the
colon and metachronous lesions of the colon. A cell proliferative
disorder of the colon can include adenoma. Cell proliferative
disorders of the colon can be characterized by hyperplasia,
metaplasia, and dysplasia of the colon. Prior colon diseases that
may predispose individuals to development of cell proliferative
disorders of the colon can include prior colon cancer. Current
disease that may predispose individuals to development of cell
proliferative disorders of the colon can include Crohn's disease
and ulcerative colitis. A cell proliferative disorder of the colon
can be associated with a mutation in a gene selected from the group
consisting of p53, ras, FAP and DCC. An individual can have an
elevated risk of developing a cell proliferative disorder of the
colon due to the presence of a mutation in a gene selected from the
group consisting of p53, ras, FAP and DCC.
[0529] A "cell proliferative disorder of the pancreas" is a cell
proliferative disorder involving cells of the pancreas. Cell
proliferative disorders of the pancreas can include all forms of
cell proliferative disorders affecting pancreatic cells. Cell
proliferative disorders of the pancreas can include pancreas
cancer, a precancer or precancerous condition of the pancreas,
hyperplasia of the pancreas, and dysaplasia of the pancreas, benign
growths or lesions of the pancreas, and malignant growths or
lesions of the pancreas, and metastatic lesions in tissue and
organs in the body other than the pancreas. Pancreatic cancer
includes all forms of cancer of the pancreas. Pancreatic cancer can
include ductal adenocarcinoma, adenosquamous carcinoma, pleomorphic
giant cell carcinoma, mucinous adenocarcinoma, osteoclast-like
giant cell carcinoma, mucinous cystadenocarcinoma, acinar
carcinoma, unclassified large cell carcinoma, small cell carcinoma,
pancreatoblastoma, papillary neoplasm, mucinous cystadenoma,
papillary cystic neoplasm, and serous cystadenoma. Pancreatic
cancer can also include pancreatic neoplasms having histologic and
ultrastructual heterogeneity (e.g., mixed cell types).
[0530] A "cell proliferative disorder of the prostate" is a cell
proliferative disorder involving cells of the prostate. Cell
proliferative disorders of the prostate can include all forms of
cell proliferative disorders affecting prostate cells. Cell
proliferative disorders of the prostate can include prostate
cancer, a precancer or precancerous condition of the prostate,
benign growths or lesions of the prostate, and malignant growths or
lesions of the prostate, and metastatic lesions in tissue and
organs in the body other than the prostate. Cell proliferative
disorders of the prostate can include hyperplasia, metaplasia, and
dysplasia of the prostate.
[0531] A "cell proliferative disorder of the skin" is a cell
proliferative disorder involving cells of the skin. Cell
proliferative disorders of the skin can include all forms of cell
proliferative disorders affecting skin cells. Cell proliferative
disorders of the skin can include a precancer or precancerous
condition of the skin, benign growths or lesions of the skin,
melanoma, malignant melanoma and other malignant growths or lesions
of the skin, and metastatic lesions in tissue and organs in the
body other than the skin. Cell proliferative disorders of the skin
can include hyperplasia, metaplasia, and dysplasia of the skin.
[0532] A "cell proliferative disorder of the ovary" is a cell
proliferative disorder involving cells of the ovary. Cell
proliferative disorders of the ovary can include all forms of cell
proliferative disorders affecting cells of the ovary. Cell
proliferative disorders of the ovary can include a precancer or
precancerous condition of the ovary, benign growths or lesions of
the ovary, ovarian cancer, malignant growths or lesions of the
ovary, and metastatic lesions in tissue and organs in the body
other than the ovary. Cell proliferative disorders of the skin can
include hyperplasia, metaplasia, and dysplasia of cells of the
ovary.
[0533] A "cell proliferative disorder of the breast" is a cell
proliferative disorder involving cells of the breast. Cell
proliferative disorders of the breast can include all forms of cell
proliferative disorders affecting breast cells. Cell proliferative
disorders of the breast can include breast cancer, a precancer or
precancerous condition of the breast, benign growths or lesions of
the breast, and malignant growths or lesions of the breast, and
metastatic lesions in tissue and organs in the body other than the
breast. Cell proliferative disorders of the breast can include
hyperplasia, metaplasia, and dysplasia of the breast.
[0534] A cell proliferative disorder of the breast can be a
precancerous condition of the breast. Compositions of the present
invention may be used to treat a precancerous condition of the
breast. A precancerous condition of the breast can include atypical
hyperplasia of the breast, ductal carcinoma in situ (DCIS),
intraductal carcinoma, lobular carcinoma in situ (LCIS), lobular
neoplasia, and stage 0 or grade 0 growth or lesion of the breast
(e.g., stage 0 or grade 0 breast cancer, or carcinoma in situ). A
precancerous condition of the breast can be staged according to the
TNM classification scheme as accepted by the American Joint
Committee on Cancer (AJCC), where the primary tumor (T) has been
assigned a stage of T0 or Tis; and where the regional lymph nodes
(N) have been assigned a stage of NO; and where distant metastasis
(M) has been assigned a stage of MO.
[0535] The cell proliferative disorder of the breast can be breast
cancer. Preferably, compositions of the present invention may be
used to treat breast cancer. Breast cancer includes all forms of
cancer of the breast. Breast cancer can include primary epithelial
breast cancers. Breast cancer can include cancers in which the
breast is involved by other tumors such as lymphoma, sarcoma or
melanoma. Breast cancer can include carcinoma of the breast, ductal
carcinoma of the breast, lobular carcinoma of the breast,
undifferentiated carcinoma of the breast, cystosarcoma phyllodes of
the breast, angiosarcoma of the breast, and primary lymphoma of the
breast. Breast cancer can include Stage I, II, IIIA, IIIB, IIIC and
IV breast cancer. Ductal carcinoma of the breast can include
invasive carcinoma, invasive carcinoma in situ with predominant
intraductal component, inflammatory breast cancer, and a ductal
carcinoma of the breast with a histologic type selected from the
group consisting of comedo, mucinous (colloid), medullary,
medullary with lymphcytic infiltrate, papillary, scirrhous, and
tubular. Lobular carcinoma of the breast can include invasive
lobular carcinoma with predominant in situ component, invasive
lobular carcinoma, and infiltrating lobular carcinoma. Breast
cancer can include Paget's disease, Paget's disease with
intraductal carcinoma, and Paget's disease with invasive ductal
carcinoma. Breast cancer can include breast neoplasms having
histologic and ultrastructual heterogeneity (e.g., mixed cell
types).
[0536] Preferably, compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph,
or solvate thereof, may be used to treat breast cancer. A breast
cancer that is to be treated can include familial breast cancer. A
breast cancer that is to be treated can include sporadic breast
cancer. A breast cancer that is to be treated can arise in a male
subject. A breast cancer that is to be treated can arise in a
female subject. Abreast cancer that is to be treated can arise in a
premenopausal female subject or a postmenopausal female subject.
Abreast cancer that is to be treated can arise in a subject equal
to or older than 30 years old, or a subject younger than 30 years
old. Abreast cancer that is to be treated has arisen in a subject
equal to or older than 50 years old, or a subject younger than 50
years old. A breast cancer that is to be treated can arise in a
subject equal to or older than 70 years old, or a subject younger
than 70 years old.
[0537] A breast cancer that is to be treated can be typed to
identify a familial or spontaneous mutation in BRCA1, BRCA2, or
p53. A breast cancer that is to be treated can be typed as having a
HER2/neu gene amplification, as overexpressing HER2/neu, or as
having a low, intermediate or high level of HER2/neu expression. A
breast cancer that is to be treated can be typed for a marker
selected from the group consisting of estrogen receptor (ER),
progesterone receptor (PR), human epidermal growth factor
receptor-2, Ki-67, CA15-3, CA 27-29, and c-Met. Abreast cancer that
is to be treated can be typed as ER-unknown, ER-rich or ER-poor. A
breast cancer that is to be treated can be typed as ER-negative or
ER-positive. ER-typing of a breast cancer may be performed by any
reproducible means. ER-typing of a breast cancer may be performed
as set forth in Onkologie 27: 175-179 (2004). A breast cancer that
is to be treated can be typed as PR-unknown, PR-rich, or PR-poor. A
breast cancer that is to be treated can be typed as PR-negative or
PR-positive. A breast cancer that is to be treated can be typed as
receptor positive or receptor negative. A breast cancer that is to
be treated can be typed as being associated with elevated blood
levels of CA 15-3, or CA 27-29, or both.
[0538] A breast cancer that is to be treated can include a
localized tumor of the breast. A breast cancer that is to be
treated can include a tumor of the breast that is associated with a
negative sentinel lymph node (SLN) biopsy. A breast cancer that is
to be treated can include a tumor of the breast that is associated
with a positive sentinel lymph node (SLN) biopsy. A breast cancer
that is to be treated can include a tumor of the breast that is
associated with one or more positive axillary lymph nodes, where
the axillary lymph nodes have been staged by any applicable method.
A breast cancer that is to be treated can include a tumor of the
breast that has been typed as having nodal negative status (e.g.,
node-negative) or nodal positive status (e.g., node-positive). A
breast cancer that is to be treated can include a tumor of the
breast that has metastasized to other locations in the body. A
breast cancer that is to be treated can be classified as having
metastasized to a location selected from the group consisting of
bone, lung, liver, or brain. A breast cancer that is to be treated
can be classified according to a characteristic selected from the
group consisting of metastatic, localized, regional,
local-regional, locally advanced, distant, multicentric, bilateral,
ipsilateral, contralateral, newly diagnosed, recurrent, and
inoperable.
[0539] A compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
may be used to treat or prevent a cell proliferative disorder of
the breast, or to treat or prevent breast cancer, in a subject
having an increased risk of developing breast cancer relative to
the population at large. A subject with an increased risk of
developing breast cancer relative to the population at large is a
female subject with a family history or personal history of breast
cancer. A subject with an increased risk of developing breast
cancer relative to the population at large is a female subject
having a germ-line or spontaneous mutation in BRCA1 or BRCA2, or
both. A subject with an increased risk of developing breast cancer
relative to the population at large is a female subject with a
family history of breast cancer and a germ-line or spontaneous
mutation in BRCA1 or BRCA2, or both. A subject with an increased
risk of developing breast cancer relative to the population at
large is a female who is greater than 30 years old, greater than 40
years old, greater than 50 years old, greater than 60 years old,
greater than 70 years old, greater than 80 years old, or greater
than 90 years old. A subject with an increased risk of developing
breast cancer relative to the population at large is a subject with
atypical hyperplasia of the breast, ductal carcinoma in situ
(DCIS), intraductal carcinoma, lobular carcinoma in situ (LCIS),
lobular neoplasia, or a stage 0 growth or lesion of the breast
(e.g., stage 0 or grade 0 breast cancer, or carcinoma in situ).
[0540] A breast cancer that is to be treated can histologically
graded according to the Scarff-Bloom-Richardson system, wherein a
breast tumor has been assigned a mitosis count score of 1, 2, or 3;
a nuclear pleiomorphism score of 1, 2, or 3; a tubule formation
score of 1, 2, or 3; and a total Scarff-Bloom-Richardson score of
between 3 and 9. A breast cancer that is to be treated can be
assigned a tumor grade according to the International Consensus
Panel on the Treatment of Breast Cancer selected from the group
consisting of grade 1, grade 1-2, grade 2, grade 2-3, or grade
3.
[0541] A cancer that is to be treated can be staged according to
the American Joint Committee on Cancer (AJCC) TNM classification
system, where the tumor (T) has been assigned a stage of TX,
T.sub.1, T1mic, T1a, T1b, T1c, T2, T3, T4, T4a, T4b, T4c, or T4d;
and where the regional lymph nodes (N) have been assigned a stage
of NX, N0, N1, N2, N2a, N2b, N3, N3a, N3b, or N3c; and where
distant metastasis (M) can be assigned a stage of MX, M0, or
M.sub.1. A cancer that is to be treated can be staged according to
an American Joint Committee on Cancer (AJCC) classification as
Stage I, Stage IIA, Stage IIB, Stage IIIA, Stage IIIB, Stage IIIC,
or Stage IV. A cancer that is to be treated can be assigned a grade
according to an AJCC classification as Grade GX (e.g., grade cannot
be assessed), Grade 1, Grade 2, Grade 3 or Grade 4. A cancer that
is to be treated can be staged according to an AJCC pathologic
classification (pN) of pNX, pNO, PN0(I-), PN0(I+), PN0 (mol-), PN0
(mol+), PN1, PN1 (mi), PN1a, PN1b, PN1c, pN2, pN2a, pN2b, pN3,
pN3a, pN3b, or pN3c.
[0542] A cancer that is to be treated can include a tumor that has
been determined to be less than or equal to about 2 centimeters in
diameter. A cancer that is to be treated can include a tumor that
has been determined to be from about 2 to about 5 centimeters in
diameter. A cancer that is to be treated can include a tumor that
has been determined to be greater than or equal to about 3
centimeters in diameter. A cancer that is to be treated can include
a tumor that has been determined to be greater than 5 centimeters
in diameter. A cancer that is to be treated can be classified by
microscopic appearance as well differentiated, moderately
differentiated, poorly differentiated, or undifferentiated. A
cancer that is to be treated can be classified by microscopic
appearance with respect to mitosis count (e.g., amount of cell
division) or nuclear pleiomorphism (e.g., change in cells). A
cancer that is to be treated can be classified by microscopic
appearance as being associated with areas of necrosis (e.g., areas
of dying or degenerating cells). A cancer that is to be treated can
be classified as having an abnormal karyotype, having an abnormal
number of chromosomes, or having one or more chromosomes that are
abnormal in appearance. A cancer that is to be treated can be
classified as being aneuploid, triploid, tetraploid, or as having
an altered ploidy. A cancer that is to be treated can be classified
as having a chromosomal translocation, or a deletion or duplication
of an entire chromosome, or a region of deletion, duplication or
amplification of a portion of a chromosome.
[0543] A cancer that is to be treated can be evaluated by DNA
cytometry, flow cytometry, or image cytometry. A cancer that is to
be treated can be typed as having 10%, 20%, 30%, 40%, 50%, 60%,
70%, 80%, or 90% of cells in the synthesis stage of cell division
(e.g., in S phase of cell division). A cancer that is to be treated
can be typed as having a low S-phase fraction or a high S-phase
fraction.
[0544] As used herein, a "normal cell" is a cell that cannot be
classified as part of a "cell proliferative disorder". A normal
cell lacks unregulated or abnormal growth, or both, that can lead
to the development of an unwanted condition or disease. Preferably,
a normal cell possesses normally functioning cell cycle checkpoint
control mechanisms.
[0545] As used herein, "contacting a cell" refers to a condition in
which a compound or other composition of matter is in direct
contact with a cell, or is close enough to induce a desired
biological effect in a cell.
[0546] As used herein, "candidate compound" refers to a compound of
the present invention, or a pharmaceutically acceptable salt,
prodrug, metabolite, polymorph or solvate thereof, that has been or
will be tested in one or more in vitro or in vivo biological
assays, in order to determine if that compound is likely to elicit
a desired biological or medical response in a cell, tissue, system,
animal or human that is being sought by a researcher or clinician.
A candidate compound is a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof. The biological or medical response can be the
treatment of cancer. The biological or medical response can be
treatment or prevention of a cell proliferative disorder. In vitro
or in vivo biological assays can include, but are not limited to,
enzymatic activity assays, electrophoretic mobility shift assays,
reporter gene assays, in vitro cell viability assays, and the
assays described herein.
[0547] As used herein, "monotherapy" refers to the administration
of a single active or therapeutic compound to a subject in need
thereof. Preferably, monotherapy will involve administration of a
therapeutically effective amount of an single active compound. For
example, cancer monotherapy with one of the compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or derivative thereof, to a subject in need of
treatment of cancer. In one aspect, the single active compound is a
compound of the present invention, or a pharmaceutically acceptable
salt, prodrug, metabolite, polymorph or solvate thereof.
[0548] As used herein, "treating" or "treat" describes the
management and care of a patient for the purpose of combating a
disease, condition, or disorder and includes the administration of
a compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
to alleviate the symptoms or complications of a disease, condition
or disorder, or to eliminate the disease, condition or
disorder.
[0549] A compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
can also be used to prevent a disease, condition or disorder. As
used herein, "preventing" or "prevent" describes reducing or
eliminating the onset of the symptoms or complications of the
disease, condition or disorder.
[0550] As used herein, the term "alleviate" is meant to describe a
process by which the severity of a sign or symptom of a disorder is
decreased. Importantly, a sign or symptom can be alleviated without
being eliminated. In a preferred embodiment, the administration of
pharmaceutical compositions of the invention leads to the
elimination of a sign or symptom, however, elimination is not
required. Effective dosages are expected to decrease the severity
of a sign or symptom. For instance, a sign or symptom of a disorder
such as cancer, which can occur in multiple locations, is
alleviated if the severity of the cancer is decreased within at
least one of multiple locations.
[0551] As used herein, the term "severity" is meant to describe the
potential of cancer to transform from a precancerous, or benign,
state into a malignant state. Alternatively, or in addition,
severity is meant to describe a cancer stage, for example,
according to the TNM system (accepted by the International Union
Against Cancer (UICC) and the American Joint Committee on Cancer
(AJCC)) or by other art-recognized methods. Cancer stage refers to
the extent or severity of the cancer, based on factors such as the
location of the primary tumor, tumor size, number of tumors, and
lymph node involvement (spread of cancer into lymph nodes).
Alternatively, or in addition, severity is meant to describe the
tumor grade by art-recognized methods (see, National Cancer
Institute, www.cancer.gov). Tumor grade is a system used to
classify cancer cells in terms of how abnormal they look under a
microscope and how quickly the tumor is likely to grow and spread.
Many factors are considered when determining tumor grade, including
the structure and growth pattern of the cells. The specific factors
used to determine tumor grade vary with each type of cancer.
Severity also describes a histologic grade, also called
differentiation, which refers to how much the tumor cells resemble
normal cells of the same tissue type (see, National Cancer
Institute, www.cancer.gov). Furthermore, severity describes a
nuclear grade, which refers to the size and shape of the nucleus in
tumor cells and the percentage of tumor cells that are dividing
(see, National Cancer Institute, www.cancer.gov).
[0552] In another aspect of the invention, severity describes the
degree to which a tumor has secreted growth factors, degraded the
extracellular matrix, become vascularized, lost adhesion to
juxtaposed tissues, or metastasized. Moreover, severity describes
the number of locations to which a primary tumor has metastasized.
Finally, severity includes the difficulty of treating tumors of
varying types and locations. For example, inoperable tumors, those
cancers which have greater access to multiple body systems
(hematological and immunological tumors), and those which are the
most resistant to traditional treatments are considered most
severe. In these situations, prolonging the life expectancy of the
subject and/or reducing pain, decreasing the proportion of
cancerous cells or restricting cells to one system, and improving
cancer stage/tumor grade/histological grade/nuclear grade are
considered alleviating a sign or symptom of the cancer.
[0553] As used herein the term "symptom" is defined as an
indication of disease, illness, injury, or that something is not
right in the body. Symptoms are felt or noticed by the individual
experiencing the symptom, but may not easily be noticed by others.
Others are defined as non-health-care professionals.
[0554] As used herein the term "sign" is also defined as an
indication that something is not right in the body. But signs are
defined as things that can be seen by a doctor, nurse, or other
health care professional.
[0555] Cancer is a group of diseases that may cause almost any sign
or symptom. The signs and symptoms will depend on where the cancer
is, the size of the cancer, and how much it affects the nearby
organs or structures. If a cancer spreads (metastasizes), then
symptoms may appear in different parts of the body.
[0556] As a cancer grows, it begins to push on nearby organs, blood
vessels, and nerves. This pressure creates some of the signs and
symptoms of cancer. If the cancer is in a critical area, such as
certain parts of the brain, even the smallest tumor can cause early
symptoms.
[0557] But sometimes cancers start in places where it does not
cause any symptoms until the cancer has grown quite large. Pancreas
cancers, for example, do not usually grow large enough to be felt
from the outside of the body. Some pancreatic cancers do not cause
symptoms until they begin to grow around nearby nerves (this causes
a backache). Others grow around the bile duct, which blocks the
flow of bile and leads to a yellowing of the skin known as
jaundice. By the time a pancreatic cancer causes these signs or
symptoms, it has usually reached an advanced stage.
[0558] A cancer may also cause symptoms such as fever, fatigue, or
weight loss. This may be because cancer cells use up much of the
body's energy supply or release substances that change the body's
metabolism. Or the cancer may cause the immune system to react in
ways that produce these symptoms.
[0559] Sometimes, cancer cells release substances into the
bloodstream that cause symptoms not usually thought to result from
cancers. For example, some cancers of the pancreas can release
substances which cause blood clots to develop in veins of the legs.
Some lung cancers make hormone-like substances that affect blood
calcium levels, affecting nerves and muscles and causing weakness
and dizziness
[0560] Cancer presents several general signs or symptoms that occur
when a variety of subtypes of cancer cells are present. Most people
with cancer will lose weight at some time with their disease. An
unexplained (unintentional) weight loss of 10 pounds or more may be
the first sign of cancer, particularly cancers of the pancreas,
stomach, esophagus, or lung.
[0561] Fever is very common with cancer, but is more often seen in
advanced disease. Almost all patients with cancer will have fever
at some time, especially if the cancer or its treatment affects the
immune system and makes it harder for the body to fight infection.
Less often, fever may be an early sign of cancer, such as with
leukemia or lymphoma.
[0562] Fatigue may be an important symptom as cancer progresses. It
may happen early, though, in cancers such as with leukemia, or if
the cancer is causing an ongoing loss of blood, as in some colon or
stomach cancers.
[0563] Pain may be an early symptom with some cancers such as bone
cancers or testicular cancer. But most often pain is a symptom of
advanced disease.
[0564] Along with cancers of the skin (see next section), some
internal cancers can cause skin signs that can be seen. These
changes include the skin looking darker (hyperpigmentation), yellow
(jaundice), or red (erythema); itching; or excessive hair
growth.
[0565] Alternatively, or in addition, cancer subtypes present
specific signs or symptoms. Changes in bowel habits or bladder
function could indicate cancer. Long-term constipation, diarrhea,
or a change in the size of the stool may be a sign of colon cancer.
Pain with urination, blood in the urine, or a change in bladder
function (such as more frequent or less frequent urination) could
be related to bladder or prostate cancer.
[0566] Changes in skin condition or appearance of a new skin
condition could indicate cancer. Skin cancers may bleed and look
like sores that do not heal. A long-lasting sore in the mouth could
be an oral cancer, especially in patients who smoke, chew tobacco,
or frequently drink alcohol. Sores on the penis or vagina may
either be signs of infection or an early cancer.
[0567] Unusual bleeding or discharge could indicate cancer. Unusual
bleeding can happen in either early or advanced cancer. Blood in
the sputum (phlegm) may be a sign of lung cancer. Blood in the
stool (or a dark or black stool) could be a sign of colon or rectal
cancer. Cancer of the cervix or the endometrium (lining of the
uterus) can cause vaginal bleeding. Blood in the urine may be a
sign of bladder or kidney cancer. A bloody discharge from the
nipple may be a sign of breast cancer.
[0568] A thickening or lump in the breast or in other parts of the
body could indicate the presence of a cancer. Many cancers can be
felt through the skin, mostly in the breast, testicle, lymph nodes
(glands), and the soft tissues of the body. A lump or thickening
may be an early or late sign of cancer. Any lump or thickening
could be indicative of cancer, especially if the formation is new
or has grown in size.
[0569] Indigestion or trouble swallowing could indicate cancer.
While these symptoms commonly have other causes, indigestion or
swallowing problems may be a sign of cancer of the esophagus,
stomach, or pharynx (throat).
[0570] Recent changes in a wart or mole could be indicative of
cancer. Any wart, mole, or freckle that changes in color, size, or
shape, or loses its definite borders indicates the potential
development of cancer. For example, the skin lesion may be a
melanoma.
[0571] A persistent cough or hoarseness could be indicative of
cancer. A cough that does not go away may be a sign of lung cancer.
Hoarseness can be a sign of cancer of the larynx (voice box) or
thyroid.
[0572] While the signs and symptoms listed above are the more
common ones seen with cancer, there are many others that are less
common and are not listed here. However, all art-recognized signs
and symptoms of cancer are contemplated and encompassed by the
instant invention.
[0573] Treating cancer can result in a reduction in size of a
tumor. A reduction in size of a tumor may also be referred to as
"tumor regression". Preferably, after treatment, tumor size is
reduced by 5% or greater relative to its size prior to treatment;
more preferably, tumor size is reduced by 10% or greater; more
preferably, reduced by 20% or greater; more preferably, reduced by
30% or greater; more preferably, reduced by 40% or greater; even
more preferably, reduced by 50% or greater; and most preferably,
reduced by greater than 75% or greater. Size of a tumor may be
measured by any reproducible means of measurement. The size of a
tumor may be measured as a diameter of the tumor.
[0574] Treating cancer can result in a reduction in tumor volume.
Preferably, after treatment, tumor volume is reduced by 5% or
greater relative to its size prior to treatment; more preferably,
tumor volume is reduced by 10% or greater; more preferably, reduced
by 20% or greater; more preferably, reduced by 30% or greater; more
preferably, reduced by 40% or greater; even more preferably,
reduced by 50% or greater; and most preferably, reduced by greater
than 75% or greater. Tumor volume may be measured by any
reproducible means of measurement.
[0575] Treating cancer results in a decrease in number of tumors.
Preferably, after treatment, tumor number is reduced by 5% or
greater relative to number prior to treatment; more preferably,
tumor number is reduced by 10% or greater; more preferably, reduced
by 20% or greater; more preferably, reduced by 30% or greater; more
preferably, reduced by 40% or greater; even more preferably,
reduced by 50% or greater; and most preferably, reduced by greater
than 75%. Number of tumors may be measured by any reproducible
means of measurement. The number of tumors may be measured by
counting tumors visible to the naked eye or at a specified
magnification. Preferably, the specified magnification is 2.times.,
3.times., 4.times., 5.times., 10.times., or 50.times..
[0576] Treating cancer can result in a decrease in number of
metastatic lesions in other tissues or organs distant from the
primary tumor site. Preferably, after treatment, the number of
metastatic lesions is reduced by 5% or greater relative to number
prior to treatment; more preferably, the number of metastatic
lesions is reduced by 10% or greater; more preferably, reduced by
20% or greater; more preferably, reduced by 30% or greater; more
preferably, reduced by 40% or greater; even more preferably,
reduced by 50% or greater; and most preferably, reduced by greater
than 75%. The number of metastatic lesions may be measured by any
reproducible means of measurement. The number of metastatic lesions
may be measured by counting metastatic lesions visible to the naked
eye or at a specified magnification. Preferably, the specified
magnification is 2.times., 3.times., 4.times., 5.times., 10.times.,
or 50.times..
[0577] Treating cancer can result in an increase in average
survival time of a population of treated subjects in comparison to
a population receiving carrier alone. Preferably, the average
survival time is increased by more than 30 days; more preferably,
by more than 60 days; more preferably, by more than 90 days; and
most preferably, by more than 120 days. An increase in average
survival time of a population may be measured by any reproducible
means. An increase in average survival time of a population may be
measured, for example, by calculating for a population the average
length of survival following initiation of treatment with an active
compound. An increase in average survival time of a population may
also be measured, for example, by calculating for a population the
average length of survival following completion of a first round of
treatment with an active compound.
[0578] Treating cancer can result in an increase in average
survival time of a population of treated subjects in comparison to
a population of untreated subjects. Preferably, the average
survival time is increased by more than 30 days; more preferably,
by more than 60 days; more preferably, by more than 90 days; and
most preferably, by more than 120 days. An increase in average
survival time of a population may be measured by any reproducible
means. An increase in average survival time of a population may be
measured, for example, by calculating for a population the average
length of survival following initiation of treatment with an active
compound. An increase in average survival time of a population may
also be measured, for example, by calculating for a population the
average length of survival following completion of a first round of
treatment with an active compound.
[0579] Treating cancer can result in increase in average survival
time of a population of treated subjects in comparison to a
population receiving monotherapy with a drug that is not a compound
of the present invention, or a pharmaceutically acceptable salt,
prodrug, metabolite, analog or derivative thereof. Preferably, the
average survival time is increased by more than 30 days; more
preferably, by more than 60 days; more preferably, by more than 90
days; and most preferably, by more than 120 days. An increase in
average survival time of a population may be measured by any
reproducible means. An increase in average survival time of a
population may be measured, for example, by calculating for a
population the average length of survival following initiation of
treatment with an active compound. An increase in average survival
time of a population may also be measured, for example, by
calculating for a population the average length of survival
following completion of a first round of treatment with an active
compound.
[0580] Treating cancer can result in a decrease in the mortality
rate of a population of treated subjects in comparison to a
population receiving carrier alone. Treating cancer can result in a
decrease in the mortality rate of a population of treated subjects
in comparison to an untreated population. Treating cancer can
result in a decrease in the mortality rate of a population of
treated subjects in comparison to a population receiving
monotherapy with a drug that is not a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or derivative thereof. Preferably, the mortality
rate is decreased by more than 2%; more preferably, by more than
5%; more preferably, by more than 10%; and most preferably, by more
than 25%. A decrease in the mortality rate of a population of
treated subjects may be measured by any reproducible means. A
decrease in the mortality rate of a population may be measured, for
example, by calculating for a population the average number of
disease-related deaths per unit time following initiation of
treatment with an active compound. A decrease in the mortality rate
of a population may also be measured, for example, by calculating
for a population the average number of disease-related deaths per
unit time following completion of a first round of treatment with
an active compound.
[0581] Treating cancer can result in a decrease in tumor growth
rate. Preferably, after treatment, tumor growth rate is reduced by
at least 5% relative to number prior to treatment; more preferably,
tumor growth rate is reduced by at least 10%; more preferably,
reduced by at least 20%; more preferably, reduced by at least 30%;
more preferably, reduced by at least 40%; more preferably, reduced
by at least 50%; even more preferably, reduced by at least 50%; and
most preferably, reduced by at least 75%. Tumor growth rate may be
measured by any reproducible means of measurement. Tumor growth
rate can be measured according to a change in tumor diameter per
unit time.
[0582] Treating cancer can result in a decrease in tumor regrowth.
Preferably, after treatment, tumor regrowth is less than 5%; more
preferably, tumor regrowth is less than 10%; more preferably, less
than 20%; more preferably, less than 30%; more preferably, less
than 40%; more preferably, less than 50%; even more preferably,
less than 50%; and most preferably, less than 75%. Tumor regrowth
may be measured by any reproducible means of measurement. Tumor
regrowth is measured, for example, by measuring an increase in the
diameter of a tumor after a prior tumor shrinkage that followed
treatment. A decrease in tumor regrowth is indicated by failure of
tumors to reoccur after treatment has stopped.
[0583] Treating or preventing a cell proliferative disorder can
result in a reduction in the rate of cellular proliferation.
Preferably, after treatment, the rate of cellular proliferation is
reduced by at least 5%; more preferably, by at least 10%; more
preferably, by at least 20%; more preferably, by at least 30%; more
preferably, by at least 40%; more preferably, by at least 50%; even
more preferably, by at least 50%; and most preferably, by at least
75%. The rate of cellular proliferation may be measured by any
reproducible means of measurement. The rate of cellular
proliferation is measured, for example, by measuring the number of
dividing cells in a tissue sample per unit time.
[0584] Treating or preventing a cell proliferative disorder can
result in a reduction in the proportion of proliferating cells.
Preferably, after treatment, the proportion of proliferating cells
is reduced by at least 5%; more preferably, by at least 10%; more
preferably, by at least 20%; more preferably, by at least 30%; more
preferably, by at least 40%; more preferably, by at least 50%; even
more preferably, by at least 50%; and most preferably, by at least
75%. The proportion of proliferating cells may be measured by any
reproducible means of measurement. Preferably, the proportion of
proliferating cells is measured, for example, by quantifying the
number of dividing cells relative to the number of nondividing
cells in a tissue sample. The proportion of proliferating cells can
be equivalent to the mitotic index.
[0585] Treating or preventing a cell proliferative disorder can
result in a decrease in size of an area or zone of cellular
proliferation. Preferably, after treatment, size of an area or zone
of cellular proliferation is reduced by at least 5% relative to its
size prior to treatment; more preferably, reduced by at least 10%;
more preferably, reduced by at least 20%; more preferably, reduced
by at least 30%; more preferably, reduced by at least 40%; more
preferably, reduced by at least 50%; even more preferably, reduced
by at least 50%; and most preferably, reduced by at least 75%. Size
of an area or zone of cellular proliferation may be measured by any
reproducible means of measurement. The size of an area or zone of
cellular proliferation may be measured as a diameter or width of an
area or zone of cellular proliferation.
[0586] Treating or preventing a cell proliferative disorder can
result in a decrease in the number or proportion of cells having an
abnormal appearance or morphology. Preferably, after treatment, the
number of cells having an abnormal morphology is reduced by at
least 5% relative to its size prior to treatment; more preferably,
reduced by at least 10%; more preferably, reduced by at least 20%;
more preferably, reduced by at least 30%; more preferably, reduced
by at least 40%; more preferably, reduced by at least 50%; even
more preferably, reduced by at least 50%; and most preferably,
reduced by at least 75%. An abnormal cellular appearance or
morphology may be measured by any reproducible means of
measurement. An abnormal cellular morphology can be measured by
microscopy, e.g., using an inverted tissue culture microscope. An
abnormal cellular morphology can take the form of nuclear
pleiomorphism.
[0587] As used herein, the term "selectively" means tending to
occur at a higher frequency in one population than in another
population. The compared populations can be cell populations.
Preferably, a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof, acts selectively on a cancer or precancerous cell
but not on a normal cell. Preferably, a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, acts selectively to
modulate one molecular target (e.g., a target protein
methyltransferase) but does not significantly modulate another
molecular target (e.g., a non-target protein methyltransferase).
The invention also provides a method for selectively inhibiting the
activity of an enzyme, such as a protein methyltransferase.
Preferably, an event occurs selectively in population A relative to
population B if it occurs greater than two times more frequently in
population A as compared to population B. An event occurs
selectively if it occurs greater than five times more frequently in
population A. An event occurs selectively if it occurs greater than
ten times more frequently in population A; more preferably, greater
than fifty times; even more preferably, greater than 100 times; and
most preferably, greater than 1000 times more frequently in
population A as compared to population B. For example, cell death
would be said to occur selectively in cancer cells if it occurred
greater than twice as frequently in cancer cells as compared to
normal cells.
[0588] A compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
can modulate the activity of a molecular target (e.g., a target
protein methyltransferase). Modulating refers to stimulating or
inhibiting an activity of a molecular target. Preferably, a
compound of the present invention, or a pharmaceutically acceptable
salt, prodrug, metabolite, polymorph or solvate thereof, modulates
the activity of a molecular target if it stimulates or inhibits the
activity of the molecular target by at least 2-fold relative to the
activity of the molecular target under the same conditions but
lacking only the presence of said compound. More preferably, a
compound of the present invention, or a pharmaceutically acceptable
salt, prodrug, metabolite, polymorph or solvate thereof, modulates
the activity of a molecular target if it stimulates or inhibits the
activity of the molecular target by at least 5-fold, at least
10-fold, at least 20-fold, at least 50-fold, at least 100-fold
relative to the activity of the molecular target under the same
conditions but lacking only the presence of said compound. The
activity of a molecular target may be measured by any reproducible
means. The activity of a molecular target may be measured in vitro
or in vivo. For example, the activity of a molecular target may be
measured in vitro by an enzymatic activity assay or a DNA binding
assay, or the activity of a molecular target may be measured in
vivo by assaying for expression of a reporter gene.
[0589] A compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
does not significantly modulate the activity of a molecular target
if the addition of the compound does not stimulate or inhibit the
activity of the molecular target by greater than 10% relative to
the activity of the molecular target under the same conditions but
lacking only the presence of said compound.
[0590] As used herein, the term "isozyme selective" means
preferential inhibition or stimulation of a first isoform of an
enzyme in comparison to a second isoform of an enzyme (e.g.,
preferential inhibition or stimulation of a protein
methyltransferase isozyme alpha in comparison to a protein
methyltransferase isozyme beta). Preferably, a compound of the
present invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, demonstrates a minimum of
a fourfold differential, preferably a tenfold differential, more
preferably a fifty fold differential, in the dosage required to
achieve a biological effect. Preferably, a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, demonstrates this
differential across the range of inhibition, and the differential
is exemplified at the IC.sub.50, i.e., a 50% inhibition, for a
molecular target of interest.
[0591] Administering a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof, to a cell or a subject in need thereof can result
in modulation (i.e., stimulation or inhibition) of an activity of a
protein methyltransferase of interest.
[0592] The present invention provides methods to assess biological
activity of a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof or methods of identifying a test compound as a
modulator (e.g., an inhibitor) of DOT1L. DOT1L polypeptides and
nucleic acids can be used to screen for compounds that bind to
and/or modulate (e.g., increase or decrease) one or more biological
activities of DOT1L, including but not limited to H3K79 HMTase
activity, SAM binding activity, histone and/or nucleosome binding
activity, AF10 binding activity, AF10-MLL or other MLL fusion
protein binding activity, and/or any other biological activity of
interest. A DOT1L polypeptide can be a functional fragment of a
full-length DOT1L polypeptide or functional equivalent thereof, and
may comprise any DOT1 domain of interest, including but not limited
to the catalytic domain, the SAM binding domain and/or the
positively charged domain, the AF10 interaction domain and/or a
nuclear export signal.
[0593] Methods of assessing DOT1L binding to histones, nucleosomes,
nucleic acids or polypeptides can be carried out using standard
techniques that will be apparent to those skilled in the art (see
the Exemplification for exemplary methods). Such methods include
yeast and mammalian two-hybrid assays and co-immunoprecipitation
techniques.
[0594] For example, a compound that modulates DOT1L H3K79 HMTase
activity can be verified by: contacting a DOT1L polypeptide with a
histone or peptide substrate comprising H3 in the presence of a
test compound; detecting the level of H3K79 methylation of the
histone or peptide substrate under conditions sufficient to provide
H3K79 methylation, wherein an elevation or reduction in H3K79
methylation in the presence of the test compound as compared with
the level of histone H3K79 methylation in the absence of the test
compound indicates that the test compound modulates DOT1L H3K79
HMTase activity.
[0595] The screening methods of the invention can be carried out in
a cell-based or cell-free system. As a further alternative, the
assay can be performed in a whole animal (including transgenic
non-human animals). Further, with respect to cell-based systems,
the DOT1L polypeptide (or any other polypeptide used in the assay)
can be added directly to the cell or can be produced from a nucleic
acid in the cell. The nucleic acid can be endogenous to the cell or
can be foreign (e.g., a genetically modified cell).
[0596] In some assays, immunological reagents, e.g., antibodies and
antigens, are employed. Fluorescence can be utilized in the
measurement of enzymatic activity in some assays. As used herein,
"fluorescence" refers to a process through which a molecule emits a
photon as a result of absorbing an incoming photon of higher energy
by the same molecule. Specific methods for assessing the biological
activity of the disclosed compounds are described in the
examples.
[0597] Administering a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof, to a cell or a subject in need thereof results in
modulation (i.e., stimulation or inhibition) of an activity of an
intracellular target (e.g., substrate). Several intracellular
targets can be modulated with the compounds of the present
invention, including, but not limited to, protein
methyltrasferase.
[0598] Activating refers to placing a composition of matter (e.g.,
protein or nucleic acid) in a state suitable for carrying out a
desired biological function. A composition of matter capable of
being activated also has an unactivated state. An activated
composition of matter may have an inhibitory or stimulatory
biological function, or both.
[0599] Elevation refers to an increase in a desired biological
activity of a composition of matter (e.g., a protein or a nucleic
acid). Elevation may occur through an increase in concentration of
a composition of matter.
[0600] As used herein, "a cell cycle checkpoint pathway" refers to
a biochemical pathway that is involved in modulation of a cell
cycle checkpoint. A cell cycle checkpoint pathway may have
stimulatory or inhibitory effects, or both, on one or more
functions comprising a cell cycle checkpoint. A cell cycle
checkpoint pathway is comprised of at least two compositions of
matter, preferably proteins, both of which contribute to modulation
of a cell cycle checkpoint. A cell cycle checkpoint pathway may be
activated through an activation of one or more members of the cell
cycle checkpoint pathway. Preferably, a cell cycle checkpoint
pathway is a biochemical signaling pathway.
[0601] As used herein, "cell cycle checkpoint regulator" refers to
a composition of matter that can function, at least in part, in
modulation of a cell cycle checkpoint. A cell cycle checkpoint
regulator may have stimulatory or inhibitory effects, or both, on
one or more functions comprising a cell cycle checkpoint. A cell
cycle checkpoint regulator can be a protein or not a protein.
[0602] Treating cancer or a cell proliferative disorder can result
in cell death, and preferably, cell death results in a decrease of
at least 10% in number of cells in a population. More preferably,
cell death means a decrease of at least 20%; more preferably, a
decrease of at least 30%; more preferably, a decrease of at least
40%; more preferably, a decrease of at least 50%; most preferably,
a decrease of at least 75%. Number of cells in a population may be
measured by any reproducible means. A number of cells in a
population can be measured by fluorescence activated cell sorting
(FACS), immunofluorescence microscopy and light microscopy. Methods
of measuring cell death are as shown in L.sub.1 et al., Proc Natl
Acad Sci USA. 100(5): 2674-8, 2003. In an aspect, cell death occurs
by apoptosis.
[0603] Preferably, an effective amount of a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, is not significantly
cytotoxic to normal cells. A therapeutically effective amount of a
compound is not significantly cytotoxic to normal cells if
administration of the compound in a therapeutically effective
amount does not induce cell death in greater than 10% of normal
cells. A therapeutically effective amount of a compound does not
significantly affect the viability of normal cells if
administration of the compound in a therapeutically effective
amount does not induce cell death in greater than 10% of normal
cells. In an aspect, cell death occurs by apoptosis.
[0604] Contacting a cell with a compound of the present invention,
or a pharmaceutically acceptable salt, prodrug, metabolite,
polymorph or solvate thereof, can induce or activate cell death
selectively in cancer cells. Administering to a subject in need
thereof a compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
can induce or activate cell death selectively in cancer cells.
Contacting a cell with a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof, can induce cell death selectively in one or more
cells affected by a cell proliferative disorder. Preferably,
administering to a subject in need thereof a compound of the
present invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, induces cell death
selectively in one or more cells affected by a cell proliferative
disorder.
[0605] The present invention relates to a method of treating or
preventing cancer by administering a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, to a subject in need
thereof, where administration of the compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, results in one or more of
the following: accumulation of cells in G1 and/or S phase of the
cell cycle, cytotoxicity via cell death in cancer cells without a
significant amount of cell death in normal cells, antitumor
activity in animals with a therapeutic index of at least 2, and
activation of a cell cycle checkpoint. As used herein, "therapeutic
index" is the maximum tolerated dose divided by the efficacious
dose.
[0606] One skilled in the art may refer to general reference texts
for detailed descriptions of known techniques discussed herein or
equivalent techniques. These texts include Ausubel et al., Current
Protocols in Molecular Biology, John Wiley and Sons, Inc. (2005);
Sambrook et al., Molecular Cloning, A Laboratory Manual (3.sup.rd
edition), Cold Spring Harbor Press, Cold Spring Harbor, N.Y.
(2000); Coligan et al., Current Protocols in Immunology, John Wiley
& Sons, N.Y.; Enna et al., Current Protocols in Pharmacology,
John Wiley & Sons, N.Y.; Fingl et al., The Pharmacological
Basis of Therapeutics (1975), Remington's Pharmaceutical Sciences,
Mack Publishing Co., Easton, Pa., 18.sup.th edition (1990). These
texts can, of course, also be referred to in making or using an
aspect of the invention
[0607] The compounds of the instant invention can also be utilized
to treat or prevent neurologic diseases or disorders. Neurologic
diseases or disorders that may be treated with the compounds of
this invention include epilepsy, schizophrenia, bipolar disorder or
other psychological and/or psychiatric disorders, neuropathies,
skeletal muscle atrophy, and neurodegenerative diseases, e.g., a
neurodegenerative disease. Exemplary neurodegenerative diseases
include: Alzheimer's, Amyotrophic Lateral Sclerosis (ALS), and
Parkinson's disease. Another class of neurodegenerative diseases
includes diseases caused at least in part by aggregation of
poly-glutamine. Diseases of this class include: Huntington's
Diseases, Spinalbulbar Muscular Atrophy (SBMA or Kennedy's Disease)
Dentatorubropallidoluysian Atrophy (DRPLA), Spinocerebellar Ataxia
1 (SCA1), Spinocerebellar Ataxia 2 (SCA2), Machado-Joseph Disease
(MJD; SCA3), Spinocerebellar Ataxia 6 (SCA6), Spinocerebellar
Ataxia 7 (SCA7), and Spinocerebellar Ataxia 12 (SCA12).
[0608] Any other disease in which epigenetic methylation, which is
mediated by DOT1, plays a role may be treatable or preventable
using compounds and methods described herein.
4. PHARMACEUTICAL COMPOSITIONS
[0609] The present invention also provides pharmaceutical
compositions comprising a compound of Formulae (I), (II), (IIIa),
(IIIb), (IIIc) and (IV) in combination with at least one
pharmaceutically acceptable excipient or carrier.
[0610] A "pharmaceutical composition" is a formulation containing
the compounds of the present invention in a form suitable for
administration to a subject. In one embodiment, the pharmaceutical
composition is in bulk or in unit dosage form. The unit dosage form
is any of a variety of forms, including, for example, a capsule, an
IV bag, a tablet, a single pump on an aerosol inhaler or a vial.
The quantity of active ingredient (e.g., a formulation of the
disclosed compound or salt, hydrate, solvate or isomer thereof) in
a unit dose of composition is an effective amount and is varied
according to the particular treatment involved. One skilled in the
art will appreciate that it is sometimes necessary to make routine
variations to the dosage depending on the age and condition of the
patient. The dosage will also depend on the route of
administration. A variety of routes are contemplated, including
oral, pulmonary, rectal, parenteral, transdermal, subcutaneous,
intravenous, intramuscular, intraperitoneal, inhalational, buccal,
sublingual, intrapleural, intrathecal, intranasal, and the like.
Dosage forms for the topical or transdermal administration of a
compound of this invention include powders, sprays, ointments,
pastes, creams, lotions, gels, solutions, patches and inhalants. In
one embodiment, the active compound is mixed under sterile
conditions with a pharmaceutically acceptable carrier, and with any
preservatives, buffers, or propellants that are required.
[0611] As used herein, the phrase "pharmaceutically acceptable"
refers to those compounds, materials, compositions, carriers,
and/or dosage forms which are, within the scope of sound medical
judgment, suitable for use in contact with the tissues of human
beings and animals without excessive toxicity, irritation, allergic
response, or other problem or complication, commensurate with a
reasonable benefit/risk ratio.
[0612] "Pharmaceutically acceptable excipient" means an excipient
that is useful in preparing a pharmaceutical composition that is
generally safe, non-toxic and neither biologically nor otherwise
undesirable, and includes excipient that is acceptable for
veterinary use as well as human pharmaceutical use. A
"pharmaceutically acceptable excipient" as used in the
specification and claims includes both one and more than one such
excipient.
[0613] A pharmaceutical composition of the invention is formulated
to be compatible with its intended route of administration.
Examples of routes of administration include parenteral, e.g.,
intravenous, intradermal, subcutaneous, oral (e.g., inhalation),
transdermal (topical), and transmucosal administration. Solutions
or suspensions used for parenteral, intradermal, or subcutaneous
application can include the following components: a sterile diluent
such as water for injection, saline solution, fixed oils,
polyethylene glycols, glycerine, propylene glycol or other
synthetic solvents; antibacterial agents such as benzyl alcohol or
methyl parabens; antioxidants such as ascorbic acid or sodium
bisulfite; chelating agents such as ethylenediaminetetraacetic
acid; buffers such as acetates, citrates or phosphates, and agents
for the adjustment of tonicity such as sodium chloride or dextrose.
The pH can be adjusted with acids or bases, such as hydrochloric
acid or sodium hydroxide. The parenteral preparation can be
enclosed in ampoules, disposable syringes or multiple dose vials
made of glass or plastic.
[0614] A compound or pharmaceutical composition of the invention
can be administered to a subject in many of the well-known methods
currently used for chemotherapeutic treatment. For example, for
treatment of cancers, a compound of the invention may be injected
directly into tumors, injected into the blood stream or body
cavities or taken orally or applied through the skin with patches.
The dose chosen should be sufficient to constitute effective
treatment but not as high as to cause unacceptable side effects.
The state of the disease condition (e.g., cancer, precancer, and
the like) and the health of the patient should preferably be
closely monitored during and for a reasonable period after
treatment.
[0615] The term "therapeutically effective amount", as used herein,
refers to an amount of a pharmaceutical agent to treat, ameliorate,
or prevent an identified disease or condition, or to exhibit a
detectable therapeutic or inhibitory effect. The effect can be
detected by any assay method known in the art. The precise
effective amount for a subject will depend upon the subject's body
weight, size, and health; the nature and extent of the condition;
and the therapeutic selected for administration. Therapeutically
effective amounts for a given situation can be determined by
routine experimentation that is within the skill and judgment of
the clinician. In a preferred aspect, the disease or condition to
be treated is cancer. In another aspect, the disease or condition
to be treated is a cell proliferative disorder.
[0616] For any compound, the therapeutically effective amount can
be estimated initially either in cell culture assays, e.g., of
neoplastic cells, or in animal models, usually rats, mice, rabbits,
dogs, or pigs. The animal model may also be used to determine the
appropriate concentration range and route of administration. Such
information can then be used to determine useful doses and routes
for administration in humans. Therapeutic/prophylactic efficacy and
toxicity may be determined by standard pharmaceutical procedures in
cell cultures or experimental animals, e.g., ED.sub.50 (the dose
therapeutically effective in 50% of the population) and LD.sub.50
(the dose lethal to 50% of the population). The dose ratio between
toxic and therapeutic effects is the therapeutic index, and it can
be expressed as the ratio, LD.sub.50/ED.sub.50. Pharmaceutical
compositions that exhibit large therapeutic indices are preferred.
The dosage may vary within this range depending upon the dosage
form employed, sensitivity of the patient, and the route of
administration.
[0617] Dosage and administration are adjusted to provide sufficient
levels of the active agent(s) or to maintain the desired effect.
Factors which may be taken into account include the severity of the
disease state, general health of the subject, age, weight, and
gender of the subject, diet, time and frequency of administration,
drug interaction(s), reaction sensitivities, and tolerance/response
to therapy. Long-acting pharmaceutical compositions may be
administered every 3 to 4 days, every week, or once every two weeks
depending on half-life and clearance rate of the particular
formulation.
[0618] The pharmaceutical compositions containing active compounds
of the present invention may be manufactured in a manner that is
generally known, e.g., by means of conventional mixing, dissolving,
granulating, dragee-making, levigating, emulsifying, encapsulating,
entrapping, or lyophilizing processes. Pharmaceutical compositions
may be formulated in a conventional manner using one or more
pharmaceutically acceptable carriers comprising excipients and/or
auxiliaries that facilitate processing of the active compounds into
preparations that can be used pharmaceutically. Of course, the
appropriate formulation is dependent upon the route of
administration chosen.
[0619] Pharmaceutical compositions suitable for injectable use
include sterile aqueous solutions (where water soluble) or
dispersions and sterile powders for the extemporaneous preparation
of sterile injectable solutions or dispersion. For intravenous
administration, suitable carriers include physiological saline,
bacteriostatic water, Cremophor EL.TM. (BASF, Parsippany, N.J.) or
phosphate buffered saline (PBS). In all cases, the composition must
be sterile and should be fluid to the extent that easy
syringeability exists. It must be stable under the conditions of
manufacture and storage and must be preserved against the
contaminating action of microorganisms such as bacteria and fungi.
The carrier can be a solvent or dispersion medium containing, for
example, water, ethanol, polyol (for example, glycerol, propylene
glycol, and liquid polyethylene glycol, and the like), and suitable
mixtures thereof. The proper fluidity can be maintained, for
example, by the use of a coating such as lecithin, by the
maintenance of the required particle size in the case of dispersion
and by the use of surfactants. Prevention of the action of
microorganisms can be achieved by various antibacterial and
antifungal agents, for example, parabens, chlorobutanol, phenol,
ascorbic acid, thimerosal, and the like. In many cases, it will be
preferable to include isotonic agents, for example, sugars,
polyalcohols such as manitol and sorbitol, and sodium chloride in
the composition. Prolonged absorption of the injectable
compositions can be brought about by including in the composition
an agent which delays absorption, for example, aluminum
monostearate and gelatin.
[0620] Sterile injectable solutions can be prepared by
incorporating the active compound in the required amount in an
appropriate solvent with one or a combination of ingredients
enumerated above, as required, followed by filtered sterilization.
Generally, dispersions are prepared by incorporating the active
compound into a sterile vehicle that contains a basic dispersion
medium and the required other ingredients from those enumerated
above. In the case of sterile powders for the preparation of
sterile injectable solutions, methods of preparation are vacuum
drying and freeze-drying that yields a powder of the active
ingredient plus any additional desired ingredient from a previously
sterile-filtered solution thereof.
[0621] Oral compositions generally include an inert diluent or an
edible pharmaceutically acceptable carrier. They can be enclosed in
gelatin capsules or compressed into tablets. For the purpose of
oral therapeutic administration, the active compound can be
incorporated with excipients and used in the form of tablets,
troches, or capsules. Oral compositions can also be prepared using
a fluid carrier for use as a mouthwash, wherein the compound in the
fluid carrier is applied orally and swished and expectorated or
swallowed. Pharmaceutically compatible binding agents, and/or
adjuvant materials can be included as part of the composition. The
tablets, pills, capsules, troches and the like can contain any of
the following ingredients, or compounds of a similar nature: a
binder such as microcrystalline cellulose, gum tragacanth or
gelatin; an excipient such as starch or lactose, a disintegrating
agent such as alginic acid, Primogel, or corn starch; a lubricant
such as magnesium stearate or Sterotes; a glidant such as colloidal
silicon dioxide; a sweetening agent such as sucrose or saccharin;
or a flavoring agent such as peppermint, methyl salicylate, or
orange flavoring.
[0622] For administration by inhalation, the compounds are
delivered in the form of an aerosol spray from pressured container
or dispenser, which contains a suitable propellant, e.g., a gas
such as carbon dioxide, or a nebulizer.
[0623] Systemic administration can also be by transmucosal or
transdermal means. For transmucosal or transdermal administration,
penetrants appropriate to the barrier to be permeated are used in
the formulation. Such penetrants are generally known in the art,
and include, for example, for transmucosal administration,
detergents, bile salts, and fusidic acid derivatives. Transmucosal
administration can be accomplished through the use of nasal sprays
or suppositories. For transdermal administration, the active
compounds are formulated into ointments, salves, gels, or creams as
generally known in the art.
[0624] The active compounds can be prepared with pharmaceutically
acceptable carriers that will protect the compound against rapid
elimination from the body, such as a controlled release
formulation, including implants and microencapsulated delivery
systems. Biodegradable, biocompatible polymers can be used, such as
ethylene vinyl acetate, polyanhydrides, polyglycolic acid,
collagen, polyorthoesters, and polylactic acid. Methods for
preparation of such formulations will be apparent to those skilled
in the art. The materials can also be obtained commercially from
Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal
suspensions (including liposomes targeted to infected cells with
monoclonal antibodies to viral antigens) can also be used as
pharmaceutically acceptable carriers. These can be prepared
according to methods known to those skilled in the art, for
example, as described in U.S. Pat. No. 4,522,811.
[0625] It is especially advantageous to formulate oral or
parenteral compositions in dosage unit form for ease of
administration and uniformity of dosage. Dosage unit form as used
herein refers to physically discrete units suited as unitary
dosages for the subject to be treated; each unit containing a
predetermined quantity of active compound calculated to produce the
desired therapeutic effect in association with the required
pharmaceutical carrier. The specification for the dosage unit forms
of the invention are dictated by and directly dependent on the
unique characteristics of the active compound and the particular
therapeutic effect to be achieved.
[0626] In therapeutic applications, the dosages of the
pharmaceutical compositions used in accordance with the invention
vary depending on the agent, the age, weight, and clinical
condition of the recipient patient, and the experience and judgment
of the clinician or practitioner administering the therapy, among
other factors affecting the selected dosage. Generally, the dose
should be sufficient to result in slowing, and preferably
regressing, the growth of the tumors and also preferably causing
complete regression of the cancer. Dosages can range from about
0.01 mg/kg per day to about 5000 mg/kg per day. In preferred
aspects, dosages can range from about 1 mg/kg per day to about 1000
mg/kg per day. In an aspect, the dose will be in the range of about
0.1 mg/day to about 50 g/day; about 0.1 mg/day to about 25 g/day;
about 0.1 mg/day to about 10 g/day; about 0.1 mg to about 3 g/day;
or about 0.1 mg to about 1 g/day, in single, divided, or continuous
doses (which dose may be adjusted for the patient's weight in kg,
body surface area in m.sup.2, and age in years). An effective
amount of a pharmaceutical agent is that which provides an
objectively identifiable improvement as noted by the clinician or
other qualified observer. For example, regression of a tumor in a
patient may be measured with reference to the diameter of a tumor.
Decrease in the diameter of a tumor indicates regression.
Regression is also indicated by failure of tumors to reoccur after
treatment has stopped. As used herein, the term "dosage effective
manner" refers to amount of an active compound to produce the
desired biological effect in a subject or cell.
[0627] The pharmaceutical compositions can be included in a
container, pack, or dispenser together with instructions for
administration.
[0628] The compounds of the present invention are capable of
further forming salts. All of these forms are also contemplated
within the scope of the claimed invention.
[0629] As used herein, "pharmaceutically acceptable salts" refer to
derivatives of the compounds of the present invention wherein the
parent compound is modified by making acid or base salts thereof.
Examples of pharmaceutically acceptable salts include, but are not
limited to, mineral or organic acid salts of basic residues such as
amines, alkali or organic salts of acidic residues such as
carboxylic acids, and the like. The pharmaceutically acceptable
salts include the conventional non-toxic salts or the quaternary
ammonium salts of the parent compound formed, for example, from
non-toxic inorganic or organic acids. For example, such
conventional non-toxic salts include, but are not limited to, those
derived from inorganic and organic acids selected from
2-acetoxybenzoic, 2-hydroxyethane sulfonic, acetic, ascorbic,
benzene sulfonic, benzoic, bicarbonic, carbonic, citric, edetic,
ethane disulfonic, 1,2-ethane sulfonic, fumaric, glucoheptonic,
gluconic, glutamic, glycolic, glycollyarsanilic, hexylresorcinic,
hydrabamic, hydrobromic, hydrochloric, hydroiodic, hydroxymaleic,
hydroxynaphthoic, isethionic, lactic, lactobionic, lauryl sulfonic,
maleic, malic, mandelic, methane sulfonic, napsylic, nitric,
oxalic, pamoic, pantothenic, phenylacetic, phosphoric,
polygalacturonic, propionic, salicyclic, stearic, subacetic,
succinic, sulfamic, sulfanilic, sulfuric, tannic, tartaric, toluene
sulfonic, and the commonly occurring amine acids, e.g., glycine,
alanine, phenylalanine, arginine, etc.
[0630] Other examples of pharmaceutically acceptable salts include
hexanoic acid, cyclopentane propionic acid, pyruvic acid, malonic
acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid,
4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,
4-toluenesulfonic acid, camphorsulfonic acid,
4-methylbicyclo-[2.2.2]-oct-2-ene-1-carboxylic acid,
3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic
acid, muconic acid, and the like. The present invention also
encompasses salts formed when an acidic proton present in the
parent compound either is replaced by a metal ion, e.g., an alkali
metal ion, an alkaline earth ion, or an aluminum ion; or
coordinates with an organic base such as ethanolamine,
diethanolamine, triethanolamine, tromethamine, N-methylglucamine,
and the like.
[0631] It should be understood that all references to
pharmaceutically acceptable salts include solvent addition forms
(solvates) or crystal forms (polymorphs) as defined herein, of the
same salt.
[0632] The compounds of the present invention can also be prepared
as esters, for example, pharmaceutically acceptable esters. For
example, a carboxylic acid function group in a compound can be
converted to its corresponding ester, e.g., a methyl, ethyl or
other ester. Also, an alcohol group in a compound can be converted
to its corresponding ester, e.g., acetate, propionate or other
ester.
[0633] The compounds of the present invention can also be prepared
as prodrugs, for example, pharmaceutically acceptable prodrugs. The
terms "pro-drug" and "prodrug" are used interchangeably herein and
refer to any compound which releases an active parent drug in vivo.
Since prodrugs are known to enhance numerous desirable qualities of
pharmaceuticals (e.g., solubility, bioavailability, manufacturing,
etc.), the compounds of the present invention can be delivered in
prodrug form. Thus, the present invention is intended to cover
prodrugs of the presently claimed compounds, methods of delivering
the same and compositions containing the same. "Prodrugs" are
intended to include any covalently bonded carriers that release an
active parent drug of the present invention in vivo when such
prodrug is administered to a subject. Prodrugs in the present
invention are prepared by modifying functional groups present in
the compound in such a way that the modifications are cleaved,
either in routine manipulation or in vivo, to the parent compound.
Prodrugs include compounds of the present invention wherein a
hydroxy, amino, sulfhydryl, carboxy or carbonyl group is bonded to
any group that may be cleaved in vivo to form a free hydroxyl, free
amino, free sulfhydryl, free carboxy or free carbonyl group,
respectively.
[0634] Examples of prodrugs include, but are not limited to, esters
(e.g., acetate, dialkylaminoacetates, formates, phosphates,
sulfates and benzoate derivatives) and carbamates (e.g.,
N,N-dimethylaminocarbonyl) of hydroxy functional groups, esters
(e.g., ethyl esters, morpholinoethanol esters) of carboxyl
functional groups, N-acyl derivatives (e.g., N-acetyl) N-Mannich
bases, Schiff bases and enaminones of amino functional groups,
oximes, acetals, ketals and enol esters of ketone and aldehyde
functional groups in compounds of the invention, and the like, See
Bundegaard, H., Design of Prodrugs, p1-92, Elesevier, New
York-Oxford (1985). The compounds, or pharmaceutically acceptable
salts, esters or prodrugs thereof, are administered orally,
nasally, transdermally, pulmonary, inhalationally, buccally,
sublingually, intraperintoneally, subcutaneously, intramuscularly,
intravenously, rectally, intrapleurally, intrathecally and
parenterally. In one embodiment, the compound is administered
orally. One skilled in the art will recognize the advantages of
certain routes of administration.
[0635] The dosage regimen utilizing the compounds is selected in
accordance with a variety of factors including type, species, age,
weight, sex and medical condition of the patient; the severity of
the condition to be treated; the route of administration; the renal
and hepatic function of the patient; and the particular compound or
salt thereof employed. An ordinarily skilled physician or
veterinarian can readily determine and prescribe the effective
amount of the drug required to prevent, counter, or arrest the
progress of the condition.
[0636] Techniques for formulation and administration of the
disclosed compounds of the invention can be found in Remington: the
Science and Practice of Pharmacy, 19.sup.th edition, Mack
Publishing Co., Easton, Pa. (1995). In an embodiment, the compounds
described herein, and the pharmaceutically acceptable salts
thereof, are used in pharmaceutical preparations in combination
with a pharmaceutically acceptable carrier or diluent. Suitable
pharmaceutically acceptable carriers include inert solid fillers or
diluents and sterile aqueous or organic solutions. The compounds
will be present in such pharmaceutical compositions in amounts
sufficient to provide the desired dosage amount in the range
described herein.
[0637] All percentages and ratios used herein, unless otherwise
indicated, are by weight. Other features and advantages of the
present invention are apparent from the different examples. The
provided examples illustrate different components and methodology
useful in practicing the present invention. The examples do not
limit the claimed invention. Based on the present disclosure the
skilled artisan can identify and employ other components and
methodology useful for practicing the present invention.
[0638] In the synthetic schemes described herein, compounds may be
drawn with one particular configuration for simplicity. Such
particular configurations are not to be construed as limiting the
invention to one or another isomer, tautomer, regioisomer or
stereoisomer, nor does it exclude mixtures of isomers, tautomers,
regioisomers or stereoisomers.
[0639] Compounds described herein are assayed for modulation of
activity, for example, histone methylation, modulation of cell
growth and/or IC.sub.50, described in the examples below. IC.sub.50
values are presented as A=<0.1 .mu.M; B=>0.1 .mu.M and <1
.mu.M; C=>1 .mu.M and <10 .mu.M; and D=>10 .mu.M and
<50 .mu.M.
TABLE-US-00002 DOT1L Compound IC.sub.50 (.mu.M) 2 0.00074 3 0.00073
5 0.00059 69 0.00251 75 0.00059 86 0.00062 87 0.0008 91 0.00218 93
0.00292
[0640] All publications and patent documents cited herein are
incorporated herein by reference as if each such publication or
document was specifically and individually indicated to be
incorporated herein by reference. Citation of publications and
patent documents is not intended as an admission that any is
pertinent prior art, nor does it constitute any admission as to the
contents or date of the same. The invention having now been
described by way of written description, those of skill in the art
will recognize that the invention can be practiced in a variety of
embodiments and that the foregoing description and examples below
are for purposes of illustration and not limitation of the claims
that follow.
5. EXAMPLES
[0641] Nuclear magnetic resonance (NMR) spectra were obtained on a
Bruker Avance 400 operating at a field strength of 400.130 MHz or a
Bruker DRX 500 MHz NMR or HNMR spectra were obtained on a 500 MHz
Bruker AVANCE III spectrometer. Common reaction solvents were
either high performance liquid chromatography (HPLC) grade or
American Chemical Society (ACS) grade, and anhydrous as obtained
from the manufacturer unless otherwise noted. LCMS was performed on
a Waters Micromass ZMD with a Waters 2795 Separations Module and
Waters 996 photodiode array detector and a Waters Micromass ZQ with
a Waters 2695 Separations Module and Waters 996 photodiode array
detector or a Waters Micromass Platform LCZ single quadrupole mass
spectrometer with a Waters 600 solvent delivery module, Waters 515
ancillary pumps, Waters 2487 UV detector and a Gilson 215
autosampler and fraction collector. Or, LCMS analysis was performed
using SQ mass spectrometer coupled to AGILENT 1200 Series HPLC.
LCMS data, where available, are provided in the examples below as
well as in Table 1. The MS data are provided using the convention
for m/z in the format, [M+H]*.
[0642] The compounds of the present invention can be prepared using
known chemical transformations adapted to the particular situation
at hand.
Preparative Example 1: Starting Materials or Intermediates
Step 1:
(1R,2S,3R,5R)-3-((5-amino-6-chloropyrimidin-4-yl)amino)-5-(hydroxy-
methyl)cyclopentane-1,2-diol
##STR00217##
[0644] A mixture of
(1R,2S,3R,5R)-3-amino-5-(hydroxymethyl)cyclopentane-1,2-diol
hydrochloride (16.9 g, 45.1 mmol) and 4,6-dichloropyrimidin-5-amine
(5.7 g, 35 mmol) in ethanol (45 mL) was evenly distributed amongst
three tubes and subjected to microwave conditions (CEM apparatus,
300 W max, 150.degree. C. max, 250 psi max, 3 min ramp, 30 min
hold) to afford brown solutions; HPLC/LC MS indicated conversion to
the desired product. The three reaction mixtures were combined and
concentrated in vacuo to afford the crude title compound as a dark
brown oil, which was concentrated from toluene (2.times.30 mL) and
carried on without purification: MS (ESI+) for
C.sub.10H.sub.15ClN.sub.4O.sub.3 m/z 275.0 (M+H).sup.+; MS (ESI-)
for C.sub.10H.sub.15ClN.sub.4O.sub.3 m/z 273.0 (M-H).sup.-.
Step 2:
((3aR,4R,6R,6aS)-6-(6-chloro-9H-purin-9-yl)-2-ethoxytetrahydro-3aH-
-cyclopenta[d][1,3]dioxol-4-yl)methanol
##STR00218##
[0646] The above crude
(1R,2S,3R,5R)-3-((5-amino-6-chloropyrimidin-4-yl)amino)-5-(hydroxymethyl)-
cyclopentane-1,2-diol was treated with ethyl orthoformate (120 mL,
720 mmol) and 10-camphorsulfonic acid (8.11 g, 34.9 mmol). The
heterogeneous brown mixture was stirred vigorously to afford a
nearly homogeneous brown solution after 10 min. At 5 h, LC MS
indicated the desired product as the major product and the reaction
was quenched with saturated aqueous NaHCO.sub.3 (120 mL). The
mixture was diluted with water (75 mL), extracted with
CH.sub.2Cl.sub.2 (3.times.200 mL), and the combined organics were
dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford the
crude title compound as a dark brown liquid, which was carried on
without further manipulation: MS (ESI+) for
C.sub.14H.sub.17ClN.sub.4O.sub.4 m/z 341.0 (M+H).sup.+.
Step 3:
((3aR,4R,6R,6aS)-6-(6-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydro-
-3aH-cyclopenta[d][1,3]dioxol-4-yl)methanol
##STR00219##
[0648] The above crude
((3aR,4R,6R,6aS)-6-(6-chloro-9H-purin-9-yl)-2-ethoxytetrahydro-3aH-cyclop-
enta[d][1,3]dioxol-4-yl)methanol was taken up in
2,2-dimethoxypropane (214 mL, 1740 mmol) and treated with
p-toluenesulfonic acid monohydrate (13.2 g, 69.5 mmol) to afford a
brown oil partially suspended in a cloudy solution, which was
stirred at rt for 1 h 20 min; HPLC/LC MS indicated complete
conversion to the desired product. The reaction was quenched by the
careful addition of sodium bicarbonate (8.76 g, 104 mmol) and a
minimal amount of water. The volatiles were removed in vacuo and
the remaining aqueous layer was diluted with water (100 mL) and
extracted with CH.sub.2Cl.sub.2 (3.times.400 mL). The combined
organics were dried (Na.sub.2SO.sub.4) and concentrated in vacuo to
afford a brown oil. Purification by column chromatography
(7.times.16 cm silica; 0-5% MeOH/CH.sub.2Cl.sub.2) afforded the
title compound (8.30 g, 74% over 3 steps) as a yellow foam: MS
(ESI+) for C.sub.14H.sub.17CN.sub.4O.sub.3 m/z 325.1 (M+H).sup.+;
MS (ESI-) for C.sub.14H17CN.sub.4O.sub.3 m/z 369.0
(M+HCO.sub.2).sup.-; HPLC purity >95 area %.
Step
4:9-((3aS,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydro-3aH-cyclo-
penta[d][1,3]dioxol-4-yl)-6-chloro-9H-purine
##STR00220##
[0650] A mixture of
((3aR,4R,6R,6aS)-6-(6-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydro-3aH-cy-
clopenta[d][1,3]dioxol-4-yl)methanol (7.9 g, 24 mmol) and
polymer-supported triphenylphosphine (3 mmol/g loading; 11 g, 34
mmol) in THE (100 mL) was cooled to 0.degree. C. (ice/brine bath)
and treated dropwise with diisopropyl azodicarboxylate (6.7 mL, 34
mmol). The tan slurry was stirred for 15 min, and treated dropwise
with a solution of diphenylphosphonic azide (7.3 mL, 34 mmol) in
THE (24 mL). The brown reaction mixture was stirred for 18.5 h as
the ice bath expired; HPLC indicated conversion to the desired
product. At 21.5 h the reaction mixture was filtered, the solids
were washed with CH.sub.2Cl.sub.2, and the filtrate was
concentrated in vacuo. The red-brown residue was taken up in
CH.sub.2Cl.sub.2 (300 mL) and washed with saturated aqueous
NaHCO.sub.3 (1.times.100 mL), water (1.times.100 mL), and brine
(1.times.150 mL). The separated organic layer was dried
(Na.sub.2SO.sub.4) and concentrated in vacuo to afford a red-orange
oil. Purification by column chromatography (7.times.16 cm silica;
0-10% acetone/CH.sub.2Cl.sub.2) afforded the title compound (4.82
g, 57%) as a yellow oil/foam: MS (ESI+) for
C.sub.14H.sub.16ClN.sub.7O.sub.2 m/z 350.1 (M+H).sup.+; MS (ESI-)
for C.sub.14H.sub.16ClN.sub.7O.sub.2 m/z 394.1 (M+HCO.sub.2).sup.-;
HPLC purity >95 area %.
Step 5:
9-((3aS,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydro-3aH-cycl-
openta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-9H-purin-6-amine
##STR00221##
[0652] A solution of
9-((3aS,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-6-chloro-9H-purine (1.29 g, 3.69 mmol) and
(2,4-dimethoxyphenyl)methanamine (0.71 mL, 4.7 mmol) in 1-butanol
(10 mL) was treated with N,N-diisopropylethylamine (0.93 mL, 5.3
mmol) and heated at 80.degree. C. for 16.5 h; HPLC/LC MS indicated
conversion to the desired product. The reaction mixture was allowed
to cool to rt and the volatiles were removed under the flow of air
to afford a brown-orange paste. Purification by column
chromatography (2.times.8 cm silica; 0-10%
acetone/CH.sub.2Cl.sub.2) afforded the title compound (1.72 g, 97%)
as a yellow-orange foam/oil: MS (ESI+) for
C.sub.23H.sub.28N.sub.8O.sub.4 m/z 481.2 (M+H).sup.+; HPLC purity
>95 area %.
Step 6:
9-((3aS,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydro-3aH-cycl-
openta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-9H-purin-6-amine
##STR00222##
[0654] A solution of
9-((3aS,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-9H-purin-6-amine (1.72
g, 3.58 mmol) in THE (16 mL) was cooled to 0.degree. C. (ice/brine
bath) and treated dropwise with a 1.0 M solution of
trimethylphosphine in THE (6.30 mL, 6.30 mmol). The cold bath was
removed after 30 min and the reaction mixture was stirred for 1.5
h; HPLC/LC MS indicated complete consumption of the starting azide.
Water (2.84 mL, 157 mmol) was added to the orange solution (gas
evolution noted) and the reaction mixture was stirred for 2.75 h at
rt; HPLC indicated complete conversion to the desired amine. The
reaction mixture was concentrated in vacuo to afford an orange oil.
The residue was taken up in CH.sub.2Cl.sub.2 (150 mL) and washed
with water (2.times.50 mL) and brine (1.times.75 mL). The separated
organic layer was dried (Na.sub.2SO.sub.4) and concentrated in
vacuo to afford the title compound (1.6 g, 98%) as a pale yellow
foam: MS (ESI+) for C.sub.23H.sub.30N.sub.6O.sub.4 m/z 455.2
(M+H).sup.+; HPLC purity >95 area %.
Step 1: ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(6-((2,4-dimethoxybenzyl)amino)-9H-purin-9-yl)--
2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)amino)cycl-
obutyl)propanoate
##STR00223##
[0656] Sodium triacetoxyborohydride (0.839 g, 3.96 mmol) was added
to a solution of
9-((3aS,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-9H-purin-6-amine (1.5
g, 3.3 mmol), ethyl 3-(3-oxocyclobutyl)propanoate (0.562 g, 3.30
mmol) and acetic acid (0.188 mL, 3.30 mmol) in 1,2-dichloroethane
(26.0 mL, 3.30E2 mmol) and the reaction was stirred at RT
overnight. The following morning the starting material was consumed
by HPLC so NaHCO.sub.3 was added and the aqueous extracted 3.times.
with DCM. Combined organics were dried with MgSO.sub.4 and purified
by FC (DCM/7N NH.sub.3 in MeOH 95:5) to yield ethyl
3-3-((((3aR,4R,6R,6aS)-6-(6-((2,4-dimethoxybenzyl)amino)-9H-purin-9-yl)-2-
,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)amino)cyclo-
butyl)propanoate (1.5 g; 75%) as a thick yellow resin/foam. MS
(ESI.sup.+) for C.sub.32H.sub.44N.sub.6O.sub.6 m/z 609.3
[M+H].sup.+.
Step 2: ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(6-((2,4-dimethoxybenzyl)amino)-9H-purin-9-yl)--
2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(isopropyl-
)amino)cyclobutyl)propanoate
##STR00224##
[0658] Ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(6-((2,4-dimethoxybenzyl)amino)-9H-purin-9-yl)--
2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)amino)cycl-
obutyl)propanoate (0.06 g, 0.1 mmol) was taken up in acetonitrile
(2.6 mL, 50 mmol) and isopropyl iodide (0.098 mL, 0.98 mmol) and
triethylamine (0.21 mL, 1.5 mmol) were added. The reaction was
heated to 80.degree. C. for 12 hours at which point the reaction
appeared to stall. Another 15 equivalents triethylamine and another
15 equivalents isopropyl iodide were added and the reaction
continued 8 hours more. Reaction appeared to have stalled again so
another 15 equivalents each of isopropyl iodide and triethylamine
were added. Upon consumption of the starting material the reaction
was concentrated and saturated Na.sub.2CO.sub.3 (20 mls) and DCM
(20 mls) was added. The residue was partitioned between the organic
layer and the aqueous layer. The aqueous layer was extracted 3
times with DCM, then the combined organics were dried and purified
by FC (DCM/7N NH.sub.3 in MeOH 97:3). Product was still
contaminated with TEA-H+I-, so to a 30 ml solution of the product
in DCM was added 20 mls saturated NaHCO.sub.3 and 10 mls 1N NaOH.
The mixture was stirred for 15 minutes then the aqueous was
extracted with DCM 3 times. The combined organics were dried with
MgSO.sub.4 and solvent removed to yield pure ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(6-((2,4-dimethoxybenzyl)amino)-9H-purin-9-yl)--
2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(isopropyl-
)amino)cyclobutyl)propanoate (0.045 g; 70%) as a brown foam/solid
with no more amine salts present by NMR. MS (ESI.sup.+) for
C.sub.35H.sub.50N.sub.6O.sub.6 m/z 651.3 [M+H].sup.+.
##STR00225##
[0659] Lithium hydroxide monohydrate (0.838 g, 20.0 mmol) was added
to a solution of ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(6-((2,4-dimethoxybenzyl)amino)-9H-purin-9-yl)--
2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(isopropyl-
)amino)cyclobutyl)propanoate (1.3 g, 2.0 mmol) in tetrahydrofuran
(30 mL, 300 mmol) and methanol (6.5 mL, 160 mmol). The reaction was
stirred overnight at RT and by next morning the starting material
was consumed and had been transformed into the acid. The reaction
was acidified with 1N HCl to pH=6. Volatiles were removed in vacuo
and remaining water removed by azeotropic distillation with ethanol
followed by 24 hours of lyophilization. The resulting brown solid
was used without further purification.
Ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo-
[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-
-4-yl)methyl)(methyl)amino)cyclobutyl)propanoate
##STR00226##
[0661] The amine ethyl
3-[3-({[(3aR,4R,6R,6aS)-6-{4-[(2,4-dimethoxybenzyl)amino]-7H-pyrrolo[2,3--
d]pyrimidin-7-yl}-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
]methyl}amino)cyclobutyl]propanoate (1.8 g, 3.0 mmol) was taken up
in methanol (20 mL, 600 mmol) and sodium cyanoborohydride (0.19 g,
3.0 mmol) was added. The pH was adjusted to ca. 6 using a 10%
solution of AcOH in MeOH, then formalin (0.29 mL, 3.9 mmol) was
added in one portion. The reaction was allowed to proceed for 3
hours at which time MS indicated complete consumption of the
starting material. NaHCO.sub.3 (saturated) added to the reaction
mixture which was then extracted 3 times with DCM. The combined
organics were dried with MgSO.sub.4 and concentrated to a yellow
resin. This residue was purified by FC (DCM/7N NH.sub.3 in MeOH
93:7) to yield ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(methyl)amino)cyclobutyl)propanoate (1.6 g; 87%) as a
colorless foam. MS (ESI.sup.+) for C.sub.34H.sub.47N.sub.5O.sub.6
m/z 622.3 [M+H].sup.+.
[0662] .sup.1H NMR (400 MHz, d.sub.3-chloroform) .delta..sub.H
8.282 (s, 1H), 7.203-7.168 (m, 1H), 6.877-6.865 (m, 1H),
6.399-6.334 (m, 2H), 6.242-6.236 (m, 1H), 5.330 (s, 1H),
4.890-4.835 (m, 2H), 4.664-4.650 (d, J=5.6 Hz, 2H), 4.391-4.354 (m,
1H), 4.067-4.000 (m, 2H), 3.757 (s, 3H), 3.710 (s, 3H), 2.864-2.784
(m, 0.5H (methine of trans isomer)), 2.553-2.474 (m, 0.5H (methine
of cis isomer), 2.432-2.370 (m, 1H), 2.322-2.278 (m, 2H),
2.212-2.089 (m, 4H), 2.022 & 2.018 (s, 3H (overlapping singlets
due to N-methyl of cis and trans isomers), 1.964-1.908 (m, 3H),
1.778-1.584 (m, 4H), 1.486 (s, 3H), 1.363-1.296 (m, 1H), 1.219 (s,
3H), 1.182-1.146 (m, 3H).
((3aR,4R,6R,6aS)-6-(6-chloro-9H-purin-9-yl)-2-ethoxytetrahydro-3aH-cyclope-
nta[d][1,3]dioxol-4-yl)methanol
##STR00227##
[0664] The above crude
(1R,2S,3R,5R)-3-((5-amino-6-chloropyrimidin-4-yl)amino)-5-(hydroxymethyl)-
cyclopentane-1,2-diol was treated with ethyl orthoformate (120 mL,
720 mmol) and 10-camphorsulfonic acid (8.11 g, 34.9 mmol). The
heterogeneous brown mixture was stirred vigorously to afford a
nearly homogeneous brown solution after 10 min. At 5 h, LC MS
indicated the desired product as the major product and the reaction
was quenched with saturated aqueous NaHCO.sub.3 (120 mL). The
mixture was diluted with water (75 mL), extracted with
CH.sub.2Cl.sub.2 (3.times.200 mL), and the combined organics were
dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford the
crude title compound as a dark brown liquid, which was carried on
without further manipulation: MS (ESI+) for
C.sub.14H.sub.17ClN.sub.4O.sub.4 m/z 341.0 (M+H).sup.+.
Step 3:
((3aR,4R,6R,6aS)-6-(6-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydro-
-3aH-cyclopenta[d][1,3]dioxol-4-yl)methanol
##STR00228##
[0666] The above crude
((3aR,4R,6R,6aS)-6-(6-chloro-9H-purin-9-yl)-2-ethoxytetrahydro-3aH-cyclop-
enta[d][1,3]dioxol-4-yl)methanol was taken up in
2,2-dimethoxypropane (214 mL, 1740 mmol) and treated with
p-toluenesulfonic acid monohydrate (13.2 g, 69.5 mmol) to afford a
brown oil partially suspended in a cloudy solution, which was
stirred at rt for 1 h 20 min; HPLC/LC MS indicated complete
conversion to the desired product. The reaction was quenched by the
careful addition of sodium bicarbonate (8.76 g, 104 mmol) and a
minimal amount of water. The volatiles were removed in vacuo and
the remaining aqueous layer was diluted with water (100 mL) and
extracted with CH.sub.2Cl.sub.2 (3.times.400 mL). The combined
organics were dried (Na.sub.2SO.sub.4) and concentrated in vacuo to
afford a brown oil. Purification by column chromatography
(7.times.16 cm silica; 0-5% MeOH/CH.sub.2Cl.sub.2) afforded the
title compound (8.30 g, 74% over 3 steps) as a yellow foam: MS
(ESI+) for C.sub.14H.sub.17CN.sub.4O.sub.3 m/z 325.1 (M+H).sup.+;
MS (ESI-) for C.sub.14H.sub.17ClN.sub.4O.sub.3 m/z 369.0
(M+HCO.sub.2).sup.-.
Step 4:
9-((3aS,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydro-3aH-cycl-
openta[d][1,3]dioxol-4-yl)-6-chloro-9H-purine
##STR00229##
[0668] A mixture of
((3aR,4R,6R,6aS)-6-(6-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydro-3aH-cy-
clopenta[d][1,3]dioxol-4-yl)methanol (7.9 g, 24 mmol) and
polymer-supported triphenylphosphine (3 mmol/g loading; 11 g, 34
mmol) in THE (100 mL) was cooled to 0.degree. C. (ice/brine bath)
and treated dropwise with diisopropyl azodicarboxylate (6.7 mL, 34
mmol). The tan slurry was stirred for 15 min, and treated dropwise
with a solution of diphenylphosphonic azide (7.3 mL, 34 mmol) in
THE (24 mL). The brown reaction mixture was stirred for 18.5 h as
the ice bath expired; HPLC indicated conversion to the desired
product. At 21.5 h the reaction mixture was filtered, the solids
were washed with CH.sub.2Cl.sub.2, and the filtrate was
concentrated in vacuo. The red-brown residue was taken up in
CH.sub.2Cl.sub.2 (300 mL) and washed with saturated aqueous
NaHCO.sub.3 (1.times.100 mL), water (1.times.100 mL), and brine
(1.times.150 mL). The separated organic layer was dried
(Na.sub.2SO.sub.4) and concentrated in vacuo to afford a red-orange
oil. Purification by column chromatography (7.times.16 cm silica;
0-10% acetone/CH.sub.2Cl.sub.2) afforded the title compound (4.82
g, 57%) as a yellow oil/foam: MS (ESI+) for
C.sub.14H16ClN.sub.7O.sub.2 m/z 350.1 (M+H).sup.+; MS (ESI-) for
C.sub.14H16ClN.sub.7O.sub.2 m/z 394.1 (M+HCO.sub.2).sup.-.
Step 5:
9-((3aS,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydro-3aH-cycl-
openta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-9H-purin-6-amine
##STR00230##
[0670] A solution of
9-((3aS,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-6-chloro-9H-purine (1.29 g, 3.69 mmol) and
(2,4-dimethoxyphenyl)methanamine (0.71 mL, 4.7 mmol) in 1-butanol
(10 mL) was treated with N,N-diisopropylethylamine (0.93 mL, 5.3
mmol) and heated at 80.degree. C. for 16.5 h; HPLC/LC MS indicated
conversion to the desired product. The reaction mixture was allowed
to cool to rt and the volatiles were removed under the flow of air
to afford a brown-orange paste. Purification by column
chromatography (2.times.8 cm silica; 0-10%
acetone/CH.sub.2Cl.sub.2) afforded the title compound (1.72 g, 97%)
as a yellow-orange foam/oil: MS (ESI+) for
C.sub.23H.sub.28N.sub.8O.sub.4 m/z 481.2 (M+H).sup.+.
Step
6:9-((3aS,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydro-3aH-cyclo-
penta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-9H-purin-6-amine
##STR00231##
[0672] A solution of
9-((3aS,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-9H-purin-6-amine (1.72
g, 3.58 mmol) in THE (16 mL) was cooled to 0.degree. C. (ice/brine
bath) and treated dropwise with a 1.0 M solution of
trimethylphosphine in THE (6.30 mL, 6.30 mmol). The cold bath was
removed after 30 min and the reaction mixture was stirred for 1.5
h; HPLC/LC MS indicated complete consumption of the starting azide.
Water (2.84 mL, 157 mmol) was added to the orange solution (gas
evolution noted) and the reaction mixture was stirred for 2.75 h at
rt; HPLC indicated complete conversion to the desired amine. The
reaction mixture was concentrated in vacuo to afford an orange oil.
The residue was taken up in CH.sub.2Cl.sub.2 (150 mL) and washed
with water (2.times.50 mL) and brine (1.times.75 mL). The separated
organic layer was dried (Na.sub.2SO.sub.4) and concentrated in
vacuo to afford the title compound (1.6 g, 98%) as a pale yellow
foam: MS (ESI+) for C.sub.23H.sub.30N.sub.6O.sub.4 m/z 455.2
(M+H).sup.+.
Step 1: ethyl
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoate
##STR00232##
[0674] A mixture of
9-((3aR,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]-
dioxol-4-yl)-9H-purin-6-amine (0.50 g, 1.6 mmol) and ethyl
3-(3-oxocyclobutyl)propanoate (0.27 g, 1.6 mmol) in methanol (10
mL) was treated with acetic acid (0.09 mL, 2 mmol) at rt and the
flask was evacuated and flushed with nitrogen (.times.3). The
reaction mixture was treated at rt with sodium cyanoborohydride
(0.26 g, 4.1 mmol), which afforded instant gas evolution and a
nearly colorless, clear solution in a few minutes. The reaction
mixture was stirred for 1 h at rt; HPLC/LC MS indicated a
.about.2:1 mixture of product to starting amine. At 1.5 h
additional ethyl 3-(3-oxocyclobutyl)propanoate (66 mg, 0.39 mmol)
in MeOH (1.0 mL) was added and the reaction mixture was stirred at
rt for 30 min; HPLC/LC MS indicated .about.70% conversion and some
dialkylation. At 2 h 15 min water (4.0 mL) was added and the
mixture was concentrated in vacuo. The residual aqueous layer was
diluted with saturated aqueous sodium bicarbonate (10 mL, to pH 9)
and extracted with CH.sub.2Cl.sub.2 (3.times.15 mL). The combined
organics were dried (Na.sub.2SO.sub.4) and concentrated in vacuo to
afford the reductive amination product as a white foam, which was
carried on without further purification: MS (ESI+) for
C.sub.22H.sub.32N.sub.6O.sub.5 m/z 461.1 (M+H)+, 483.1 (M+Na)+.
Step 2: ethyl
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoate
##STR00233##
[0676] The above crude secondary amine was taken up in methanol (10
mL) and treated with sodium cyanoborohydride (0.30 g, 4.8 mmol). A
solution of 10% v/v acetic acid in methanol was added to adjust the
pH to .about.6, followed by the dropwise addition of 37% aqueous
formaldehyde (0.65 mL, 6.3 mmol), which afforded gas evolution. The
reaction mixture was stirred at rt for 1 h; HPLC/LC MS indicated
complete conversion to the desired product. At 1.5 h, water (5.0
mL) was added and the reaction mixture was concentrated in vacuo.
The residue was diluted with saturated aqueous NaHCO.sub.3 (10 mL,
to pH .about.9) and extracted with CH.sub.2Cl.sub.2 (3.times.15
mL). The combined organics were diluted with a small amount of EtOH
to afford a clear solution, dried (Na.sub.2SO.sub.4), and
concentrated in vacuo to afford a nearly colorless oil.
Purification by column chromatography (4.times.17 cm silica; 0-5% 7
N methanolic NH.sub.3/CH.sub.2Cl.sub.2) afforded the title compound
(0.50 g, 60%) as a white foam/colorless oil: MS (ESI+) for
C.sub.23H.sub.34N.sub.6O.sub.5 m/z 475.1 (M+H)+, 497.1 (M+Na)+.
Step 1: ethyl
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoate
##STR00234##
[0678] A mixture of
9-((3aR,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]-
dioxol-4-yl)-9H-purin-6-amine (2.04 g, 6.66 mmol) and ethyl
3-(3-oxocyclobutyl)propanoate (1.2 g, 7.0 mmol) in methanol (41 mL)
was treated with acetic acid (0.37 mL, 6.5 mmol) at rt and the
flask was evacuated and flushed with nitrogen (.times.3). The
reaction mixture was treated at rt with sodium cyanoborohydride
(1.0 g, 16 mmol), which afforded instant gas evolution and a nearly
colorless, clear solution in a few minutes. The reaction mixture
was stirred for 1 h at rt; HPLC/LC MS indicated starting material
remained. At 1 h 20 min additional ethyl
3-(3-oxocyclobutyl)propanoate (0.50 g, 2.93 mmol) in MeOH (3 mL)
was added. The reaction mixture was stirred for 30 min and treated
with water (12 mL). The mixture was concentrated in vacuo and the
residual aqueous layer was diluted with saturated aqueous sodium
bicarbonate (40 mL, to pH 9) and extracted with CH.sub.2Cl.sub.2
(3.times.60 mL). The combined organics were dried
(Na.sub.2SO.sub.4) and concentrated in vacuo to afford the crude
title compound as a white foam/very pale yellow oil, which was
carried on without further purification: MS (ESI+) for
C.sub.22H.sub.32N.sub.6O.sub.5 m/z 461.2 (M+H).sup.+ and 483.1
(M+Na).sup.+.
Step 2: ethyl
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)propanoate
##STR00235##
[0680] A solution of the above crude ethyl
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoate in
acetonitrile (30 mL) was treated with potassium carbonate (6.3 g,
46 mmol) and isopropyl iodide (3.9 mL, 39 mmol). The reaction
mixture in a sealed tube was heated at 90.degree. C. for 6.5 h;
HPLC indicated a 4:1 mixture of product to starting material. The
reaction mixture was stirred overnight (17.5 h) at rt, treated with
additional isopropyl iodide (2.0 mL, 20 mmol), and heated at
90.degree. C. for 3 h; HPLC/LC MS indicated nearly complete
conversion. The reaction mixture was cooled to rt and the solids
were removed by vacuum filtration, rinsing with CH.sub.3CN, and the
filtrate was concentrated in vacuo to afford a dull orange oil with
precipitate. Purification by column chromatography (5.times.14.5 cm
silica; 0-10% 7 N methanolic NH.sub.3/CH.sub.2Cl.sub.2) afforded
the title compound (0.49 g, 15%) as a white foam/colorless oil. The
mixed fractions containing product were repurified by column
chromatography (4.times.10.5 cm silica; 0-5% 7 N methanolic
NH.sub.3/CH.sub.2Cl.sub.2) to afford the title compound (1.66 g,
40%) as a white foam/colorless oil contaminated with the
bisreductive amination byproduct from Step 1: MS (ESI+) for
C.sub.25H.sub.38N.sub.6O.sub.5 m/z 503.2 (M+H).sup.+.
ethyl
3-(3-((((1R,2R,3S,4R)-4-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2-
,3-d]pyrimidin-7-yl)-2,3-dihydroxycyclopentyl)methyl)amino)cyclobutyl)prop-
anoate
##STR00236##
[0682] Sodium triacetoxyborohydride (2.43 g, 115 mmol) was added to
a solution of
(1S,2R,3R,5R)-3-(aminomethyl)-5-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrol-
o[2,3-d]pyrimidin-7-yl)cyclopentane-1,2-diol (2.6 g, 5.7 mmol) and
ethyl 3-(3-oxocycobutyl)propanoate (0.976 g, 5.73 mmol) and Acetic
acid (0.326 ml, 5.73 mmol) in 1,2-Dichloroethane (20 ml) and
reaction was stirred at RT overnight. NaHCO.sub.3 was added and the
aqueous layer was extracted 3.times. with DCM. The combined
organics were dried with MgSO.sub.4, filtered, concentrated and
purified by flash chromatogrpahy (DCM/7N NH.sub.3 in MeOH 90:10) to
give de desired compound (1.8 g) as a thick yellow resin.
N-(2,4-dimethoxybenzyl)-7-((3aS,4R,6R,6aR)-6-((isopropylamino)methyl)-2,2--
dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyr-
imidin-4-amine
##STR00237##
[0684] A solution of
7-((3aS,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-a-
mine (7.50 g, 16.5 mmol) in 1,2-Dichloroethane (140 mL, 1800 mmol)
was treated with Acetone (1.34 mL, 18.2 mmol;) and Acetic acid
(0.94 mL, 16 mmol) dropwise followed by Sodium
triacetoxyborohydride (4.20 g, 19.8 mmol) and the mixture was
stirred at RT for 4 h. HPLC analysis indicated the reaction was
complete. The reaction mixture was diluted with 200 mL
CH.sub.2Cl.sub.2 and washed with 150 mL sat NaHCO.sub.3. The
aqueous phase was washed with 100 mL CH.sub.2Cl.sub.2 and the
combined organic phase was dried over Na.sub.2SO.sub.4, filtered
and concentrated to yield a an oil that produced a stiff foam when
placed under high vac. The crude material (9.3 g) was carried on
directly to the next step.
ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo-
[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-
-4-yl)methyl)(isopropyl)amino)cyclobutyl)propanoate
##STR00238##
[0686] A solution of
N-(2,4-dimethoxybenzyl)-7-((3aS,4R,6R,6aR)-6-((isopropylamino)methyl)-2,2-
-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]py-
rimidin-4-amine (9.50 g, 15.3 mmol) in 1,2-Dichloroethane (75 mL,
950 mmol) was treated with ethyl 3-(3-oxocyclobutyl)propanoate
(3.92 g, 23.0 mmol) and Acetic acid (1.0 mL, 18 mmol;) dropwise
followed by Sodium triacetoxyborohydride (4.58 g, 21.6 mmol) and
the mixture was stirred at RT for 6 days. The reaction mixture was
diluted with 150 mL CH.sub.2Cl.sub.2 and washed with 100 mL sat
NaHCO.sub.3. The aqueous phase was washed with 100 mL
CH.sub.2Cl.sub.2 and the combined organic phase was dried over
Na.sub.2SO.sub.4, filtered and concentrated to yield a light brown
viscous glass.
[0687] The crude material was purified by flash chromatography
(SiO.sub.2 eluting with 2-3% 7N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2) to yield a slightly glass/stiff foam
(7.10 g).
ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo-
[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-
-4-yl)methyl)amino)cyclobutyl)propanoate
##STR00239##
[0689] A solution of
7-((3aS,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-a-
mine (8.00 g, 15.2 mmol) in 1,2-Dichloroethane (119.5 mL, 1517
mmol) was treated with ethyl 3-(3-oxocyclobutyl)propanoate (2.58 g,
15.2 mmol;) and Acetic acid (0.86 mL, 15 mmol) dropwise followed by
Sodium triacetoxyborohydride (3.86 g, 18.2 mmol) and the mixture
was stirred at RT for 19 h. The reaction mixture was diluted with
150 mL CH.sub.2Cl.sub.2 and washed with 150 mL sat NaHCO.sub.3. The
aqueous phase was washed with 70 mL CH.sub.2Cl.sub.2 and the
combined organic phase was dried over Na.sub.2SO.sub.4, filtered
and concentrated to yield a tan glass that produced a sticky foam
when placed under high vac. The crude material was purified by
flash chromatography (SiO.sub.2 eluting with 3-4% 7N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2) to yield a light yellow viscous oil
that producted a sticky foam under high vacuum (5.03 g). MS 608.3
(M+H).
Example 1: Synthesis of
1-((3-((((2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yl)-3,4-dihydroxytetrahydrof-
uran-2-yl)methyl)(methyl)amino)cyclobutyl)methyl)-3-(4-(tert-butyl)phenyl)-
urea (Compound 110)
Step 1: Synthesis of methyl 3-oxocyclobutanecarboxylate
##STR00240##
[0691] To a solution of DCC (5.96 g, 28.95 mmol) in DCM (20 ml) was
added dropwise a mixture of 3-oxocyclobutanecarboxylic acid (3.0 g,
26.31 mmol), MeOH (1.68 g, 52.62 mmol) and DMAP (2.57 g, 21.05
mmol) in DCM (30 ml). The reaction mixture was stirred at RT
overnight. The mixture was filtrated. The filtrate was washed with
0.5 M HCl solution (50 ml). The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified by SGC
(PE:EA=5:1) to obtain the title compound (4.0 g). .sup.1H N.R (500
MHz, CDCl.sub.3): .delta. 3.77 (s, 3H), 3.42-3.26 (m, 5H) ppm.
Step 2: Synthesis of methyl
3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutanecarboxylate
##STR00241##
[0693] A solution of methyl 3-oxocyclobutanecarboxylate (1.28 g
crude),
9-((3aR,4R,6R,6aR)-2,2-dimethyl-6-((methylamino)methyl)tetrahydrofuro[3,4-
-d][1,3]dioxol-4-yl)-9H-purin-6-amine (2.0 g, 6.25 mmol) (Townsend
et al Org Lett 2009, 11, 2976-2679) and Ti(iPrO).sub.4 (1.78 g,
6.25 mmol) in MeOH (50 mL) was stirred at 45.degree. C. for 2 h,
then NaCNBH.sub.3 (0.79 g, 12.50 mmol) was added. The reaction was
stirred at RT overnight. The reaction was quenched with aq. sat.
NaHCO.sub.3 (40 mL), filtered, extracted with DCM (40 mL.times.3),
dried over Na.sub.2SO.sub.4 and concentrated. The residue was
purified by SGC (DCM:MeOH=12:1) to obtain title compound (1.7 g,
Yield 63%). .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.28-8.27
(m, 1H), 8.21 (s, 1H), 6.20-6.18 (m, 1H), 5.52 (dd, J=1.5, 6.0 Hz,
1H), 5.00 (dd, J=3.0, 6.0 Hz, 1H), 5.33 (brs, 1H), 3.65-3.63 (m,
3H), 2.77-2.55 (m, 4H), 2.19-2.11 (m, 5H), 2.00-1.82 (m, 2H), 1.59
(s, 3H), 1.38 (s, 3H) ppm; ESI-MS (m/z): 433.2[M+1].sup.+.
Step 3: Synthesis of
(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofur-
o[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)methanol
##STR00242##
[0695] To a solution of methyl
3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutanecarboxylate
(1.0 g, 2.31 mmol) in THE (40 ml) was added LiAlH.sub.4 (0.53 g,
13.89 mmol) at 0.degree. C. and the mixture was stirred overnight.
Water (1.0 g) and 15% NaOH solution (3.0 g) were added slowly to
the mixture and upon stirring for 15 min, the mixture was filtered.
The filtrate was concentrated to obtain the crude title compound
which was used directly in the next step.
Step 4: Synthesis of
(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofur-
o[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)methyl
methanesulfonate
##STR00243##
[0697] To a solution of
(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofur-
o[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)methanol
taken directly from the previous step in DCM (25 ml) was added
Et.sub.3N (467 mg, 4.62 mmol) and MsCl (264 mg, 2.31 mmol) as a
solution in DCM (5 ml). The mixture was stirred for 2 h. Water (20
ml) and DCM (30 ml.times.2) was added. The organic layer was dried
over Na.sub.2SO.sub.4 and concentrated, purified with Prep-TLC
(DCM:MeOH=10:1) to give the title compound (390 mg, Yield 35% for
two steps). .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.27 (s,
1H), 8.21 (s, 1H), 6.203-6.200 (m, 1H), 5.529 (dd, J=2.0, 7.0 Hz,
1H), 5.010 (dd, J=3.0, 6.0 Hz, 1H), 4.354 (dd, J=3.5, 8.0 Hz, 1H),
4.197-4.182 (m, 2H), 3.585 (brs, 1H), 3.073-2.948 (m, 5H),
2.595-2.513 (m, 2H), 2.394 (brs, 1H), 2.207 (brs, 1H), 2.107 (s,
3H), 2.030-1.989 (m, 1H), 1.840-1.811 (m, 2H), 1.586 (s, 3H), 1.390
(brs, 1H), 1.329-1.280 (m, 6H), 0.905-0.878 (m, 1H) ppm; ESI-MS
(m/z): 483.3[M+1].sup.+.
Step 5: Synthesis of
9-((3aR,4R,6R,6aR)-6-(((3-(azidomethyl)cyclobutyl)(methyl)amino)methyl)-2-
,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine
##STR00244##
[0699] To a solution of
(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofur-
o[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)methyl
methane sulfonate (150 mg, 0.31 mmol) in DMF (3 ml) was added
NaN.sub.3 (81 mg, 1.24 mmol). The mixture was heated at 70.degree.
C. for 3 h. Water (30 ml) was added and the mixture was extracted
with ethyl acetate (20 ml.times.3). The combined organic layers
were dried over Na.sub.2SO.sub.4 and concentrated. The residue was
purified by Prep-TLC (DCM:MeOH=30:1) to obtain the title compound
(90 mg, Yield 67%). ESI-MS (m/z): 430.2[M+1].sup.+.
Step 6: Synthesis of
9-((3aR,4R,6R,6aR)-6-(((3-(aminomethyl)cyclobutyl)(methyl)amino)methyl)-2-
,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine
##STR00245##
[0701] Pd/C (10 mg) was added to a solution of
9-((3aR,4R,6R,6aR)-6-(((3-(azidomethyl)cyclobutyl)(methyl)amino)methyl)-2-
,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine
(90 mg, 0.21 mmol) in MeOH (6 ml). The mixture was stirred at RT
overnight under an atmosphere of H2. The mixture was filtered and
the filtrate was concentrated to obtain the title compound which
was used directly in the next step.
Step 7: Synthesis of
1-((3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydro-
furo[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)methyl)-3-(4-(-
tert-butyl)phenyl)urea
##STR00246##
[0703] To a solution of
9-((3aR,4R,6R,6aR)-6-(((3-(aminomethyl)cyclobutyl)(methyl)amino)methyl)-2-
,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine
in DCM (4 ml) was added 1-tert-butyl-4-isocyanatobenzene (37 mg).
The mixture was stirred at RT for 1 h. The mixture was concentrated
and purified via preparative-TLC (DCM:MeOH=10:1) to give the title
compound (55 mg, Yield 45% for two steps). ESI-MS (m/z):
578.3[M+1].
Step 8: Synthesis of Compound 110
[0704] A solution of
1-((3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydro-
furo[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)methyl)-3-(4-(-
tert-butyl)phenyl)urea (55 mg) in HCl/MeOH (2.5 mol/L) (2 mL) was
stirred at RT for 2 h and then concentrated to dryness.
K.sub.2CO.sub.3 (52 mg) in water (0.5 mL) and MeOH (5 mL) was
added. The resulting mixture was stirred for another 10 min at RT,
filtered and the filtrate was concentrated. The residue was
purified by preparative-HPLC to give Compound 110 (10 mg, yield:
25%) as a white solid. .sup.1HNMR (500 MHz, MeOD): .delta..sub.H
8.26 (s, 1H), 8.18 (s, 1H), 7.26-7.19 (m, 4H), 5.96 (d, J=4.5 Hz,
1H), 4.674-4.655 (m, 1H), 4.24-4.16 (m, 2H), 3.15 (d, J=5.0 Hz,
2H), 2.83-2.73 (m, 3H), 2.20-1.59 (m, 8H), 1.26 (s, 9H) ppm; ESI-MS
(m/z): 539.3 [M+1].sup.+.
Example 2: Synthesis of
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((((1r,3S)-3-(2-(5-(tert-butyl)-
-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)tetrahy-
drofuran-3,4-diol (Compound 2)
Step 1: Synthesis of cis and trans methyl
3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)methyl)amino)cyclobutanecarboxylate
##STR00247##
[0706] A solution of methyl 3-oxocyclobutanecarboxylate (4.60 g,
35.94 mmol),
9-((3aR,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydrofuro[3,4--
d][1,3]dioxol-4-yl)-9H-purin-6-amine (11.0 g, 35.94 mmol) and
Ti(iPrO).sub.4 (4.0 g, 14.08 mmol) in MeOH (80 mL) was stirred at
45.degree. C. for 2 h, then NaCNBH.sub.3 (4.5 g, 71.87 mmol) was
added. The reaction was stirred at RT overnight. The reaction was
quenched with aq. sat. NaHCO.sub.3 (40 mL) and filtered, extracted
with DCM (80 mL.times.3), dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified by preparative-HPLC to
obtain the title compound (6.2 g, Yield 41%). .sup.1H NMR (500 MHz,
CDCl.sub.3): .delta..sub.H 8.38-8.34 (m, 1H), 7.90 (s, 1H), 5.98
(d, J=3.0 Hz, 1H), 5.75 (br s, 2H), 5.48-5.46 (m, 1H), 5.03-5.01
(m, 1H), 4.35-4.33 (m, 1H), 3.69-3.66 (m, 3H), 3.50-3.17 (m, 1H),
3.05-2.73 (m, 3H), 2.48-2.44 (m, 2H), 1.95-1.91 (m, 2H), 1.62 (s,
3H), 1.39 (s, 3H) ppm; ESI-MS (m/z): 419.2[M+1].sup.+.
[0707] The cis/trans mixture of methyl
3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)methyl)amino)cyclobutanecarboxylate (6.2 g)
was separated via chiral HPLC (CHIRALCEL AD-H 20*250 mm, Sum
(Daicel), Column temperature: 35.degree. C., Mobile phase:
CO2/Methanol (0.1% DEA)=70/30, Flow rate: 50 g/min) to give the
pure cis product (3.5 g) and pure trans product (1.7 g).
Step 2: Synthesis of (1S,3s)-methyl
3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutanecarboxylate
##STR00248##
[0709] To a solution of cis methyl
3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)methyl)amino)cyclobutanecarboxylate (2.0 g,
4.78 mmol) in CH.sub.3CN (15 ml) was added 2-iodopropane (4.0 g,
23.92 mmol) and K.sub.2CO.sub.3 (1.0 g, 7.18 mmol). The reaction
was heated to 95.degree. C. overnight in a sealed tube. The mixture
was filtered, the filtrate was concentrated and purified by SGC
(DCM:MeOH=12:1) to obtain the title compound (1.9 g, Yield 86%).
.sup.1H NMR (500 MHz, CDCl.sub.3): .delta..sub.H 8.37 (s, 1H), 7.89
(s, 1H), 6.03 (d, J=1.5 Hz, 1H), 5.53-5.48 (m, 3H), 5.00 (br s,
1H), 4.25 (brs, 1H), 3.66 (s, 3H), 3.19-3.18 (m, 1H), 2.96 (brs,
1H), 2.80-2.78 (m, 1H), 2.67-2.58 (m, 2H), 2.20-2.12 (m, 4H), 1.62
(s, 3H), 1.39 (s, 3H), 1.00 (d, J=6.0 Hz, 3H), 0.84 (d, J=6.0 Hz,
3H) ppm; ESI-MS (m/z): 461.4[M+1].sup.+.
Step 3: Synthesis of
(1S,3s)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrah-
ydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutanecarbald-
ehyde
##STR00249##
[0711] To a solution of (1S,3s)-methyl
3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutanecarboxylate
(1.2 g, 2.60 mmol) in DCM (50 ml) was added DIBAL-H dropwise at
-78.degree. C. until all the starting material was consumed as
determined by TLC. MeOH (2 ml) was added and the mixture was
stirred to RT for 30 min. upon which water (50 ml) was added and
the mixture was extracted with DCM (50 ml.times.2). The organic
layer was dried over Na.sub.2SO.sub.4 and concentrated to obtain
crude title compound (1.0 g which was used) directly in the next
step. .sup.1H NMR (500 MHz, CDCl.sub.3): .delta..sub.H 9.56 (d,
J=2.5 Hz, 1H), 8.36 (s, 1H), 7.88 (s, 1H), 6.03 (d, J=2.5 Hz, 1H),
5.66 (br s, 2H), 5.50 (dd, J=2.0, 6.5 Hz, 1H), 5.01 (dd, J=3.5, 6.5
Hz, 1H), 3.331-3.337 (m, 1H), 2.96-2.97 (m, 1H), 2.77-2.59 (m, 3H),
2.14-2.05 (m, 4H), 1.60 (s, 3H), 1.39 (s, 3H), 1.01 (d, J=6.5 Hz,
3H), 0.85 (d, J=6.0 Hz, 3H) ppm.
Step 4: Synthesis of (E)-ethyl
3-((1S,3s)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)acry-
late
##STR00250##
[0713] To a solution of
(1S,3s)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrah-
ydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutane
carbaldehyde (930 mg, 2.16 mmol) in CH.sub.3CN:DCM=5:1 (50 ml) was
added ethyl 2-(diethoxyphosphoryl)acetate (484 mg, 2.16 mmol), DBU
(328 mg, 2.16 mmol) and LiCl (91 mg, 2.16 mmol). The mixture was
stirred at RT for 1 h and then concentrated. Water (20 ml) was
added and the mixture extracted with DCM (25 ml.times.3). The
combined organic layers were dried over Na.sub.2SO.sub.4,
concentrated and the residue was purified by SGC (DCM:MeOH=30:1) to
obtain title compound (900 mg, Yield 83%). .sup.1H NMR (500 MHz,
CDCl.sub.3): .delta..sub.H 8.36 (s, 1H), 7.89 (s, 1H), 6.94-6.90
(m, 1H), 6.03 (s, 1H), 5.72-5.89 (m, 1H), 5.57 (s, 2H), 5.52 (d,
J=4.5 Hz, 1H), 5.00 (dd, J=3.5, 6.0 Hz, 1H), 4.25 (d, J=3.0 Hz,
1H), 4.21-4.17 (m, 2H), 3.14 (brs, 1H), 2.961-2.936 (m, 1H),
2.74-2.52 (m, 3H), 2.22-2.14 (m, 2H), 1.79-1.76 (m, 2H), 1.60 (s,
3H), 1.40 (s, 3H), 1.30-1.27 (m, 3H), 1.00 (d, J=7.0 Hz, 3H), 0.82
(d, J=6.5 Hz, 3H) ppm; ESI-MS (m/z): 501.4[M+1].sup.+.
Step 5: Synthesis of ethyl
3-((1S,3r)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)prop-
anoate
##STR00251##
[0715] To a solution of (E)-ethyl
3-((1S,3s)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)
acrylate (900 mg, 1.8 mmol) in MeOH (50 ml) was added Pd/C (20 mg).
The mixture was stirred at RT overnight under an atmosphere of
hydrogen. The mixture was filtered and the filtrate was
concentrated to obtain title compound (700 mg, Yield 78%). .sup.1H
NMR (500 MHz, CDCl.sub.3): .delta..sub.H 8.36 (s, 1H), 7.89 (s,
1H), 6.03 (d, J=2.5 Hz, 1H), 5.69 (s, 2H), 5.51 (dd, J=2.5, 8.0 Hz,
1H), 4.99 (dd, J=4.0, 7.5 Hz, 1H), 4.26 (brs, 1H), 4.13-4.08 (m,
2H), 2.99-2.92 (m, 2H), 2.706-2.655 (m, 1H), 2.539-2.486 (m, 1H),
2.18-2.02 (m, 4H), 1.76 (brs, 1H), 1.65-1.60 (m, 5H), 1.43-1.37 (m,
5H), 1.26-1.23 (m, 2H), 0.97 (d, J=9.0 Hz, 3H), 0.79 (d, J=8.5 Hz,
3H) ppm; ESI-MS (m/z): 503.4[M+1].sup.+.
Step 6: Synthesis of
3-((1S,3r)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)prop-
anoic acid
##STR00252##
[0717] To a solution of ethyl
3-((1S,3r)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)
(isopropyl)amino)cyclobutyl)propanoate (650 mg, 1.29 mmol) in
THE:MeOH=5:1 (30 ml) was added LiOH.H.sub.2O (543 mg, 1.29 mmol).
The mixture was stirred at RT overnight, concentrated and then
taken up in MeOH (10 ml). 1M HCl solution was added dropwise at
0.degree. C. until pH=7. The mixture was concentrated and purified
with preparative-HPLC to give title compound (170 mg).
Step 7: Synthesis of
N-(2-amino-4-(tert-butyl)phenyl)-3-((1S,3r)-3-((((3aR,4R,6R,6aR)-6-(6-ami-
no-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(isopropyl)amino)cyclobutyl)propanamide
##STR00253##
[0719] To a solution of
3-((1S,3r)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)prop-
anoic acid (170 mg, 0.36 mmol) in DCM (15 ml) was added
4-tert-butylbenzene-1,2-diamine (117 mg, 0.72 mmol), EDCI (137 mg,
0.72 mmol), HOBT (97 mg, 0.72 mmol) and TEA (217 mg, 2.15 mmol).
The mixture was stirred at RT overnight and concentrated. Saturated
NaHCO.sub.3 solution (20 ml) was added and the mixture extracted
with DCM (20 ml.times.3). The organic layers were dried over
Na.sub.2SO.sub.4 and concentrated. The crude was purified with
preparative-TLC (DCM:MeOH=12:1) to afford the title compound (110
mg crude).
Step 8: Synthesis of
9-((3aR,4R,6R,6aR)-6-((((1r,3S)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol--
2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltetrahydrofuro[-
3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine
##STR00254##
[0721] A solution of
N-(2-amino-4-(tert-butyl)phenyl)-3-((1S,3r)-3-((((3aR,4R,6R,6aR)-6-(6-ami-
no-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(isopropyl)amino)cyclobutyl)propanamide (110 mg) in AcOH (10 ml)
was heated to 65.degree. C. overnight. The mixture was
concentrated, saturated NaHCO.sub.3 solution (20 ml) was added and
the mixture extracted with DCM (20 ml.times.3). The combined
organic layers were dried over Na.sub.2SO.sub.4 and concentrated to
give the title compound (105 mg crude). .sup.1H NMR (500 MHz,
CDCl.sub.3): .delta..sub.H 8.36 (s, 1H), 7.89 (s, 1H), 7.48-7.24
(m, 3H), 6.01 (d, J=1.5 Hz, 1H), 5.60-5.53 (m, 3H), 4.98 (dd,
J=3.0, 6.5 Hz, 1H), 4.22 (brs, 1H), 2.97 (brs, 1H), 2.874-2.847 (m,
1H), 2.56-2.50 (m, 3H), 1.87-1.78 (m, 2H), 1.70-1.54 (m, 7H),
1.35-1.17 (m, 14H), 0.90 (d, J=6.5 Hz, 3H), 0.80 (d, J=6.5 Hz, 3H)
ppm; ESI-MS (m/z): 603.5[M+1].sup.+.
Step 9: Synthesis of Compound 2
[0722] A solution of
9-((3aR,4R,6R,6aR)-6-((((r,3S)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-
-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3-
,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine (105 mg) in HCl/MeOH (2.5
mol/L) (10 mL) was stirred at RT for 2 h, then concentrated to
dryness. K.sub.2CO.sub.3 (96 mg) in water (0.5 mL) and MeOH (5 mL)
were added and the resulting mixture was stirred for another 10 min
at RT and then filtered. The filtrate was concentrated and the
residue was purified by preparative-HPLC (xbridge 30 mm*150 mm,
Mobile phase: A: water (10 mM NH4HCO3) B: CAN, Gradient: 35-45% B
in 10 min, 45-45% B in 6 min, stop at 20 min, Flow rate: 50 ml/min)
to give Compound 2 (50 mg, yield: 51%) as a white solid. .sup.1HNMR
(500 MHz, MeOD): .delta..sub.H 8.29 (s, 1H), 8.20 (s, 1H),
7.47-7.39 (m, 3H), 5.96 (d, J=4.0 Hz, 1H), 4.70-4.75 (m, 1H),
4.26-4.27 (m, 1H), 4.05-4.06 (m, 1H), 3.140-3.155 (m, 1H),
3.00-2.76 (m, 5H), 2.18-2.16 (m, 2H), 1.87-1.85 (m, 2H), 1.57-1.55
(m, 2H), 1.36 (s, 9H), 1.01 (d, J=6.5 Hz, 3H), 0.94 (d, J=6.5 Hz,
3H) ppm; ESI-MS (m/z): 563.4 [M+1].sup.+.
Example 3: Synthesis of
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((((1s,3R)-3-(2-(5-(tert-butyl)-
-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)tetrahy-
drofuran-3,4-diol (Compound 3)
Step 1: Synthesis of (1R,3r)-methyl
3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutanecarboxylate
##STR00255##
[0724] To a solution of (1R,3r)-methyl
3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)methyl)amino)cyclobutanecarboxylate (1.7 g,
4.07 mmol) in CH.sub.3CN (15 ml) was added 2-iodopropane (3.5 g,
20.3 mmol) and K.sub.2CO.sub.3 (0.84 g, 6.10 mmol). The reaction
was heated to 95.degree. C. overnight in a sealed tube. The mixture
was filtered and the filtrate was and concentrated and purified by
SGC (DCM:MeOH=12:1) to obtain title compound (1.35 g, Yield 72%).
.sup.1H NMR (500 MHz, CDCl.sub.3): .delta..sub.H 8.36 (s, 1H), 7.88
(s, 1H), 6.03 (d, J=2.0 Hz, 1H), 5.55 (m, 2H), 5.49 (dd, J=1.5, 6.0
Hz, 1H), 5.01 (dd, J=3.5, 6.0 Hz, 1H), 4.254-4.247 (m, 1H), 3.68
(s, 3H), 3.60-3.50 (m, 1H), 2.930-2.917 (m, 1H), 2.79-2.74 (m, 2H),
2.59-2.57 (m, 1H), 2.25-2.12 (m, 4H), 1.60 (s, 3H), 1.39 (s, 3H),
1.00 (d, J=6.5 Hz, 3H), 0.83 (d, J=7.0 Hz, 3H) ppm; ESI-MS (m/z):
461.3[M+1].sup.+.
Step 2: Synthesis of
(1R,3r)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrah-
ydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutanecarbald-
ehyde
##STR00256##
[0726] To a solution of (1R,3r)-methyl
3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutanecarboxylate
(1.35 g, 2.93 mmol) in DCM (50 ml) at
-78.degree. C. was added DiBAL-H dropwise until the starting
material was completely consumed as determined by TLC. MeOH (2 ml)
was added and the mixture was stirred at room temperature (RT) for
30 min. Water (50 ml) was added and the mixture was extracted with
DCM (50 ml.times.2). The combined organic layers were dried over
Na.sub.2SO.sub.4 and concentrated to obtain crude title compound
(1.1 g) which was used directly in the next step. .sup.1H NMR (500
MHz, CDCl.sub.3): .delta..sub.H 9.80 (s, 1H), 8.35 (s, 1H), 7.88
(s, 1H), 6.04 (s, 1H), 5.56 (s, 2H), 5.50 (d, J=6.5 Hz, 1H),
5.028-5.026 (m, 1H), 4.26 (brs, 1H), 3.33-3.30 (m, 1H), 2.956-2.930
(m, 1H), 2.80-2.55 (m, 3H), 2.27-2.07 (m, 4H), 1.60 (s, 3H), 1.39
(s, 3H), 1.00 (d, J=7.0 Hz, 3H), 0.82 (d, J=6.5 Hz, 3H) ppm.
Step 3: Synthesis of (E)-ethyl
3-((1R,3r)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)acry-
late
##STR00257##
[0728] To a solution of
(1R,3r)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrah-
ydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutanecarbald-
ehyde (1.1 g, 2.56 mmol) in CH.sub.3CN:DCM=5:1 (50 ml) was added
ethyl 2-(diethoxyphosphoryl)acetate (573 mg, 2.56 mmol), DBU (389
mg, 2.56 mmol) and LiCl (107 mg, 2.56 mmol). The mixture was
stirred at RT for 1 h and concentrated, upon which water (20 ml)
was added and the mixture was extracted with DCM (25 ml.times.3).
The combined organic layers were dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified by SGC (DCM:MeOH=30:1) to
obtain the title compound (1.0 g, Yield 78%). .sup.1H NMR (500 MHz,
CDCl.sub.3): .delta..sub.H 8.35 (s, 1H), 7.89 (s, 1H), 7.16-7.11
(m, 1H), 6.03 (d, J=2.0 Hz, 1H), 5.79-5.76 (m, 1H), 5.56 (s, 2H),
5.51 (dd, J=1.5, 6.0 Hz, 1H), 5.02 (dd, J=3.0, 6.0 Hz, 1H), 4.25
(d, J=8.0 Hz, 1H), 4.22-4.17 (m, 2H), 3.44 (brs, 1H), 2.93 (brs,
1H), 2.78-2.56 (m, 3H), 2.27-2.16 (m, 2H), 1.93-1.91 (m, 2H), 1.60
(s, 3H), 1.40 (s, 3H), 1.31-1.27 (m, 3H), 0.98 (d, J=6.5 Hz, 3H),
0.82 (d, J=6.5 Hz, 3H) ppm; ESI-MS (m/z): 501.4[M+1].sup.+.
Step 4: Synthesis of ethyl
3-((1R,3s)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)prop-
anoate
##STR00258##
[0730] To a mixture of (E)-ethyl
3-((1R,3r)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)acry-
late (1.0 g, 2.0 mmol) and 10% Pd/C (30 mg) in MeOH (50 ml) was
added Pd/C (30 mg). The mixture was stirred at RT overnight under a
hydrogen atmosphere. The resulting mixture was filtered and the
filtrate was concentrated to obtain the title compound (1.0 g,
Yield 100%). .sup.1H NMR (500 MHz, CDCl.sub.3): .delta..sub.H 8.36
(s, 1H), 7.89 (s, 1H), 6.03 (d, J=2.5 Hz, 1H), 5.58 (s, 2H), 5.51
(dd, J=2.0, 6.5 Hz, 1H), 5.00 (dd, J=3.5, 6.0 Hz, 1H), 4.276-4.269
(m, 1H), 4.13-4.09 (m, 2H), 3.38-3.37 (m, 1H), 2.94-2.54 (m, 3H),
2.22-1.97 (m, 5H), 1.79-1.62 (m, 4H), 1.60 (s, 3H), 1.40 (s, 3H),
1.28-1.23 (m, 2H), 0.97 (d, J=7.0 Hz, 3H), 0.79 (d, J=7.0 Hz, 3H)
ppm; ESI-MS (m/z): 503.4[M+1].sup.+.
Step 5: Synthesis of
3-((1R,3s)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)prop-
anoic acid
##STR00259##
[0732] To a solution of ethyl
3-((1R,3s)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)prop-
anoate (360 mg, 0.72 mmol) in THE:MeOH=5:1 (30 ml) was added
LiOH.H.sub.2O (301 mg, 7.20 mmol). The mixture was stirred at RT
overnight, concentrated then dissolved in MeOH (10 ml). 1 M HCl
solution was added dropwise at 0.degree. C. until pH=7. The mixture
was concentrated to give the title compound crude and was used
directly in the next step.
Step 6: Synthesis of
N-(2-amino-4-(tert-butyl)phenyl)-3-((1R,3s)-3-((((3aR,4R,6R,6aR)-6-(6-ami-
no-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(isopropyl)amino)cyclobutyl)propanamide
##STR00260##
[0734] To a solution of
3-((1R,3s)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)prop-
anoic acid in DMF (5 ml) was added
4-(tert-butyl)benzene-1,2-diamine (235 mg, 1.43 mmol), EDCI (274
mg, 1.43 mmol), HOBT (193 mg, 1.43 mmol) and TEA (435 mg, 4.30
mmol). The mixture was heated to 45.degree. C. overnight and
concentrated. Saturated NaHCO.sub.3 solution (20 ml) was added and
the mixture was extracted with DCM (20 ml.times.3). The organic
layers were dried over Na.sub.2SO.sub.4 and concentrated. The crude
was purified with preparative-TLC (DCM:MeOH=12:1) to afford the
title compound (110 mg), which was carried forward into the next
step without further purification.
Step 7: Synthesis of
9-((3aR,4R,6R,6aR)-6-((((1s,3R)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol--
2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltetrahydrofuro[-
3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine
##STR00261##
[0736] A solution of
N-(2-amino-4-(tert-butyl)phenyl)-3-((1R,3s)-3-((((3aR,4R,6R,6aR)-6-(6-ami-
no-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(isopropyl)amino)cyclobutyl)propanamide (110 mg) in AcOH (15 ml)
was heated at 65.degree. C. overnight. The mixture was
concentrated, saturated NaHCO.sub.3 solution (20 ml) was added and
the mixture was extracted with DCM (20 ml.times.3). The combined
organic layers were dried over Na.sub.2SO.sub.4 and concentrated to
give the title compound (100 mg crude). .sup.1H NMR (500 MHz,
CDCl.sub.3): .delta..sub.H 8.36 (s, 1H), 7.94 (s, 1H), 7.48-7.27
(m, 3H), 6.07 (d, J=1.5 Hz, 1H), 5.64-5.58 (m, 3H), 5.02 (dd,
J=3.0, 6.0 Hz, 1H), 4.30 (brs, 1H), 3.38-3.37 (m, 1H), 2.97-2.95
(m, 1H), 2.76-2.55 (m, 3H), 1.97-1.74 (m, 5H), 1.67-1.57 (m, 5H),
1.45-1.40 (m, 12H), 0.99 (d, J=6.5 Hz, 3H), 0.83 (d, J=6.5 Hz, 3H)
ppm; ESI-MS (m/z): 603.5[M+1].sup.+.
Step 8: Synthesis of Compound 3
[0737] A solution of
9-((3aR,4R,6R,6aR)-6-((((1s,3R)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol--
2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltetrahydrofuro[-
3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine (190 mg) in HCl/MeOH (2.5
mol/L) (15 mL) was stirred at RT for 2 h and concentrated to
dryness. K.sub.2CO.sub.3 (161 mg) in water (0.5 mL) and MeOH (5 mL)
were added. The resulting mixture was stirred for another 10 min at
RT then filtered. The filtrate was concentrated and the residue was
purified by preparative HPLC (xbridge 30 mm*150 mm, Mobile phase:
A: water (10 mM NH4HCO3) B: CAN, Gradient: 35-45% B in 10 min,
45-45% B in 6 min, stop at 20 min, Flow rate: 50 ml/min) to give
Compound 3 (65 mg, yield: 70%) as a white solid. .sup.1HNMR (500
MHz, MeOD): .delta..sub.H 8.29 (s, 1H), 8.19 (s, 1H), 7.47-7.28 (m,
3H), 5.95 (d, J=4.5 Hz, 1H), 4.744-4.724 (m, 1H), 4.27-4.26 (m,
1H), 4.07-4.06 (m, 1H), 3.56 (brs, 1H), 3.01-2.78 (m, 5H), 2.17
(brs, 2H), 2.00-1.93 (m, 2H), 1.80-1.79 (m, 2H), 1.36 (s, 9H), 1.02
(d, J=5.5 Hz, 3H), 0.95 (d, J=6.0 Hz, 3H) ppm; ESI-MS (m/z): 563.5
[M+1].sup.+.
Example 4: Synthesis of
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((((1s,3R)-3-(2-(5-chloro-6-(tr-
ifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)-
methyl)tetrahydrofuran-3,4-diol (Compound 4)
Step 1: Synthesis of
N-(2-amino-4-chloro-5-(trifluoromethyl)phenyl)-3-((1R,3s)-3-((((3aR,4R,6R-
,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]diox-
ol-4-yl)methyl)(isopropyl)amino)cyclobutyl)propanamide
##STR00262##
[0739] To a solution of
3-((1R,3s)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)prop-
anoic acid (250 mg, 0.53 mmol) in DCM (30 ml) was added
4-chloro-5-(trifluoromethyl)benzene-1,2-diamine (221 mg, 1.05
mmol), EDCI (201 mg, 1.05 mmol), HOBT (142 mg, 1.05 mmol) and TEA
(320 mg, 3.15 mmol). The mixture was stirred at RT overnight, upon
which saturated NaHCO.sub.3 solution (20 ml) was added and the
mixture was extracted with DCM (20 ml.times.3). The combined
organic layers were dried over Na.sub.2SO.sub.4, filtered and
concentrated. The crude was purified via preparative-TLC
(DCM:MeOH=12:1) to afford the title compound (250 mg crude).
Step 2: Synthesis of
9-((3aR,4R,6R,6aR)-6-((((1s,3R)-3-(2-(5-chloro-6-(trifluoromethyl)-1H-ben-
zo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethylt-
etrahydrofuro[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine
##STR00263##
[0741] A solution of
N-(2-amino-4-chloro-5-(trifluoromethyl)phenyl)-3-((1R,3s)-3-((((3aR,4R,6R-
,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]diox-
ol-4-yl)methyl)(isopropyl)amino)cyclobutyl)propanamide (250 mg) in
AcOH (15 ml) was heated to 65.degree. C. overnight. The mixture was
concentrated, saturated NaHCO.sub.3 solution (20 ml) was added and
the mixture was extracted with DCM (20 ml.times.3). The combined
organic layers were dried over Na.sub.2SO.sub.4 and concentrated to
give the title compound (200 mg crude).
Step 3: Synthesis of Compound 4
[0742] A solution of
9-((3aR,4R,6R,6aR)-6-((((1s,3R)-3-(2-(5-chloro-6-(trifluoromethyl)-1H-ben-
zo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethylt-
etrahydrofuro[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine (200 mg) in
HCl/MeOH (2.5 mol/L) (15 mL) was stirred at RT for 2 h upon which
it was concentrated to dryness. K.sub.2CO.sub.3 (166 mg) in water
(0.5 mL) and MeOH (5 mL) were added and the resulting mixture was
stirred for another 10 min at RT. The mixture was filtered and the
filtrate was concentrated. The residue was purified by
preparative-HPLC to give Compound 4 (80 mg, yield: 43%) as a white
solid. .sup.1HNMR (500 MHz, MeOD): .delta..sub.H 8.29 (s, 1H), 8.19
(s, 1H), 7.88 (s, 1H), 7.68 (s, 1H), 5.96 (d, J=4.0 Hz, 1H),
4.748-4.730 (m, 1H), 4.284-4.263 (m, 1H), 4.09 (br s, 1H),
3.65-3.50 (m, 1H), 3.03-2.85 (m, 5H), 2.191-2.176 (m, 2H),
2.03-2.00 (m, 2H), 1.80 (brs, 2H), 1.02 (d, J=6.0 Hz, 3H), 0.96 (d,
J=6.6 Hz, 3H) ppm; ESI-MS (m/z): 609.2 [M+1].sup.+.
Example 5: Synthesis of
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((((1r,3S)-3-(2-(5-chloro-6-(tr-
ifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)-
methyl)tetrahydrofuran-3,4-diol (Compound 5)
Step 1: Synthesis of
N-(2-amino-4-chloro-5-(trifluoromethyl)phenyl)-3-((1S,3r)-3-((((3aR,4R,6R-
,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]diox-
ol-4-yl)methyl)(isopropyl)amino)cyclobutyl)propanamide
##STR00264##
[0744] To a solution of
3-((1S,3r)-3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltet-
rahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)prop-
anoic acid in DCM:DMF=15:1 (30 ml) was added
4-chloro-5-(trifluoromethyl)benzene-1,2-diamine (334 mg, 1.60
mmol), EDCI (304 mg, 1.60 mmol), HOBT (215 mg, 1.60 mmol) and TEA
(483 mg, 4.80 mmol). The mixture was stirred overnight at RT. The
mixture was concentrated, saturated NaHCO.sub.3 solution (20 ml)
was added and the resultant mixture was extracted with DCM (20
ml.times.3). The combined organic layers were dried over
Na.sub.2SO.sub.4 and concentrated. The crude residue was purified
via preparative TLC (DCM:MeOH=12:1) to afford the title compound
(220 mg).
Step 2: Synthesis of
9-((3aR,4R,6R,6aR)-6-((((r,3S)-3-(2-(5-chloro-6-(trifluoromethyl)-1H-benz-
o[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethylte-
trahydrofuro[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine
##STR00265##
[0746] A solution of
N-(2-amino-4-chloro-5-(trifluoromethyl)phenyl)-3-((1S,3r)-3-((((3aR,4R,6R-
,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]diox-
ol-4-yl)methyl)(isopropyl)amino)cyclobutyl)propanamide (220 mg) in
AcOH (15 ml) was heated at 65.degree. C. overnight. The mixture was
concentrated, saturated NaHCO.sub.3 solution (20 ml) was added, and
the mixture was extracted with DCM (20 ml.times.3). The combined
organic layers were dried over Na.sub.2SO.sub.4 and concentrated to
give the title compound (190 mg).
Step 3: Synthesis of Compound 5
[0747] A solution of
9-((3aR,4R,6R,6aR)-6-((((r,3S)-3-(2-(5-chloro-6-(trifluoromethyl)-1H-benz-
o[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethylte-
trahydrofuro[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine (190 mg) in
HCl/MeOH (2.5 mol/L) (15 mL) was stirred at RT for 2 h then it was
concentrated to dryness. K.sub.2CO.sub.3 (161 mg) in water (0.5 mL)
and MeOH (5 mL) were added and the resulting mixture was stirred
for another 10 min at RT. The mixture was filtered and the filtrate
was concentrated. The residue was purified by preparative-HPLC to
give Compound 5 (90 mg, yield: 51%) as a white solid. .sup.1HNMR
(500 MHz, MeOD): .delta..sub.H 8.29 (s, 1H), 8.19 (s, 1H), 7.88 (s,
1H), 7.67 (s, 1H), 5.95 (d, J=5.0 Hz, 1H), 4.736-4.716 (m, 1H),
4.268-4.246 (m, 1H), 4.070-4.051 (m, 1H), 3.15 (brs, 1H), 3.00-2.71
(m, 5H), 2.17 (brs, 2H), 1.93-1.88 (m, 2H), 1.58-1.56 (m, 2H), 1.01
(d, J=5.5 Hz, 3H), 0.95 (d, J=6.0 Hz, 3H) ppm; ESI-MS (m/z): 609.2
[M+1].sup.+.
Example 6: Synthesis of
(2R,3R,4S,5R)-2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-((5-(ter-
t-butyl)-1H-benzo[d]imidazol-2-yl)methyl)cyclobutyl)(methyl)amino)methyl)t-
etrahydrofuran-3,4-diol (Compound 6)
Step 1: Synthesis of
7-((3aR,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]-
dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
##STR00266##
[0749] A solution of
((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrim-
idin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methanol
(2.83 g, 6.20 mmol) and triphenylphosphine (2.28 g, 8.68 mmol) in
dry tetrahydrofuran (32 mL) was cooled at 0.degree. C. in an
ice/water bath. Diisopropyl azodicarboxylate (1.71 mL, 8.68 mmol)
was added dropwise, followed by a solution of diphenylphosphonic
azide (1.87 mL, 8.68 mmol) in tetrahydrofuran (5.3 mL, 66 mmol).
Upon addition of the DPPA solution, a white milky precipitate
formed. After about 30 minutes, the reaction mixture was allowed to
warm to room temperature and stir overnight. After 24 h, HPLC
indicated that all the starting material had been consumed. The
reaction mixture was concentrated to about 1/2 the original volume
and purified by flash chromatography (175 g silica gel, 10-55%
EA/hept) to yield the title compound (2.49 g, 83%) as a slightly
yellow stiff foam: MS (ESI+) for C.sub.23H.sub.27N.sub.7O.sub.5 m/z
482.2 (M+H).sup.+; (ESI-) for C.sub.23H.sub.27N.sub.7O.sub.5 m/z
480.1 (M+H), m/z 526.1 (M+CO.sub.2H); HPLC purity 97%.
Step 2: Synthesis of
7-((3aR,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]-
dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
##STR00267##
[0751] A solution of
((3aR,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]di-
oxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
(2.49 g, 5.17 mmol) in tetrahydrofuran (50 mL, 600 mmol) was
treated dropwise with a solution of 1.0 M of trimethylphosphine in
tetrahydrofuran (7.24 mL, 7.24 mmol) and the mixture was stirred
for 20 h. The reaction mixture was treated with water (1.80 mL,
99.9 mmol) and stirred at RT for 2 h. The reaction mixture was
concentrated, the crude product was taken up in 90 mL
CH.sub.2Cl.sub.2 and washed with four 30 mL portions of H.sub.2O
and 15 mL brine. The solution was dried over Na.sub.2SO.sub.4,
filtered and concentrated. The crude material was purified by flash
chromatography (120 g silica gel, 3-10% 7N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2) to yield the title compound (1.76 g,
75%) as a foam: MS (ESI+) for C.sub.23H.sub.29N.sub.5O.sub.5 m/z
456.2 (M+H).sup.+; (ESI-) for C.sub.26H.sub.35N.sub.5O.sub.5 m/z
454.1 (M-H).sup.+; HPLC purity 92% (ret. time, 2.65 min).
Step 3: Synthesis of methyl
2-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)amino)cyclobutyl)acetate
##STR00268##
[0753] A solution
7-((3aR,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]-
dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
(400 mg, 0.88 mmol) and methyl 2-(3-oxocyclobutyl)acetate (100 mg,
0.70 mmol) [prepared using the procedure found in US Patent
Application Publication 2009/0118287] in 1,2-dichloroethane (12 mL)
was treated dropwise with acetic acid (50 uL, 0.88 mmol). The
solution was treated with sodium triacetoxyborohydride (260 mg, 1.2
mmol) in one portion and allowed to stir at room temperature until
complete by HPLC. After 4 h, HPLC indicated the reaction was about
80% complete. An additional 20 mg of ketone was added and stirring
was continued for 2.5 h. The reaction mixture was diluted with 30
mL CH.sub.2Cl.sub.2 and washed with 15 mL sat NaHCO.sub.3. The
aqueous phase was washed with 15 mL CH.sub.2Cl.sub.2 and the
combined organic phase was dried over Na.sub.2SO.sub.4. The organic
phase was filtered and concentrated to yield a light yellow glass
that was purified by flash chromatography (70 g silica gel; 2% 7N
NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2) to yield the title
compound (270 mg, 66%) as a colorless glass: MS (ESI+) for
C.sub.3H39N507 m/z 582.2 (M+H).sup.+; HPLC purity >95% (ret.
time, 2.88 min).
Step 4: Synthesis of methyl
2-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(methyl)amino)cyclobutyl)acetate
##STR00269##
[0755] A solution of methyl
2-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)amino)cyclobutyl)acetate (267 mg, 0.459 mmol) in methanol (12 mL)
was treated with sodium cyanoborohydride (380 mg, 6.1 mmol). The pH
of the solution was adjusted to .about.6 by the dropwise addition
of a 10% (v/v) solution of glacial acetic acid in methanol. The
mixture was treated with 37% formaldehyde (0.57 mL, 7.6 mmol)
dropwise and the mixture was stirred at room temperature for 1 h at
which time, HPLC indicated the starting material was consumed. The
reaction mixture was concentrated to remove the methanol. The
aqueous solution that remained was diluted with 25 mL NaHCO.sub.3
and the aqueous phase was extracted with three 20 mL portions of
CH.sub.2Cl.sub.2. The organic phase was washed with 20 mL sat
NaHCO.sub.3, dried over Na.sub.2SO.sub.4, filtered and concentrated
to yield the title compound (272 mg, 100%) as a colorless stiff
foam which was found to be of sufficient purity for use in the next
step: MS (ESI+) for C.sub.31H.sub.41N.sub.5O.sub.7 m/z 596.5
(M+H).sup.+; HPLC purity >95% (ret. time, 2.89 min).
Step 5: Synthesis of
2-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(methyl)amino)cyclobutyl)acetic acid
##STR00270##
[0757] A solution of methyl
2-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(methyl)amino)cyclobutyl)acetate (270 mg, 0.453 mmol) in methanol
(8.6 mL) was treated dropwise with a solution of sodium hydroxide
(36 mg, 0.91 mmol) in water (0.9 mL, 50 mmol) and the mixture was
heated at 50.degree. C. After 17 h, HPLC indicated the reaction was
complete. The reaction mixture was cooled to room temperature and
treated with 0.91 mL 1.0N HCl to adjust the pH to .about.7. The
solution was concentrated to remove the methanol and the resulting
aqueous suspension was lyophilized to yield a white solid. The
material was used as is in the next step, assuming a quantitative
recovery: MS (ESI+) for C.sub.3H.sub.39N.sub.5O.sub.7 m/z 582.4
(M+H).sup.+; MS (ESI-) for C.sub.30H.sub.39N.sub.5O.sub.7 m/z 580.4
(M-H).sup.-; HPLC purity >95% (ret. time, 2.72 min).
Step 6: Synthesis of
N-(2-amino-4-(tert-butyl)phenyl)-2-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimet-
hoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)acetamide
##STR00271##
[0759] A solution of
2-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(methyl)amino)cyclobutyl)acetic acid and
4-tert-butylbenzene-1,2-diamine (89.4 mg, 0.545 mmol) in
N,N-dimethylformamide (4.5 mL) was treated with
N,N-diisopropylethylamine (0.261 mL, 1.50 mmol) dropwise followed
by N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
hexafluorophosphate (259 mg, 0.681 mmol). The solution was allowed
to stir at room temperature for 18 h during which time LCMS
indicated the starting material had been consumed. The reaction
mixture was concentrated under high vac. The residue was taken up
in 30 mL ethyl acetate and 20 mL 1/1 H.sub.2O/sat NaHCO.sub.3
solution. The mixture was extracted and the aqueous phase was
washed with 35 mL ethyl acetate. The combined organic phase was
washed with two 20 mL portions of H.sub.2O, and 20 mL brine. The
organic phase was dried over Na.sub.2SO.sub.4, filtered and
concentrated to yield a tannish brown glass/stiff foam. The crude
material was purified by flash chromatography (35 g silica gel; 4%
7N NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2) to yield the title
compound (272 mg, 82%) as a light tan glass/stiff foam which was a
mixture of regioisomeric amides: MS (ESI+) for
C.sub.41H.sub.53N.sub.7O.sub.6 m/z 728.8 (M+H).sup.+; MS (ESI-) for
C.sub.41H.sub.53N.sub.7O.sub.6 m/z 726.9 (M-H).sup.+; HPLC purity
>95%, (ret. time, 3.14, 3.17 min) two peaks observed due to
amide regioisomers.
Step 7: Synthesis of
7-((3aR,4R,6R,6aR)-6-(((3-((5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)methy-
l)cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]d-
ioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
##STR00272##
[0761]
N-(2-amino-4-(tert-butyl)phenyl)-2-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-
-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetra-
hydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)acetamide
(272 mg, 0.374 mmol) was taken up in acetic acid (7.2 mL) and the
solution was heated at 65.degree. C. After 1.5 h, HPLC indicated
the reaction was complete. The reaction was cooled to room
temperature and the solvent removed under high vac. The residue was
taken up in 35 mL CH.sub.2Cl.sub.2 and the organic phase was washed
with 25 mL sat NaHCO.sub.3 solution and 20 mL 2% Na.sub.2CO.sub.3
solution. The organic phase was dried over Na.sub.2SO.sub.4,
filtered and concentrated to yield a light tan glass/stiff foam.
The crude material was purified by flash chromatography (30 g
silica gel; 4% 7N NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2) to yield
the title compound (224 mg, 84%) as a light tan glass which was a
mixture of cis and trans diastereomers about the cyclobutyl ring:
MS (ESI+) for C.sub.40H.sub.51N.sub.7O.sub.5 m/z 710.6 (M+H).sup.+;
MS (ESI-) for C.sub.41H.sub.51N.sub.7O.sub.5 m/z 708.7 (M-H).sup.-;
HPLC purity >95% (ret. time, 3.29, 3.33 min), two peaks observed
due to diastereomers about the cyclobutyl ring.
Step 8: Synthesis of Compound 6
[0762]
7-((3aR,4R,6R,6aR)-6-(((3-((5-(tert-butyl)-1H-benzo[d]imidazol-2-yl-
)methyl)cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3,4-d]-
[1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ami-
ne (170 mg, 0.24 mmol) was dissolved in a mixture of
trifluoroacetic acid (5.0 mL) and water (0.5 mL) which had been
precooled at 0.degree. C. in an ice bath. The solution was stirred
at 0.degree. C. for 30 minutes, and warmed to room temperature.
After 5 h at room temperature, the now very pink reaction mixture
was concentrated. The residue was taken up in 10 mL MeOH and
concentrated. This procedure was repeated twice and the residue
placed on high vac for 1 h. The material was taken up in 7 mL MeOH
and was treated with 130 mg K.sub.2CO.sub.3 and five drops of
water. The mixture was allowed to stir for 1 hr, during which time
the solution was found to be basic. The mixture was filtered
through a fine frit, the solids were washed with 10 mL MeOH and the
filtrate was concentrated to yield a nearly colorless solid. The
crude material was purified by flash chromatography (30 g silica
gel; 12% 7N NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2) to yield
Compound 6
(81 mg, 65%) as a colorless glass/stiff foam: MS (ESI+) for
C.sub.28H.sub.37N.sub.7O.sub.3 m/z 520.4 (M+H).sup.+; MS (ESI-) for
C.sub.28H.sub.37N.sub.7O.sub.3 m/z 518.5 (M-H).sup.-; HPLC purity
>95% (ret. time, 2.51 min); .sup.1H NMR (400 MHz, d4-MeOH)
.delta..sub.H ppm 8.08 (s, 1H), 7.48 (br. s., 1H), 7.39 (d, J=8.50
Hz, 1H), 7.29 (dd, J=8.40, 4.87 Hz, 1H), 6.63 (m, 1H), 6.12 (d,
J=4.15 Hz, 1H), 4.40 (m, 1H), 4.09 (m, 2H), 3.15 (m, 0.5H), 3.02
(d, J=8.09 Hz, 1H), 2.92 (d, J=7.26 Hz, 1H), 2.84 (m, 0.5H), 2.65
(m, 2H), 2.43 (m, 1H), 2.29 (m, 1H), 2.20 (d, J=5.80 Hz, 3H), 2.13
(m, 1H), 1.99 (br. s., 1H), 1.67 (m, 1H), 1.37 (d, J=3.94 Hz, 9H),
1.30 (dd, J=13.99, 4.66 Hz, 1H).
Example 7: Synthesis of
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(6-ch-
loro-5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl-
)amino)methyl)cyclopentane-1,2-diol (Compound 7)
Step 1: Synthesis of
4,6-dichloro-5-(2,2-diethoxyethyl)pyrimidine
##STR00273##
[0764] Title compound was prepared by the method of Montgomery,
see: Montgomery, J. A.; Hewson, K. J. Med. Chem. 10, 665
(1967).
Step 2: Synthesis of
(1R,2S,3R,5R)-3-((6-chloro-5-(2,2-diethoxyethyl)pyrimidin-4-yl)amino)-5-(-
hydroxymethyl)cyclopentane-1,2-diol
##STR00274##
[0766] A mixture of 4,6-dichloro-5-(2,2-diethoxyethyl)pyrimidine
(5.35 g, 20.2 mmol) and
(1R,2S,3R,4R)-2,3-dihydroxy-4-(hydroxymethyl)cyclopentanaminium
chloride (9.29 g, 24.3 mmol) was taken up in ethanol (236 mL),
treated with Et.sub.3N (11.2 mL, 80.8 mmol) and heated at reflux
for 23 h; HPLC/LC MS indicated consumption of starting materials
and presence of product. The reaction mixture was concentrated to
afford a tan slurry, which was carried on crude: MS (ESI+) for
C.sub.16H.sub.26ClN.sub.3O.sub.5 m/z 376.2 (M+H).sup.+; MS (ESI-)
for C.sub.16H26ClN.sub.3O.sub.5 m/z 374.2 (M-H).sup.-; HPLC purity
>95% (ret. time, 2.436 min). Variation on route from J. Med.
Chem. 10, 665 (1967).
Step 3: Synthesis of
(1R,2S,3R,5R)-3-(4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(hydroxymeth-
yl)cyclopentane-1,2-diol
##STR00275##
[0768] A suspension of crude
(1R,2S,3R,5R)-3-((6-chloro-5-(2,2-diethoxyethyl)pyrimidin-4-yl)amino)-5-(-
hydroxymethyl)cyclopentane-1,2-diol in 1,4-dioxane (160 mL) was
treated with a 1 M aqueous solution of HCl (30 mL, 30 mmol) and
stirred at RT for 69.5 h; HPLC indicated clean conversion to one
product, LC MS showed mass for desired product. The reaction
mixture was neutralized with concentrated aqueous NH.sub.40H (to pH
7) and the volatiles were removed in vacuo to afford a brown
slurry, which was carried on without further purification: MS
(ESI+) for C.sub.12H.sub.14ClN.sub.3O.sub.3 m/z 284.1 (M+H).sup.+;
MS (ESI-) for C.sub.12H.sub.14ClN.sub.3O.sub.3 m/z 282.2
(M-H).sup.-, 328.2 (M+HCO.sub.2).sup.-; HPLC purity >95% (ret.
time, 1.947 min). Variation on route from J. Med. Chem. 10, 665
(1967).
Step 4: Synthesis of
((3aR,4R,6R,6aS)-6-(4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethy-
ltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methanol
##STR00276##
[0770] A mixture of crude
(1R,2S,3R,5R)-3-(4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(hydroxymeth-
yl)cyclopentane-1,2-diol (10 g, .about.20 mmol, 54% pure by NMR)
and 2,2-dimethoxypropane (100 mL, 800 mmol) was treated with
p-toluenesulfonic acid monohydrate (7.28 g, 38.3 mmol) and the
yellow-brown reaction mixture was stirred vigorously for 1.25 h, at
which time the only solids were a fine tan precipitate. HPLC
indicated nearly complete consumption of the starting material. The
reaction mixture was diluted with water (30 mL) and neutralized
with solid NaHCO.sub.3 (4.80 g, 57.1 mmol). The volatiles were
carefully removed in vacuo and the resulting brown aqueous solution
was extracted with EtOAc (3.times.100 mL). The combined organics
were dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford a
tan paste. Purification by column chromatography (4.times.22 cm
silica; 0-66% EtOAc/Hex) afforded the title compound (4.38 g, 70%,
one step) as a colorless foam/glass: MS (ESI+) for
C.sub.15H.sub.18ClN.sub.3O.sub.3 m/z 324.2 (M+H).sup.+; MS (ESI-)
for C.sub.15H.sub.18ClN.sub.3O.sub.3 m/z 368.2 (M+HCO.sub.2).sup.-;
HPLC purity >95% (ret. time, 3.034 min).
Step 5: Synthesis of
7-((3aS,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-4-chloro-7H-pyrrolo[2,3-d]pyrimidine
##STR00277##
[0772]
((3aR,4R,6R,6aS)-6-(4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-d-
imethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methanol (2.68
g, 8.28 mmol) was dissolved in THE (32 mL), treated with PPh.sub.3
(3.05 g, 11.6 mmol), and the reaction vessel was cooled in an
ice-brine bath. Diisopropyl azodicarboxylate [DIAD] (2.3 mL, 12
mmol) was added dropwise via syringe and the mixture was stirred
for 10 min. A solution of diphenylphosphonic azide [DPPA] (2.50 mL,
11.6 mmol) in THE (7.8 mL) was added dropwise via syringe to afford
a off-white mixture, which was stirred for 21 h, allowing the ice
bath to warm to RT; HPLC/LC MS indicated complete consumption of
starting material and formation of product. At 22.5 h the reaction
mixture was concentrated in vacuo and purified by column
chromatography (4.times.22 cm silica; 0-25% EtOAc/Hex) to afford
the title compound (2.27 g, 78%) as a clear, colorless oil: MS
(ESI+) for C.sub.15H.sub.17ClN.sub.6O.sub.2 m/z 349.2 (M+H).sup.+;
MS (ESI-) for C.sub.15H17ClN.sub.6O.sub.2 m/z 393.2
(M+HCO.sub.2).sup.-; HPLC purity >95% (ret. time, 4.169
min).
Step 6: Synthesis of
7-((3aS,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-a-
mine
##STR00278##
[0774] A solution of
7-((3aS,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-4-chloro-7H-pyrrolo[2,3-d]pyrimidine and
2,4-dimethoxybenzylamine (1.2 mL, 7.8 mmol) in 1-butanol (18.6 mL)
was treated with N,N-diisopropylethylamine (1.4 mL, 7.8 mmol) and
heated at 80.degree. C. for 22 h; HPLC/LC MS indicated 90%
conversion to the desired product. The volatiles were removed and
the yellow-brown paste was taken up in CH.sub.2Cl.sub.2 (90 mL) and
washed with water (2.times.30 mL) and brine (1.times.45 mL). The
separated organic layer was dried (Na.sub.2SO.sub.4) and
concentrated in vacuo to afford an orange oil. Purification by
column chromatography (2.times.22 cm silica; 0-50% EtOAc/Hex)
afforded the title compound (2.23 g, 72%) as a pale yellow
glass/foam: MS (ESI+) for C.sub.24H.sub.29N.sub.7O.sub.4 m/z 480.5
(M+H).sup.+; MS (ESI-) for C.sub.24H.sub.29N.sub.7O.sub.4 m/z 524.3
(M+HCO.sub.2).sup.-; HPLC purity >95% (ret. time, 3.551
min).
Step 7: Synthesis of
7-((3aS,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-a-
mine
##STR00279##
[0776] A solution of
7-((3aS,4R,6R,6aR)-6-(azidomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-a-
mine (2.23 g, 4.65 mmol) in THE (33 mL, 410 mmol) was cooled to
0.degree. C. and treated dropwise with a 1.0 M solution of
trimethylphosphine in THE (9.3 mL, 9.3 mmol). The cold bath was
removed and the reaction mixture was allowed to warm to RT with
stirring for 1 h; no starting material remained by HPLC. At 1.5 h,
water (4.3 mL, 240 mmol) was added and the reaction mixture was
stirred for 1 h 15 min; TLC indicated one product. The reaction
mixture was concentrated in vacuo to afford a light orange paste.
The residue was diluted with CH.sub.2Cl.sub.2 (120 mL) and washed
with water (2.times.40 mL) and brine (1.times.40 mL). The organic
layer was dried (Na.sub.2SO.sub.4) and concentrated in vacuo to
afford an orange oil. Purification by column chromatography
(2.times.22 cm silica; 0-5% 7 N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2) afforded the title compound (1.97 g,
53% over 3 steps) as a colorless foam: MS (ESI+) for
C.sub.24H.sub.31N.sub.5O.sub.4 m/z 454.3 (M+H).sup.+; HPLC purity
>95% (ret. time, 2.541 min).
Step 8: Synthesis of ethyl
3-(2,2-dichloro-3-oxocyclobutyl)propanoate
##STR00280##
[0778] A mixture of 4-pentenoic acid ethyl ester (7.07 g, 55.2
mmol) and zinc-copper couple (10.2 g, 140 mmol) in diethyl ether
(170 mL) and 1,2-dimethoxyethane (25 mL) was treated dropwise with
trichloroacetyl chloride (25 g, 140 mmol). The mixture was stirred
at room temperature for 3 days. The reddish heterogeneous reaction
mixture was filtered through a pad of celite and the pad was washed
with 300 mL Et.sub.2O. The filtrate was concentrated to about one
half the original volume and the organic phase was washed with two
150 mL portions of H.sub.2O and one 150 mL portion of sat
NaHCO.sub.3. The organic phase was dried over MgSO.sub.4, filtered
and concentrated to yield a brown liquid. The material was purified
by vacuum distillation (90-100.degree. C. @0.044 torr) to yield the
title compound (10.49 g, 80%) as a light yellow liquid: GC purity
95.8% (ret. time, 4.92 min).
Step 9: Synthesis of ethyl 3-(3-oxocyclobutyl)propanoate
##STR00281##
[0780] A solution of ethyl
3-(2,2-dichloro-3-oxocyclobutyl)propanoate (10.49 g, 43.87 mmol)
and ammonium chloride (12 g, 220 mmol) in methanol (310 mL, 7600
mmol) was treated in small portions with zinc powder (14 g, 220
mmol). The reaction mixture was heated at reflux for 3 h after
which time GC indicated the reaction was complete. The reaction
mixture was cooled to room temperature and was filtered through
Celite, washing the pad with Et.sub.2O. The filtrate was
concentrated in vacuo to afford pale yellow solution. The solution
was diluted with 200 mL Et.sub.2O and washed with 100 mL water. The
separated aqueous layer was back extracted with 100 mL Et.sub.2O
and the combined organic phase was washed with 100 mL 1:1
water/brine, 50 mL water, and 150 mL saturated aqueous NaHCO.sub.3.
The organic layer was dried over MgSO.sub.4, filtered and
concentrated in vacuo to afford the title compound (4.49 g, 60%) as
a pale yellow oil which was of sufficient purity for use in the
next step: GC purity >95% (ret. time, 4.24 min).
Step 10: Synthesis of 3-(3-oxocyclobutyl)propanoic acid
##STR00282##
[0782] A solution of ethyl 3-(3-oxocyclobutyl)propanoate (200 mg,
1.18 mmol) in methanol (4 mL) was treated with water (0.75 mL) and
a 2N solution of sodium hydroxide (0.75 mL, 1.41 mmol) and the
solution was heated at 55.degree. C. till the starting material was
consumed by TLC (25% EA/hept). After 1 h the starting material was
found to be consumed. The reaction mixture was cooled to room
temperature and concentrated to remove the MeOH. The aqueous phase
was diluted with 2 mL H.sub.2O and made acidic to pH-2 with 1N HCl.
The solution was saturated with NaCl and extracted with three 10 mL
portions of ethyl acetate. The organic phase was dried over
Na.sub.2SO.sub.4, filtered and concentrated to yield the title
compound (157 mg, 94%) as a light orange viscous oil that was used
as is in the next step: GC purity 63.2% (ret. time, 4.27 min).
Step 11: Synthesis of
N-(2-amino-4-chloro-5-(trifluoromethyl)phenyl)-3-(3-oxocyclobutyl)propana-
mide
##STR00283##
[0784] A solution of 3-(3-oxocyclobutyl)propanoic acid (157 mg,
0.696 mmol) and 4-chloro-5-(trifluoromethyl)benzene-1,2-diamine
(146 mg, 0.696 mmol) in N,N-dimethylformamide (2.5 mL) was cooled
at 0.degree. C. The solution was treated with
N,N-diisopropylethylamine (0.364 mL, 2.09 mmol) dropwise followed
by N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
hexafluorophosphate (291 mg, 0.765 mmol) in one portion. The
solution was allowed to stir and slowly warm to room temperature.
After 40 h, the reaction mixture was concentrated partially under
high vac. The remaining brown liquid was taken up in 25 mL EA and
15 mL 1/1 sat NaHCO.sub.3/H.sub.2O and extracted. The aqueous phase
was washed with two 15 mL portions of ethyl acetate and the
combined organic phase was washed with 30 mL portions of H.sub.2O
and brine. The organic phase was dried over MgSO.sub.4, filtered
and concentrated to yield a tannish brown viscous oil/glass. The
crude material was purified by flash chromatography (40 g silica
gel, 50-80% EA/hept) to yield the title compound (72 mg, 31%) as a
slightly tan glass/stiff foam: MS (ESI+) for
C.sub.14H.sub.14ClF.sub.3N.sub.2O.sub.2 m/z 335.2 (M+H).sup.+; MS
(ESI-) for C.sub.14H.sub.14ClF.sub.3N.sub.2O.sub.2 m/z 333.3
(M-H).sup.-; HPLC purity 78.2% (ret. time, 3.56 min).
Step 12: Synthesis of
3-(2-(6-chloro-5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobu-
tanone
##STR00284##
[0786]
N-(2-amino-4-chloro-5-(trifluoromethyl)phenyl)-3-(3-oxocyclobutyl)p-
ropanamide (72 mg, mmol) was taken up in acetic acid (3.2 mL) and
the solution was heated at 65.degree. C. for 26 h, upon which HPLC
indicated the starting material was consumed and a new product had
formed. The reaction mixture was cooled and the solvent was removed
under high vac. The light brown residue was taken up in 20 mL ethyl
acetate and the organic phase was washed with 10 mL portions of sat
NaHCO.sub.3 and H.sub.2O. The organic phase was dried over
Na.sub.2SO.sub.4, filtered and concentrated to yield a light brown
glass. The crude material was purified by prep TLC (20 cm.times.20
cm.times.1.0 mm prep TLC plate, 3% MeOH/EA) to yield the title
compound (45 mg, 66%) as a tan glass: MS (ESI+) for
C.sub.14H12ClF.sub.3N20 m/z 317.2 (M+H).sup.+; MS (ESI-) for
C.sub.14H.sub.12ClF.sub.3N.sub.2O m/z 315.2 (M-H).sup.-; HPLC
purity 84.1% (ret. time, 2.98 min).
Step 13: Synthesis of
7-((3aS,4R,6R,6aR)-6-(((3-(2-(6-chloro-5-(trifluoromethyl)-1H-benzo[d]imi-
dazol-2-yl)ethyl)cyclobutyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclo-
penta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimid-
in-4-amine
##STR00285##
[0788] A solution of
7-((3aS,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-a-
mine (80 mg, 0.18 mmol) and
3-(2-(6-chloro-5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobu-
tanone (45 mg, 0.14 mmol) in 1,2-dichloroethane (2.4 mL) was
treated dropwise with acetic acid (10 uL, 0.18 mmol). The solution
was treated with sodium triacetoxyborohydride (53 mg, 0.25 mmol) in
one portion and allowed to stir at room temperature till complete
by HPLC. After 4 h, the reaction mixture was diluted with 10 mL
CH.sub.2Cl.sub.2 and washed with 10 mL sat NaHCO.sub.3. The aqueous
phase was washed with 10 mL CH.sub.2Cl.sub.2 and the combined
organic phase was dried over Na.sub.2SO.sub.4. The solution was
filtered and concentrated to yield a light tan glass/stiff foam.
The crude material was purified by flash chromatography (25 g
silica gel; 5% 7N NH.sub.3 in CH.sub.3OH/CHCl.sub.3) to yield the
title compound (76 mg, 71%) as a colorless glass/stiff foam: MS
(ESI+) for C.sub.38H43ClF.sub.3N704 m/z 754.3 (M+H).sup.+; MS
(ESI-) for C.sub.38H43ClF.sub.3N.sub.7O.sub.4 m/z 752.3
(M-H).sup.-; HPLC purity 90.5% (ret. time, 3.24 min).
Step 14: Synthesis of
7-((3aS,4R,6R,6aR)-6-(((3-(2-(6-chloro-5-(trifluoromethyl)-1H-benzo[d]imi-
dazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydro-3-
aH-cyclopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d-
]pyrimidin-4-amine
##STR00286##
[0790] A solution of
7-((3aS,4R,6R,6aR)-6-(((3-(2-(6-chloro-5-(trifluoromethyl)-1H-benzo[d]imi-
dazol-2-yl)ethyl)cyclobutyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclo-
penta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimid-
in-4-amine (76 mg, 0.10 mmol) in methanol (2.5 mL) was treated with
sodium cyanoborohydride (84 mg, 1.3 mmol). The pH of the solution
was adjusted to .about.6 by the dropwise addition of a 10% (v/v)
solution of glacial acetic acid in methanol. The mixture was
treated with 37% aqueous formaldehyde (0.12 mL, 1.7 mmol) dropwise
and the mixture was stirred at room temperature till complete by
LCMS. After 2 h, the reaction was complete and the reaction mixture
was concentrated to remove the methanol. The aqueous solution that
remained was diluted with 7 mL NaHCO.sub.3 and the aqueous phase
was extracted with three 10 mL portions of CH.sub.2Cl.sub.2. The
organic phase was dried over Na.sub.2SO.sub.4, filtered, and
concentrated to yield a colorless stiff foam/glass. The crude
material was purified by flash chromatography (20 g silica gel; 4%
7N NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2) to yield the title
compound (62 mg, 80%) as a colorless glass: MS (ESI+) for
C.sub.39H.sub.45ClF.sub.3N.sub.7O.sub.4 m/z 768.0 (M+H).sup.+; MS
(ESI-) for C.sub.39H.sub.45ClF.sub.3N.sub.7O.sub.4 m/z 766.3
(M-H).sup.-; HPLC purity 92.1% (ret. time, 3.29 min).
Step 15: Synthesis of Compound 7
[0791]
7-((3aS,4R,6R,6aR)-6-(((3-(2-(6-chloro-5-(trifluoromethyl)-1H-benzo-
[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrah-
ydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo-
[2,3-d]pyrimidin-4-amine (60 mg, 0.078 mmol) was dissolved in a
mixture of trifluoroacetic acid (3.6 mL) and water (0.4 mL) which
had been precooled at 0.degree. C. in an ice bath. The solution was
stirred at 0.degree. C. for 30 minutes, and then warmed to room
temperature. After 3 h at room temperature, HPLC indicated the
reaction was complete. The now very pink reaction mixture was
concentrated. The residue was taken up in 10 mL MeOH and
concentrated. This procedure was repeated twice and the residue
placed on high vac for 1 h. The material was taken up in 7 mL MeOH
and was treated with 120 mg K.sub.2CO.sub.3 and ten drops of water.
The mixture was allowed to stir for 1 hr. The mixture was filtered
through a fine frit, the solids were washed with 10 mL MeOH and the
filtrate was concentrated to yield a nearly colorless solid. The
crude material was purified by flash chromatography (30 g silica
gel; 10-15% 7N NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2) to yield
Compound 7 (31 mg, 69%) as a colorless glass/stiff foam: MS (ESI+)
for C.sub.27H.sub.31ClF.sub.3N.sub.7O.sub.2 m/z 578.3 (M+H).sup.+;
MS (ESI-) for C.sub.27H.sub.31ClF.sub.3N.sub.7O.sub.2 m/z 576.4
(M-H).sup.-; HPLC purity >95% (ret. time, 2.57 min); 1H NMR (400
MHz, d4-MeOD) .delta..sub.H 8.06 (s, 1H), 7.89 (d, J=2.07 Hz, 1H),
7.69 (d, J=2.28 Hz, 1H), 7.21 (dd, J=3.42, 1.76 Hz, 1H), 6.59 (d,
J=3.52 Hz, 1H), 4.33 (t, J=6.84 Hz, 1H), 3.89 (q, J=5.25 Hz, 1H),
3.03 (m, 0.5H), 2.89 (m, 2H), 2.70 (m, 0.5H), 2.49 (m, 1H), 2.40
(m, 2H), 2.27 (br. s., 2H), 2.16 (d, J=7.26 Hz, 4H), 2.06 (m, 2H),
1.91 (m, 2H), 1.62 (m, 1H), 1.52 (m, 1H).
Example 8: Synthesis of Compounds 8-140
[0792] Compounds 8-140 were synthesized by methods similar to those
described for Examples 1-7 or by reaction schemes depicted in the
general schemes. Detailed descriptions of how some of them were
prepared are provided below. The MS and NMR data of Compounds 2-140
are provided in Table 1 or Examples provided herein.
Compound 8:
1-(3-((((2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yl)-3,4-dihydroxytetrahydrofu-
ran-2-yl)methyl)(methyl)amino)cyclobutyl)-3-(4-(tert-butyl)phenyl)urea
Benzyl (3-oxocyclobutyl)carbamate
[0793] To a solution of 3-oxocyclobutanecarboxylic acid (1.0 g,
8.77 mmol) and DIEA (1.92 g, 14.92 mmol) in toluene (8 mL) was
added DPPA (2.89 g, 10.52 mmol) at rt. The mixture was heated to
60.degree. C. under Argon for 3 h, then benzyl alcohol (1.14 g,
10.52 mmol) was added. The mixture was stirred at 60.degree. C.
overnight. The reaction was concentrated, the residue was purified
by SGC (PE:EA=8:1) to afford the desired compound (240 mg, yield
50%). .sup.1H NMR (500 MHz, CDCl.sub.3). .delta..sub.H 7.38-7.33
(m, 5H), 5.12 (d, J=7.5 Hz, 2H), 4.34-4.33 (brs, 1H), 3.44-3.39 (m,
2H), 3.10-3.07 (brs, 2H) ppm; ESI-MS (m/z): 220.2 [M+1].sup.+.
benzyl
(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahy-
drofuro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)carbamate
[0794] To a solution of
9-((3aR,4R,6R,6aR)-2,2-dimethyl-6-((methylamino)methyl)tetrahydrofuro[3,4-
-d][1,3]dioxol-4-yl)-9H-purin-6-amine (190 mg, 0.59 mmol) and
benzyl (3-oxocyclobutyl)carbamate (240 mg, 1.37 mmol) in MeOH (5
mL) was added Ti[OCH(CH.sub.3).sub.2].sub.4 (216 mg, 0.59 mmol).
The mixture was stirred at rt for 1 h. Then NaCNBH.sub.3 (95 mg,
1.52 mmol) was added, the reaction was stirred at rt overnight. The
reaction was filtered and evaporated, the residue was purified by
prep-TLC (DCM:MeOH=20:1) to obtain the desired compound (90 mg,
yield 29%).
[0795] .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.28 (s, 1H),
8.21 (s, 1H), 7.33-7.28 (m, 5H), 6.19 (d, J=2.0 Hz, 1H), 5.52-5.51
(m, 1H), 5.03 (s, 1H), 5.00-4.98 (m, 1H), 4.34 (t, J=3.5 Hz, 1H),
3.70 (m, 1H), 2.58-2.47 (m, 4H), 2.38-2.26 (m, 2H), 2.09 (s, 3H),
1.69-1.67 (m, 1H), 1.58 (s, 3H), 1.37 (s, 3H) ppm; ESI-MS (m/z):
524.3 [M+1].sup.+.
N1-(((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[-
3,4-d][1,3]dioxol-4-yl)methyl)-N1-methylcyclobutane-1,3-diamine
[0796] To a solution of benzyl
(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofur-
o[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)carbamate
(190 mg, 0.17 mmol) and Pd(OH).sub.2 (14 mg, 0.1 mmol) in MeOH (5
mL) was charged with H.sub.2. The reaction was stirred at
35.degree. C. for 5 h. The reaction was filtered with Celite and
concentrated to dryness. The residue was purified by prep-TLC
(DCM:MeOH=10:1) to the desired compound (28 mg, yield 42%). .sup.1H
NMR (500 MHz, MeOD): .delta..sub.H 8.29 (s, 1H), 8.21 (s, 1H), 6.19
(d, J=2.0 Hz, 1H), 5.51 (dd, J=6.5 and 2.0 Hz, 1H), 5.00 (dd, J=6.0
and 3.5 Hz, 1H), 4.34 (d, J=8.5 Hz, 1H), 3.14-3.11 (m, 1H),
2.60-2.57 (m, 1H), 2.52-2.48 (m, 2H), 2.34-2.31 (m, 2H), 2.10 (s,
3H), 1.66 (q, J=10.0 Hz, 1H), 1.58 (s, 3H), 1.48 (q, J=10.0 Hz,
1H), 1.37 (s, 3H) ppm; ESI-MS (m/z): 390.2[M+1].sup.+.
1-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofu-
ro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)-3-(4-(tert-buty-
l)phenyl)urea
[0797] A solution of
N1-(((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)methyl)-N1-methylcyclobutane-1,3-diamine
(28 mg, 0.072 mmol) and TEA (22 mg, 0.22 mmol) in THE (3 mL) was
added dropwise 1-tert-butyl-4-isocyanatobenzene (18 mg, 0.11 mmol)
in DCM (0.5 mL). The reaction was stirred for 1 hour at room
temperature. The reaction was concentrated and purified by Prep-TLC
(twice, DCM:MeOH:NH.sub.40H=300:30:8, V/V) to obtain the desired
compound (28 mg, Yield: 88%) as pale white solid. .sup.1H NMR (500
MHz, MeOD): .delta..sub.H 8.29 (s, 1H), 8.24 (s, 1H), 7.28-7.22 (m,
4H), 6.25 (d, J=2.5 Hz, 1H), 5.52-5.50 (m, 1H), 5.07-5.05 (m, 1H),
4.46-4.44 (m, 1H), 3.88-3.85 (m, 1H), 2.97 (brs, 1H), 2.80-2.78 (m,
2H), 2.48-2.42 (m, 2H), 2.30 (s, 3H), 1.84-1.82 (m, 1H), 1.60 (s,
3H), 1.58-1.56 (m, 1H), 1.39 (s, 3H), 1.28 (s, 9H) ppm; ESI-MS
(m/z): 565.3 [M+1].sup.+.
1-(3-((((2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yl)-3,4-dihydroxytetrahydrofur-
an-2-yl)methyl)(methyl)amino)cyclobutyl)-3-(4-(tert-butyl)phenyl)urea
##STR00287##
[0799] A solution of
1-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)-3-(4-(tert-but-
yl)phenyl)urea (125 mg, 0.23 mmol) in TFA (0.90 mL) and 0.10 mL of
water were stirred for 1 hour at room temperature. The reaction was
concentrated to dryness, dissolved in MeOH (5 mL) and
K.sub.2CO.sub.3 (60 mg) in 0.5 mL of water was added dropwise. The
reaction was stirred at rt for 0.5 h and concentrated to obtain the
residue which was purified by prep-TLC
(DCM:MeOH:NH.sub.4OH=300:30:8, V/V) to obtain the desired compound
(75 mg, Yield: 65%) as pale white solid. .sup.1H NMR (500 MHz,
MeOD): .delta..sub.H 8.27 (s, 1H), 8.21 (s, 1H), 7.28-7.20 (m, 4H),
6.00-5.99 (m, 1H), 4.77-4.75 (m, 1H), 4.28-4.23 (m, 2H), 3.92-3.88
(m, 1H), 2.92 (brs, 1H), 2.83-2.81 (m, 2H), 2.59-2.56 (m, 2H), 2.32
(s, 3H), 1.74-1.64 (m, 2H), 1.27 (s, 9H) ppm; ESI-MS (m/z): 525.3
[M+1].sup.+.
Compounds 9 and 12
##STR00288## ##STR00289##
[0800] Compound 9:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(6-chloro-5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl-
)(methyl)amino)methyl)cyclopentane-1,2-diol
[0801] .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.07 (s, 1H),
7.90 (s, 1H), 7.69 (s, 1H), 7.22 (d, J=3.5 Hz, 1H), 6.61 (d, J=3.5
Hz, 1H), 4.34 (t, J=6.5 Hz, 1H), 3.89 (t, J=5.0 Hz, 1H), 2.88 (t,
J=7.0 Hz, 2H), 2.74-2.68 (m, 1H), 2.55-2.49 (m, 1H), 2.46-2.35 (m,
2H), 2.32-2.22 (m, 3H), 2.17 (s, 3H), 2.00-1.90 (m, 3H), 1.68-1.60
(m, 1H), 1.58-1.48 (m, 2H) ppm; LC-MS (m/z): 578.3 [M+1].sup.+.
Compound 12:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3--
(2-(6-chloro-5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl-
)(methyl)amino)methyl)cyclopentane-1,2-diol
[0802] .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.07 (s, 1H),
7.91 (s, 1H), 7.70 (s, 1H), 7.22 (d, J=3.5 Hz, 1H), 6.61 (d, J=4.0
Hz, 1H), 4.34 (dd, J=7.0 and 6.0 Hz, 1H), 3.90 (t, J=5.0 Hz, 1H),
3.05-3.00 (m, 1H), 2.92 (t, J=7.5 Hz, 2H), 2.55-2.49 (m, 1H),
2.47-2.35 (m, 2H), 2.32-2.22 (m, 1H), 2.20-2.02 (m, 8H), 1.93-1.86
(m, 2H), 1.70-1.60 (m, 1H) ppm; LC-MS (m/z): 578.3 [M+1].sup.+.
Compounds 10 and 11
##STR00290## ##STR00291##
[0803] Compound 10:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl((1r,-
3S)-3-(2-(5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)am-
ino)methyl)cyclopentane-1,2-diol
[0804] .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.06 (s, 1H),
7.79 (s, 1H), 7.62 (d, J=9.0 Hz, 1H), 7.47 (d, J=8.5 Hz, 1H), 7.20
(d, J=3.5 Hz, 1H), 6.59 (d, J=3.0 Hz, 1H), 4.33-4.30 (m, 1H),
3.88-3.86 (m, 1H), 3.32-3.31 (m, 1H), 2.89-2.86 (m, 2H), 2.67-2.66
(m, 1H), 2.48-2.26 (m, 6H), 2.14 (s, 3H), 1.95-1.93 (m, 3H),
1.62-1.48 (m, 3H) ppm; LC-MS (m/z): 544.3 [M+1].sup.+.
Compound 11:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl((1s,-
3R)-3-(2-(5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)am-
ino)methyl)cyclopentane-1,2-diol
[0805] .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.06 (s, 1H),
7.79 (s, 1H), 7.62 (d, J=8.5 Hz, 1H), 7.47 (d, J=8.5 Hz, 1H), 7.20
(d, J=3.0 Hz, 1H), 6.59 (d, J=3.5 Hz, 1H), 4.33-4.31 (m, 1H),
3.90-3.87 (m, 1H), 3.01-3.0 (m, 1H), 2.92-2.89 (m, 2H), 2.48-2.03
(m, 13H), 1.93-1.89 (m, 2H), 1.63-1.61 (m, 1H) ppm; LC-MS (m/z):
544.3 [M+1].sup.+.
Compound 13:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl(3-(2-
-(5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)meth-
yl)cyclopentane-1,2-diol
N-(2-amino-4-(trifluoromethyl)phenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-d-
imethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahy-
dro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)prop-
anamide
##STR00292##
[0807] N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (0.44 g, 1.2 mmol) was added to a solution of
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(methyl)amino)cyclobutyl)propanoic acid (460 mg, 0.77
mmol), N,N Diisopropylethylamine (0.44 mL, 2.6 mmol) in
N,N-Dimethylformamide (5 mL). The reaction was stirred overnight at
RT, partially concentrated then NaHCO.sub.3 (saturated) was added.
The aqueous layer was extracted 3.times. with EtOAc and the
combined organics were dried with MgSO.sub.4, filtered,
concentrated and purified by flash chromatography (DCM/7N NH.sub.3
in MeOH 95:5) to give the desired compound (0.34 g) as a solid.
N-(2,4-dimethoxybenzyl)-7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(5-
-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)-
tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-
-amine
##STR00293##
[0809]
N-(2-amino-4-(trifluoromethyl)phenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4--
((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyl-
tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobut-
yl)propanamide (0.38 g, 0.50 mmol) and Acetic acid (5 ml) were
stirred overnight at 65.degree. C. The volatiles were removed in
vacuo and remaining water was removed by azeotropic distillation
with ethanol followed by 1 hour on high vacuum. The resulting
residue was partitioned between NaHCO.sub.3 (saturated) and DCM.
The aqueous layer was extracted (3.times.) and combined organics
were dried with MgSO.sub.4, filtered, concentrated and then
purified by flash chromatography (DCM/7N NH.sub.3 in MeOH 93:7) to
yield an off white foam.
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl(3-(2--
(5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methy-
l)cyclopentane-1,2-diol
##STR00294##
[0811] Trifluoroacetic Acid (5 ml) added to a mixture of Water (0.5
ml)
N-(2,4-dimethoxybenzyl)-7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(-
5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl-
)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin--
4-amine (0.31 g, 0.42 mmol) at RT. The reaction was allowed to
proceed overnight when it was quenched with Triethylsilane (0.13
ml, 0.84 mmol). The volatiles were removed in vacuo and resulting
residue was partitioned between saturated NaHCO.sub.3 and DCM/MeOH
(10:1). The aqueous layer was extracted (3.times.) more with
DCM/MeOH (10:1) and the combined organics were dried over
MgSO.sub.4, filtered and concentrated. The residue was purified by
flash chromatography (DCM/7N NH.sub.3 in MeOH 87:13) to give the
desired compound (0.12 g) as an off-white foam/gum. MS (ESI.sup.+)
for C.sub.27H.sub.32F.sub.3N.sub.7O.sub.2 m/z 544.5 [M+H].sup.+; MS
(ESI.sup.-) for C.sub.27H.sub.32F.sub.3N.sub.7O.sub.2 m/z 542.3
[M-H].sup.-; HPLC purity >85% (ret. time, 2.418 min.) .sup.1H
NMR (400 MHz, d.sub.4-MeOH) .delta..sub.H 8.078 (s, 1H), 7.812 (s,
1H), 7.654-7.634 (m, 1H), 7.507-7.487 (m, 1H), 7.229-7.214 (m, 1H),
6.617-6.608 (d, J 3.6 Hz, 1H), 4.361-4.322 (m, 1H), 3.927-3.887 (m,
1H), 3.062-3.024 (m, 0.5H (methine of trans isomer)), 2.944-2.873
(m, 2H), 2.758-2.554 (m, 0.5H (methine of cis isomer)), 2.554-2.507
(m, 1H), 2.447-2.351 (m, 2H), 2.291-2.263 (m, 2H), 2.194-2.054 (m,
6H), 1.960-1.887 (m, 3H), 1.686-1.480 (m, 2H). Retention time:
2.418 HPLC Conditions: Agilent Zorbax Exlipse XDB-C18 column,
4.6.times.50 mm (1.8 um packing), Solvent A-Water (0.1% TFA),
Solvent B-Acetonitrile (0.07% TFA).6 min gradient from 5 to 95% B;
1 min hold; then recycle.
Compound 14:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5-(t-
ert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)-
cyclopentane-1,2-diol
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimeth-
oxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3-
aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanami-
de
##STR00295##
[0813] N,N,N'N'-Tetramethyl-O-(7-azabenzotriazol-1-yl}uronium
Hexafluorophosphate (0.44 g, 1.2 mmol) added to a solution of
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(methyl)amino)cyclobutyl)propanoic acid (460 mg, 0.77 mmol)
and N,N-Diisopropylethylamine (0.44 mL, 2.6 mmol) and
4-tert-butylbenzene-1,2-diamine (0.15 g, 0.93 mmol) in
N,N-Dimethylformamide (5 mL, 60 mmol). The reaction was stirred
overnight at RT, partially concentrated to ca. 2 m is and then
NaHCO.sub.3 (saturated) was added. The mixture was extracted with
EtOAc (3.times.) and the combined organics were dried with
MgSO.sub.4 and concentrated. Purified by flash chromatography
(DCM/7N NH.sub.3 in MeOH 95:5) to yield a solid (0.24 g).
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethy-
l)cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d-
][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-am-
ine
##STR00296##
[0815] A solution of
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimet-
hoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro--
3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanam-
ide (0.24 g, 0.32 mmol) in Acetic acid (5 ml, 90 mmol) was stirred
overnight at 60.degree. C. The volatiles were removed in vacuo and
remaining residue partitioned between Na.sub.2CO.sub.3 (2N) and
DCM. The aqueous layer was extracted 3.times. with DCM and the
combined organics dried with MgSO.sub.4, filtered and concentrated.
The residue was purified by flash chromatography (DCM/7N NH.sub.3
in MeOH 94:6) to yield the desired compound (0.20 g) as a
solid.
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5-(te-
rt-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)c-
yclopentane-1,2-diol
##STR00297##
[0817] Trifluoroacetic Acid (5 ml) added to a mixture of Water (0.5
ml) and
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl-
)ethyl)cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclope-
nta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-
-4-amine (0.20 g, 0.28 mmol) at RT. The reaction was allowed to
proceed overnight upon which it was quenched with Triethylsilane
(0.088 ml, 0.55 mmol). The volatiles were removed in vacuo and
resulting residue was partitioned between saturated NaHCO.sub.3 and
DCM/MeOH (10:1). Aqueous extracted 3.times. more with DCM/MeOH
(10:1) and combined organics were dried over MgSO.sub.4, filtered,
concentrated and purified by flash chromatography (DCM/7N NH.sub.3
in MeOH 87:13) to give the desired product as an off-white foam
(0.060 g). MS (ESI.sup.+) for C.sub.30H.sub.41N.sub.7O.sub.2 m/z
532.3 [M+H].sup.+; MS (ESI-) for C.sub.30H.sub.41N.sub.7O.sub.2 m/z
530.4 [M-H].sup.-; HPLC purity >94% (ret. time, 2.723 min.)
.sup.1H NMR (400 MHz, d4-MeOH) .delta..sub.H 8.079 (s, 1H), 7.500
(s, 1H), 7.418-7.398 (m, 1H), 7.310-7.307 (m, 1H), 7.230-7.216 (m,
1H), 6.619-6.610 (m, 1H), 4.355-4.316 (m, 1H), 3.926-3.887 (m, 1H),
3.088-3.017 (m, 0.5H (methine of trans isomer)), 2.879-2.809 (m,
2H), 2.745-2.685 (m, 0.5H (methine of cis isomer), 2.532-2.512 (m,
1H), 2.446-2.373 (m, 2H), 2.294-2.276 (m, 2H), 2.202-2.012 (m, 5H),
1.685-1.603 (m, 1H), 1.545-1.504 (m, 1H), 1.383 (s, 1H). Retention
time. 2.723 mins HPLC Conditions: Agilent Zorbax Exlipse XDB-C18
column 4.6.times.50 mm (1.8 urn packing), Solvent A-Water (0.1%
TFA), Solvent B-Acetonitrile (0.07% TFA) 6 min gradient from 5 to
95% B; 1 min hold; then recycle.
Compound 15:
(1R,2S,3R,5R)-3-{4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl}-5-{[propan-2-yl-
({4-[5-(trifluoromethyl)-1H-1,3-benzodiazol-2-yl]butyl})amino]methyl}cyclo-
pentane-1,2-diol
Step 1:
7-[(3aS,4R,6R,6aR)-2,2-dimethyl-6-{[propan-2-yl({4-[5-(trifluorome-
thyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazol-2-yl]butyl})-
amino]methyl}-hexahydrocyclopenta[d][1,3]dioxol-4-yl]-7H-pyrrolo[2,3-d]pyr-
imidin-4-amine
##STR00298##
[0819] A solution of
3-{[5-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benzo-
diazol-2-yl]methyl}cyclobutane-1-carbaldehyde (243 mg, 0.59 mmol),
7-[(3aS,4R,6R,6aR)-2,2-dimethyl-6-[(propan-2-ylamino)methyl]-hexahydrocyc-
lopenta[d][1,3]dioxol-4-yl]-7H-pyrrolo[2,3-d]pyrimidin-4-amine (170
mg, 0.49 mmol) and MgSO.sub.4 (710 mg, 5.90 mmol) in DCE (10 ml)
was stirred for 15 min. STAB (175 mg, 0.83 mmol) was then added to
the reaction mixture and stirred for 1 h at RT. The reaction was
monitored by LCMS, no amine was seen after 1 h. Sat. NaHCO.sub.3
(20 ml) was added to the reaction mixture and stirred for 5 mins.
Brine (10 ml) was then added to the reaction mixture. The product
was extracted with DCM (2.times.30 ml), dried over
Na.sub.2SO.sub.4, filtered and evaporated. Purification by silica
gel column chromatography, eluting with 7N NH.sub.3 in MeOH:DCM
(1:99-4:96) gave the desired product (170 mg, 47%) as an oil; MS
(ESI.sup.+) for C.sub.38H.sub.54F.sub.3N.sub.7O.sub.3Si m/z 742.40
[M+H].sup.+; HPLC purity 100% (ret. time, 1.78 min); .sup.1H NMR
(500 MHz, CHLOROFORH-d) .delta.H ppm -0.25-0.11 (9H, m), 0.66-1.04
(8H, m), 1.17-1.48 (5H, m), 1.49-1.61 (3H, m), 1.77-2.04 (2H, m),
2.19-2.37 (5H, m), 2.37-2.51 (2H, m), 2.52-2.80 (2H, m), 2.81-2.91
(1H, m), 2.91-3.22 (2H, m), 3.34-3.79 (2H, m), 4.29-4.52 (1H, m),
4.78-5.12 (2H, m), 5.28-5.70 (4H, m), 6.34 (1H, d, J=3.63 Hz),
6.80-7.16 (1H, m), 7.29-7.71 (2H, m), 7.71-8.11 (1H, m), 8.11-8.46
(1H, m)
Step 2.
(1R,2S,3R,5R)-3-{4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl}-5-{[prop-
an-2-yl({4-[5-(trifluoromethyl)-1H-1,3-benzodiazol-2-yl]butyl})amino]methy-
l}cyclopentane-1,2-diol
##STR00299##
[0821] 12N HCl (3 ml) was added slowly to a solution of 7-(3
aS,4R,6R,6aR)-2,2-dimethyl-6-({propan-2-yl[(3-{[5-(trifluoromethyl)-1-{[2-
-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazol-2-yl]methyl}cyclobutyl)-
methyl]amino}methyl)-hexahydrocyclopenta[d][1,3]dioxol-4-yl]-7H-pyrrolo[2,-
3-d]pyrimidin-4-amine (170 mg, 0.23 mmol) in MeOH (3 ml) and
stirred at 40.degree. C. for 2.5 h. The reaction was monitored by
LCMS, no starting material seen after 2.5 h. The reaction mixture
was concentrated in vacuo, then basified with 7N NH.sub.3 in MeOH.
This was then evaporated to dryness. Purification by silica gel
column chromatography, eluting with 7N NH.sub.3 in MeOH:DCM (1:9)
gave the desired product (100 mg, 76%) as a white solid; MS
(ESI.sup.+) for C.sub.29H.sub.36F.sub.3N.sub.7O.sub.3 m/z 572.40
[M+H].sup.+; HPLC purity 99% (ret. time, 2.17 min); .sup.1H NMR
(500 MHz, CHLOROFORH-d) .delta..sub.H ppm 0.78-1.19 (6H, m),
1.35-1.69 (2H, m), 1.80-2.04 (1H, m), 2.11-2.87 (10H, m), 2.88-3.18
(3H, m), 3.79-4.06 (1H, m), 4.15-4.47 (1H, m), 4.82-5.12 (1H, m),
6.43-6.77 (1H, m), 7.19 (1H, d, J=3.47 Hz), 7.47 (1H, d, J=8.35
Hz), 7.63 (1H, d, J=8.51 Hz), 7.79 (1H, s), 7.95-8.24 (1H, m).
Compound 18:
(1R,2S,3R,5R)-3-{4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl}-5-({methyl[(3-{-
[5-(trifluoromethyl)-1H-1,3-benzodiazol-2-yl]methyl}cyclobutyl)methyl]amin-
o}methyl)cyclopentane-1,2-diol
Step 1: 3-[2-(benzyloxy)-2-oxoethylidene]cyclobutane-1-carboxylic
acid
##STR00300##
[0823] A mixture of cyclobutaneone-3-carboxylic acid (5 g, 43.82
mmol), benzyl-2-(dimethoxyphosphoryl)acetate (13.58 g, 52.59 mmol),
LiOH (4.20 g, 23.95 mmol) and 3.ANG. activated molecular sieves (25
g, powder form) in THE (250 ml) was heated to reflux under nitrogen
for 4 h. The reaction was allowed to cool to RT and EtOAc (100 ml)
followed by HCl (1N, 100 ml) were added. This mixture was filtered
through celite. The phases were separated and the aqueous layer was
extracted with EtOAc (4.times.50 ml). The combined organic layers
were dried over Na.sub.2SO.sub.4, filtered and concentrated to
yield a colorless oil. Dry flash chromatography over SiO.sub.2,
eluting with Hept:EtOAc from 7:3 to 1:1 gave the desired product as
a colorless oil (5.2 g, 39%); MS (ESI.sup.+) for C.sub.14H1404 m/z
269.05 [M+Na].sup.+; MS (ESI.sup.-) for C.sub.14H1404 m/z 245.15
[M-H].sup.-; HPLC purity 81% (ret. time, 1.85 min); .sup.1H NMR
(250 MHz, CHLOROFORM-d) .delta..sub.H ppm 2.95-3.62 (5H, m),
4.96-5.31 (2H, m), 5.75 (1H, t, J=2.21 Hz), 7.27-7.45 (5H, m).
Step 2. benzyl
2-{3-[methoxy(methyl)carbamoyl]cyclobutylidene}acetate
##STR00301##
[0825] To an ice cold solution of
3-[2-(benzyloxy)-2-oxoethylidene]cyclobutane-1-carboxylic acid (2.0
g, 8.12 mmol), N-Methyl-morpholine (2.70 ml, 24.36 mmol) in DCM (50
ml) was added isobutyl chloroformate (1.70 ml, 12.99 mmol)
drop-wise over 5 min. After an additional 5 min,
methoxy(methyl)amine hydrochloride (1.58 g, 16.24 mmol) was added
and the mixture was stirred overnight whislt allowing to warm to
RT. The reaction mixture was then diluted with DCM (30 ml), washed
with 0.1N HCl (50 ml) then sat. NaHCO.sub.3 (50 ml), dried over
Na.sub.2SO.sub.4, filtered and evaporated. Purification by silica
gel column chromatography, eluting with EtOAc:heptanes from 1:9 to
3:7 gave the desired product (1.54 g, 65%); MS (ESI.sup.+) for
C.sub.16H.sub.19NO.sub.4 m/z 290.10 [M+H].sup.+; HPLC purity 100%
(ret. time, 1.86 min); .sup.1H NMR (500 MHz, CHLOROFORHM-d)
.delta..sub.H ppm 2.96 (1H, ddd, J=16.98, 8.79, 1.81 Hz), 3.17-3.30
(4H, m), 3.30-3.48 (2H, m), 3.51-3.63 (1H, m), 3.64-3.75 (3H, m),
5.15 (2H, s), 5.73 (1H, quin, J=2.25 Hz), 7.29-7.42 (5H, m).
Step 3. 2-{3-[methoxy(methyl)carbamoyl]cyclobutyl}acetic acid
##STR00302##
[0827] Palladium on charcoal (10%, 0.1 g) was added to a solution
of benzyl 2-{3-[methoxy(methyl)carbamoyl]cyclobutylidene}acetate
(1.54 g, 5.32 mmol) in EtOH (20 ml) and stirred under an atmosphere
of hydrogen at RT for 6 h. The reaction mixture was filtered
through celite and evaporated to dryness to give a colorless oil
(1.04 g, 89%); MS (ESI.sup.+) for C.sub.9H.sub.15NO.sub.4 m/z
202.00 [M+H].sup.+; MS (ESI).sup.- for C.sub.9H.sub.15NO.sub.4 m/z
200.05 [M-H].sup.+; HPLC purity 92% (ret. time, 1.10 min); .sup.1H
NMR (500 MHz, MeOD) .delta..sub.H ppm 1.84-2.08 (2H, m), 2.28-2.53
(4H, m), 2.56-2.72 (1H, m), 3.06-3.22 (3H, m), 3.38-3.56 (1H, m),
3.68 (3H, d, J=8.04 Hz).
Step 4i.
3-({[2-amino-5-(trifluoromethyl)phenyl]carbamoyl}methyl)-N-methox-
y-N-methylcyclobutane-1-carboxamide
##STR00303##
[0829] TEA (1.49 ml, 10.70 mmol) was added to a suspension of
2-{3-[methoxy(methyl)carbamoyl]cyclobutyl}acetic acid, EDC.HCl 1.18
g, 6.20 mmol), HOBt.xH.sub.2O (0.77 g, 5.69 mmol) in DCM (20 ml) at
0.degree. C. and stirred for 5 min before the addition of
4-(trifluoromethyl)benzene-1,2-diamine (1.04 g, 10.34 mmol). This
was stirred for a further 20 min at 0.degree. C. then allowed to
warm to RT. The reaction was monitored by LC MS, after 2 hours the
reaction mixture was washed with 1N HCl (50 ml) then sat.
NaHCO.sub.3 (50 ml). This was dried over Na.sub.2SO.sub.4, filtered
and evaporated to dryness. Purification by silica gel column
chromatography, eluting with EtOAc gave the desired product (0.83
g, 44%) as a beige solid; MS (ESI.sup.+) for
C.sub.16H.sub.20N.sub.3O.sub.3 m/z 360.00 [M+H].sup.+; HPLC purity
90% (ret. time, 1.64 min); .sup.1H NMR (500 MHz, MeOD)
.delta..sub.H ppm 2.02-2.16 (2H, m), 2.31-2.68 (4H, m), 2.69-2.88
(1H, m), 3.18 (3H, d, J=4.41 Hz), 3.36-3.59 (1H, m), 3.65-3.76 (3H,
m), 6.81-6.97 (1H, m), 7.02-7.27 (1H, m), 7.28-7.48 (1H, m).
Step 4ii.
N-methoxy-N-methyl-3-{[5-(trifluoromethyl)-1H-1,3-benzodiazol-2--
yl]methyl}-cyclobutane-1-carboxamide
##STR00304##
[0831] A solution of the
3-({[2-amino-5-(trifluoromethyl)phenyl]carbamoyl}methyl)-N-methoxy-N-meth-
ylcyclobutane-1-carboxamide (0.82 g, 2.29 mmol) in AcOH (10 ml) was
heated to reflux (.about.125.degree. C.) whilst stirring for 2.5 h.
The reaction was monitored by LC MS. The reaction mixture was
allowed to cool to RT and then evaporated in vacuo. The residue was
dissolved in DCM (30 ml) and washed with sat. NaHCO.sub.3 (50 ml),
dried over Na.sub.2SO.sub.4, filtered and evaporated. The crude was
purified by silica gel column chromatography, eluting with MeOH:DCM
(2:98-5:95) to give a yellow-brown oil (0.75 g, 94%); MS
(ESI.sup.+) for C.sub.16H.sub.18F.sub.3N.sub.3O.sub.2 m/z 342.10
[M+H].sup.+; HPLC purity 97% (ret. time, 1.40 min); .sup.1H NMR
(500 MHz, MeOD) .delta..sub.H ppm 1.96-2.17 (2H, m), 2.27-2.55 (2H,
m), 2.66-2.92 (1H, m), 2.97-3.15 (2H, m), 3.15-3.22 (3H, m),
3.32-3.62 (1H, m), 3.63-3.73 (3H, m), 7.37-7.53 (1H, m), 7.53-8.00
(2H, m).
Step 5.
N-methoxy-N-methyl-3-{[5-(trifluoromethyl)-1-{[2-(trimethylsilyl)e-
thoxy]methyl}-1H-1,3-benzodiazol-2-yl]methyl}cyclobutane-1-carboxamide
##STR00305##
[0833] K.sub.2CO.sub.3 (381 mg, 2.76 mmol), followed by SEM-Cl (430
.mu.l, 2.43 mmol) was added to a solution of the
N-methoxy-N-methyl-3-{[5-(trifluoromethyl)-1H-1,3-benzodiazol-2-yl]methyl-
}cyclobutane-1-carboxamide (754 mg, 2.21 mmol) in DMF (10 ml) at RT
and stirred overnight. The reaction was monitored by LCMS. The
reaction mixture was diluted with water (3 ml), brine (30 ml) and
then extracted with EtOAc (2.times.50 ml). This was dried over
Na.sub.2SO.sub.4, filtered and evaporated to give a clear orange
oil. Purification by silica gel column chromatography, eluting with
EtOAc:heptanes (1:1-1) gave the desired products as a beige oils as
a single regioisomer (217 mg, 21%); MS (ESI.sup.+) for
C.sub.22H.sub.32F.sub.3N.sub.3O.sub.3Si m/z 472.55 [M+H].sup.+;
HPLC purity 99% (ret. time, 2.37 min); .sup.1H NMR (250 MHz,
CHLOROFORM-d) .delta..sub.H ppm -0.31-0.22 (9H, m), 0.83-1.00 (2H,
m), 2.03-2.25 (2H, m), 2.30-2.74 (2H, m), 2.85-3.14 (3H, m), 3.18
(3H, s), 3.32-3.60 (3H, m), 3.61-3.72 (3H, m), 5.52 (2H, s), 7.51
(1H, dd, J=8.38, 1.22 Hz), 7.70 (1H, s), 7.79 (1H, d, J=8.53 Hz)
and as a mixture of regioisomers (280 mg, 27%);
C.sub.22H.sub.32F.sub.3N.sub.3O.sub.3Si m/z 472.55 [M+H].sup.+;
HPLC purity 72% & 21% (ret. time, 2.39 & 2.36 min); .sup.1H
NMR (250 MHz, CHLOROFORM-d) .delta. ppm -0.08--0.01 (9H, m),
0.81-0.99 (2H, m), 1.94-2.24 (2H, m), 2.38-2.74 (2H, m), 2.90-3.14
(3H, m), 3.14-3.22 (3H, m), 3.33-3.63 (3H, m), 3.63-3.70 (3H, m),
5.40-5.64 (2H, m), 7.40-7.74 (2H, m), 7.75-8.14 (1H, m).
Step 6.
3-{[5-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-1,-
3-benzodiazol-2-yl]methyl}cyclobutane-1-carbaldehyde
##STR00306##
[0835] DIBAL (0.69 ml, 0.69 mmol, 1M in THF) was added dropwise to
a solution of
N-methoxy-N-methyl-3-{[5-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]m-
ethyl}-1H-1,3-benzodiazol-2-yl]methyl}cyclobutane-1-carboxamide in
THE at -10.degree. C. whilst stirring. The reaction was continued
for 3 hours at -10.degree. C. The reaction mixture was poured onto
sat. aq. Rochelle's salt (20 ml), diluted with Et.sub.2O (50 ml)
and stirred for 30 min. This was then separated and the organic
layer was washed with Rochelle's salt (30 ml), sat. NaHCO.sub.3,
(30 ml) and brine (30 ml). This was dried over Na.sub.2SO.sub.4,
filtered and evaporated to give a colorless gum (190 mg, 87%); MS
(ESI.sup.+) for C.sub.20H.sub.27F.sub.3N.sub.2O.sub.2Si m/z 413.6
[M+H].sup.+; HPLC purity 87% (ret. time, 2.25 min); .sup.1H NMR
(500 MHz, CHLOROFORM-d) .delta..sub.H ppm -0.10-0.09 (9H, m),
0.88-1.00 (4H, m), 1.63-1.97 (2H, m), 2.07-2.23 (2H, m), 2.24-2.37
(0H, m), 2.37-2.42 (1H, m), 2.43-2.66 (2H, m), 2.93-3.34 (4H, m),
5.49-5.61 (2H, m), 7.16-7.31 (2H, m), 7.32-7.32 (2H, m), 7.49-7.61
(1H, m), 7.73 (1H, s), 7.83 (1H, d, J=8.35 Hz).
Step 7.
7-[(3aS,4R,6R,6aR)-2,2-dimethyl-6-({[(3-{[5-(trifluoromethyl)-1-{[-
2-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazol-2-yl]methyl}cyclobutyl-
)methyl]amino}methyl)-hexahydrocyclopenta[d][1,3]dioxol-4-yl]-7H-pyrrolo[2-
,3-d]pyrimidin-4-amine
##STR00307##
[0837] A solution of
3-{[5-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benzo-
diazol-2-yl]methyl}cyclobutane-1-carbaldehyde (190 mg, 0.46 mmol),
(1R,2S,3R,5R)-3-{4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl}-5-(aminomethyl)-
cyclopentane-1,2-diol (140 mg, 0.46 mmol) and MgSO.sub.4 (554 mg,
4.61 mmol) in DCE (10 ml) was stirred for 15 min. STAB (137 mg,
0.645 mmol) was then added to the reaction mixture and stirred. The
reaction was monitored by LCMS, after which no amine was seen. Sat.
NaHCO.sub.3 (20 ml) was added to the reaction mixture and stirred
for 5 mins. Brine (10 ml) was added to the reaction mixture and the
product was extracted with DCM (2.times.30 ml), dried over
Na.sub.2SO.sub.4, filtered and evaporated. Purification by silica
gel column chromatography, eluting with 7N NH.sub.3 in MeOH:DCM
(1:99-5:95) gave the desired product as a pink foamy solid. The
mixed fractions were combined and purified on a prep.TLC plate
eluting with 7N NH.sub.3 in MeOH:DCM (2.times.4:96) to give a total
of 170 mg, 53%. MS (ESI.sup.+) for
C.sub.35H.sub.48F.sub.3N.sub.7O.sub.3Si m/z 700 [M+H].sup.+; HPLC
purity 100% (ret. time, 1.79 min); .sup.1H NMR (250 MHz,
CHLOROFORH-d) .delta..sub.H ppm -0.26-0.16 (9H, m), 0.72-0.97 (2H,
m), 1.30 (3H, s), 1.38-1.60 (5H, m), 1.91-2.59 (8H, m), 2.60-3.27
(6H, m), 3.35-3.70 (2H, m), 4.52 (1H, t, J=5.94 Hz), 4.78-5.20 (4H,
m), 5.39-5.66 (2H, m), 6.36 (1H, d, J=3.65 Hz), 7.03 (1H, d, J=3.65
Hz), 7.42-7.59 (1H, m), 7.69 (1H, s), 7.79 (1H, d, J=8.53 Hz), 8.32
(1H, s).
Step
8.7-[(3aS,4R,6R,6aR)-2,2-dimethyl-6-({methyl[(3-{[5-(trifluoromethyl)-
-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazol-2-yl]methyl}cyclo-
butyl)methyl]amino}methyl)-hexahydrocyclopenta[d][1,3]dioxol-4-yl]-7H-pyrr-
olo[2,3-d]pyrimidin-4-amine
##STR00308##
[0839] Formaldehyde (36 .mu.l, 0.49 mmol, 37% (aq) was added to a
solution of
7-[(3aS,4R,6R,6aR)-2,2-dimethyl-6-({[(3-{[5-(trifluoromethyl)-1-{[2-(t-
rimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazol-2-yl]methyl}cyclobutyl)met-
hyl]amino}methyl)-hexahydrocyclopenta[d][1,3]dioxol-4-yl]-7H-pyrrolo[2,3-d-
]pyrimidin-4-amine in MeOH (5 ml) and THE (5 ml), and the reaction
was stirred at RT for 30 mins. NaCNBH.sub.3 (18 mg, 0.29 mmol) was
added portionwise, and the reaction stirred for a further 2 hours
at RT after which LC MS showed the reaction to be complete. The
reaction mixture was concentrated under in vacuo, and the residue
partitioned between water (20 ml) and DCM (20 ml) and the layers
were separated. The aqueous layer was extracted with DCM
(2.times.20 ml), the combined organics were then dried over
Na.sub.2SO.sub.4, and concentrated. Purification by prep. TLC,
eluting with 7N NH.sub.3 in MeOH:DCM (5:95) gave the desired
product (114 mg, 66%) as a colourless oil; MS (ESI.sup.+) for
C.sub.36H.sub.50F.sub.3N.sub.7O.sub.3Si m/z 714.45 [M+H].sup.+;
HPLC purity 100% (ret. time, 1.77 min); .sup.1H NMR (500 MHz,
CHLOROFORM-d) .delta..sub.H ppm -0.14-0.05 (9H, m), 0.85-0.99 (2H,
m), 1.18-1.35 (3H, m), 1.37-1.53 (2H, m), 1.52-1.64 (3H, m),
1.89-2.15 (3H, m), 2.18-2.28 (2H, m), 2.29-2.70 (6H, m), 2.72-2.96
(1H, m), 2.97-3.20 (2H, m), 3.48 (3H, s), 3.50-3.58 (2H, m),
4.37-4.58 (1H, m), 4.85-5.05 (2H, m), 5.11-5.38 (2H, m), 5.41-5.57
(2H, m), 6.35 (1H, d, J=3.63 Hz), 6.84-7.19 (1H, m), 7.50 (1H, d,
J=8.35 Hz), 7.68 (1H, s), 7.78 (1H, d, J=8.35 Hz), 8.13-8.52 (1H,
m)
Step 9.
(1R,2S,3R,5R)-3-{4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl}-5-({meth-
yl[(3-{[5-(trifluoromethyl)-1H-1,3-benzodiazol-2-yl]methyl}cyclobutyl)meth-
yl]amino}methyl)cyclopentane-1,2-diol
##STR00309##
[0841] A solution of HCl in MeOH (1:1, 3 ml) was added to
7-[(3aS,4R,6R,6aR)-2,2-dimethyl-6-({methyl[(3-{[5-(trifluoromethyl)-1-{[2-
-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazol-2-yl]methyl}cyclobutyl)-
methyl]amino}methyl)-hexahydrocyclopenta[d][1,3]dioxol-4-yl]-7H-pyrrolo[2,-
3-d]pyrimidin-4-amine at 0.degree. C. and stirred. This was then
immediately stirred at 40.degree. C. for 4 h (reaction monitored by
LCMS) The reaction was continued for a further 1 h. LCMS still
showed 30% acetal deprotected SM. A further 1 ml of HCl (36% aq)
was added to the reaction mixture and continued at 40.degree. C.
for a further 1 h. The reaction mixture was then evaporated in
vacuo. The residue was dissolve in DCM (100 ml)+MeOH (1 ml) and
washed with sat. NaHCO.sub.3 (2.times.50 ml), dried over
Na.sub.2SO.sub.4, filtered and evaporated. Purification by
prep.TLC, eluting with 7N NH.sub.3 in MeOH:DCM (1:9); MS
(ESI.sup.+) for C.sub.27H.sub.32F.sub.3N.sub.7O.sub.2 m/z 544
[M+H].sup.+; HPLC purity 96% (ret. time, 2.03 min); .sup.1H NMR
(500 MHz, MeOD) .delta..sub.H ppm 1.47-1.58 (1H, m), 1.58-1.69 (1H,
m), 1.87-2.12 (1H, m), 2.12-2.54 (10H, m), 2.53-2.91 (3H, m),
2.93-3.15 (2H, m), 3.88-4.08 (1H, m), 4.32 (1H, dd, J=7.57, 6.15
Hz), 4.88-5.01 (1H, m), 6.59 (1H, d, J=3.63 Hz), 7.20 (1H, d,
J=3.63 Hz), 7.42-7.53 (1H, m), 7.63 (1H, d, J=8.20 Hz), 7.79 (1H,
s), 7.95-8.22 (1H, m).
Compound 19:
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)eth-
yl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclopen-
ta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin--
4-amine
[0842] A solution of
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(isopropyl)amino)cyclobutyl)propanoic acid (490 mg, 0.79
mmol) and 4-tert-butylbenzene-1,2-diamine (155 mg, 0.946 mmol) in
N,N-Dimethylformamide (8.1 ml) was treated with
N,N-Diisopropylethylamine (0.453 ml, 2.60 mmol) dropwise followed
by N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (449 mg, 1.18 mmol) in one portion. The
reaction mixture was stirred at RT overnight. The reaction mixture
was concentrated under high vacuum and the residue was partitioned
between 50 ml EtOAc and 50 ml 1/1 H.sub.2O/sat NaHCO.sub.3. The
aqueous phase was extracted with 30 ml EtOAc and the combined
organic phase was washed with 30 ml portions of H.sub.2O and brine.
The organic phase was dried over Na.sub.2SO.sub.4, filtered and
concentrated to give a glass/stiff foam. The crude material was
purified by flash chromatography (SiO.sub.2, eluting with 4% 7N N
NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2) to give the desired
intermediate as a mixture of amide regioisomers (400 mg). A
solution of intermediate (0.40 g) in Acetic acid (15 ml) was heated
at 65.degree. C. for 2.5 h, the reaction mixture was cooled and
placed under high vacuum to remove the acetic acid. The residue was
taken up in 60 ml CH.sub.2Cl.sub.2 and washed with 40 ml portions
of sat NaHCO.sub.3 and 2% Na.sub.2CO.sub.3 solution. The organic
phase was dried over Na.sub.2SO.sub.4, filtered and concentrated to
yield a glass/stiff foam. The material was placed on high vacuum
and used directly in the next step (380 mg)
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5-(te-
rt-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methy-
l)cyclopentane-1,2-diol
##STR00310##
[0844]
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2--
yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cy-
clopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyri-
midin-4-amine (390 mg, 0.52 mmol) was dissolved in a mixture of
Trifluoroacetic Acid (7.2 ml) and Water (0.8 ml) which had been
pre-cooled at 0.degree. C. in an ice bath. The solution was stirred
at 0.degree. C. for 30 minutes, then warmed to RT After 2.5 at RT
the residue was taken up in 15 ml MeOH and concentrated. This
procedure was repeated twice and the residue placed on high vacuum.
The material was taken up in 15 ml MeOH (gave a slurry) and was
treated with 500 mg K.sub.2CO.sub.3 and 8 drops of water. The
mixture was allowed to stir for 1 hr, during which time the
solution was found to be basic. The mixture was filtered through a
finefrit, the solids were washed with 10 ml MeOH and the filtrate
was concentrated to yield an off white solid. The material was left
on high vacuum overnight. The crude material was purified by flash
chromatography (SiO.sub.2, eluting with 8-10% 7N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2) to give a glass/stiff foam (0.22 g).
.sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.06 (s, 1H), 7.48
(br. s., 1H), 7.39 (m, 1H), 7.27 (m, 1H), 7.20 (d, J=3.52 Hz, 1H),
6.60 (m, 1H), 4.32 (t, J=6.43 Hz, 1H), 3.93 (t, J=5.29 Hz, 1H),
3.54 (m, 0.2H), 3.11 (t, J=9.33 Hz, 1H), 3.02 (m, 1H), 2.82 (m,
2H), 2.66 (dd, J=1 3.68, 8.09 Hz, 1H), 2.46 (m, 1H), 2.36 (m, 1H),
2.23 (m, 3H), 2.05 (m, 1H), 1.91 (m, 3H), 1.59 (m, 3H), 1.36 (s,
9H), 1.02 (m, 6H).
Compound 20:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(6-ch-
loro-5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopro-
pyl)amino)methyl)cyclopentane-1,2-diol
7-((3aS,4R,6R,6aR)-6-(((3-(2-(6-chloro-5-(trifluoromethyl)-1H-benzo[d]imid-
azol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltetrahydro-
-3aH-cyclopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-
-d]pyrimidin-4-amine
##STR00311##
[0846] A solution of
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(isopropyl)amino)cyclobutyl)propanoic acid (78.5 mg, 0.126
mmol) and 4-chloro-5-(trifluoromethyl)benzene-1,2-diamine (31.9 mg,
0.151 mmol) in N,N-Dimethylformamide (1.3 ml) was treated with
N,N-Diisopropylethylamine (72.5 ul, 0.416 mmol dropwise followed by
N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (72.0 mg, 0.189 mmol) in one portion. The
reaction mixture was stirred at RT for 7.5 h. The reaction mixture
was placed in the fridge overnight. The reaction mixture was
concentrated under high vacuum and the residue was partitioned
between 20 ml EtOAc and 20 ml 1/1 H.sub.2O/sat NaHCO.sub.3. The
aqueous phase was extracted with 10 ml EtOAc and the combined
organic phase was washed with 10 ml portions of H.sub.2O and brine.
The organic phase was dried over Na.sub.2SO.sub.4, filtered and
concentrated. The crude material was purified by flash
chromatography (SiO.sub.2, eluting with 3% 7N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2) to give the desired intermediate as a
glass/stiff foam (regiosomeric amides and the cis/trans
diastereomers, 87 mg). The intermediate (0.087 g) was taken up in
in Acetic acid (4.5 ml,) was heated at 65.degree. C. for 6 h, it
was cooled to RT and stirred at RT for 48 hr. The reaction was
heated at 65.degree. C. for 8 hr, then at RT o/n then at 65.degree.
C. for a further 6.5 h. The mixture was cooled and placed under
high vacuum to remove the acetic acid. The residue was taken up in
15 ml CH.sub.2Cl.sub.2 and washed with 10 ml portions of sat.
NaHCO.sub.3 and 2% Na.sub.2CO.sub.3 solution. The organic phase was
dried over Na.sub.2SO.sub.4, filtered and concentrated to yield a
glass/stiff foam. The material was placed on high vacuum overnight.
The crude material was purified by prep TLC on a 20 cm.times.20
cm.times.1.0 mm prep TLC plate eluting twice with 4% 7N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2. The product band was isolated to yield
the product as a white solid (48 mg).
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(6-chl-
oro-5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isoprop-
yl)amino)methyl)cyclopentane-1,2-diol
##STR00312##
[0848]
7-((3aS,4R,6R,6aR)-6-(((3-(2-(6-chloro-5-(trifluoromethyl)-1H-benzo-
[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltet-
rahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrr-
olo[2,3-d]pyrimidin-4-amine (525 mg, 0.683 mmol) was dissolved in a
mixture of Trifluoroacetic Acid (9 ml) and Water (1 ml) which had
been pre-cooled at 0.degree. C. in an ice bath. The solution was
stirred at 0.degree. C. for 30 minutes, then warmed to RT. After 4
h at RT, the mixture was concentrated. The residue was taken up in
20 ml MeOH and concentrated. This procedure was repeated twice and
the residue placed on high vacuum. The material was taken up in 15
ml MeOH (gave a slurry) and was treated with 500 mg K.sub.2CO.sub.3
and 15 drops of water. The mixture was allowed to stir for 1 hr,
during which time the solution was found to be basic. The mixture
was filtered through a fine frit, the solids were washed with 10 ml
MeOH and the filtrate was concentrated to yield an off white solid.
The material was left on high vacuum overnight. The crude material
was purified by flash chromatography (SiO.sub.2, eluting with 12%
7N NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2) to give a colorless
glass/stiff foam. .sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm
8.06 (s, 1H), 7.89 (d, J=2.07 Hz, 1H), 7.69 (d, J=2.28 Hz, 1H),
7.21 (dd, J=3.42, 1.76 Hz, 1H), 6.59 (d, J=3.52 Hz, 1H), 4.33 (t,
J=6.84 Hz, 1H), 3.89 (q, J=5.25 Hz, 1H), 3.03 (m, 0.5H), 2.89 (m,
2H), 2.70 (m, 0.5H), 2.49 (m, 1H), 2.40 (m, 2H), 2.27 (br. s., 2H),
2.16 (d, J=7.26 Hz, 4H), 2.06 (m, 2H), 1.91 (m, 2H), 1.62 (m, 1H),
1.52 (m, 1H).
Compound 21:
(1R,2S,3R,5R)-3-{4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl}-5-({[(3-{[6-chl-
oro-5-(trifluoromethyl)-1H-1,3-benzodiazol-2-yl]methyl}cyclobutyl)methyl](-
propan-2-yl)amino}methyl)cyclopentane-1,2-diol
Step 1:
7-[(3aS,4R,6R,6aR)-6-({[(3-{[6-chloro-5-(trifluoromethyl)-1-{[2-(t-
rimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazol-2-yl]methyl}cyclobutyl)met-
hyl](propan-2-yl)amino}methyl)-2,2-dimethyl-hexahydrocyclopenta[d][1,3]dio-
xol-4-yl]-7H-pyrrolo[2,3-d]pyrimidin-4-amine
##STR00313##
[0850] A solution of
3-{[6-chloro-5-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H--
1,3-benzodiazol-2-yl]methyl}cyclobutane-1-carbaldehyde (223 mg,
0.50 mmol),
7-[(3aS,4R,6R,6aR)-2,2-dimethyl-6-[(propan-2-ylamino)methyl]-hexah-
ydrocyclopenta[d][1,3]dioxol-4-yl]-7H-pyrrolo[2,3-d]pyrimidin-4-amine
(172 mg, 0.50 mmol) and MgSO.sub.4 (600 mg, 5.00 mmol) in DCE (10
ml) was stirred for 15 min. STAB (148 mg, 0.70 mmol) was then added
to the reaction mixture and stirred for 1 h at RT. The reaction was
monitored by LCMS, no amine was seen after 2.5 h. Sat. NaHCO.sub.3
(20 ml) was added to the reaction mixture and stirred for 5 mins.
Brine (20 ml) was then added to the reaction mixture. The product
was extracted with DCM (2.times.30 ml), dried over
Na.sub.2SO.sub.4, filtered and evaporated. Purification by prep.
HPLC gave the desired product (174 mg, 45%) as a white solid; MS
(ESI.sup.+) for C.sub.38H.sub.53ClF.sub.3N.sub.7O.sub.3Si m/z
776.30 [M+H].sup.+; HPLC purity 100% (ret. time, 1.68 min); .sup.1H
NMR (500 MHz, MeOD) .delta..sub.H ppm -0.25-0.11 (9H, m), 0.90 (2H,
td, J=7.88, 3.47 Hz), 1.30 (3H, s), 1.37 (6H, t, J=7.49 Hz), 1.55
(3H, s), 1.63-1.84 (1H, m), 1.99-2.37 (2H, m), 2.37-3.03 (6H, m),
3.04-3.28 (3H, m), 3.34-3.50 (1H, m), 3.61 (2H, td, J=7.92, 2.29
Hz), 3.79 (1H, br. s.), 4.68 (1H, t, J=6.70 Hz), 4.95-5.27 (2H, m),
5.55-5.81 (1H, m), 6.88 (1H, d, J=3.63 Hz), 7.06-7.62 (2H, m),
7.61-7.94 (1H, m), 7.94-8.16 (1H, m), 8.22 (1H, s)
Step 2.
(1R,2S,3R,5R)-3-{4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl}-5-({[(3--
{[6-chloro-5-(trifluoromethyl)-1H-1,3-benzodiazol-2-yl]methyl}cyclobutyl)m-
ethyl](propan-2-yl)amino}methyl)cyclopentane-1,2-diol
##STR00314##
[0852] 12N HCl (1.5 ml) was added slowly to a solution of
7-[(3aS,4R,6R,6aR)-6-({[(3-{[6-chloro-5-(trifluoromethyl)-1-{[2-(trimethy-
lsilyl)ethoxy]methyl}-1H-1,3-benzodiazol-2-yl]methyl}cyclobutyl)methyl](pr-
opan-2-yl)amino}methyl)-2,2-dimethyl-hexahydrocyclopenta[d][1,3]dioxol-4-y-
l]-7H-pyrrolo[2,3-d]pyrimidin-4-amine (174 mg, 0.22 mmol) in MeOH
(1.5 ml) and stirred at 40.degree. C. for 6 h. The reaction was
monitored by LCMS, no starting material seen after 2.5 h. The
reaction mixture was concentrated in vacuo, then basified with 7N
NH.sub.3 in MeOH (5 ml). This was then evaporated to dryness.
Purification by silica gel column chromatography, eluting with 7N
NH.sub.3 in MeOH:DCM (1:9) gave the desired product (36 mg, 27%) as
a white solid; MS (ESI.sup.+) for
C.sub.29H.sub.35ClF.sub.3N.sub.7O.sub.2 m/z 606.30 [M+H].sup.+;
HPLC purity 100% (ret. time, 2.59 min); .sup.1H NMR (500 MHz,
MeOD).sup.6H ppm 0.93-1.09 (6H, m), 1.41-1.64 (2H, m), 1.76-2.06
(1H, m), 2.15-2.65 (9H, m), 2.65-2.88 (1H, m), 2.91-3.13 (3H, m),
3.80-4.09 (1H, m), 4.19-4.46 (1H, m), 4.88-4.94 (1H, m), 6.46-6.77
(1H, m), 7.19 (1H, d, J=3.63 Hz), 7.69 (1H, s), 7.89 (1H, s), 8.06
(1H, s)
Compound 22:
(1R,2S,3R,5R)-3-{4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl}-5-{[({3-[(5-ter-
t-butyl-1H-1,3-benzodiazol-2-yl)methyl]cyclobutyl}methyl)(propan-2-yl)amin-
o]methyl}cyclopentane-1,2-diol
Stage 1:
3-{[(2-amino-4-tert-butylphenyl)carbamoyl]methyl}-N-methoxy-N-met-
hylcyclobutane-1-carboxamide
##STR00315##
[0854] N,N-Diisopropylethylamine (5.19 ml, 29.82 mmol) was added to
a suspension of 2-{3-[methoxy(methyl)carbamoyl]cyclobutyl}acetic
acid (3 g, 14.91 mmol), 4-tBu phenylene diamine (2.69 g, 16.4 mmol)
and
(1-cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbeni-
um hexafluorophosphate (7.02 g, 16.4 mmol) in dichloromethane (60
ml) at 0.degree. C. and stirred for 20 mins before being allowed to
warm to RT. The reaction was left at RT for 4 h. The reaction
mixture was concentrated, and the residue redissolved in EtOAc (60
ml). The solution was washed with water (3.times.60 ml), then brine
(60 ml), dried (MgSO.sub.4) and concentrated under reduced
pressure. The crude material was purified by dry flash
chromatography, eluting with 100% EtOAc to afford the title
compound (3.74 g, 46%) as a brown oil: MS (ESI.sup.+) for
C.sub.19H.sub.29N.sub.3O.sub.3 m/z 348.5 [M+H].sup.+; LC purity 26%
and 44% (UV), 18% and 68% (ELS), (ret. time, 1.65 and 1.71
min).
Stage 2:
3-[(5-tert-butyl-1H-1,3-benzodiazol-2-yl)methyl]-N-methoxy-N-meth-
ylcyclobutane-1-carboxamide
##STR00316##
[0856] A stirred solution of
3-{[(2-amino-4-tert-butylphenyl)carbamoyl]methyl}-N-methoxy-N-methylcyclo-
butane-1-carboxamide (70%, 2.62 g, 5.28 mmol) in acetic acid (25
ml) was heated to reflux for 1 h. The reaction mixture was
concentrated under reduced pressure, and the residue partitioned
between sat. NaHCO.sub.3 (aq) (25 ml) and EtOAc (25 ml), and the
layers separated. The aqueous layer was extracted with EtOAc
(2.times.25 ml), the combined organics were washed with brine (50
ml), dried (MgSO.sub.4) and concentrated. The crude material was
purified by silica flash column chromatography, eluting with 1-10%
2M NH.sub.3 in MeOH in DCM to afford the title compound (2.02 g,
93%) as a yellow gum: MS (ESI.sup.+) for
C.sub.19H.sub.27N.sub.3O.sub.2 m/z 330.5 [M+H].sup.+; LC purity 80%
(UV), 100% (ELS), (ret. time, 1.51 min); 1H NMR (500 MHz,
CHLOROFORM-d) .delta..sub.H ppm 7.56 (br. s., 1H), 7.48 (d, J=8.5
Hz, 1H), 7.30 (dd, J=8.5, 1.7 Hz, 1H), 3.80-3.90 (m, 1H), 3.65 (s,
3H), 3.47-3.57 (m, 1H), 3.20 (s, 3H), 2.94-3.13 (m, 2H), 2.88 (dt,
J=16.1, 8.1 Hz, 1H), 2.32-2.60 (m, 2H), 2.01-2.17 (m, 2H), 1.38 (s,
9H).
Stage 3:
3-[(5-tert-butyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benz-
odiazol-2-yl)methyl]-N-methoxy-N-methylcyclobutane-1-carboxamide
##STR00317##
[0858] To a solution of
3-[(5-tert-butyl-1H-1,3-benzodiazol-2-yl)methyl]-N-methoxy-N-methylcyclob-
utane-1-carboxamide (80%, 2.02 g, 4.91 mmol) in
N,N-dimethylformamide (40 ml) under N2 was added potassium
carbonate (1.36 g, 9.81 mmol) and the reaction was stirred for 1 h.
2-(Trimethylsilyl)ethoxymethyl chloride (1.31 ml, 7.36 mmol) was
added slowly, and stirring maintained for 20 h. The reaction
mixture was filtered and concentrated under reduced pressure. The
residue was taken up in EtOAc (50 ml) and washed with water
(3.times.50 ml), then brine (50 ml) before being dried (MgSO.sub.4)
and concentrated. The crude material was purified by silica flash
column chromatography, eluting with 50-100% EtOAc in heptane to
afford the title compound (1.21 g, 54%) as a colourless gum: MS
(ESI.sup.+) for C.sub.25H41N303Si m/z 461.0 [M+H].sup.+; LC purity
97% (UV), 100% (ELS), (ret. time, 1.98 min); .sup.1H NMR (500 MHz,
CHLOROFORM-d) .delta. ppm 7.58-7.89 (m, 1H), 7.33 (s, 2H),
5.39-5.53 (m, 2H), 3.65 (s, 3H), 3.49-3.58 (m, 2H), 3.41 (br. s.,
1H), 3.10-3.23 (m, 3H), 3.03 (s, 3H), 2.32-2.63 (m, 2H), 2.00-2.18
(m, 2H), 1.32-1.45 (m, 9H), 0.91 (td, J=8.1, 4.9 Hz, 2H),
-0.13-0.07 (m, 9H).
Stage 4:
3-[(5-tert-butyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benz-
odiazol-2-yl)methyl]cyclobutane-1-carbaldehyde
##STR00318##
[0860] To a solution of
3-[(5-tert-butyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazol--
2-yl)methyl]-N-methoxy-N-methylcyclobutane-1-carboxamide (0.61 g,
1.32 mmol) in tetrahydrofuran (15 ml) was added dropwise 1M
diisobutylaluminum hydride solution in toluene (3.29 ml) under N2
at -10.degree. C. The reaction was stirred at this temperature for
2.5 h before being quenched by the addition of methanol (2 ml) and
stirred for 5 mins. The solution was poured onto saturated aq.
Rochelle's salt (20 ml), diluted with Et.sub.2O (30 ml) and stirred
for 30 min. This was then separated and the organic layer was
washed with Rochelle's salt (30 ml), sat. NaHCO.sub.3 (30 ml), and
brine (30 ml). This was dried (MgSO.sub.4) and concentrated to
afford the title compound (0.69 g, 130%) as a colourless gum which
was used crude in the next reaction.
Stage 5:
7-[(3aS,4R,6R,6aR)-6-{[({3-[(5-tert-butyl-1-{[2-(trimethylsilyl)e-
thoxy]methyl}-1H-1,3-benzodiazol-2-yl)methyl]cyclobutyl}methyl)(propan-2-y-
l)amino]methyl}-2,2-dimethyl-hexahydrocyclopenta[d][1,3]dioxol-4-yl]-7H-py-
rrolo[2,3-d]pyrimidin-4-amine
##STR00319##
[0862]
3-[(5-Tert-butyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benzod-
iazol-2-yl)methyl]cyclobutane-1-carbaldehyde (282.3 mg, 0.7 mmol),
7-[(3aS,4R,6R,6aR)-2,2-dimethyl-6-[(propan-2-ylamino)methyl]-hexahydrocyc-
lopenta[d][1,3]dioxol-4-yl]-7H-pyrrolo[2,3-d]pyrimidin-4-amine
(243.41 mg, 0.7 mmol) and magnesium sulfate (127.22 mg, 1.06 mmol)
were stirred in 1,2-dichloroethane (10 ml) for 15 mins. Sodium
triacetoxyborohydride (179.21 mg, 0.85 mmol) was added, and the
reaction stirred overnight. The reaction was quenched by the
addition of sat. Na.sub.2CO.sub.3 (10 ml), and the solution was
extracted with DCM (3.times.10 ml). The combined organics were
dried (MgSO.sub.4) and concentrated. The crude material was
purified by mass directed prep-HPLC (acidic method). After
combining the fractions, a small amount of 7M NH.sub.3 in MeOH was
added to basify the solution. After concentration, the residue was
partitioned between water (5 ml) and DCM (5 ml), and the layers
separated. The aqueous layer was extracted with DCM (2.times.3 ml)
and the combined organics were dried (MgSO.sub.4) and concentrated
to afford the title compound (58.7 mg, 11%) as a colourless gum: MS
(ESI.sup.+) for C.sub.41H.sub.63N.sub.7O.sub.3Si m/z 730.2
[M+H].sup.+; LC purity 95% (UV), 100% (ELS), (ret. time, 1.65 min);
1H NMR (500 MHz, CHLOROFORM-d) .delta..sub.H 8.30 (s, 1H), 7.75 (s,
1H), 7.32 (s, 2H), 7.04 (d, J=3.6 Hz, 1H), 6.37 (d, J=3.6 Hz, 1H),
5.40-5.50 (m, 2H), 5.24 (br. s., 2H), 4.89-5.04 (m, 2H), 4.45 (d,
J=5.2 Hz, 1H), 3.50 (s, 3H), 2.11-3.09 (m, 13H), 2.01 (s, 6H),
1.34-1.49 (m, 6H), 1.29 (s, 3H), 0.85-1.02 (m, 8H), -0.11-0.05 (m,
9H).
Stage 6:
(1R,2S,3R,5R)-3-{4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl}-5-{[({3-
-[(5-tert-butyl-1H-1,3-benzodiazol-2-yl)methyl]cyclobutyl}methyl)(propan-2-
-yl)amino]methyl}cyclopentane-1,2-diol
##STR00320##
[0864]
7-[(3aS,4R,6R,6aR)-6-{[({3-[(5-tert-butyl-1-{[2-(trimethylsilyl)eth-
oxy]methyl}-1H-1,3-benzodiazol-2-yl)methyl]cyclobutyl}methyl)(propan-2-yl)-
amino]methyl}-2,2-dimethyl-hexahydrocyclopenta[d][1,3]
dioxol-4-yl]-7H-pyrrolo[2,3-d]pyrimidin-4-amine (58.7 mg, 0.08
mmol) was dissolved in conc. HCl solution (5 ml) and methanol (5
ml) and heated to 40.degree. C. for 2 h. The reaction mixture was
concentrated under reduced pressure, and the residue partitioned
between sat. NaHCO.sub.3 (aq) (10 ml) and EtOAc (10 ml). The layers
were separated and the aqueous layer was extracted with EtOAc
(2.times.10 ml), the combined organics were then dried (MgSO.sub.4)
and concentrated. The crude material was purified by prep TLC,
eluting with 10% 2M NH.sub.3 in MeOH in DCM to afford the title
compound (24.7 mg, 55%) as a colourless gum: MS (ESI.sup.+) for
C.sub.32H.sub.45N.sub.7O.sub.2 m/z 560.4 [M+H].sup.+; LC purity
100% (UV), (ret. time, 5.10 min); .sup.1H NMR (500 MHz, Acetone)
.delta..sub.H 8.12 (s, 1H), 7.30-7.60 (m, 2H), 7.13-7.28 (m, 2H),
6.54 (d, J=3.5 Hz, 1H), 6.33 (br. s., 1H), 4.85-5.07 (m, 1H), 4.36
(t, J=6.2 Hz, 1H), 4.08 (t, J=5.2 Hz, 1H), 3.01 (d, J=7.7 Hz, 1H),
2.93 (d, J=7.4 Hz, 2H), 2.78 (br. s., 2H), 2.59-2.73 (m, 2H),
2.15-2.56 (m, 7H), 1.68 (dt, J=12.4, 9.7 Hz, 1H), 1.44-1.62 (m,
2H), 1.35 (s, 9H), 0.90-1.07 (m, 6H).
Compound 23:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5,6--
dichloro-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)cy-
clopentane-1,2-diol
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5,6-dichloro-1H-benzo[d]imidazol-2-yl)ethyl)-
cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][-
1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amin-
e
##STR00321##
[0866] A solution of
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(methyl)amino)cyclobutyl)propanoic acid (450 mg, 0.76 mmol)
and 4,5-Dichloro-1,2-phenylenediamine (161 mg, 0.910 mmol) in
N,N-Dimethylformamide (7.8 ml) was treated with
N,N-Diisopropylethylamine (0.44 ml, 2.5 mmol) dropwise followed by
N,N,N',N'-Tetramethyl-0-(7-azabenzotriazol-1-yl}uronium
Hexafluorophosphate (432 mg, 1.14 mmol) in one portion. The
reaction mixture was stirred at RT 5.5 h. The reaction mixture was
concentrated under high vac and the residue was partitioned between
50 ml EtOAC and 50 ml 1/1 H.sub.2O/sat NaHCO.sub.3. The aqueous
phase was extracted with 30 ml EtOAc and the combined organic phase
was washed with 30 ml portions of H.sub.2O and brine. The organic
phase was dried over Na.sub.2SO.sub.4, filtered and concentrated to
a foam. The crude material was purified by flash chromatography
(SiO.sub.2 eluting with 5% 7N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2) to give the intermediate amide (as a
mixture of amide regioisomers, 520 mg).
[0867] The intermediate amide (0.52 g) in Acetic acid (16 ml) was
heated at 65.degree. C. for 5.5 h, the reaction mixture was cooled
and placed under high vac to remove the acetic acid. The residue
was taken up in 70 ml CH.sub.2Cl.sub.2 and washed with 50 ml
portions of sat NaHCO.sub.3 and 2% Na.sub.2CO.sub.3 solution. The
organic phase was dried over Na.sub.2SO.sub.4, filtered and
concentrated to yield a foam. The material was placed on high
vacuum overnight and the residue was purified twice by flash
chromatography (SiO.sub.2, eluting with 4% 7NH NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2, 2.sup.nd column eluting with 2-6% EtOH
sat w/NH.sub.3/CH.sub.2Cl.sub.2) to give the desired compound (377
mg) .sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.10 (s, 1H),
7.63 (d, J=1.66 Hz, 2H), 7.21 (m, 1H), 7.13 (d, J=8.29 Hz, 1H),
6.62 (d, J=3.32 Hz, 1H), 6.54 (d, J=2.07 Hz, 1H), 6.43 (dd, J=8.40,
2.38 Hz, 1H), 4.96 (m, 2H), 4.65 (s, 2H), 4.51 (m, 1 H), 3.84 (d,
J=1.04 Hz, 3H), 3.76 (s, 3H), 2.99 (m, 0.5H), 2.83 (m, 2H), 2.66
(m, 0.5H), 2.38 (m, 4H), 2.22 (m, 1H), 2.15 (d, J=7.67 Hz, 3H),
2.08 (m, 2H), 2.00 (m, 2H), 1.90 (m, 1H), 1.84 (m, 1H), 1.53 (s,
3H), 1.46 (m, 1H), 1.29 (s, 3H)
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5,6-d-
ichloro-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)cyc-
lopentane-1,2-diol
##STR00322##
[0869]
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5,6-Dichloro-1H-benzo[d]imidazol-2-yl-
)ethyl)cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclope-
nta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-
-4-amine (377 mg, 0.513 mmol) was dissolved in a mixture of
Trifluoroacetic Acid (7.1 ml) and Water (0.8 ml) which had been
pre-cooled at 0.degree. C. in an ice bath. The solution was stirred
at 0.degree. C. for 30 minutes, then warmed to and stirring was
continued for 3 h at RT. The suspension was concentrated and the
residue was taken up in 15 ml MeOH and concentrated. This procedure
was repeated twice and the residue placed on high vacuum. The
material was taken up in 10 ml MeOH (gave a slurry) and was treated
with 500 mg K.sub.2CO.sub.3 and 0.2 ml of water. The mixture was
allowed to stir for 1.5 hr, during which time the pH of the
solution was .about.9. The mixture was filtered through a fine
frit, the solids were washed with 20 ml MeOH and the filtrate was
concentrated to yield an off white solid. The material was left on
high vacuum overnight and purified by flash chromatography
(SiO.sub.2, eluting with 10-12% 7N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2) to give the desired product (227 mg).
.sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.06 (s, 1H), 7.63
(m, 2H), 7.21 (dd, J=3.63, 2.38 Hz, 1H), 6.59 (d, J=3.52 Hz, 1H),
4.32 (m, 1H), 3.88 (m, 1H), 3.01 (m, 0.5H), 2.84 (m, 2H), 2.70 (m,
0.5H), 2.51 (m, 1H), 2.40 (m, 2H), 2.26 (m, 2H), 2.17 (d, J=7.05
Hz, 3H), 2.11 (m, 2H), 2.02 (m, 1H), 1.90 (m, 3H), 1.62 (m, 1H),
1.49 (m, 1H).
Compound 24:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl(3-(2-
-(5-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)met-
hyl)cyclopentane-1,2-diol
N-(2-amino-4-(trifluoromethoxy)phenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4--
dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrah-
ydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)pro-
panamide
##STR00323##
[0871] N N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (1.2 g, 3.2 mmol) added to a solution
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(methyl)amino)cyclobutyl)propanoic acid (1.25 g, 210 mmol)
and N,N-Diisopropylethylamine (1.2 mL, 6.9 mmol) and
4-(trifluoromethoxy)benzene-1,2-diamine (0.48 g, 2.5 mmol) in
N,N-Dimethylformamide (10 mL). The mixture was stirred overnight at
RT, partially concentrated to ca 2 mls and then NaHCO.sub.3
(saturated) was added. The mixture was extracted with EtOAc
(3.times.) and the combined organics were dried with MgSO.sub.4,
filtered and concentrated. The residue was purified by flash
chromatography (DCM/7N N13 in MeOH 95:5) to give the desired
compound (2 g) as an oil.
N-(2,4-dimethoxybenzyl)-7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(5-
-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl-
)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin--
4-amine
##STR00324##
[0873] A solution of
N-(2-amino-4-(trifluoromethoxy)phenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-
-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetra-
hydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)pr-
opanamide (2 g, 2 mmol) in Acetic acid (4 ml), was stirred
overnight at 60 The volatiles removed in vacuo and remaining
residue purified by flash chromatography (DCM/7N NH.sub.3 in MeOH
92:8) to give the desired compound (1 g) as a solid.
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl(3-(2--
(5-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)meth-
yl)cyclopentane-1,2-diol
##STR00325##
[0875] Trifluoroacetic Acid (20 ml) added to a mixture of Water (2
ml) and
N-(2,4-dimethoxybenzyl)-7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(-
5-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methy-
l)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-
-4-amine (1 g, 1 mmol) at RT.
[0876] The reaction was stirred for 1.5 hours then quenched with
Triethylsilane (0.43 ml, 2.7 mmol). The volatiles were removed in
vacuo and resulting residue was purified twice by flash
chromatography (DCM/7N NH.sub.3 in MeOH 87:13) to give the desired
product (0.28 g) as a foam. MS (ESI.sup.+) for
C.sub.27H.sub.32F.sub.3N.sub.7O.sub.3 m/z 560.2 [M+H].sup.+; MS
(ESI.sup.-) for C.sub.27H.sub.32F.sub.3N.sub.7O.sub.3 m/z 558.2
[M-H].sup.-; HPLC purity >93% (ret. time, 2.504 min.) .sup.1H
NMR (400 MHz, d4-MeOH) .delta..sub.H 8081 (s, 1H), 7.551-7,530 (m,
1H). 7.414 (s, 1H), 7.230-7.216 (m, 1H), 7.156-7134 m, 1H),
6.621-6.613 (d, J=3.2 Hz, 1H), 4.362-4.326 (m, 1H), 3.952-3.915 (m,
1H), 3.237-3.182 (m, 0.5H (methine of trans isomer), 2.927-2.857
(m, 2.5H (contains methine of cis isomer)), 2.697-2.646 (m, 1H),
2.590-2.515 (m, 1H), 2.479-2.408 (m, 1N), 2.343-2.305 (m, 5H),
2.210-2.174 (m, 2H), 2.086-1.949 (m, 4H), 1.721-1.545 (m, 2H).
Retention time: 2.52 min 1HPLC Conditions:Agilent Zorbax Exlipse
XDB-C18 column, 4.6.times.50 mm (1.8 urn packing), Solvent A-Water
(0.1% TFA), Solvent B-Acetonitrile (0.07% TFA) 6 min gradient from
5 to 95% B; min hold; then recycle.
Compound 25:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5-(t-
ert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(ethyl)amino)methyl)c-
yclopentane-1,2-diol
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimeth-
oxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3-
aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(ethyl)amino)cyclobutyl)propanamid-
e
##STR00326##
[0878] N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (0.84 g, 2.2 mmol) added to a solution
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(ethyl)amino)cyclobutyl)propanoic acid (0.89 g, 1.5 mmol)
and N,N-Diisopropylethylamine (0.84 ml, 4.8 mmol) and
4-tert-butylbenzene-1,2-diamine (0.29 g, 1.8 mmol) in
N,N-Dimethylformamide (9 ml). The reaction was stirred overnight at
RT, partially concentrated to ca. 2 ml and then NaHCO.sub.3
(saturated) was added. The mixture was extracted with EtOAc
(3.times.) and the combined organics were dried with MgSO.sub.4,
filtered and concentrated. The residue was purified by flash
chromatography (DCM I 7N NH.sub.3 in MeOH 94:6) to give the desired
compound (0.88 g) as a colorless solid. Retention time C: 3.363
minutes.
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethy-
l)cyclobutyl)(ethyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d]-
[1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ami-
ne
##STR00327##
[0880] A solution of
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimet-
hoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro--
3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(ethyl)amino)cyclobutyl)propanami-
de (0.88 g, 2 mmol) in Acetic acid (3 mL) was stirred overnight at
60.degree. C., the volatiles removed in vacuo and remaining residue
purified by flash chromatography (DCM/7N NH.sub.3 in MeOH 93:7) to
give the desired compound (0.85 g) as a foam.
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5-(te-
rt-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(ethyl)amino)methyl)cy-
clopentane-1,2-diol
##STR00328##
[0882] Trifluoroacetic Acid (20 mL) was added to a mixture of Water
(2 mL) and
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl-
)ethyl)cyclobutyl)(ethyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclopen-
ta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin--
4-amine (0.85 g, 12 mmol) at RT. The reaction was allowed to
proceed for one hour at which time Triethylsilane (0.37 mL, 2.3
mmol) was added. The volatiles were removed in vacuo and resulting
residue was taken up in MeOH (3 mls), 1 ml of K.sub.2CO.sub.3
(saturated) was added and reaction stirred at RT for 1 hour. The
mixture was partitioned between H.sub.2O and DCM/MeOH (9:1). The
aqueous layer was extracted (3.times.) and the combined organics
dried with MgSO.sub.4, filtered and concentrated. The resulting
residue was purified by flash chromatography (DCM/7N NH.sub.3 in
MeOH 90:10) to yield the desired product (0.150 g) as an off white
foam. MS (ESI.sup.+) for C.sub.31H.sub.43N.sub.7O.sub.2 n 546.3
[M+H].sup.+; MS (ESI.sup.-) for C.sub.31H.sub.43N.sub.7O.sub.2 m/z
544.3 [M-H].sup.-; HPLC purity >91% (ret. time, 2.734 min.)
.sup.1H NMR (400 MHz, d4-MeOH) .delta..sub.H 8.081 (s, 1H), 7.493
(s, 1H), 7.414-7.393 (m, 1H), 7.291-7.266 (m, 1H), 7.215-7.202 (m,
1H), 6.619-6.609 (d, J=4.0 Hz, 1H), 4.345-4.312 (m, 14),
3.923-3.885 (m, 1H), 2.994-2.915 (m, 0.5H (methine of trans
isomer)), 2.860-2.793 (m, 2H), 2.701-2.578 (m, 3H), 2.501-2.380 (m,
2H), 2.259-2.234 (m, 2H), 2.109-2.008 (m, 3H), 1.920-1.880 (m, 3H),
1.658-1.499 (m, 2H), 1.364 (s, 9H), 1.036-0.991 (n, 31-1).
Retention time: 2.734 minutes. HPLC Conditions:Agilent Zorbax
Exlipse XDB-C18 column, 4.6.times.50 mm (1.8 um packing), Solvent
A-Water (0.1% TFA), Solvent B-Acetonitrile (0.07% TFA). 6 min
gradient from 5 to 95% B min hod; then recycle
Compound 26:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5-br-
omo-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)cyclope-
ntane-1,2-diol
N-(2-amino-4-bromophenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenz-
yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cycl-
openta[d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanamide
##STR00329##
[0884] N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (1.20 g, 3.16 mmol) was added to a solution
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(methyl)amino)cyclobutyl)propanoic acid (1.25 g, 2.10 mmol)
and N,N-Diisopropylethylamine (1.21 ml, 6.95 mmol) and
4-bromobenzene-1,2-diamine (0.472 g, 2.53 mmol) in
N,N-Dimethylformamide (13.0 ml). The reaction was stirred overnight
at RT, partially concentrated to ca. 2 m is and then NaHCO.sub.3
(saturated) was added. The mixture was extracted with EtOAc
(3.times.) and the combined organics were dried with MgSO.sub.4,
filtered and concentrated. The residue was purified by flash
chromatography (DCM/7N NH.sub.3 in MeOH 95:5) to give the desired
compound (1.2 g) as a solid.
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-bromo-1H-benzo[d]imidazol-2-yl)ethyl)cyclo-
butyl)(methyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]d-
ioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
##STR00330##
[0886] A solution of
N-(2-amino-4-bromophenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxyben-
zyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyc-
lopenta[d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanamide
(1.2 g, 1.6 mmol) in Acetic acid (4 ml, 70 mmol) was stirred
overnight at 60.degree. C., the volatiles were removed in vacuo and
remaining residue purified directly by flash chromatography (DCM/7N
NH.sub.3 in MeOH 91:9) to give (0.9 g) as a foam.
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5-bro-
mo-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)cyclopen-
tane-1,2-diol
##STR00331##
[0888] Trifluoroacetic Acid (20 ml) added to a mixture of Water (2
ml) and
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-bromo-1H-benzo[d]imidazol-2-yl)ethyl)cycl-
obutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]-
dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
(0.9 g, 1 mmol) at RT. The reaction mixture was stirred for one
hour, Triethylsilane (0.39 ml, 2.4 mmol) was added. The volatiles
were removed in vacuo and resulting residue was purified twice by
flash chromatography (DCM/7N NH.sub.3 in MeOH 87:13). The residue
was taken up in MeOH/H.sub.2O (5:0.5 ml) and K.sub.2CO.sub.3 (100
mg) added. The mixture was stirred for 1 hour, then concentrated
and purified by flash chromatography (DCM/7N NH.sub.3 in MeOH
87:13) to give the desired product (0.15 g) as an off-white
foam/gum.
cyclobutyl)(methyl)amino)methyl)cyclopentane-1,2-diol (0.15 g; 20%)
as an off-white foam/gum. MS (ESI.sup.+) for
C.sub.26H.sub.32BrN.sub.7O.sub.2 m/z 554.1 [M+H].sup.+; MS
(ESI.sup.-) for C.sub.26H.sub.32BrN.sub.7O.sub.2 m/z 552.1
[M-H].sup.-; HPLC purity >90% (ret. time, 2.298 min.) .sup.1H
NMR (400 MHz, d.sub.4-MeOH) .delta..sub.H 8.080 (s, 1H), 7.652 (s,
1H), 7.425-7.403 (m, 1H), 7.342-7.7.312 (m, 1H), 7.232-7.217 (m,
1H), 6.620-6.611 (d, J=3.6 Hz, 1H), 4.358-4.318 (m, 1H),
3.928-3.888 (m, 1H), 3.099-3.039 (m, 0.5H (methine of trans
isomer)), 2.898-2.829 (m, 2H), 2.777-2.726 (m, 0.5H (methine of cis
isomer)), 2.580-2.529 (m, 1H), 2.453-2.397 (m, 2H), 2.307-2.141
(7H), 2.068-2.017 (m, 1H), 1.955-1.891 (m, 3H), 1.695-1.506 (m,
2H).
Compound 27:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((isopropyl(3-(2-(5-(1-methylcy-
clobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydr-
ofuran-3,4-diol
9-((3aR,4R,6R,6aR)-6-((isopropyl(3-(2-(5-(1-methylcyclobutyl)-1H-benzo[d]i-
midazol-2-yl)ethyl)cyclobutyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3,4-
-d][1,3]dioxol-4-yl)-9H-purin-6-amine
##STR00332##
[0890] A solution of
5'-{[3-(2-carboxyethyl)cyclobutyl](isopropyl)amino}-5'-deoxy-2',3'-O-isop-
ropylideneadenosine (0.463 g, 0.976 mmol) and
4-(1-methylcyclobutyl)benzene-1,2-diamine (0.184 g, 1.04 mmol) in
N,N-Dimethylformamide (10 ml,) was cooled at 0.degree. C. The
[0891] solution was treated with N,N-Diisopropylethylamine (0.462
ml, 2.65 mmol) dropwise followed by
N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (0.367 g, 0.965 mmol) in one portion. The
mixture was stirred at 0.degree. C. for 1 h then slowly warmed to
RT. After 5 h at RT the reaction mixture was stored in the fridge
overnight, the reaction mixture was placed under high vacuum. The
resultant glass was taken up in 30 ml H.sub.2O and extracted with
30 ml portion of 10% MeOH/EtOAc. The aqueous phase was further
extracted with 30 ml portion of EtOAc. The combined organic phases
were washed with 25 ml portions of sat NaHCO.sub.3 and brine and
dried over Na.sub.2SO.sub.4. The mixture was filtered and
concentrated to yield a glass/foam (700 mg). The crude material was
purified by flash chromatography (SiO.sub.2, 4-5% 7N NH.sub.3 in
MeOH/CH.sub.2Cl.sub.2) to give the intermediate amide (.about.80%
purity, 390 mg).
[0892] The intermediate amide (110 mg, 0.174 mmol) was taken up in
4 ml acetic acid and the solution was heated at 65.degree. C. The
reaction mixture was cooled and the volatiles were removed under
high vacuum to yield a glass. The crude product was taken up in 25
ml CH.sub.2Cl.sub.2 and washed with 20 ml sat NaHCO.sub.3 and 2%
Na.sub.2CO.sub.3 solution, dried over Na.sub.2SO.sub.4, filtered
and concentrated to yield a glass/stiff foam. The crude material
was purified by prep TLC (SiO.sub.2, eluting with 7% 7N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2) to give the desired product as a stiff
foam (54 mg). This above procedure was repeated (except
purification was by flash chromatography, SiO.sub.2 eluting with
4-5% 7N NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2) on a further batch
of the intermediate amide (389 mg) to yield a further 368 mg of the
desired compound which was combined with the above
benzimidazole.
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((isopropyl(3-(2-(5-(1-methylcyc-
lobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydro-
furan-3,4-diol
##STR00333##
[0894]
9-((3aR,4R,6R,6aR)-6-((isopropyl(3-(2-(5-(1-methylcyclobutyl)-1H-be-
nzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine 422 mg, 0.686 mmol)
was dissolved in a mixture of Trifluoroacetic Acid (6.3 ml, 82
mmol) and water (0.7 ml, 40 mmol) which had been pre-cooled at
0.degree. C. in an ice bath. The solution was stirred at 0.degree.
C. for 30 minutes, upon which the ice bath was removed and the
mixture was warmed to RT. The mixture was stirred at RT for 2.5 h
upon which the residue was taken up in 12 ml MeOH, concentrated to
dryness. This was repeated twice, the resultant glass was placed
under high vacuum. The crude residue was diluted with 11 ml MeOH,
treated with 600 mg K.sub.2CO.sub.3 and 0.5 ml H.sub.2O and allowed
to stir at RT till the solution was basic as determined by pH
paper. The mixture was filtered and the solids were washed with 20
ml MeOH. The solution was concentrated to a residue that was placed
under high vacuum. The crude material was purified by flash
chromatography (SiO.sub.2, eluting with 10-12% 7N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2) to give the desired compound as a
stiff foam/glass (256 mg). .sup.1H NMR (400 MHz, MeOD)
.delta..sub.H ppm 8.30 (m, 1H), 8.20 (d, J=1.04 Hz, 1H), 7.38 (d,
J=8.09 Hz, 1H), 7.22 (br. s., 1H), 6.98 (dd, J=8.50, 1.66 Hz, 1H),
5.96 (m, 1H), 4.74 (t, J=4.87 Hz, 1H), 4.27 (d, J=3.11 Hz, 1H),
4.08 (m, 1H), 3.56 (m, 1H), 3.13 (m, 1H), 3.00 (m, 1H), 2.90 (dd,
J=14.51, 4.35 Hz, 1H), 2.75 (m, 3H), 2.41 (m, 2H), 2.12 (m, 5H),
2.00 (m, 1H), 1.84 (m, 3H), 1.58 (m, 1H), 1.46 (s, 3H), 1.02 (m,
3H), 0.95 (d, J=6.63 Hz, 3H).
Compound 28
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((isopropyl((1r,3S)-3-(2-(5-(1--
methylcyclobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)t-
etrahydrofuran-3,4-diol
[0895] The diastereomers were separated by SFC. The material was
taken up in MeOH/H.sub.2O and lyophilized to a white powder (132
mg). .sup.1H NMR (400 MHz, MeOD) d ppm 8.30 (s, 1H), 8.20 (s, 1H),
7.38 (d, J=8.09 Hz, 1H), 7.22 (s, 1H), 6.99 (dd, J=8.40, 1.55 Hz,
1H), 5.96 (d, J=4.56 Hz, 1H), 4.73 (m, 1H), 4.26 (t, J=5.29 Hz,
1H), 4.07 (m, 1H), 3.13 (m, 1H), 3.00 (m, 1H), 2.90 (dd, J=14.41,
4.46 Hz, 1H), 2.76 (t, J=7.15 Hz, 2H), 2.70 (m, 1H), 2.42 (m, 2H),
2.18 (m, 2H), 2.11 (m, 3H), 1.85 (m, 4H), 1.57 (q, J=8.85 Hz, 2H),
1.47 (s, 3H), 1.02 (d, J=6.84 Hz, 3H), 0.95 (d, J=6.63 Hz, 3H).
Compound 29:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl(3-(2-
-(5-(1-methylcyclobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)m-
ethyl)cyclopentane-1,2-diol
(1-methylcyclobutyl)benzene
##STR00334##
[0897] A stirred mixture of benzene (5.0 ml, 56 mmol) and Sulfuric
acid (1.17 ml, 21.9 mmol) was cooled to 0.degree. C. and treated
dropwise with a solution of Methylenecyclobutane (1.00 ml, 10.8
mmol) in Benzene (3.0 ml, 34 mmol) over -1 h. Upon completion of
the addition, the reaction mixture was stirred an additional 1 h
while being warmed to RT. The mixture extracted with 15 ml of
hexane. The organic phase was washed with 10 ml H.sub.2O and 10 ml
sat NaHCO.sub.3, dried over Na.sub.2SO.sub.4, filtered and
concentrated to yield a colorless liquid. The liquid was purified
by kugelrohr distillation (5-10 torr) to yield the desired compound
as a colourless liquid. First fraction was collected at
75-85.degree. C. as a colorless liquid (330 mg) of product as a
colorless liquid. 1-(1-methylcyclobutyl)-4-nitrobenzene
##STR00335##
[0898] 70% Nitric acid (7:3, Nitric acid:Water, 0.375 ml, 5.92
mmol) was added dropwise over 60 minutes to a solution of
(1-methylcyclobutyl)benzene (346 mg. 2.37 mmol) in Acetic anhydride
(1.4 mL, 15 mmol) cooled at 0.degree. C. The temperature of the
solution was maintained below 5.degree. C. during the addition.
Upon completion of the addition, the reaction was stirred for 60
minutes with cooling. The reaction mixture was poured into 40 ml
ice water and the ice was allowed to melt. The aqueous phase was
extracted with three .times.20 ml portions of Et.sub.2O and the
combined organic phase was washed with 25 ml H.sub.2O followed by
two 20 ml portions of sat NaHCO.sub.3 solution. The organic phase
was dried over Na.sub.2SO.sub.4, filtered and concentrated to yield
a light the product as an oil (418 mg) which was used as is in the
next step. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.16 (d,
J=8.71 Hz, 2H), 7.29 (d, J=8.71 Hz, 2H), 2.41 {m, 2H), 2.15 (m,
3H), 1.88 (m, 1H). 1.48 (s, 3H)
4-(1-methylcyclobutyl)aniline
##STR00336##
[0900] A solution of 1-(1-methylcyclobutyl)-4-nitrobenzene (708 mg,
3.70 mmol) in Ethanol (24 mL, 410 mmol) was treated carefully with
5% Pd on Carbon (87 mg. 0.041 mmol). The reaction flask was
evaculated and filled with hydrogen gas three times and the
reaction mixture was stirred under an atmosphere of Hydrogen for 19
h. The reaction mixture was filtered
[0901] through a pad of Solka Floc.RTM. and the pad was washed with
25 mL EtOH. The solvent was removed to yield an oil that was placed
under high vacuum briefly to yield the desired compound (609 mg)
which was used directly in the next step. .sup.1H NMR (400 MHZ,
CDCI3) o ppm 6.99 (m, 2H), 6.66 (m, 2H), 3.39 (br. s., 2H), 2.35
(m, 2H), 2.05 (m, 3H), 1.82 (m, 1H), 1.42 (s, 3H).
2,2,2-trifluoro-N-(4-(1-methylcyclobutyl)-2-nitrophenyl)acetamide
##STR00337##
[0903] 4-(1-Methylcyclobutyl)aniline (500 mg, 2.79 mmol) and
Ammonium Nitrate (220 mg, 2.8 mmol) were treated with
Trifluoroacetic anhydride (1.97 mL, 14.0 mmol) followed by
Chloroform (10 mL, 120 mmol). The reaction mixture was allowed to
stir at RT till for 5 h upon which all the solids had dissolved.
The reaction mixture was poured into 50 ml H.sub.2O and extracted
with three 25 ml portions of CH.sub.2Cl.sub.2. The combined organic
phase was washed with 10 mL sat. NaHCO.sub.3, dried over
Na.sub.2SO.sub.4, filtered and concentrated to an oil. The crude
material was purified by flash chromatography (SiO.sub.2, eluting
with 2.5-3.5% ethyl ether/hex.) to give the desired compound (800
mg).
4-(1-methylcyclobutyl)-2-nitroaniline
##STR00338##
[0905] A solution of
2,2,2-trifluoro-N-[4-(1-methylcyclobutyl)-2-nitrophenyl]acetamide
(580 mg. 1.9 mmol) in Methanol (18 ml, 440 mmol) was treated with a
solution of Potassium carbonate (788 mg, 5.70 mmol) in Water (4.5
ml, 250 mmol) and the mixture was heated at 45.degree. C. for 50
minutes The reaction mixture was cooled to RT and the methanol was
removed in vacuo. The remaining aqueous phase was diluted with 10
ml H.sub.2O and extracted with three 20 ml portions of EtOAc. The
combined organic phase was dried over Na.sub.2SO.sub.4, filtered
and concentrated to yield an oil. The material was placed on high
vacuum where upon it solidified giving the desired compound (400
mg). The material was used directly in the next step .sup.1H NMR
(400 MHz. CDCI3) d ppm 7.90 (d, J=2.07 Hz, 1H), 7.23 (dd, J=8.50,
2.28 Hz, 1H), 6.77 (d, J=8.71 Hz, 1H), 5.96 (br. s., 2H), 2.33 (m,
2H), 2.13 (m, 1H), 2.04 (m, 2H), 1.84 (m, 1H), 1.43 (s, 3H).
4-(1-methylcyclobutyl)benzene-1,2-diamine
##STR00339##
[0907] A solution of 4-(1-methylcyclobutyl)-2-nitroaniline (138 mg.
0.668 mmol) in ethanol (8.5 mL, 140 mmol) was carefully treated
with 10% Palladium on Carbon (14.2 mg, 0.0134 mmol) as a slurry in
ethanol. The reaction flask was evacuated and filled with hydrogen
gas three times and the reaction mixture was stirred under an
atmosphere of Hydrogen for 4 h. The reaction mixture was filtered
through a pad of Solka Floc.RTM. and the pad was washed with 20 ml
MeOH. The filtrate was concentrated to yield an oil which was
placed under high vacuum yielding the desired compound as a solid
(119 mg) which was used directly in the next step. .sup.1H NMR (400
MHZ. CDCl.sub.3) .delta..sub.H ppm 6.66 (m, 1H), 6.53 (m, 2H), 3.34
(br. s., 4H), 2.33 (m, 2H), 2.08 (m, 1H), 1.99 (m. 2H). 1.80 (m,
1H), 1.42 (s, 3H).
N-(2,4-dimethoxybenzyl)-7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(5-
-(1-methylcyclobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)meth-
yl)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimidi-
n-4-amine
##STR00340##
[0909] A solution of
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(methyl)amino)cyclobutyl)propanoic acid (387 mg, 0.652
mmol) and [8]4-(1-methylcyclobutyl)benzene-1,2-diamine (120 mg,
0.68 mmol) in N,N-Dimethylformamide
[0910] (6.7 ml, 87 mmol) was treated with N,N-Diisopropylethylamine
(0.38 ml, 2.2 mmol) dropwise followed by
N,N,N',N'-Tetramethyl-0-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (372 mg, 0.978 mmol) in one portion. The
reaction mixture was stirred at RT for 2.5 hr, the reaction mixture
was then concentrated under high vacuum. The residue was
partitioned between 40 ml EtOAc (some CH.sub.2Cl.sub.2 was added to
aid in solublizing the product) and 40 ml 1/1 H.sub.2O/sat
NaHCO.sub.3. The aqueous phase was extracted with 30 ml 1/1
EA/CH.sub.2Cl.sub.2 and the combined organic phase was dried over
Na.sub.2SO.sub.4, filtered and concentrated. The crude material was
purified by flash chromatography (SiO.sub.2, eluting with 5-6% 7N
NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2) to give the desired
intermediate as a mixture of amide regioisomers.
[0911] The intermediate was taken up in Acetic acid (5.4 ml, 95
mmol) and the solution was heated at 65.degree. C. for 3 hours. The
acetic acid was removed under high vacuum with the aid of a warm
water bath. The crude product was taken up in 30 ml
CH.sub.2Cl.sub.2 and the organic phase was washed with 10 ml
portions of sat NaHCO.sub.3 and 2% K.sub.2CO.sub.3 solutions, dried
over Na.sub.2SO.sub.4, filtered and concentrated to a glass that
produced a foam under high vacuum. The crude material was purified
by flash chromatography (SiO.sub.2 eluting with 5% 7N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2 to yield the desired product (140
mg).
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl(3-(2--
(5-(1-methylcyclobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)me-
thyl)cyclopentane-1,2-diol
##STR00341##
[0913]
N-(2,4-dimethoxybenzyl)-7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-((methyl(-
3-(2-(5-(1-methylcyclobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)ami-
no)methyl)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]p-
yrimidin-4-amine (128 mg, 0.174 mmol) was dissolved in a mixture of
Trifluoroacetic Acid (3.60 ml, 46.7 mmol) and Water (0.4
[0914] ml, 20 mmol) which had been pre-cooled at 0.degree. C. in an
ice bath. The solution was stirred at 0.degree. C. for 30 minutes,
then the ice bath was removed and the mixture was warmed to RT at
which this temperature was maintained for a further 2.5 hours. The
reaction mixture was concentrated in vacuo. The residue was taken
up in 3 ml MeOH and concentrated and the process was repeated
twice. The resultant white residue was placed on high vacuum. The
crude residue was combined with another batch of crude material
(prepared identically, .about.1/3 of the amount used in this
reaction), diluted with 5 mL MeOH, treated with 140 mg
K.sub.2CO.sub.3, 10 drops of H.sub.2O and allowed to stir at RT
till the solution was basic by pH paper. The mixture was filtered
and the solids were washed with 15 ml MeOH. The solution was
concentrated to an oil that was placed on high vacuum. The crude
material was purified by flash chromatography (SiO.sub.2, eluting
with 10-15% 7N NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2 to yield the
desired product as glass/stiff foam (68 mg). .sup.1H NMR (400 MHz,
MeOD) .delta..sub.H ppm 8.06 (s, 1H), 7.39 (d, J=7.88 Hz, 1H), 7.23
(s, 1H), 7.21 (dd, J=3.63, 1.76 Hz, 1H), 6.99 (m, 1H), 6.60 (d,
J=3.52 Hz, 1H), 4.93 (m, 1H), 4.32 (m, 1H), 3.89 (m, 1H), 3.03 (m,
1H), 2.83 (m, 2H), 2.70 (q, J=8.15 Hz, 1H), 2.52 (m, 1H), 2.40 (m,
4H), 2.27 (m, 2H), 2.18 (d, J=6.22 Hz, 3H), 2.11 (m, 4H), 2.03 (m,
1H), 1.86 (m, 4H), 1.62 (m, 1H), 1.51 (m, 1H), 1.47 (s, 3H).
Compound 30:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((methyl((1r,3S)-3-(2-(5-(1-met-
hylcyclobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetr-
ahydrofuran-3,4-diol
##STR00342##
[0916] The diastereomers were separated by SFC. The material was
taken up in MeOH/H.sub.2O and lyophilized to a white powder (67
mg). .sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.27 (s, 1H),
8.20 (s, 1H), 7.38 (d, J=8.29 Hz, 1H), 7.22 (br. s., 1H), 6.99 (dd,
J=8.40, 1.55 Hz, 1H), 5.97 (d, J=4.15 Hz, 1H), 4.69 (dd, J=5.18,
4.15 Hz, 1H), 4.22 (t, J=5.60 Hz, 1H), 4.16 (m, 1H), 2.77 (m, 2H),
2.72 (d, J=8.09 Hz, 1H), 2.67 (m, 2H), 2.42 (m, 2H), 2.21 (m, 2H),
2.15 (s, 3H), 2.10 (m, 3H), 1.85 (m, 4H), 1.47 (s, 3H), 1.46 (m,
2H).
Compound 31:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((isopropyl((1s,3R)-3-(2(5-(1
methylcyclobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)-
tetrahydrofuran-3,4-diol
##STR00343##
[0918] The diastereomers were separated by SFC. The material was
found to be the trans diastereomer by NMR. The material was taken
up in MeOH/H.sub.2O and lyophilized to a white powder. (63 mg).
.sup.1H NMR (400 MHz MeOD) .delta..sub.H ppm 8.31 (s, 1H), 8.2 (s,
1H), 7.38 (d, J=8.29 Hz, 1H) 7.22 (s, 1H), 6.99 (dd, J=8.29, 1.66
Hz, 1H), 5.97 (d, J=4.56 Hz, 1H), 4.74 (m, 1H), 4.27 (J=5.39 Hz,
1H), 4.09 (m, 1H), 353 (m, 1H), 3.1 (m, 1H), 2.93 (dd, J=14.72,
4.35 Hz, 1H), 2.80 (t=7.46 Hz, 2H), 2.73 (dd, J=1 4.51, 7.46 Hz,
1H), 2.42 (m, 2H), 213 (m, 5H), 2.01 (m, 3H), 1.82 (m, 3H), 1.47 s,
3H), 1.02 (d, J=6.63 Hz, 3H), 0.95 (d, J=6.63 Hz, 3H).
Compound 32:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino-
)methyl)cyclopentane-1,2-diol
##STR00344##
[0920] The disatereosisomers were separated by SFC. (conditions
listed below) yielded 120 mg. Preparative Method: IC (2.times.15
cm), 35% isopropanol (0.2% DEA))/CO.sub.2, 100 bar, 60 mL/min, 220
nm., inj vol.: 0.75 mL, 4 mg/mL methanol Peak 1: 5.27 minutes.
.sup.1H NMR (400 MHz, d.sub.4-MeOH) .delta..sub.H 8.080 (s, 1H),
7.495 (s, 1H), 7.417-7.395 (m, 1H), 7.303-7.281 (m, 1H),
7.220-7.212 (m, 1H), 6.619-6.610 (m, 1H), 4.349-4.315 (m, 1H),
2.837-2.802 (m, 2H), 2.718-2.641 (m, 1H), 2.508-2.365 (m, 3H),
2.284-2.258 (m, 3H), 2.156 (s, 3H), 1.954-1.906 (m, 1H),
1.549-1.460 (m, 2H), 1.375 (s, 9H).
Compound 33:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((methyl(3-(2-(5-(1-methylcyclo-
butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydrofu-
ran-3,4-diol
##STR00345##
[0921]
9-((3aR,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(5-(1-methylcyclobu-
tyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine
##STR00346##
[0923] A solution of
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoic
acid (0.461 g, 1.03 mmol) and
4-(1-methylcyclobutyl)benzene-1,2-diamine (0.150 g, 0.851 mmol) in
N,N-Dimethylformamide (11 ml, 140 mmol) was cooled at 0.degree. C.
The solution was treated with N,N-Diisopropylethylamine (0.489 ml,
2.81 mmol) dropwise followed by
N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (0.388 g, 1.02 mmol) in one portion. The
mixture was stirred at 0.degree. C. for 30 minutes then slowly
warmed to RT, stirring was continued at RT for 6 h. The reaction
mixture was diluted with 30 ml, H.sub.2O and extracted with 25 ml
portions of 10% MeOH/EtOAc. The aqueous phase was further extracted
with two 20 ml portions of EtOAc. The combined organic phases were
washed with 25 ml portions of sat. NaHCO.sub.3, brine and dried
over Na.sub.2SO.sub.4. The solution was filtered and concentrated
to yield a glass. The crude material was purified by flash
chromatography (SiO.sub.2, eluting with 5% 7N NH.sub.3 in
MeOH/CH.sub.2Cl.sub.2.
[0924] The intermediate amide was taken up in Acetic acid (7.0 ml,
120 mmol) and the solution was heated at 65.degree. C. for 2.5 h,
the reaction mixture was cooled and the acetic acid was removed
under high vacuum to yield a glass. The crude product was taken up
in 25 ml CH.sub.2Cl.sub.2 and washed with 20 ml sat NaHCO.sub.3, 2%
Na.sub.2CO.sub.3 solution, dried over Na.sub.2SO.sub.4, filtered
and concentrated in vacuo to yield a glass/stiff foam. The crude
material was purified by flash chromatography (SiO.sub.2, eluting
with 5-7% 7N NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2) to give the
desired compound (214 mg).
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((methyl(3-(2-(5-(1-m-
ethylcyclobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)te-
trahydrofuran-3,4-diol
##STR00347##
[0925]
9-((3aR,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(5-(1-methylcyclobu-
tyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine (188 mg, 0.320 mmol) was
dissolved in a mixture of Trifluoroacetic Acid (4.00 ml, 51.9 mmol)
and Water (0.4 ml, 20 mmol) which had been pre-cooled at 0.degree.
C. in an ice bath. The solution was stirred at 0.degree. C. The
reaction was stirred for 30 minutes at 0.degree. C., then the ice
bath was removed and the mixture was warmed to RT where stirring
was continued for a further 2 h. The reaction mixture was
concentrated in vacuo. The residue was taken up in 10 ml MeOH and
concentrated and the process was repeated twice. The resultant
glass was placed under high vacuum for 1 h. The crude residue was
diluted with 7 ml MeOH, treated with 150 mg K.sub.2CO.sub.3 and 10
drops of H.sub.2O and allowed to stir at RT till the solution was
basic by pH paper. The mixture was filtered and the solids were
washed with 10 ml MeOH. The solution was concentrated to a residue
that was placed under high vacuum. The crude material was purified
by flash chromatography (SiO.sub.2, eluting with 10-15% 7N NH.sub.3
in CH.sub.3OH/CH.sub.2Cl.sub.2) to give the desired compound as a
glass/stiff foam (66%). .sup.1H NMR (400 MHz, MeOD) .delta..sub.H
ppm 8.28 (m, 1H), 8.20 (m, 1H), 7.38 (d, J=8.29 Hz, 1H), 7.23 (s,
1H), 7.00 (dd, J=8.40, 1.55 Hz, 1H), 5.98 (t, J=3.21 Hz, 1H), 4.70
(m, 1H), 4.24 (q, J=5.18 Hz, 1H), 4.17 (m, 1H), 3.10 (m, 0.4H),
2.80 (m, 3H), 2.71 (d, J=5.60 Hz, 2H), 2.43 (m, 2H), 2.23 (dd,
J=11.71, 6.12 Hz, 1H), 2.19 (m, 3H), 2.12 (m, 4H), 1.99 (m, 1H),
1.85 (m, 4H), 1.48 (s, 3H), 1.48 (m, 1H).
Compound 34:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((methyl((1s,3R)-3-(2-(5-(1-met-
hylcyclobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetr-
ahydrofuran-3,4-diol
##STR00348##
[0927] The diastereomers were separated by SFC. The material was
taken up in MeOH/H.sub.2O and lyophilized to a white powder (30 mg)
.sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.28 (s, 1H), 8.19
(s, J=4.15 Hz, 1H), 4.69 (m, 1H), 4.23 (t, J=5.49 Hz, 1H), 4.17 (m,
1H), 3.07 (m, 1H), 2.81 (t, J=7.57 Hz, 2H), 2.68 (m, 2H), 2.42 (m,
2H), 2.17 (s, 3H), 2.09 (m, 6H), 1.97 (m, 2H), 1.84 (m, 3H), 1.47
(s, 3H).
35:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-
-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)am-
ino)methyl)cyclopentane-1,2-diol
##STR00349##
[0929] The diastereoisomers were separated by SFC The following SFC
separation (120 mg).
[0930] Preparative Method: IC (2.times.15 cm), 35% isopropanol
(0.2% DEA))/CO.sub.2, 100 bar, 60 mL/min, 220 nm., inj vol.: 0.75
mL, 4 mg/mL methanol. Peak 2: 6.24 minutes. .sup.1H NMR (400 MHz,
d.sub.4-MeOH) .delta..sub.H 8.078 (s, 1H), 7.501 (s, 1H),
7.422-7.401 (m, 1H), 7.310-7.285 (m, 1H), 7.228-7.219 (m, 1H),
6.618-6.609 (m, 1H), 4.355-4.320 (m, 1H), 3.053-2.977 (m, 1H),
2.874-2.836 (m, 2H), 2.535-2.268 (m, 4H), 2.177-2.003 (m, 8H),
1.909-1.869 (m, 2H), 1.677-1.595 (m, 1H), 1.381 (s, 9H).
Compound 36:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl((1r,-
3S)-3-(2-(5-(1-methylcyclobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl-
)amino)methyl)cyclopentane-1,2-diol
##STR00350##
[0932] The diastereoisomers were separated by SFC. The material was
taken up in MeOH/H.sub.2O and lyophilized to a white solid (15 mg).
.sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.06 (s, 1H), 7.38
(d, J=8.09 Hz, 1H), 7.23 (br s., 1H), 7.20 (d, J=3.32 Hz, 11H), 699
(n, 1H), 6.60 (d, J=3.52 Hz, 1H), 4.95 (m, 1H), 4.31 (t, J=6.74 Hz,
1H), 3.88 (m, 1H), 2.81 (m, 2H), 266 (m, 1H), 2.40 (m, 5H), 2.25
(m, 3H), 2.14 (br s., 3H), 2.11 (m, 3H), 1.91 (m, 2H), 1.83 (m,
1H), 1.61 (m, 1H), 1.51 (m, 1H), 1.47 (s, 3H).
Compound 37:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl((1r,-
3S)-3-(2-(5-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)a-
mino)methyl)cyclopentane-1,2-diol
##STR00351##
[0934] The diastereoisomers were separated by SFC. Lyophilization
gave the desired product as a colorless solid (0.060 g). .sup.1H
NMR (400 MHz, d.sub.4-MeOH) .delta..sub.H 8.079 (s, 1H),
7.546-7.524 (m, 1H), 7.409 (s, 1H), 7.226-7.217 (m, 1H),
7.150-7.124 (m, 1H), 6.619-6.610 (m, 1H), 4.355-4.320 (m, 1H),
3.912-3.885 (m, 1H), 2.881-2.845 (m, 2H), 2.727-2.674 (m, 1H),
2.538-2.262 (m, 6H), 2.172 (s, 3H), 1.955-1.916 (m, 3H),
1.670-1.492 (m, 3H).
Compound 38:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(ethyl)amino)-
methyl)cyclopentane-1,2-diol
##STR00352##
[0936] The diastereosiomers were separated by SFC. After
lyophilization the desired product was recovered as a colorless
solid (32 mg).
[0937] Preparative Method: Lux-3 (2.times.15 cm), 30% ethanol (0.2%
DEA))/CO.sub.2, 100 bar, 65 mL/min, 220 nm., inj vol.: 0.4 mL, 6.2
mg/mL methanol. .sup.1H NMR (400 MHz, d4-MeOH) .delta..sub.H 8.082
(s, 1H), 7.493 (s, 1H), 7.415-7.393 (m, 1H), 7.295-7.270 (m, 1H),
7.215-7.206 (m, 1H), 6.621-6.612 (m, 1H), 4.343-4.309 (m, 1H),
3.924-3.897 (m, 1H), 3.044-2.962 (m, 1H), 2.834-2.798 (m, 2H),
2.728-2.695 (m, 1H), 2.660-2.607 (m, 2H), 2.543-2.380 (m, 2H),
2.281-2.257 (m, 3H), 1.932-1.906 (m, 3H), 1.660-1.523 (m, 3H),
1.368 (s, 9H), 1.050-1.014 (t, J=7.2 Hz, 3H).
Compound 39:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl((1s,-
3R)-3-(2-(5-(1-methylcyclobutyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl-
)amino)methyl)cyclopentane-1,2-diol
##STR00353##
[0939] The diastereoisomers were separated by SFC. The material was
taken up in MeOH/H2O and lyophilized to a white powder (19 mg).
.sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.06 (s, 1H), 7.39
(d. J=8.50 Hz, 1H), 7.21 (m, 2H), 6.99 (dd, J=8.29, 1.45 Hz, 1H),
6.59 (d, J=3.52 Hz, 1H), 4.94 (m, 1H), 4.32 (dd, J=7.77, 5.91 Hz,
1H), 3.89 (m, 1H), 3.00 (m, 1H), 2.83 (t, J=7.57 Hz, 2H), 2.49 (m,
1H), 2.38 (m, 4H), 2.23 (m, 1H), 2.16 (s. 3H), 2.11 (m, 5H), 2.01
(m, 2H), 1.84 (m. 3H), 1.61 (m, 1H), 1.46 (s, 3H).
Compound 40:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclopropylm-
ethyl)amino)methyl)cyclopentane-1,2-diol
##STR00354##
[0941] The diastereoisomers were separated by SFC. The material was
taken up in MeOH/H.sub.2O and lyophilized to yield a white powder
(45 mg). .sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.06 (s,
1H), 7.48 (br. s., 1H), 7.39 (d, J=8.29 Hz, 1H), 7.27 (m, 1H), 7.20
(d, J=3.73 Hz, 1H), 6.60 (d, J=3.52 Hz, 1H), 4.32 (dd, J=7.36, 6.12
Hz, 1H), 3.90 (m, 1H), 3.06 (m, 1H), 2.81 (t, J=6.84 Hz, 2H), 2.74
(m, 1H), 2.55 (dd, J=12.75, 7.77 Hz, 1H), 2.41 (m, 1H), 2.37 (d,
J=6.84 Hz, 2H), 2.29 (m, 3H), 1.91 (m, 3H), 1.60 (m, 1H), 1.50 (m,
2H), 1.36 (s, 9H), 0.87 (m, 1H), 0.48 (d, J=8.09 Hz, 2H), 0.10 (m,
2H).
Compound 41:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)am-
ino)methyl)cyclopentane-1,2-diol
[0942] The diastereoisomers were separated by SFC followed by
lyophilization from H.sub.2O/MeOH/CH.sub.3CN to yield a white
powder (100 mg). .sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.06
(s, 1H), 7.48 (br. s., 1H), 7.39 (m, 1H), 7.27 (m, 1H), 7.20 (d,
J=3.52 Hz, 1H), 6.60 (m, 1H), 4.32 (t, J=6.43 Hz, 1H), 3.93 (t,
J=5.29 Hz, 1H), 3.54 (m, 0.2H), 3.11 (t, J=9.33 Hz, 1H), 3.02 (m,
1H), 2.82 (m, 2H), 2.66 (dd, J=13.68, 8.09 Hz, 1H), 2.46 (m, 1H),
2.36 (m, 1H), 2.23 (m, 3H), 2.05 (m, 1H), 1.91 (m, 3H), 1.59 (m,
3H), 1.36 (s, 9H), 1.02 (m, 6H).
Compound 42:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclobutylme-
thyl)amino)methyl)cyclopentane-1,2-diol
Step 1: ethyl
3-((1S,3r)-3-((cyclobutylmethyl)(((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxyben-
zyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyc-
lopenta[d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoate
##STR00355##
[0944] The amine ethyl
3-[3-({[(3aR,4R,6R,6aS)-6-{4-[(2,4-dimethoxybenzyl)amino]-7H-pyrrolo[2,3--
d]pyrimidin-7-yl}-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
]methyl}amino)cyclobutyl]propanoate (1.8 g, 3.0 mmol) was taken up
in methanol (20 mL, 600 mmol) and sodium cyanoborohydride (0.37 g,
5.9 mmol) was added. The pH was adjusted to ca. 6 using a 10%
solution of AcOH in methanol, then cyclobutanecarboxaldehyde (0.32
g, 3.8 mmol) was added in one portion. The reaction was allowed to
proceed for 5 hours at which time HPLC indicated the reaction had
stalled. Another 1.3 equivalents of cyclobutanecarboxaldehyde was
added and the reaction continued overnight. NaHCO.sub.3 (saturated)
was added to the reaction mixture which was then extracted 3 times
with DCM. The combined organics were dried with MgSO.sub.4 and
concentrated to a yellow resin. Cis and trans isomers were
separable on silica. Purification by FC (DCM/7N NH.sub.3 in MeOH
96:4) yielded 2 separate batches of product, each enriched in one
respective isomer to about 90%. Top isomer: 0.38 g (11:1 mixture,
cis) Bottom isomer: 0.31 g (6:1 mixture, trans). MS (ESI.sup.+) for
C.sub.35H.sub.49N.sub.5O.sub.6 m/z 676.7 [M+H].sup.+; HPLC purity
>69% (ret. time, 3.791).
Step 2:
N-(2-amino-5-(tert-butyl)phenyl)-3-((1S,3r)-3-((cyclobutylmethyl)(-
((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimi-
din-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)-
amino)cyclobutyl)propanamide
##STR00356##
[0946] Top Isomer (cis): Lithium hydroxide monohydrate (0.236 g,
5.62 mmol) was added to a solution of ethyl
3-((1S,3r)-3-((cyclobutylmethyl)(((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxyben-
zyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyc-
lopenta[d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoate (6
mL, 70 mmol) and methanol (1.5 mL, 37 mmol). The reaction was
stirred overnight at room temperature and by the next morning the
starting material was consumed and had been transformed into the
acid. The reaction was acidified with 1N HCl to pH=6. The volatiles
were removed in vacuo and the remaining water was removed by
azeotropic distillation with ethanol followed by 72 hours on the
lyophilizer. The resulting off white solid was used without further
purification. Retention time: 3.330 minutes MS (ESI.sup.+) for
C.sub.36H.sub.49N.sub.5O.sub.6 m/z 648.4 [M+H].sup.+; MS
(ESI.sup.-) for C.sub.36H.sub.49N.sub.5O.sub.6 m/z 646.4
[M-H].sup.-; HPLC purity >97% (ret. time, 3.329).
[0947] N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
hexafluorophosphate (0.334 g, 0.880 mmol) was added to a solution
of
3-{cis-3-[(cyclobutylmethyl){[(3aR,4R,6R,6aS)-6-{4-[(2,4-dimethoxybenzyl)-
amino]-7H-pyrrolo[2,3-d]pyrimidin-7-yl}-2,2-dimethyltetrahydro-3aH-cyclope-
nta[d][1,3]dioxol-4-yl]methyl}amino]cyclobutyl}propanoic acid (0.38
g, 0.59 mmol) and N,N-diisopropylethylamine (0.337 mL, 1.94 mmol)
and 4-tert-butylbenzene-1,2-diamine (0.116 g, 0.704 mmol) in
N,N-dimethylformamide (3.63 mL, 46.9 mmol). The reaction was
stirred overnight at room temperature and by the next morning the
starting material was consumed. The reaction was partially
concentrated to ca. 2 mLs and then NaHCO.sub.3 (saturated) was
added. The mixture was extracted with EtOAc 3 times and the
combined organics were dried with MgSO.sub.4 and concentrated. The
resulting residue was purified by FC (DCM/7N NH.sub.3 in MeOH 95:5)
to yield
N-(2-amino-5-(tert-butyl)phenyl)-3-((1S,3r)-3-((cyclobutylmethyl)(((3aR,4-
R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-y-
l)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)amino)c-
yclobutyl)propanamide (0.30 g; 64%) as a purple-brown amorphous
solid. HPLC purity >19% (ret. time, 3.574 min.)
Step 3:
7-((3aS,4R,6R,6aR)-6-((((1r,3S)-3-(2-(5-(tert-butyl)-1H-benzo[d]im-
idazol-2-yl)ethyl)cyclobutyl)(cyclobutylmethyl)amino)methyl)-2,2-dimethylt-
etrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-py-
rrolo[2,3-d]pyrimidin-4-amine
##STR00357##
[0949] A solution of
N-(2-amino-4-tert-butylphenyl)-3-{cis-3-[(cyclobutylmethyl){[(3aR,4R,6R,6-
aS)-6-{4-[(2,4-dimethoxybenzyl)amino]-7H-pyrrolo[2,3-d]pyrimidin-7-yl}-2,2-
-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl]methyl}amino]cyclobu-
tyl}propanamide (0.3 g, 0.4 mmol) in acetic acid (1.0 mL, 20 mmol)
was stirred overnight at 65.degree. C. and by next morning the
starting material was consumed. The volatiles were removed in vacuo
and the resulting residue purified by FC (DCM/7N NH.sub.3 in MeOH
93:7) to yield
7-((3aS,4R,6R,6aR)-6-((((1r,3S)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol--
2-yl)ethyl)cyclobutyl)(cyclobutylmethyl)amino)methyl)-2,2-dimethyltetrahyd-
ro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2-
,3-d]pyrimidin-4-amine as a off white solid. MS (ESI.sup.+) for
C.sub.46H.sub.61N.sub.7O.sub.4 m/z 777.7 [M+H].sup.+; HPLC purity
>64% (ret. time, 3.690 min.).
Step 4:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r-
,3S)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclo-
butylmethyl)amino)methyl)cyclopentane-1,2-diol
##STR00358##
[0951] Trifluoroacetic acid (5 mL, 60 mmol) was added to a mixture
of water (0.5 mL, 20 mmol) and
7-((3aS,4R,6R,6aR)-6-((((1r,3S)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol--
2-yl)ethyl)cyclobutyl)(cyclobutylmethyl)amino)methyl)-2,2-dimethyltetrahyd-
ro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2-
,3-d]pyrimidin-4-amine (0.2 g, 0.2 mmol) at room temperature. The
reaction was allowed to proceed overnight at which time the bright
pink suspension was quenched with triethylsilane (0.082 mL, 0.52
mmol). The volatiles were removed in vacuo and the resulting
residue was taken up in methanol (15 mls). 500 mgs of
K.sub.2CO.sub.3 and 8 drops H.sub.2O were added and the reaction
was stirred at room temperature for 1 hour. The mixture was
filtered and the filter cake washed with 10 mLs methanol. The
filtrate was concentrated and the resulting residue was purified by
FC (DCM/7N NH.sub.3 in MeOH 90:10) to yield
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclobutylme-
thyl)amino)methyl)cyclopentane-1,2-diol (0.037 g; 20%) as a
colorless solid. MS (ESI.sup.+) for C.sub.34H.sub.47N.sub.7O.sub.2
m/z 586.3 [M+H].sup.+; HPLC purity >89% (ret. time, 2.970 min.)
.sup.1H NMR (400 MHz, d.sub.4-MeOH) .delta..sub.H 8.083 (s, 1H),
7.498 (s, 1H), 7.417-7.396 (m, 1H), 7.302-7.277 (m, 1H),
7.206-7.197 (m, 1H), 6.621-6.612 (m, 1H), 4.347-4.314 (m, 1H),
3.912-3.885 (m, 1H), 2.973-2.922 (m, 1H), 2.836-2.800 (m, 2H),
2.662-2.366 (m, 6H), 2.282-2.241 (m, 3H), 2.061-2.034 (m, 2H),
1.912-1.494 (m, 10H), 1.374 (s, 9H).
Compound 43:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5-(t-
ert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclobutyl)amino)met-
hyl)cyclopentane-1,2-diol
##STR00359##
[0952] ethyl
3-(3-(cyclobutyl(((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-py-
rrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]d-
ioxol-4-yl)methyl)amino)cyclobutyl)propanoate
##STR00360##
[0954] Ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)amino)cyclobutyl)propanoate (0.84 g, 1.4 mmol) was taken up
in methanol (10 ml) and Sodium cyanoborohydride (0.087 g, 1.4 mmol)
was added. The pH was adjusted to ca 6 using a 10% solution of AcOH
in MeOH, then Cyclobutanone (0.15 ml, 21 mmol) added in one
portion. The reaction was stirred at RT for 3 days. NaHCO.sub.3
(sat'd) added to the reaction mixture which was then extracted
(3.times.) with DCM. The combined organics were dried with
MgSO.sub.4, filtered and concentrated. The material was used
without further purification.
3-(3-(cyclobutyl(((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyr-
rolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]di-
oxol-4-yl)methyl)amino)cyclobutyl)propanoic acid
##STR00361##
[0956] Lithium hydroxide monohydrate (0.58 g, 14 mmol) added to a
solution of ethyl
3-(3-(cyclobutyl(((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)ami-
no)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta-
[d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoate (0.91 g, 1.4
mmol) in Tetrahydrofuran (12 ml, 150 mmol) and Methanol (3 ml, 60
mmol). The reaction mixture was stirred overnight at RT, upon which
it was acidified with 1 N HCl to pH=6 The volatiles were removed in
vacuo and the remaining water removed by azeotropic distillation
with ethanol followed by 18 hours on lyophilizer. The resulting off
white solid was used without further purification.
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-(cyclobutyl(((3aR,4R,6R,6aS)-6-(4-((-
2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethylte-
trahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propana-
mide
##STR00362##
[0958] N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (0.783 g, 2.06 mmol) was added to a solution of
3-(3-(cyclobutyl(((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-py-
rrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]d-
ioxol-4-yl)methyl)amino)cyclobutyl)propanoic acid (0.87 g, 1.4
mmol) and N,N-Diisopropylethylamine (0.789 ml, 4.53 mmol) and
[8)4-tert-butylbenzene-1,2-diamine (0.270 g, 1.65 mmol) in
N,N-Dimethylformamide (8.50 m) The reaction was stirred overnight
at RT, upon which the mixture was partially concentrated to ca 2 ms
and then NaHCO.sub.3 (saturated) was added. The mixture was
extracted with EtOAc (3.times.) and the combined organics were
dried with MgSO.sub.4, filtered and concentrated. The residue was
purified by flash chromatography (DCM/7NNH.sub.3 in MeOH 95:5) to
give the desired compound (0.76 g) as a solid.
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethy-
l)cyclobutyl)(cyclobutyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclopen-
ta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin--
4-amine
##STR00363##
[0960] A solution of
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-(cyclobutyl(((3aR,4R,6R,6aS)-6-(4-(-
(2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethylt-
etrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propan-
amide (0.76 g, 097 mmol) in Acetic acid (2 ml) was stirred
overnight at 60.degree. C. The volatiles were removed in vacuo and
the remaining residue purified directly by flash chromatography
(DCM/7N NH.sub.3 in MeOH 91:9) to give the desired compound (0.61
g) as a foam.
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5-(te-
rt-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclobutyl)amino)meth-
yl)cyclopentane-1,2-diol
##STR00364##
[0962] Trifluoroacetic Acid (10 ml, 200 mmol) added to a mixture of
Water (1 ml, 80 mmol) and
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)eth-
yl)cyclobutyl)(cyclobutyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclope-
nta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-
-4-amine (0.61 g, 0.80 mmol) at RT. The reaction was stirred o/n at
RT and was quenched by the addition of Triethylsilane (0.26 ml, 1.6
mmol). The volatiles were removed in vacuo and resulting and the
residue was taken up in MeOH (15 mls), 500 mg K.sub.2CO.sub.3 and 8
drops of water were added and the reaction was stirred at RT for 1
hour. The mixture was filtered and the filter cake was washed with
MeOH (10 ml). The filtrate was concentrated and the resulting
residue purified by flash chromatogrpahy (DCM/7N N3 in MeOH 90:10)
to give the desired product (0.13 g) as a colorless foam. MS
(ESI.sup.+) for C.sub.33H.sub.45N.sub.7O.sub.2 m/z 572.2
[M+H].sup.+; MS (ESI.sup.-) for C.sub.33H.sub.45N.sub.7O.sub.2 m/z
570.2 [M-H].sup.-; HPLC purity >90% (ret. time, 2.850 min.)
1H1NMR (400 MHz, di-MeOH) .delta..sub.H 8.083 (s, 1H), 7.492 (s,
1H), 7.412-7.392 (m, 1H), 7.309-7.286 (m, 1H), 7.220-7.205 (m, 1H),
6.620-6.610 (d, J=4.0 Hz, 1H), 4.321-4.283 (m, 1H), 3.888-3.848 (m,
1H), 3.505-3.417 (m, 0.5H (methine of trans isomer)), 3.231-3.147
(m, 0.5H) (methine of cis isomer)), 3.051-2.953 (m, 1H),
2.871-2.732 (m, 3H), 2.583-2.501 (m, 1H), 2.441-2.368 (m, 1H),
2.244-2.205 (m, 3H), 2.170-1.833 (m, 9H), 1.695-1.560 (m, 4H),
1.384 (s, 9H).
Compound 44:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5-(t-
ert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclopropylmethyl)am-
ino)methyl)cyclopentane-1,2-diol
ethyl
3-(3-((cyclopropylmethyl)(((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzy-
l)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclo-
penta[d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoate
##STR00365##
[0964] The amine ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-1-cyclopenta[d][1,3]dioxol-4--
yl)methyl)amino)cyclobutyl)propanoate (0.90 g, 1.5 mmol) was taken
up in Methanol (10 mL) and Sodium cyanoborohydride (0.093 g, 1.5
mmol) was added. The pH was adjusted to ca. 6 using a 10% solution
of AcOH in MeOH. The reaction was stirred o/n at RT. NaHCO.sub.3
(sat'd) was added to r.times.n mixture which was then extracted
(3.times.) with DCM. The combined organics were dried with
MgSO.sub.4, filtered and concentrated. The material was used
without further purification.
3-(3-((cyclopropylmethyl)(((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amin-
o)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[-
d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoic acid
##STR00366##
[0966] Lithium hydroxide, monohydrate (0.62 g, 15 mmol) added to a
solution of ethyl
3-(3-((cyclopropylmethyl)(((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)ami-
no)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta-
[d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoate (0.98 g, 1.5
mmol) in Tetrahydrofuran (13 ml) and Methanol (3 ml). The reaction
was stirred for 24 hours at RT, acidified with 1 N HCl to pH==6.
The volatiles removed in vacuo and remaining water removed by
azeotropic distillation with ethanol followed by 18 hours on
lyophilizer. The resulting off white solid was used without further
purification.
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((cyclopropylmethyl)(((3aR,4R,6R,6aS-
)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-d-
imethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)amino)cyclobuty-
l)propanamide
##STR00367##
[0968] N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (0.846 g, 2.22 mmol) added to a solution of
3-(3-((cyclopropylmethyl)(((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)ami-
no)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta-
[d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoic acid (0.94 g,
1.5 mmol) and N,N-Diisopropylethylamine (0.852 mL, 4.89 mmol) and
[8]4-tert-butylbenzene-1,2-diamine (0.292 g, 1.78 mmol) in
N,N-Dimethylformamide (9.19 mL, 119 mmol). The reaction was stirred
overnight at RT, partially concentrated to ca. 2 ms and NaHCO.sub.3
(saturated) was added. The mixture extracted with EtOAc (3.times.)
and the combined organics were dried with MgSO.sub.4, filtered and
concentrated. The residue was purified by flash chromatography (DCM
7N NH.sub.3 in MeOH 95:5) to give the desired compound (0.92 g) as
a solid.
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethy-
l)cyclobutyl)(cyclopropylmethyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-c-
yclopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyr-
imidin-4-amine
##STR00368##
[0970]
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((cyclopropylmethyl)(((3aR,4R-
,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl-
)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)amino)cy-
clobutyl)propanamide (1.1 g, 14 mmol) in Acetic acid (5 ml) was
heated at 60.degree. C. overnight. The solution was concentrated
and purified by flash chromatography (DCM/7N NH.sub.3 ion MeOH
93:7) to yield the desired compound (0.57 g) as a colorless
foam.
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5-(te-
rt-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclopropylmethyl)ami-
no)methyl)cyclopentane-1,2-diol
##STR00369##
[0972] Trifluoroacetic Acid (10 mL) added to a mixture of Water (I
mL) and
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)eth-
yl)cyclobutyl)(cyclopropylmethyl)amino)methyl)-2,2-dimethyltetrahydro-3aH--
cyclopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]py-
rimidin-4-amine (0.52 g, 0.68 mmol) at RT. The reaction was stirred
overnight at RT and Triethylsilane (0.22 mL, 1.4 mmol) was added.
The volatiles were removed in vacuo and resulting residue was taken
up in MeOH (15 mls), 500 mgs of K.sub.2CO.sub.3 and 8 drops
H.sub.2O were added and the reaction stirred at RT for 1 hour. The
mixture was filtered and the filter cake washed with 10 ml MeOH.
The filtrate was concentrated and the resulting residue purified by
flash chromatography (DCM I 7N NH.sub.3 in MeOH 90:10) to give the
desired product (0.196 g) as an off white foam. MS (ESI.sup.+) for
C.sub.33H.sub.45N.sub.7O.sub.2 m/z 572.6 [M+H].sup.+; MS
(ESI.sup.-) for C.sub.33H.sub.45N.sub.7O.sub.2 m/z 570.3
[M-H].sup.-; HPLC purity >90% (ret. time, 2.850 min.) .sup.1H
NMR (400 MHz, d.sub.4-MeOH) .delta..sub.H 7.944 (s, 1H), 7.361 (s,
1H), 7.280-7.259 (m, 1H), 7.172-7.150 (m, 1H), 7.092-7.078 (m, 1H),
6.484-6.475 (d, J=3.6 Hz, 1H), 4.222-4.185 (m, 1H), 3.815-3.779 (m,
1H), 3.329 (m, 0.51-1 (methine of trans isomer)), 2.961 (m, 0.5H
(methine of cis isomer), 2.745-2.627 (m, 3H), 2.503-2.450 (m, 1H),
2.301-2.187 (m, 5H), 2.036-1.890 (m, 21), 1.793-1.776 (m, 31-1),
1.529-1.385 (m, 2H), 1.246 (s, 9H), 0.808-0.739 (m, 1H),
0.394-0.362 (m, 2H), 0.012-0.013 (m, 2H). Retention time: 2.850
minutes. HPLC Conditions:Agilent Zorbax Exlipse XDB-C18 column,
4.6.times.50 mm (1.8 urn packing), Solvent A-Water (0.1% TFA),
Solvent B-Acetonitrile (0.07% TFA) 6 min gradient from 5 to 95% B;
1 min hold; then recycle.
Compound 45: ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(isobutyl)amino)cyclobutyl)propanoate
[0973] The amine ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)amino)cyclobutyl)propanoate (1.7 g, 2.8 mmol) was taken up
in Methanol (2.0 ml) and Sodium cyanoborohydride (0.35 g, 5.6 mmol)
was added. The pH was adjusted to ca 6 using a 10% solution of AcOH
in MeOH, then isobutyraldehyde (0.33 ml, 3.6 mmol) added in one
portion. The reaction stirred at RT for 3 hours. Another 1.3 eq. of
isobutyraldehyde was added and stirring was continued overnight.
NaHCO.sub.3 (sat'd) was added to reaction mixture which was then
extracted (3.times.) with DCM. The combined organics was dried with
MgSO.sub.4 and concentrated. The residue was purified by flash
chromatography (DCM 7N N-13 in MeOH 97:3) to give the desired
compound (1.75 g) as a colorless foam.
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d-
]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-
methyl)(isobutyl)amino)cyclobutyl)propanoic acid
##STR00370##
[0975] Lithium hydroxide, monobydrate (1.11 g, 26.4 mmol) was added
to a solution ethyl
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(isobutyl)amino)cyclobutyl)propanoate (1.75 g, 2.64 mmol)
in Tetrahydrofuran (13 ml, 160 mmol) and Methanol (3 ml, 70 mmol).
The reaction was stirred for 24 hours at RT, acidified with 1 N HC
to pH=6, the volatiles removed in vacuo and remaining water removed
by azeotropic distillation with ethanol followed by 18 hours on
lyophilizer. The resulting off white solid was used without further
purification,
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimeth-
oxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3-
aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(isobutyl)amino)cyclobutyl)propana-
mide
##STR00371##
[0977] N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (1.52 g, 4.01 mmol) added to a solution of
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(isobutyl)amino)cyclobutyl)propanoic acid (1.7 g, 2.7 mmol)
and N,N-Diisopropylethylamine (1.54 ml, 8.82 mmol) and
4-tert-butylbenzene-1,2-diamine (0.527 g, 3.21 mmol) in
N,N-Dimethylformamide (16.6 ml). The reaction was stirred overnight
at RT, partially concentrated to ca. 2 mls and then NaHCO.sub.3
(saturated) added. Them mixture was extracted with EtOAc 3.times.
and the combined organics were dried with MgSO.sub.4 filtered,
concentrated and purified by flash chromatography (DCM/7N NH.sub.3
in MeOH 95:5) to yield the desired amide (1.71 g) as a solid.
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethy-
l)cyclobutyl)(isobutyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclopenta-
[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4--
amine
##STR00372##
[0979] A solution of
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimet-
hoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro--
3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(isobutyl)amino)cyclobutyl)propan-
amide (1.71 g 219 mmol) in Acetic acid (6 ml) was stirred overnight
at 60.degree. C., the volatiles were removed in vacuo and the
remaining residue was purified by flash chromatography (SiO.sub.2,
DCM I 7N NH.sub.3 in MeOH 94:6) to yield the desired compound (0.9
g) as a foam.
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-(((3-(2-(5-(te-
rt-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isobutyl)amino)methyl-
)cyclopentane-1,2-diol
##STR00373##
[0981] Trifluoroacetic Acid (20 ml, 300 mmol) added to a mixture of
Water (2 ml, 100 mmol) and
7-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)eth-
yl)cyclobutyl)(isobutyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclopent-
a[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-
-amine (0.9 g 1 mmol) at RT The reaction was stirred overnight and
Triethylsilane (0.38 ml, 2.4 mmol) was added. The volatiles were
removed in vacuo and resulting residue was taken up in MeOH (15
ml), 500 mgs of K.sub.2CO.sub.3 and 8 drops H.sub.2O were added and
reaction stirred at RT for 1 hour. The mixture was filtered and
filter cake washed with 10 ml MeOH. The filtrate was concentrated
and the resulting residue purified by flash chromatography (DCM/7N
NH.sub.3 in MeOH 90:10) to yield the desired product (0.274 g) as
an off white foam. MS (ESI.sup.+) for
C.sub.33H.sub.47N.sub.7O.sub.2 m/z 574.6 [M+H].sup.+; MS
(ESI.sup.-) for C.sub.33H.sub.45N.sub.7O.sub.2 m/z 572.4
[M-H].sup.+; HPLC purity >86% (ret. time, 2.918 min.) .sup.1H
NMR (400 MHz, d4-MeOH) .delta..sub.H 8.078 (s, 1H), 7.497 (s, 1H),
7.416-7.396 (m, 1H), 7.305-7.284 (m, 1H), 7.216-7.200 (m, 1H),
6.621-6.612 (d, J=3.6 Hz, 1H), 4.368-4.334 (m, 1H), 3.930-3.894 (m,
1H), 2.934-2.918 (m, 1H), 2.866-2.797 (m, 2H), 2.652-2.583 (m, 1H),
2.444-2.361 (m, 2H), 2.287-2.199 (m, 2H), 2.166-2.119 (m, 3.5H
(contains methine of trans isomer)), 2.048-2.012 (m, 1H),
1.921-1.748 (m, 3.5H (contains methine of cis isomer)), 1.622-1.494
(m, 2H), 1.380 (s, 9H), 1.269-1.252 (m, 1H), 0.932-0.879 (m,
6H).
Compound 46:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclobutyl)a-
mino)methyl)cyclopentane-1,2-diol
[0982] The diastereoisomers were separated by SFC. The material was
taken up in MeOH/H.sub.2O and lyophilized to yield a white powder
(23.7 mg). .sup.1H NMR (400 MHZ, MeOD) .delta..sub.H ppm 8.06 (s,
1H), 7.48 (br. s., 1H), 7.39 (d, J=8.71 Hz, 1H), 7.28 (dd, J=8.60,
1.76 Hz, 1H), 7.19 (d, J=3.52 Hz, 1H), 6.60 (d, J=3.52 Hz, 1H),
4.84 (m, 1H), 4.28 (dd, J=7.26, 6.22 Hz, 1H), 3.84 (t, J=5.70 Hz,
1H), 3.16 (m, 1H), 2.99 (m, 1H), 2.80 (t, J=7.15 Hz, 2H), 2.73 (dd,
J=13.68, 6.22 Hz, 1H), 2.49 (dd, J=13.68, 7.67 Hz, 1H), 2.38 (m,
1H), 2.22 (m, 3H), 2.00 (m, 4H), 1.91 (m, 3H), 1.60 (m, 5H), 1.36
(s, 9H).
Compound 47:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5-bromo-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl-
)cyclopentane-1,2-diol
[0983] Diastereoisomers were separated by SFC. The material was
taken up in MeOH/H.sub.2O and lyophilized to yield a white powder
(21 mg). .sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.06 (s.
1H), 7.63 (br. s., 1H), 7.39 (m, 1H), 7.30 (dd, J=8.50, 1.66 Hz,
1H), 7.20 (d, J=3.52 Hz, 1H), 6.60 (d, J=3.52 Hz, 1H), 4.32 (dd,
J=7.67, 6.01 Hz, 1H). 3.88 (m, 1H), 2.82 (t, J=7.15 Hz, 2H), 2.71
(m, 1 H), 2.52 (m, 1H), 2.41 (m, 2H), 2.25 (m, 2H), 2.18 (s, 3H),
2.03 (m, 1H), 1.92 (m, 3H), 1.62 (m, 1H), 1.51 (m, 2H).
Compound 48:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isobutyl)ami-
no)methyl)cyclopentane-1,2-diol
[0984] Diastereoisomers separated by SFC. After lyophilization (78
mg) obtained of a colorless solid.
Compound 49:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(ethyl)amino)-
methyl)cyclopentane-1,2-diol
[0985] The diastereoisomers were separated by SFC.
[0986] Lux-3 (2.times.15 cm), 30% ethanol (0.2% DEA))/CO.sub.2, 100
bar, 65 mL/min, 220 nm. Inj vol.: 0.4 mL, 6.2 mg/mL methanol.
.sup.1H NMR (400 MHz, d.sub.4-MeOH) .delta..sub.H 8.081 (s, 1H),
7.499 (s, 1H), 7.416-7.395 (m, 1H), 7.308-7.282 (m, 1H),
7.226-7.216 (m, 1H), 6.619-6.610 (m, 1H), 4.344-4.310 (m, 1H),
3.922-3.895 (m, 1H), 3.410-3.329 (m, 1H), 2.875-2.837 (m, 2H),
2.738-2.689 (m, 1H), 2.659-2.607 (m, 2H), 2.535-2.483 (m, 1H),
2.452-2.380 (m, 1H), 2.311-2.224 (m, 1H), 2.158-2.121 (m, 3H),
2.061-2.030 (m, 2H), 1.913-1.863 (m, 2H), 1.674-1.590 (m, 1H),
1.381 (s, 9H), 1.056-1.020 (t, J=7.2 Hz, 3H).
Compound 50:
(1R,2S,3R,5R)-3-(6-amino-9H-purin-9-yl)-5-(((3-(2-(5-(tert-butyl)-1H-benz-
o[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)cyclopentane-1,-
2-diol
ethyl
3-(3-((((1R,2R,3S,4R)-4-(6-((2,4-dimethoxybenzyl)amino)-9H-purin-9-y-
l)-2,3-dihydroxycyclopentyl)methyl)(isopropyl)amino)cyclobutyl)propanoate
##STR00374##
[0988] The amine ethyl
3-(3-((((1R,2R,3S,4R)-4-(6-((2,4-dimethoxybenzyl)amino)-9H-purin-9-yl)-2,-
3-dihydroxycyclopentyl)methyl)amino)cyclobutyl)propanoate (1.5 g,
2.5 mmol) was taken up in Acetonitrile (66 mL) and Isopropyl iodide
(2.5 mL, 25 mmol) and Triethylamine (5.2 mL, 37 mmol) were added.
The reaction was heated to 80.degree. C. for 12 hours. Another 15
eq. TEA and another 15 eq iPrI were added and the reaction was
continued for a further 8 hours. Another 15 equivalents each of
iPrI and TEA were added and heating was continued overnight. The
reaction was concentrated and saturated Na2CO3 (20 ml) and DCM (20
ml) were added. The layers were separated and the aqueous layer was
further extracted 3 more times, the combined organics were dried
and purified by flash chromatography (SiO.sub.2, DCM 7N NH.sub.3 in
MeOH 97:3).
[0989] The residue obtained was dissolved in 30 ml DCM and washed
with 20 ml saturated NaHCO.sub.3 and 10 ms 1 N NaOH. The aqueous
was extracted with DCM 3 times, the combined organics were dried
over MgSO.sub.4 and solvent removed to yield the desired product
(1.3 g) as a foam/solid.
(3-((((1R,2R,3
S,4R)-4-(6-((2,4-dimethoxybenzyl)amino)-9H-purin-9-yl)-2,3-dihydroxycyclo-
pentyl)methyl)(isopropyl)amino)cyclobutyl)propanoic acid
##STR00375##
[0991] Lithium hydroxide, monohydrate (0.838 g, 20.0 mmol) added to
a solution of ethyl 3-(3
(3-((((1R,2R,3S,4R)-4-(6-((2,4-dimethoxybenzyl)amino)-9H-purin-9-yl)-2,3--
dihydroxycyclopentyl)methyl)(isopropyl)amino)cyclobutyl)propanoic
acid (1.3 g, 2.0 mmol) in Tetrahydrofuran (30 ml, 300 mmol) and
Methanol (6.5 ml, 160 mmol). The reaction was stirred overnight at
RT, acidified with 1 N HCl to pH=6. The volatiles were removed in
vacuo and remaining water removed by azeotropic distillation with
ethanol followed by lyophilization. The resulting solid was used
without further purification.
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((1R,2R,3S,4R)-4-(6-((2,4-dimethox-
ybenzyl)amino)-9H-purin-9-yl)-2,3-dihydroxycyclopentyl)methyl)(isopropyl)a-
mino)cyclobutyl)propanamide
##STR00376##
[0993] N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (1.19 g, 3.13 mmol) added to a solution of
3-{3-[{[(3aR,4R,6R,6aS)-6-{6-[(2,4-dimethoxybenzyl)amino]-9H-purin-9-yl}--
2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl]methyl}{isopropyl-
)amino]cyclobutyl}propanoic acid (1.30 g, 2.09 mmol) and
N,N-Diisopropylethylamine (1.20 ml, 6.89 mmol) and
4-tert-butylbenzene-1,2-diamine (0.411 g, 2.50 mmol) in
N,N-Dimethylformamide (12.9 ml). The reaction was stirred for 2
hours, NaHCO.sub.3 (saturated) was added and the mixture extracted
with EtOAc (3.times.) and the combined organics was dried over
MgSO.sub.4 filtered and concentrated. The residue was purified by
flash chromatography (DCM->DCM 7N NH.sub.3 in MeOH 95:5) to
yield the desired amide (1.4 g) as a solid.
9-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethy-
l)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclopent-
a[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-9H-purin-6-amine
##STR00377##
[0995]
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((1R,2R,3S,4R)-4-(6-((2,4-d-
imethoxybenzyl)amino)-9H-purin-9-yl)-2,3-dihydroxycyclopentyl)methyl)(isop-
ropyl)amino)cyclobutyl)propanamide (1.4 g, 1.8 mmol) in Acetic acid
(5 ml, 90 mmol) stirred overnight at 60.degree. C. The reaction was
concentrated and purified by flash chromatography (DCM->DCM/7N
NH.sub.3 in MeOH 94:6) to yield the desired compound (0.91 g) as a
foam.
(1R,2S,3R,5R)-3-(6-amino-9H-purin-9-yl)-5-(((3-(2-(5-(tert-butyl)-1H-benzo-
[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)cyclopentane-1,2-
-diol
##STR00378##
[0997] Trifluoroacetic Acid (10 ml, 100 mmol) added to a mixture of
Water (1 ml, 60 mmol) and
9-((3aS,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)eth-
yl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyclopen-
ta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-9H-purin-6-amine
(0.91 g, 1.2 mmol) at RT. The reaction was stirred overnight at RT.
The reaction was then heated to 35.degree. C. and triethylsilane
(0.39 ml, 2.4 mmol) was added. The reaction was stirred at
35.degree. C. for a further 2 days. The volatiles were removed in
vacuo and resulting residue was taken up in MeOH (15 mls). 500 mgs
of K.sub.2CO.sub.3 and 8 drops H.sub.2O were added and reaction
stirred at RT for 1 hour. The mixture was filtered and filter cake
washed with 10 mls MeOH. The filtrate was concentrated and the
resulting residue was purified by flash chromatography (DCM/7N
NH.sub.3 in MeOH 90:10) to yield the desired product (0.142 g) as a
colorless solid after several days of lyophilization.
Compound 51:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclobutyl)a-
mino)methyl)cyclopentane-1,2-diol
[0998] The diastereoisomers were separated by SFC (25 mg). .sup.1H
NMR (400 MHz, MeOD) .delta..sub.H ppm 8.06 (s, 1H), 7.48 (br. s.,
1H), 7.38 (d, J=7.88 Hz, 1H), 7.27 (dd, J=8.60, 1.55 Hz, 1H), 7.19
(d, J=3.52 Hz, 1H), 6.60 (d, J=3.73 Hz, 1H), 4.85 (m, 1H), 4.29 (m,
1H), 3.85 (t, J=5.60 Hz, 1H), 3.41 (m, 1H), 3.17 (m, 1H), 2.83 (t,
J=7.36 Hz, 2H), 2.74 (dd, J=13.68, 6.63 Hz, 1H), 2.51 (dd, J=13.79,
7.57 Hz, 1H), 2.38 (m, 1H), 2.18 (m, 3H), 2.09 (m, 1H), 2.02 (m,
5H), 1.83 (m, 2H), 1.60 (m, 3H), 1.36 (s, 9H).
Compound 52:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5,6-dichloro-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)m-
ethyl)cyclopentane-1,2-diol
[0999] The diastereoisomers were separated by SFC. The material was
taken up in MeOH/H.sub.2O and lyophilized to yield a cream colored
solid (64 mg). .sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.06
(s, 1H), 7.63 (s, 2H), 7.20 (d, J=3.52 Hz, 1H), 6.59 (d, J=3.73 Hz,
1H), 4.32 (dd. J=7.88, 6.01 Hz. 1H), 3.88 (dd, J=5.60, 4.77 Hz,
1H), 2.82 (t, J=7.26 Hz, 2H), 2.69 (m, 1H), 2.48 (m, 1H), 2.38 (m,
2H), 2.24 (m, 3H), 2.15 (s, 3H), 1.91 (m, 3H), 1.61 (m, 1H), 1.50
(m, 2H).
Compound 53:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isobutyl)ami-
no)methyl)cyclopentane-1,2-diol
[1000] Diastereisomers separated by SFC. After lyophilization of a
colorless solid (85 mg) was recovered.
Compound 54:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclopropylm-
ethyl)amino)methyl)cyclopentane-1,2-diol
[1001] The diastereosiomers were separated by SFC. The material was
taken up in MeOH/H.sub.2O and lyophilized to yield a white powder
(53 mg). .sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.06 (s,
1H), 7.48 (br. s., 1H), 7.39 (d, J=8.50 Hz, 1H), 7.28 (dd, J=8.60,
1.76 Hz, 1H), 7.21 (d, J=3.52 Hz, 1H), 6.60 (d, J=3.52 Hz, 1H),
4.32 (dd, J=7.67, 5.80 Hz, 1H), 3.91 (m, 1H), 3.44 (m, 1H), 2.84
(t, J=7.57 Hz, 2H), 2.78 (dd, J=13.27, 7.05 Hz, 1H), 2.58 (dd,
J=13.06, 7.67 Hz, 1H), 2.40 (m, 3H), 2.11 (t, J=6.22 Hz, 3H), 2.02
(m, 2H). 1.87 (m, 2H), 1.62 (m, 1H), 1.37 (s, 9H), 0.87 (m, 1H),
0.49 (m, 2H), 0.12 (m, 2H).
Compound 55:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3--
(2-(5-bromo-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl-
)cyclopentane-1,2-diol
[1002] Diastereoisomers were separated SFC .sup.1H NMR (400 MHZ,
MeOD) .delta..sub.H ppm 8.05 (s, 1H), 7.63 (s. 1H), 7.39 (m, 1H),
7.30 (dd, J=8.50, 1.66 Hz, 1H), 7.20 (d, J=3.52 Hz. 1H), 6.59 (d,
J=3.73 Hz, 1H), 4.32 (dd, J=7.77, 5.91 Hz, 1H), 3.88 (dd, J=5.70,
4.66 Hz, 1H), 2.99 (m. 1H), 2.84 (t, J=7.57 Hz, 2H). 2.48 (m, 1H),
2.41 (dd, J=7.98, 4.87 Hz, 1H), 2.34 (m, 1H), 2.24 (m, 1H), 2.15
(s, 3H). 2.10 (m. 3H), 2.01 (m, 2H), 1.86 (t. J=8.19 Hz, 2H), 1.61
(m, 1H).
Compound 56:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((isopropyl(3-
-(2-(5-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)-
methyl)cyclopentane-1,2-diol
N-(2-amino-4-(trifluoromethoxy)phenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4--
dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrah-
ydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)-
propanamide
##STR00379##
[1004] N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (1.19 g, 314 mmol) added to a solution of
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(isopropyl)amino)cyclobutyl)propanoic acid (1.3 g, 2.1
mmol) and N,N-Diisopropylethylamine (120 mL, 6.90 mmol) and
4-(trifluoromethoxy)benzene-1,2-diamine (0.482 g, 2.51 mmol) in
N,N-Dimethylformamide (13.0 mL, 167 mmol). The reaction was stirred
overnight at RT and was partially concentrated to ca 2 mls and then
NaHCO.sub.3 (saturated) was added. The mixture was extracted with
EtOAc (3.times.) and the combined organics were dried with
MgSO.sub.4 filtered and concentrated. The residue was purified by
flash chromatogrpahy (DCM/7N NH.sub.3 in MeOH 95:5) to the desired
amide (1.4 g) as a solid.
N-(2,4-dimethoxybenzyl)-7-((3aS,4R,6R,6aR)-6-((isopropyl(3-(2-(5-(trifluor-
omethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)-2,2-dime-
thyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimid-
in-4-amine
##STR00380##
[1006]
N-(2-amino-4-(trifluoromethoxy)phenyl)-3-(3-((((3aR,4R,6R,6aS)-6-(4-
-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethy-
ltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cycl-
obutyl)propanamide (1.4 g, 1.8 mmol) was heated in AcOH at
60.degree. C. overnight. The reaction mixture was concentrated in
vacuo giving the crude product.
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((isopropyl(3--
(2-(5-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)m-
ethyl)cyclopentane-1,2-diol
##STR00381##
[1008] Trifluoroacetic Acid (10 mL, 100 mmol) added to a mixture of
Water (1 mL, 60 mmol) and
N-(2,4-dimethoxybenzyl)-7-((3aS,4R,6R,6aR)-6-((isopropyl(3-(2-(5-(trifluo-
romethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)-2,2-dim-
ethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimi-
din-4-amine (0.91 g 12 mmol) at RT. The reaction was stirred
overnight at RT then quenched by the addition of Triethylsilane
(0.37 mL, 2.3 mmol). The volatiles were removed in vacuo and
resulting residue was taken up in MeOH (15 mls) 500 mgs of
K.sub.2CO.sub.3 and 8 drops H.sub.2O were added and reaction
stirred at RT for 1 hour. Mixture was filtered and filter cake
washed with 10 mls MeOH. The filtrate was concentrated and the
resulting residue purified by flash chromatography (DCM/7N NH.sub.3
in MeOH 90:10) to yield the desired product (0.232 g) as an off
white foam. MS (ESI.sup.+) for
C.sub.29H.sub.36F.sub.3N.sub.7O.sub.3 m/z 588.2 [M+H].sup.+; MS
(ESI.sup.-) for C.sub.29H.sub.36F.sub.3N.sub.7O.sub.3 m/z 586.2
[M-H].sup.+; HPLC purity >90% (ret. time, 2.570 min.) .sup.1H
NMR (400 MHz, d4-MeOH) .delta..sub.H 8.082 & 8.079 (s, 11-,
overlapping peaks due to cis and trans isomers), 7.554-7.524 (m,
1H), 7.414 (s, 1H), 7.225-7.209 (m, 1H), 7.155-7.127 (m, 1H),
6.618-6.609 (m, 1H), 4.363-4.323 (m, 1H), 3.976-3.932 (m, 1H),
3.606-3.524 (m, 0.5H (methine from trans isomer)), 3.156-3.110 (m,
0.5H (methine from cis isomer), 3.089-3.006 (m, 1H), 2.731-2.679
(m, 1H), 2.544-2360 (m, 2H), 2.256-2.239 (m, 3H), 2.093-2.061 (m,
2H), 1.987-1.861 (m, 3H), 1.648-1.568 (m, 2H), 1.072-1.006 (m,
6H).
Compound 57:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl((1s,-
3R)-3-(2-(5-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)a-
mino)methyl)cyclopentane-1,2-diol
[1009] The diastereoisomers were separated by SFC. The material was
lyophilized to give a solid (78 mg). .sup.1H NMR (400 MHz, d4-MeOH)
.delta..sub.H 8.076 (s, 1H), 7.548-7.527 (m, 1H), 7.414 (s, 1H),
7.227-7.218 (m, 1H), 7.148-7.123 (m, 1H), 6.616-6.607 (m, 1H),
4.361-4.327 (m, 1H), 3.926-3.899 (m, 1H), 3.037-3.000 (m, 1H),
2.907-2.870 (m, 2H), 2.538-2.283 (m, 4H), 2.178-2.013 (m, 8H),
1.913-1.872 (m, 2H), 1.680-1.599 (m, 1H).
Compound 58:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3--
(2-(5,6-dichloro-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)m-
ethyl)cyclopentane-1,2-diol
[1010] The diastereomers were separated by SFC. The material was
taken up in MeOH/H.sub.2O and lyophilized to yield a tan powder (73
mg). .sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.06 (s, 1H),
7.64 (s, 2H), 7.21 (d, J=3.73 Hz, 1H), 6.59 (d, J=3.52 Hz, 1H),
4.32 (dd, J=7.77, 6.12 Hz, 1H), 3.89 (m, 1H), 3.01 (m, 1H), 2.86
(t, J=7.67 Hz, 2H), 2.51 (m, 1H), 2.40 (m, 2H), 2.27 (m, 1H), 2.18
(s, 3H), 2.11 (m, 3H), 2.02 (q, J=6.43 Hz. 2H), 1.88 (t, J=8.19 Hz,
2H), 1.63 (m, 1H).
Compound 59:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclobutylme-
thyl)amino)methyl)cyclopentane-1,2-diol
Step 1: ethyl
3-((1R,3s)-3-((cyclobutylmethyl)(((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxyben-
zyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyc-
lopenta[d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoate
##STR00382##
[1012] The amine ethyl
3-[3-({[(3aR,4R,6R,6aS)-6-{4-[(2,4-dimethoxybenzyl)amino]-7H-pyrrolo[2,3--
d]pyrimidin-7-yl}-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
]methyl}amino)cyclobutyl]propanoate (1.8 g, 3.0 mmol) was taken up
in methanol (20 mL, 600 mmol) and sodium cyanoborohydride (0.37 g,
5.9 mmol) was added. The pH was adjusted to ca. 6 using a 10%
solution of AcOH in methanol, then cyclobutanecarboxaldehyde (0.32
g, 3.8 mmol) was added in one portion. The reaction was allowed to
proceed for 5 hours at which time HPLC indicated the reaction had
stalled. Another 1.3 equivalents of cyclobutanecarboxaldehyde was
added and the reaction continued overnight. NaHCO.sub.3 (saturated)
was added to the reaction mixture which was then extracted 3 times
with DCM. The combined organics were dried with MgSO.sub.4 and
concentrated to a yellow resin. Cis and trans isomers were
separable on silica. Purification by FC (DCM/7N NH.sub.3 in MeOH
96:4) yielded 2 separate batches of product, each enriched in one
respective isomer to about 90%. Top isomer: 0.38 g (5:1 mixture,
cis) Bottom isomer: 0.31 g (7:1 mixture, trans). MS (ESI.sup.+) for
C.sub.35H.sub.49N.sub.5O.sub.6 m/z 676.7 [M+H].sup.+; HPLC purity
>69% (ret. time, 3.791).
Step 2:
N-(2-amino-5-(tert-butyl)phenyl)-3-((1R,3s)-3-((cyclobutylmethyl)(-
((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimi-
din-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)-
amino)cyclobutyl)propanamide
##STR00383##
[1014] Bottom Isomer (trans): Lithium hydroxide monohydrate (0.192
g, 4.59 mmol) was added to a solution of ethyl
3-((1R,3s)-3-((cyclobutylmethyl)(((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxyben-
zyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyc-
lopenta[d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoate (0.31
g, 0.46 mmol) in tetrahydrofuran (6 mL, 70 mmol) and methanol (1.5
mL, 37 mmol). The reaction was stirred overnight at room
temperature and by the next morning the starting material was
consumed and had been transformed into the acid. The reaction was
acidified with 1N HCl to pH=6. The volatiles were removed in vacuo
and the remaining water removed by azeotropic distillation with
ethanol followed by 24 hours of lyophilization. The resulting off
white solid was used without further purification. HPLC purity
>94% (ret. time, 3.344).
[1015] N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
hexafluorophosphate (0.273 g, 0.718 mmol) added to a solution of
3-{trans-3-[(cyclobutylmethyl){[(3aR,4R,6R,6aS)-6-{4-[(2,4-dimethoxybenzy-
l)amino]-7H-pyrrolo[2,3-d]pyrimidin-7-yl}-2,2-dimethyltetrahydro-3aH-cyclo-
penta[d][1,3]dioxol-4-yl]methyl}amino]cyclobutyl}propanoic acid
(0.31 g, 0.48 mmol) and N,N-diisopropylethylamine (0.275 mL, 1.58
mmol) and 4-tert-butylbenzene-1,2-diamine (0.0943 g, 0.574 mmol) in
N,N-dimethylformamide (2.96 mL, 38.3 mmol). The reaction was
stirred overnight at room temperature and by the next morning the
starting material was consumed. The reaction was partially
concentrated to ca. 2 mls and then NaHCO.sub.3 (saturated) was
added. The mixture was extracted with EtOAc 3 times and the
combined organics were dried with MgSO.sub.4 and concentrated. The
resulting residue was purified by FC (DCM/7N NH.sub.3 in MeOH 95:5)
to yield
N-(2-amino-5-(tert-butyl)phenyl)-3-((1R,3s)-3-((cyclobutylmethyl)(((3aR,4-
R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-y-
l)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)amino)c-
yclobutyl)propanamide (0.29 g; 76%) as a purple-brown amorphous
solid. HPLC purity >20% (ret. time, 3.650 min.)
Step 3:
7-((3aS,4R,6R,6aR)-6-((((1s,3R)-3-(2-(5-(tert-butyl)-1H-benzo[d]im-
idazol-2-yl)ethyl)cyclobutyl)(cyclobutylmethyl)amino)methyl)-2,2-dimethylt-
etrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-py-
rrolo[2,3-d]pyrimidin-4-amine
##STR00384##
[1017] A solution of
N-(2-amino-5-(tert-butyl)phenyl)-3-((1R,3s)-3-((cyclobutylmethyl)(((3aR,4-
R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-y-
l)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)amino)c-
yclobutyl)propanamide (0.3 g, 0.4 mmol) in acetic acid (1.0 mL, 20
mmol) was stirred overnight at 65.degree. C. and by next morning
the starting material was consumed. The volatiles were removed in
vacuo and the resulting residue purified by FC (DCM/7N NH.sub.3 in
MeOH 93:7) to yield
7-((3aS,4R,6R,6aR)-6-((((1s,3R)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol--
2-yl)ethyl)cyclobutyl)(cyclobutylmethyl)amino)methyl)-2,2-dimethyltetrahyd-
ro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2-
,3-d]pyrimidin-4-amineas an off white solid. HPLC purity >73%
(ret. time, 3.709 min.).
Step 4:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s-
,3R)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclo-
butylmethyl)amino)methyl)cyclopentane-1,2-diol
##STR00385##
[1019] Trifluoroacetic acid (5 mL, 70 mmol) was added to a mixture
of water (0.5 mL, 30 mmol) and
7-((3aS,4R,6R,6aR)-6-((((1s,3R)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol--
2-yl)ethyl)cyclobutyl)(cyclobutylmethyl)amino)methyl)-2,2-dimethyltetrahyd-
ro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2-
,3-d]pyrimidin-4-amine (0.23 g, 0.30 mmol) at room temperature. The
reaction was allowed to proceed overnight at which time the bright
pink suspension was quenched with triethylsilane (0.095 mL, 0.59
mmol). The volatiles were removed in vacuo and the resulting
residue was taken up in MeOH (15 mls). 500 mgs of K.sub.2CO.sub.3
and 8 drops H.sub.2O were added and reaction stirred at room
temperature for 1 hour. The mixture was filtered and the filter
cake was washed with 10 mLs methanol. The filtrate was concentrated
and the resulting residue purified by FC (DCM/7N NH.sub.3 in MeOH
90:10) to yield
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(cyclobutylme-
thyl)amino)methyl)cyclopentane-1,2-diol (0.018 g, 10%) as a
colorless solid. MS (ESI.sup.+) for C.sub.34H.sub.47N.sub.7O.sub.2
m/z 586.4 [M+H].sup.+; HPLC purity >93% (ret. time, 2.070 min.)
.sup.1H NMR (400 MHz, d.sub.4-MeOH) .delta..sub.H 8.083 (s, 1H),
7.501 (s, 1H), 7.421-7.400 (m, 1H), 7.315-7.290 (m, 1H),
7.218-7.209 (m, 1H), 6.621-6.612 (m, 1H), 4.350-4.317 (m, 1H),
3.930-3.903 (m, 1H), 3.403-3.367 (m, 1H), 2.880-2.843 (m, 2H),
2.722-2.360 (m, 6H), 2.323-2.241 (m, 2H), 2.173-1.606 (m, 13H),
1.387 (s, 9H).
Compound 60:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((isopropyl((-
1r,3S)-3-(2-(5-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobuty-
l)amino)methyl)cyclopentane-1,2-diol
[1020] Diastereoisomers separated by SFC. After lyophilization, of
a colorless solid (62 n) was recovered.
Compound 61:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((isopropyl((-
1s,3R)-3-(2-(5-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobuty-
l)amino)methyl)cyclopentane-1,2-diol
[1021] Diastereosiomers were separated SFC. The material was taken
up in MeOH/H.sub.2O and lyophilized to yield an off white powder
(89 mg). .sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.06 (s,
1H), 7.52 (d, J=8.71 Hz, 1H), 7.40 (s, 1H), 7.19 (d, J=3.52 Hz,
1H), 7.12 (m, 1H), 6.59 (d,
[1022] J=3.52 Hz, 1H), 4.88 (m, 1H), 4.33 (m, 1H), 3.94 (t, J=5.39
Hz, 1H), 3.52 (m, 1H), 3.01 (m, 1H), 2.87 (t, J=7.15 Hz, 2H), 2.68
(dd, J=13.48, 7.88 Hz, 1H), 2.47 (dd, J=13.27, 7.46 Hz, 1H), 2.37
(m, 1H), 2.21 (m, 3H), 2.04 (m, 3H), 1.84 (m, 2H), 1.58 (m, 1H),
1.02 (d, J=6.63 Hz, 3H), 0.98 (d, J=6.43 Hz, 3H).
Compound 62:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl(3-(2-
-(5-(oxetan-3-yl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)c-
yclopentane-1,2-diol
N-(4-(oxetan-3-yl)phenyl)acetamide
##STR00386##
[1024] (4-Acetamidophenyl)boronic acid {670 mg, 3.7 mmol), Nickel
(II) iodide (35 mg, 0.11 mmol), trans-2-aminocyclohexanol (17 mg,
0.11 mmol), and Sodium hexamethyldisilazane (690 mg, 3.7 mmol) were
weighed into a microwave reaction vial. A septum was placed over
the top, nitrogen was purged and Isopropyl alcohol (5.7 ml, 75
mmol) was added. The vial was purged with nitrogen for 10 minutes
and 3-iodooxetane (344 mg, 1.87 mmol) was added in 0.75 ml
isopropyl alcohol. The septum was replaced with a microwave vial
cap and the mixture was heated in a microwave reactor (microwave
conditions: CEM Discovery Explorer microwave reactor; Ramp time: 10
min; 80.degree. C. for 30 min; power: 300 W). The crude reaction
mixture was diluted with 8 ml EtOH and the suspension was filtered
through a pad of Solka Floc.RTM.. The pad was washed with 35 ml
EtOH and the filtrate was concentrated. The crude material was
purified by flash chromatography (SiO.sub.2, eluting with 40-60%
EtOAc/CH.sub.2Cl.sub.2) to give the desired product as an oil (200
mg).
N-(2-nitro-4-(oxetan-3-yl)phenyl)acetamide
##STR00387##
[1026] Sulfuric acid (9.4 ml, 180 mmol) was added carefully to 70%
nitric acid (7:3, Nitric acid:Water, 11 ml, 170 mmol) which was
cooled at 0.degree. C. over about 5-10 minutes. The mixture was
stirred for 10 minutes at 0.degree. C., then allowed to warm to RT
by removal of the ice bath. The acid solution was transfered to a
separatory funnel and Methylene chloride (20 mL, 300 mmol) was
added. The funnel was shaken for 5 minutes, and the phases were
allowed to separate.
[1027] The organic phase (upper phase) was isolated and the process
was repeated with an additional 20 ml CH.sub.2Cl.sub.2. The organic
extracts were combined, it was assumed the organic phase contained
about 5 g (-80 mmol) of anhydrous HNO.sub.3. Using a 50 fold
excess, this required about 25 ml of solution. The nitric acid
solution was cooled in a ice bath. The
N-(4-oxetan-3-ylphenyl)acetamide (21.0 mg, 0.70 mmol) was treated
with 25 ml of the chilled HNO.sub.3/CH.sub.2Cl.sub.2 solution and
was allowed to stir about 30 minutes. The reaction mixture was
carefully poured into 45 ml 10% NH.sub.4OH solution and carefully
shaken. The phases were separated and the aqueous phase was washed
with 20 ml CH.sub.2Cl.sub.2. The combined organic phase was dried
over Na.sub.2SO.sub.4, filtered and concentrated. The crude
material was purified
[1028] by flash chromatography (SiO.sub.2, eluting with 25-35%
EtOAc/CH.sub.2Cl.sub.2) to yield the desired product as a solid
(170 mg).
2-nitro-4-(oxetan-3-yl)aniline
##STR00388##
[1030] A suspension of N-(2-nitro-4-oxetan-3-ylphenyl)acetamide
(125 mg, 0.529 mmo) in aqueous hydrazine (8 ml, 160 mmol) was
heated at 70.degree. C. for 2 h, the reaction mixture was cooled to
45.degree. C. and the hydrazine was removed in vacuo to yield a
solid. The crude material was purified by flash chromatography
(SiO.sub.2, eluting with 20% EtOAc/CH.sub.2Cl.sub.2 to yield the
desired product (71 mg).
4-(oxetan-3-yl)benzene-1,2-diamine
##STR00389##
[1032] A solution of 2-nitro-4-oxetan-3-ylaniline (91 mg, 0.47
mmol) in Ethanol
[1033] (6.1 ml) was carefully treated with 10% Palladium on carbon
(10 mg, 0.009 mmol) as a slurry in ethanol. The reaction flask was
evaculated and filled with hydrogen gas three times
[1034] and the reaction was allowed to stir under an atmosphere of
hydrogen for 2 h. The reaction mixture was filtered through a pad
of Solka Floc.RTM. and the pad was washed with 25 ml MeOH. The
filtrated was concentrated to yield an oil that solidified under
high vacuum overnight to give the desired compound (72 mg). The
material was used as is in the next step. 1H NMR (400 MHZ,
CDCl.sub.3) .delta..sub.H ppm 6.81 (d, J=1.52 Hz, 1H), 6.70 (m,
2H), 5.02 (dd, J=8.34, 5.81 Hz, 2H), 4.74 (m, 2H), 4.09 (m, 1H),
3.45 (br. s., 2H), 3.37 (br. s., 2H).
N-(2,4-dimethoxybenzyl)-7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(5-
-(oxetan-3-yl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetr-
ahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-ami-
ne
##STR00390##
[1036] A solution of
3-(3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl-
)methyl)(methyl)amino)cyclobutyl)propanoic acid (250 mg. 0.42 mmol)
and 4-oxetan-3-ylbenzene-1,2-diamine (72 mg, 0.44 mmol) in
N,N-Dimethylformamide (4.3 ml, 56 mmol) was treated with
N,N-Diisopropylethylamine (0.24 ml, 1.4 mmol) dropwise followed by
N,N,N',N'-Tetramethyl-0-(7-azabenzotriazol-1-yl)uronium
Hexafluorophosphate (240 mg, 0.632 mmol) in one portion. The
reaction mixture was stirred at RT for 6 hours, upon which the
reaction mixture was concentrated under high vacuum. The residue
was partitioned between 30 ml EtOAc (some MeOH was added to aid in
solublizing the product) and 30 ml 1/1 H.sub.2O/sat NaHCO.sub.3.
The aqueous phase was extracted with 30 mL EtOAc and the combined
organic phase was dried over Na.sub.2SO.sub.4, filtered and
concentrated to a glass/stiff foam. The crude material was purified
by flash chromatography (SiO.sub.2, eluting with 6-7% 7N NH.sub.3
in CH.sub.3OH/CH.sub.2Cl.sub.2. Two sets of products were found, a
less polar pair and a more polar pair corresponding to the 2 aide
regio-siomers. Each regio-siomer was processed separately in the
next step.
[1037] The amide (130 mg) was taken up in 5 ml glacial acetic acid
and heated at 65.degree. C. for (2.25 h, the reaction was cooled
and placed in the fridge overnight. The acetic acid was removed
under high vacuum with the aid of a warm water bath. The two
batches of crude product was taken up in 30 ml CH.sub.2Cl.sub.2 and
the organic phase was washed with 10 ml portions of sat NaHCO.sub.3
and 2% Na.sub.2CO.sub.3 solutions, dried over Na.sub.2SO.sub.4,
filtered and concentrated. The crude material was purified by flash
chromatography (SiO.sub.2, eluting with 5.5-6.5% 7N NH.sub.3 in
CH.sub.3OH/CH.sub.2Cl.sub.2 to give the desired compound (140
mg).
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((methyl(3-(2--
(5-(oxetan-3-yl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)cy-
clopentane-1,2-diol
##STR00391##
[1039]
N-(2,4-dimethoxybenzyl)-7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-((methyl(-
3-(2-(5-(oxetan-3-yl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)meth-
yl)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimidi-
n-4-amine (115 mg, 0.159 mmol) was dissolved in a mixture of
Trifluoroacetic Acid (4.00 ml, 51.9 mmol) and Water (0.40 ml, 22
mmol) which had been pre-cooled at 0.degree. C. in an ice bath. The
solution was stirred at 0.degree. C. for 2 h, the reaction mixture
was allowed to warm to RT. After 1 the reaction mixture was
concentrated in vacuo. The residue was taken up in 6 ml MeOH,
concentrated and the process was repeated twice. The resultant
residue was placed on high vacuum. The crude residue was diluted
with 2 ml MeOH, treated with 140 mg K.sub.2CO.sub.3 and 10 drops of
H.sub.2O and allowed to stir at RT until the solution was basic by
pH paper. The solution was filtered through a fine frit and the
solids washed with MeOH. The filtrate was concentrated to a solid
that was placed on high vacuum overnight. The crude material was
purified by prep TLC on two 20 cm.times.20 cm.times.1.0 mm prep TLC
plate, eluting with 14% 7N NH.sub.3 in CH.sub.3OH/CH.sub.2Cl.sub.2
to ive the product as a colorless glass (37 mg) .sup.1H NMR (400
MHz, MeOD) .delta..sub.H ppm 8.06 (s, 1H), 7.52 (br. s., 1H), 7.48
(d, J=8.29 Hz, 1H), 7.28 (m, 1H), 7.20 (t, J=3.42 Hz, 1H), 6.60 (d,
J=3.52 Hz, 1H), 5.12 (m, 2H), 4.80 (m, 2H), 4.38 (m, 1H), 4.32 (m,
1H), 3.89 (q, J=5.60 Hz, 1H), 3.04 (m, 1H), 2.85 (m, 2H), 2.70 (m,
1H), 2.52 (m, 1H), 2.41 (m. 2H), 2.27 (dd, J=10.99, 6.63 Hz, 2H),
2.19 (s, 3H), 2.17 (s, 3H), 2.14 (m, 1H), 2.03 (d, J=7.88 Hz, 1H),
1.91 (m, 3H), 1.62 (m, 1H), 1.51 (m, 1H).
Compound 63:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((methyl((1r,3S)-3-(2-(5-(oxeta-
n-3-yl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydrof-
uran-3,4-diol
[1040] The diastereomers were separated by SFC. The material was
taken up in MeOH/H.sub.2O and lyophilized to a tan powder (58 mg).
.sup.1H NMR (400 MHz, MeOD) H ppm 8.26 (s, 1H), 8.19 (s, 1H), 7.51
(s, 1H), 7.47 (d, J=8.29 Hz, 1H), 7.26 (dd, J=8.29, 1.45 Hz, 1H),
5.97 (d, J=3.94 Hz, 1H), 5.11 (dd, J=8.29, 6.01 Hz, 2H), 4.79 (t,
J=6.32 Hz, 2H), 4.69 (m, 1H), 4.36 (m, 1H), 4.22 (t, J=5.60 Hz,
1H), 4.15 (m, 1H), 2.79 (t, J=7.15 Hz, 2H), 2.72 (m, 1H), 2.66 (m,
2H), 2.21 (m, 2H), 2.14 (s, 3H), 1.88 (m, 3H), 1.45 (m, 2H).
Compound 64:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((methyl(3-(2-(5-(oxetan-3-yl)--
1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydrofuran-3,4-
-diol
Step 1:
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetr-
ahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoi-
c acid
##STR00392##
[1042] A solution of ethyl
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoate
(0.39 g, 0.82 mmol) in methanol (14 mL) was treated with a 1 M
aqueous solution of sodium hydroxide (1.56 mL, 1.56 mmol) and the
reaction mixture was heated at 50.degree. C. with stirring for 3.5
h; HPLC/LC MS indicated conversion to the desired product. The
reaction mixture was concentrated in vacuo and the aqueous residue
was diluted with water (10 mL) and extracted with CH.sub.2Cl.sub.2
(3.times.5 mL). The aqueous layer was treated with a 1 M aqueous
solution of hydrogen chloride (1.44 mL, 1.44 mmol) to adjust to pH
7. The clear, colorless solution was lyophilized to afford the
crude title compound (0.487 g, 110%) as a slightly off-white solid,
yield accounts for 1.56 mmol NaCl (91 mg): MS (ESI+) for
C.sub.21H.sub.30N.sub.6O.sub.5 m/z 447.1 (M+H).sup.+; MS (ESI-) for
C.sub.21H.sub.30N.sub.6O.sub.5 m/z 445.2 (M-H).sup.-; HPLC purity
>95% (ret. time, 1.949 min).
Step 2:
N-(2-amino-5-(oxetan-3-yl)phenyl)-3-(3-((((3aR,4R,6R,6aR)-6-(6-ami-
no-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(methyl)amino)cyclobutyl)propanamide
##STR00393##
[1044] A suspension of the above crude
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoic
acid and 4-(oxetan-3-yl)benzene-1,2-diamine (0.135 g, 0.822 mmol)
in methylene chloride (8.0 mL) was treated with
N,N-diisopropylethylamine (0.716 mL, 4.11 mmol) and cooled to
-5.degree. C. (ice/brine).
N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium
hexafluorophosphate [HATU] (0.469 g, 1.23 mmol) was added and the
reaction mixture was stirred for 5.25 h, warming to 15.degree. C.;
HPLC/LC MS indicated complete conversion. The reaction mixture was
concentrated in vacuo and diluted with CH.sub.2Cl.sub.2 (15 mL) and
water (7.5 mL). The separated aqueous layer was extracted with
CH.sub.2Cl.sub.2 (2.times.10 mL). The combined organics were dried
(Na.sub.2SO.sub.4) and concentrated in vacuo to afford a
brown-purple semi-opaque oil/foam. Purification by column
chromatography (2.times.8 cm silica; 0-5% 7 N methanolic
NH.sub.3/CH.sub.2Cl.sub.2) afforded both amide regioisomers of the
title compound (0.45 g, 82%) as a semi-opaque pink foam: MS (ESI+)
for C.sub.30H.sub.40N.sub.5O.sub.5 m/z 593.3 (M+H).sup.+; MS (ESI-)
for C.sub.30H.sub.40N.sub.8O.sub.5 m/z 591.3 (M-H).sup.- and 637.4
(M+HCO.sub.2).sup.-; HPLC purity 90% (ret. time, 2.097 min).
Step
3:9-((3aR,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(5-(oxetan-3-yl)-1H-
-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydrofuro[3,4-d]-
[1,3]dioxol-4-yl)-9H-purin-6-amine
##STR00394##
[1046]
N-(2-amino-5-(oxetan-3-yl)phenyl)-3-(3-((((3aR,4R,6R,6aR)-6-(6-amin-
o-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)-
(methyl)amino)cyclobutyl)propanamide (0.446 g, 0.752 mmol) was
taken up in acetic acid (7.7 mL, 140 mmol) and heated at 65.degree.
C. for 3.5 h; HPLC/LC MS indicated complete conversion. At 3.75 h
the acetic acid was removed by distillation with minimal warming to
afford an orange oil, which was taken up in CH.sub.2Cl.sub.2 (45
mL) and washed with saturated aqueous NaHCO.sub.3 (2.times.30 mL).
The aqueous layer was treated with NaCl until saturated and
extracted with CH.sub.2Cl.sub.2 (2.times.20 mL). The combined
organic layers were dried (Na.sub.2SO.sub.4) and concentrated in
vacuo to afford a light orange oil. Purification by column
chromatography (2.times.8 cm silica; 0-5% 7 N methanolic
NH.sub.3/CH.sub.2Cl.sub.2) afforded the title compound (0.28 g,
65%) as a light orange foam: MS (ESI+) for
C.sub.30H.sub.35N.sub.5O.sub.4 m/z 575.3 (M+H).sup.+; MS (ESI-) for
C.sub.30H.sub.3NO.sub.4 m/z 573.3 (M-H).sup.-; HPLC purity >95%
(ret. time 2.142 min).
Step 4:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((methyl(3-(2-(5-(oxetan-
-3-yl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydrofu-
ran-3,4-diol
##STR00395##
[1048] To a cooled (ice bath) flask containing
9-((3aR,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(5-(oxetan-3-yl)-1H-benzo-
[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydrofuro[3,4-d][1,3]d-
ioxol-4-yl)-9H-purin-6-amine (0.28 g, 0.42 mmol) was added a
precooled (ice bath) solution of trifluoroacetic acid (6.4 mL, 84
mmol) in water (0.75 mL, 42 mmol). The reaction mixture was stirred
for 5.75 h at 0.degree. C.; HPLC/LC MS indicated nearly complete
consumption of starting material. At 6 h the flask was removed from
the cold bath and the volatiles were removed by distillation at rt.
The residue was diluted with MeOH (15 mL) and treated with
potassium carbonate (0.32 g, 2.3 mmol) and water (1 mL) and the
mixture was stirred for 20 min at rt; pH 2. Additional potassium
carbonate (0.20 g, 1.4 mmol) was added and the mixture was stirred
for 20 min; pH 8-9. The solution was filtered through a fine frit,
rinsing with MeOH, and the filtrate was concentrated in vacuo to
afford a tan semi-solid. Purification by column chromatography
(3.times.8 cm silica; 10-20% 7 N methanolic
NH.sub.3/CH.sub.2Cl.sub.2) afforded the title compound (123 mg,
55%) as a nearly colorless glass: MS (ESI+) for
C.sub.27H.sub.34N.sub.8O.sub.4 m/z 535.3 (M+H).sup.+; MS (ESI-) for
C.sub.27H.sub.34N.sub.8O.sub.4 m/z 533.3 (M-H).sup.-; HPLC purity
>95% (ret. time 1.765 min); .sup.1H NMR (400 MHz, d4-MeOH)
mixture of cis/trans isomers .delta..sub.H 8.29-8.25 (m, 1H),
8.21-8.17 (m, 1H), 7.51 (s, 1H), 7.47 (d, J=8.3 Hz, 1H), 7.26 (dd,
J=1.6, 8.3 Hz, 1H), 6.00-5.96 (m, 1H), 5.11 (dd, J=5.8, 8.3 Hz,
2H), 4.81-4.76 (m, 2H), 4.73-4.68 (m, 1H), 4.40-4.31 (m, 1H),
4.27-4.13 (series of m, 2H), 3.13-3.03 (m, 0.4H), 2.86-2.66 (series
of m, 4.6H), 2.30-1.80 (series of m, 8.6H), 1.55-1.40 (m,
1.4H).
Compound 65:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-methyl((1s,3R)-3-(2-(5-(oxetan-3--
yl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydrofuran-
-3,4-diol
[1049] The diastereomers were separated at by SFC The material was
taken up in MeOH/H.sub.2O and lyophilized to a white powder (35 g).
.sup.1H NMR (400 MHz, MeOD) .delta..sub.H ppm 8.28 (s, 1H), 8.19
(s, 1H), 7.51 (s, 1H, 7.48 (df, 1 Hz, 1H1), 7.27 (dd, J=8.29, 1.45
Hz, 1H), 5.98 (d, J=4.15 Hz, 1H), 5.12 (dd, J=8405.91 Hz, 2H), 4.79
(t, J=6.32 Hz, 2H), 4.69 (dd, J=5.39, 4.15 Hz, 1H), 436 (m, 1) 4.23
(t, J=5.60 Hz H, 4.17 (m, H), 305 (m. 1H) 0.83 (t, J=7.46 Hz, 2H),
2.67 (m. 2H), 2.16 (s, 3H), 2.08 (m, 2H), 1.98 (m, 3H), 1.83 (m,
2H).
Compound 67:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-[({3-[2-(5-cyclobutyl-1H-1,3-be-
nzodiazol-2-yl)ethyl]cyclobutyl}(propan-2-yl)amino)methyl]oxolane-3,4-diol
Step 1: Benzyl
3-[3-({[(3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tetrahydro-
-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}amino)cyclobutyl]propanoate
##STR00396##
[1051] A suspension of the
9-[(3aR,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyl-tetrahydro-2H-furo[3,4-d]-
[1,3]dioxol-4-yl]-9H-purin-6-amine (1.45 g, 4.736 mmol), benzyl
3-(3-oxocyclobutyl)propanoate (1.21 g, 5.209 mmol) and acetic acid
(246.45 .mu.l, 4.31 mmol) in DCE:iPrOH (4:1, 50 ml) was stirred at
RT for 1 h. A further aliquot of DCE (40 ml) and iPrOH (5 ml) was
added to the reaction mixture and continued for 1 hour. STAB (1.28
g, 6.03 mmol) was then added and the reaction mixture was stirred
for 18 hours. The reaction mixture was quenched with 1N
Na.sub.2CO.sub.3 (10 ml), and the product was extracted with DCM
(2.times.30 ml). This was dried over Na.sub.2SO.sub.4, filtered and
evaporated to dryness. Purification by silica gel column
chromatography, eluting with 7N NH.sub.3 in MeOH:DCM (1:99-3:97)
gave the desired product as a colourless oil, 1.51 g (58%); MS
(ESI.sup.+) for C.sub.27H.sub.34N.sub.6O.sub.5 m/z 523.65
[M+H].sup.+; HPLC purity 97% (ret. time, 1.43 min); H NMR (500 MHz,
CHLOROFORM-d) .delta. ppm 8.35 (d, J=5.3 Hz, 1H), 7.87 (d, J=33.8
Hz, 1H), 7.40-7.29 (m, 5H), 6.08-5.94 (m, 1H), 5.59-5.42 (m, 3H),
5.10 (d, J=4.0 Hz, 2H), 5.03-4.96 (m, 1H), 4.33 (dq, J=7.3, 3.9 Hz,
1H), 3.12 (ddd, J=23.0, 14.6, 7.5 Hz, 1H), 2.85-2.77 (m, 1H), 2.74
(dd, J=12.5, 6.6 Hz, 1H), 2.39-2.07 (m, 4H), 1.90-1.64 (m, 5H),
1.61 (s, 4H), 1.38 (s, 3H), 1.28-1.06 (m, 1H).
Step 2. Benzyl
3-[3-({[(3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tetrahydro-
-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}(propan-2-yl)amino)cyclobutyl]propa-
noate
##STR00397##
[1053] K.sub.2CO.sub.3 (528.92 mg, 3.83 mmol) was added to a
solution of benzyl
3-[3-({[(3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tet-
rahydro-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}amino)cyclobutyl]propanoate
(1.00 g, 1.91 mmol) and 2-iodopropane (0.57 ml, 5.74 mmol) in MeCN
and stirred at 95.degree. C. in a sealed tube for 18 hours. The
reaction mixture was diluted with EtOAc (20 ml), filtered and
evaporated to dryness. Purification by silica gel chromatography,
eluting with 7N NH.sub.3 in MeOH:DCM (1:99-5:95) gave the desired
product as a colourless oil, 700 mg (65%); MS (ESI.sup.+) for
C.sub.30H.sub.40N.sub.6O.sub.5 m/z 565.70 [M+H].sup.+; HPLC purity
96% (ret. time, 1.48 min); .sup.1H NMR (500 MHz, CHLOROFORM-d)
.delta..sub.H ppm 8.35 (d, J=3.5 Hz, 1H), 7.88 (d, J=3.2 Hz, 1H),
7.44-7.29 (m, 5H), 6.03 (t, J=2.2 Hz, 1H), 5.62-5.42 (m, 3H), 5.10
(d, J=3.3 Hz, 2H), 5.06-4.92 (m, 1H), 4.26 (dt, J=9.9, 3.4 Hz, 1H),
3.46-2.84 (m, 2H), 2.88-2.61 (m, 1H), 2.51 (ddd, J=14.0, 9.1, 7.5
Hz, 1H), 2.33-2.15 (m, 2H), 2.50-2.13 (m, 2H), 2.16-1.74 (m, 4H),
1.60 (s, 3H), 1.43-1.35 (m, 4H), 0.96 (d, J=6.7 Hz, 3H), 0.79 (d,
J=6.6 Hz, 3H).
Step 3.
3-[3-({[(3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tet-
rahydro-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}(propan-2-yl)amino)cyclobuty-
l]propanoic acid
##STR00398##
[1055] 10% Pd-C (70 mg) was added to a solution of benzyl
3-[3-({[(3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tetrahydro-
-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}(propan-2-yl)amino)cyclobutyl]propa-
noate (790 mg, 1.40 mmol) in EtOH (20 ml) and stirred under an
atmosphere of hydrogen for 18 hours at RT. A further aliquot of 10%
Pd-C (70 mg) was added and the reaction was continued stirring
under hydrogen for 4 hours. This was filtered and evaporated in
vacuo, and then evaporated from DCM (2.times.20 ml) to give 680 mg
(quant.) of a white foamy solid; MS (ESI.sup.+) for
C.sub.23H.sub.34N.sub.6O.sub.5 m/z 475.20 [M+H].sup.+; HPLC purity
100% (ret. time, 1.11 min); .sup.1H NMR (500 MHz, CHLOROFORH-d)
.delta..sub.H ppm 8.29 (d, J=16.3 Hz, 1H), 7.97 (d, J=16.2 Hz, 1H),
6.86 (s, 2H), 6.05 (dd, J=4.3, 1.7 Hz, 1H), 5.66-5.43 (m, 1H), 5.00
(ddd, J=19.3, 6.3, 3.2 Hz, 1H), 4.30 (s, 1H), 3.47-2.85 (m, 2H),
2.60 (ddd, J=38.8, 24.1, 13.5 Hz, 2H), 2.19 (ddd, J=14.7, 11.9, 7.1
Hz, 2H), 2.07-1.94 (m, 2H), 1.81 (dd, J=65.1, 6.9 Hz, 3H),
1.66-1.46 (m, 5H), 1.45-1.22 (m, 4H), 1.00 (d, J=6.4 Hz, 3H), 0.89
(dd, J=12.2, 6.6 Hz, 3H).
Step 4.
3-[3-({[(3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tet-
rahydro-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}(propan-2-yl)amino)cyclobuty-
l]-N-(2-amino-4/5-cyclobutylphenyl)propanamide
##STR00399##
[1057] TEA (0.54 ml, 3.90 mmol) was added to a solution of
3-[3-({[(3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tetrahydro-
-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}(propan-2-yl)amino)cyclobutyl]propa-
noic acid (308.46 mg, 0.65 mmol), 4-cyclobutylbenzene-1,2-diamine
(210.90 mg, 1.30 mmol), ethyl (2E)-cyano(hydroxyimino)ethanoate
(184.75 mg, 1.30 mmol), and EDC.HCl (249.21 mg, 1.30 mmol) in DCM
(15 ml) at RT and stirred for two hours. The reaction mixture was
concentrated in vacuo, then DCM (50 ml) was added. This was washed
with sat. NaHCO.sub.3 (2.times.30 ml). The aqueous was extracted
with DCM (50 ml). The combined organics was dried over
Na.sub.2SO.sub.4, filtered and evaporated. The product was purified
by silica gel column chromatography eluting with EtOAc, and then 7N
NH.sub.3 in MeOH:DCM (5:95), to give a grey oil, 468 mg (93%); MS
(ESI.sup.+) for C.sub.33H.sub.46N.sub.8O.sub.4 m/z 619.35
[M+H].sup.+; HPLC purity 80% (ret. time, 1.44 min).
Step 5.
9-[(3aR,4R,6R,6aR)-6-[({3-[2-(5-cyclobutyl-1H-1,3-benzodiazol-2-yl-
)ethyl]cyclobutyl}(propan-2-yl)amino)methyl]-2,2-dimethyl-tetrahydro-2H-fu-
ro[3,4-d][1,3]dioxol-4-yl]-9H-purin-6-amine
##STR00400##
[1059] AcOH (10 ml) was added to
3-[3-({[(3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tetrahydro-
-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}(propan-2-yl)amino)cyclobutyl]-N-(2-
-amino-4/5-cyclobutylphenyl)propanamide (468 mg, 0.61 mmol) and
heated to 65.degree. C. whilst stirring for 4 hours. The reaction
mixture was concentrated in vacuo, then dissolved in DCM (100 ml)
and washed with sat. NaHCO.sub.3 (2.times.80 ml), dried over
Na.sub.2SO.sub.4, filtered and evaporated. Purification by flash
silica gel chromatography (Biotage, Isolera, 25 g cartridge),
eluting with 3N ammonia in MeOH:DCM (O-1:9) gave the desired
product with purity approx 80%. Further purification by preparative
HPLC afforded the desired product as a grey oil, 120 mg (27%); MS
(ESI.sup.+) for C.sub.33H.sub.44N.sub.8O.sub.3 m/z 601 [M+H].sup.+;
HPLC purity 100% (ret. time, 1.43 min); .sup.1H NMR (500 MHz,
CHLOROFORH-d) .delta..sub.H ppm 8.62 (d, J=53.1 Hz, 6H), 8.57 (s,
2H), 8.25 (d, J=19.3 Hz, 1H), 7.90 (d, J=20.3 Hz, 1H), 7.50 (dd,
J=8.3, 3.9 Hz, 1H), 7.41 (d, J=4.5 Hz, 1H), 7.18-7.06 (m, 1H), 6.52
(d, J=96.3 Hz, 2H), 6.07 (dd, J=10.2, 1.3 Hz, 1H), 5.52-5.39 (m,
1H), 5.07 (dd, J=6.2, 3.4 Hz, 1H), 4.44 (td, J=9.3, 5.1 Hz, 1H),
3.59 (dq, J=17.4, 8.7 Hz, 1H), 3.36-3.10 (m, 2H), 3.08-2.92 (m,
2H), 2.87-2.72 (m, 2H), 2.43-2.27 (m, 2H), 2.24-1.65 (m, 10H), 1.57
(s, 4H), 1.37 (s, 3H), 1.10 (d, J=6.6 Hz, 3H), 0.91 (dd, J=8.9, 6.8
Hz, 3H).
Step 6.
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-[({3-[2-(5-cyclobutyl-1H-
-1,3-benzodiazol-2-yl)ethyl]cyclobutyl}(propan-2-yl)amino)methyl]oxolane-3-
,4-diol
##STR00401##
[1061] 12N HCl (24 mmol, 2 ml) was added dropwise to a solution of
9-[(3aR,4R,6R,6aR)-6-[({3-[2-(5-cyclobutyl-1H-1,3-benzodiazol-2-yl)ethyl]-
cyclobutyl}(propan-2-yl)amino)methyl]-2,2-dimethyl-tetrahydro-2H-furo[3,4--
d][1,3]dioxol-4-yl]-9H-purin-6-amine (120 mg, 0.164 mmol) in MeOH
(2 ml) at 0.degree. C. whilst stirring. This was then allowed to
warm to RT and continued for 6 hours. The reaction mixture was
cooled to 0.degree. C. and basified with 7N NH.sub.3 in MeOH (10
ml). This was then evaporated in vacuo. The crude product was
absorbed onto silica gel (1 ml), placed onto an isolute flash Si
cartridge (10 g) and purified, eluting with 7N NH.sub.3 in MeOH:DCM
(1:9) to give a white solid, 36 mg (38%); MS (ESI.sup.+) for
C.sub.30H.sub.40NO.sub.3 m/z 561.45 [M+H].sup.+; HPLC purity 100%
(ret. time, 1.13 min); .sup.1H NMR (500 MHz, CHLOROFORH-d)
.delta..sub.H ppm 8.29 (d, J=4.6 Hz, 1H), 8.20 (d, J=1.6 Hz, 1H),
7.50-7.18 (m, 2H), 7.06 (ddd, J=8.3, 4.6, 1.3 Hz, 1H), 6.01-5.90
(m, 1H), 4.73 (dd, J=9.8, 5.1 Hz, 1H), 4.26 (q, J=5.4 Hz, 1H),
4.14-4.03 (m, 1H), 3.60-3.15 (m, 2H), 3.07-2.86 (m, 2H), 2.84-2.67
(m, 3H), 2.42-2.31 (m, 2H), 2.25-2.11 (m, 4H), 2.10-1.95 (m, 2H),
1.92-1.74 (m, 4H), 1.57 (dd, J=12.2, 6.2 Hz, 1H), 1.02 (dd, J=6.6,
4.0 Hz, 3H), 0.95 (dd, J=6.6, 2.3 Hz, 3H).
Compound 68:
(2R,3R,4S,5R)-2-(6-Amino-9H-purin-9-yl)-5-{[(3-{2-[5-(1-methoxy-2-methylp-
ropan-2-yl)-1H-1,3-benzodiazol-2-yl]ethyl}cyclobutyl)(methyl)amino]methyl}-
oxolane-3,4-diol
Step 1: Benzyl
3-[3-({[(3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tetrahydro-
-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}amino)cyclobutyl]propanoate
##STR00402##
[1063] A suspension of
9-[(3aR,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyl-tetrahydro-2H-furo[3,4-d]-
[1,3]dioxol-4-yl]-9H-purin-6-amine (5.00 g, 16.3 mmol), benzyl
3-(3-oxocyclobutyl)-propanoate (4.17 g, 18.0 mmol) and acetic acid
(0.85 ml, 14.8 mmol) in DCE:iPrOH (7:2) (90 ml) was stirred at r.t.
for 2 h. Sodium triacetoxyborohydride (4.40 g, 20.8 mmol) was added
in portions and the mixture left to stir for 18 h at r.t. The
reaction mixture was quenched with 1M Na.sub.2CO.sub.3 solution (10
ml) and the product was extracted with DCM (3.times.30 ml). The
combined organic layers were dried over Na.sub.2SO.sub.4, filtered
and evaporated to dryness. Purification by silica gel flash column
chromatography eluting with 1% 7M NH.sub.3 in MeOH:99% DCM gave the
product as a yellow oil (5.25 g, 55%, 89% pure): MS (ESI.sup.+) for
C.sub.27H.sub.34N.sub.6O.sub.5 m/z 523.6 [M+H].sup.+; LC purity 89%
(ret. time, 1.60 min); .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta..sub.H 8.35 (d, J=6.0 Hz, 1H), 7.91 (s, 1H), 7.30-7.39 (m,
5H), 6.01 (dd, J=3.0 Hz, 1.6, 1H), 5.72 (br. s., 2H), 5.50 (dt,
J=6.4 Hz, 3.3, 1H), 5.10 (d, J=3.8 Hz, 2H), 4.98-5.04 (m, 1H),
4.31-4.38 (m, 1H), 2.97-3.34 (m, 1H), 2.72-2.85 (m, 2H), 2.22-2.35
(m, 3H), 2.14 (td, J=8.3, 4.3 Hz, 1H), 1.73-1.90 (m, 4H), 1.64-1.71
(m, 1H,), 1.62 (d, J=1.4 Hz, 3H), 1.39 (s, 3H), 1.10-1.27 (m,
1H).
Step 2: Benzyl
3-[3-({[(3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tetrahydro-
-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}(methyl)amino)cyclobutyl]propanoate
##STR00403##
[1065] Benzyl
3-[3-({[(3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tetrahydro-
-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}amino)cyclobutyl]propanoate
(3.2 g, 6.12 mmol) was dissolved in methanol (32 ml). Formaldehyde
in water (37%) (0.92 ml, 12.3 mmol) was added and stirred for 45
min before adding sodium cyanoborohydride (0.54 g, 8.57 mmol)
portionwise. The reaction was stirred for 2 h at r.t. before adding
water (1 ml) and evaporating off the solvent at r.t. The residue
was purified by chromatography with 7M ammonia in methanol/DCM to
give the desired compound as a yellow oil (mix of diastereomers)
(2.05 g, 62%, 82% pure): MS (ESI.sup.+) for
C.sub.28H.sub.36N.sub.6O.sub.5 m/z 537.6 [M+H].sup.+; LC purity 82%
(ret. time, 1.60 min); .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta..sub.H 8.32-8.38 (m, 1H), 7.91-7.97 (m, 1H), 7.32-7.39 (m,
5H), 6.05-6.10 (m, 1H), 5.61 (br. s., 2H), 5.53 (ddd, J=16.5, 6.4,
1.8 Hz, 1H), 5.11 (m, 2H), 4.93-5.00 (m, 1H), 4.32-4.40 (m, 1H),
2.50-2.88 (m, 1H), 2.37-2.49 (m, 2H), 2.21-2.30 (m, 2H), 2.10 (m,
3H), 1.91-2.04 (m, 1H), 1.73-1.79 (m, 2H), 1.64-1.72 (m, 2H),
1.56-1.63 (m, 4H), 1.41 (s, 3H), 1.15 (q, J=9.7 Hz, 1H).
Step
3:3-[3-({[(3aR,4R,6R,6aR)-6-(6-Amino-9H-purin-9-yl)-2,2-dimethyl-tetr-
ahydro-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}(methyl)amino)cyclobutyl]prop-
anoic acid
##STR00404##
[1067] Benzyl
3-[3-({[(3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tetrahydro-
-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}(methyl)amino)cyclobutyl]propanoate
(1.19 g, 2.22 mmol) was dissolved in ethanol (24 ml) and 10%
palladium on charcoal (50% wet paste) (0.24 g) added. The
suspension was stirred under an atmosphere of hydrogen for 18 h. It
was then filtered and the solid washed with ethanol (10 ml). As the
reaction was incomplete, further palladium on charcoal (0.21 g) was
added and the reaction continued under hydrogen for a further 24 h.
Filtered through double glass fibre, washed with ethanol and
evaporated to dryness to give the desired compound as a white foam
(mix of diastereomers) (0.85 g, 86%): MS (ESI.sup.+) for
C.sub.21H.sub.30N.sub.6O.sub.5 m/z 447.5 [M+H].sup.+; LC purity 86%
(ret. time, 1.08 min); .sup.1H NMR (500 MHz, d4-MeOD) .delta..sub.H
8.18-8.35 (m, 2H), 6.14-6.30 (m, 1H), 5.25-5.56 (m, 1H), 5.01-5.11
(m, 1H), 4.35-4.52 (m, 1H), 3.63-3.81 (m, 1H), 3.24-3.30 (m, 1H),
2.98-3.16 (m, 1H), 2.89 (ddd, J=17.2, 13.4, 3.7 Hz, 1H), 2.36 (m,
2H), 2.17-2.26 (m, 1H), 1.98-2.17 (m, 3H), 1.67-1.92 (m, 3H),
1.49-1.64 (m, 4H), 1.39 (s, 3H), 1.20-1.33 (m, 1H).
Step 4: Methyl 2-(4-fluorophenyl)-2-methylpropanoate
##STR00405##
[1069] Sodium hydride (60% suspension in mineral oil) (2.64 g, 66
mmol) was washed with heptanes (2.times.20 ml) and suspended in THF
(40 ml). A solution of methyl 2-(4-fluorophenyl)acetate (5.05 g, 30
mmol) in THF (10 ml) was added and stirred for 30 mins. Methyl
iodide (5.6 ml, 90 mmol) was added in 1 ml portions over 30 mins,
initially with cooling to 10.degree. C. then gentle warming to
50.degree. C. as the gas evolution ceased. After 4.5 h, water (50
ml) was added and the mixture extracted with EtOAc (2.times.50 ml).
The combined organic phases were washed with brine (30 ml) and
dried over MgSO.sub.4 before filtering and evaporating to dryness
to leave an orange oil (5.08 g, 79%, 91% pure by .sup.1H NMR): MS
(ESI.sup.+) for C.sub.11H.sub.13FO.sub.2 m/z 196.2 [M+H].sup.+; LC
purity 80% (ret. time, 1.94 min); .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta..sub.H 7.29-7.34 (m, 2H), 6.98-7.05 (m, 2H), 3.66 (s, 3H),
1.58 (s, 6H).
Step 5: 2-(4-Fluorophenyl)-2-methylpropan-1-ol
##STR00406##
[1071] Methyl 2-(4-fluorophenyl)-2-methylpropanoate (5.08 g, 26.9
mmol) was dissolved in THE (51 ml) and cooled to 0.degree. C.
before adding a solution of lithium aluminium hydride (1M in THF)
(38.8 ml, 38.8 mmol) dropwise over 30 mins. When the addition was
complete, the reaction was warmed to r.t. and stirred for 3 h.
After recooling on ice, water (1.35 ml) was added cautiously
followed by 15% NaOH in water (1.35 ml) and more water (4.05 ml).
The suspension was stirred at r.t. for 30 mins before the solid was
filtered off and washed with THE (2.times.30 ml). The solvent was
evaporated and the product purified by chromatography with
EtOAc/heptanes to give a clear oil (3.48 g, 80%): MS (ESI.sup.+)
for C.sub.10H.sub.13FO m/z 168.2 [M+H].sup.+; LC purity 94% (ret.
time, 1.76 min); .sup.1H NMR (500 MHz, CDCl.sub.3) .delta..sub.H
7.41-7.32 (m, 2H), 7.15-7.00 (m, 2H), 3.62 (d, J=6.4 Hz, 2H), 1.35
(s, 6H).
Step 6:1-Fluoro-4-(1-methoxy-2-methylpropan-2-yl)benzene
##STR00407##
[1073] Sodium hydride (1.664 g, 41.6 mmol, 60% dispersion in
mineral oil) was suspended in dry THE (18 ml) under N2 and
2-(4-fluorophenyl)-2-methylpropan-1-ol (3.500 g, 20.8 mmol) in dry
THE (18 ml) was slowly added to the suspension at 0.degree. C.
After complete addition the reaction was warmed to r.t. and left
for 1 h. Iodomethane (6.5 ml, 0.104 mmol) was slowly added at r.t.
and the reaction left for 3 h. The reaction was quenched by slow
addition of H.sub.2O (35 ml). The layers were separated and the
aqueous layer was extracted with EtOAc (3.times.35 ml). The
combined organic layers were dried over MgSO.sub.4, filtered and
concentrated in vacuo to give the crude product. The product was
purified by silica flash column chromatography using between 100%
heptane to 10% EtOAc:90% heptane as eluent to give the product as a
colourless oil (2.991 g, 79%): LC purity 98% (ret. time, 2.16 min);
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta..sub.H 7.40-7.32 (m, 2H),
7.11-6.96 (m, 2H), 3.39 (s, 2H), 3.33 (s, 3H), 1.33 (s, 6H).
Step
7:1-Fluoro-4-(1-methoxy-2-methylpropan-2-yl)-2-nitrobenzene
##STR00408##
[1075] Fluoro-4-(1-methoxy-2-methylpropan-2-yl)benzene (2.987 g,
16.4 mmol) was cooled in a salt ice/water bath to -20.degree. C.
and sulfuric acid (27 ml) was slowly added dropwise with stirring.
On addition of sulfuric acid the solution turned a bright orange.
Nitric acid (3 ml) was slowly added dropwise over 15-20 mins. On
addition of nitric acid the solution turned dark yellow/brown and
some white solid precipitated. The reaction was left for 30 mins
then poured over ice (450 g). The mixture was extracted with DCM
(2.times.225 ml) and the combined organic extracts were dried over
MgSO.sub.4, filtered and concentrated in vacuo to give the crude
product. The product was purified by silica flash column
chromatography using between 100% heptanes to 20% EtOAc:80%
heptanes to give the product as a yellow oil (2.239 g, 60%): LC
purity 96% (ret. time, 2.18 min); .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta..sub.H 8.08 (dd, J=7.1, 2.5 Hz, 1H), 7.67 (ddd, J=8.7, 4.1,
2.5 Hz, 1H), 7.23 (dd, J=10.6, 8.8 Hz, 1H), 3.41 (s, 2H), 3.33 (s,
3H), 1.37 (s, 6H).
Step 8:1-Azido-4-(1-methoxy-2-methylpropan-2-yl)-2-nitrobenzene
##STR00409##
[1077] Fluoro-4-(1-methoxy-2-methylpropan-2-yl)-2-nitrobenzene
(2.227 g, 9.80 mmol) was dissolved in DMF (25 ml) and sodium azide
(1.274 g, 19.6 mmol) was added at r.t. and the reaction stirred
overnight. The reaction was quenched with water (75 ml) and the
mixture was extracted with TBME (3.times.75 ml). The combined
organic layers were dried over MgSO.sub.4, filtered and
concentrated in vacuo to give the crude product. The product was
purified by silica flash column chromatography using between 100%
heptane to 15% EtOAc:85% heptane as eluent to give the product as a
yellow oil (2.301 g, 75%, 80% pure): LC purity 79% (ret. time, 2.15
min); .sup.1H NMR (500 MHz, CDCl.sub.3) .delta..sub.H 7.97 (d,
J=2.2 Hz, 1H), 7.66 (dd, J=8.5, 2.2 Hz, 1H), 7.33-7.22 (m, 1H),
3.40 (s, 2H), 3.33 (s, 3H), 1.36 (s, 6H).
Step 9:4-(1-Methoxy-2-methylpropan-2-yl)benzene-1,2-diamine
##STR00410##
[1079] Azido-4-(1-methoxy-2-methylpropan-2-yl)-2-nitrobenzene
(1.102 g, 3.52 mmol) was dissolved in EtOH (30 ml) and Pd/C (10%
wt.) (0.110 g, 10% wt.) was added. The reaction was purged 3 times
with N2 then 3 times with H2 and the reaction stirred at r.t.
overnight. The mixture was filtered through Celite and the filtrate
was concentrated in vacuo to give the crude product. The product
was purified by silica flash column chromatography using between
100% heptane to 100% EtOAc as eluent to give the product as a pale
brown oil which solidified into a dark orange solid (0.481 g, 63%,
90% pure): MS (ESI.sup.+) for C.sub.11H.sub.18N.sub.2O m/z 195.1
[M+H].sup.+; LC purity 87% (ret. time, 0.99 min); .sup.1H NMR (500
MHz, CDCl.sub.3) .delta..sub.H 6.65 (dd, J=11.2, 1.9 Hz, 2H), 6.58
(d, J=7.9 Hz, 1H), 3.26 (s, 2H), 3.24 (s, 3H), 1.20 (s, 6H).
Step 10:
3-[3-({[(3aR,4R,6R,6aR)-6-(6-Amino-9H-purin-9-yl)-2,2-dimethyl-te-
trahydro-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}(methyl)amino)cyclobutyl]-N-
-[2-amino-4-(1-methoxy-2-methylpropan-2-yl)phenyl]propanamide
##STR00411##
[1081]
3-[3-({[(3aR,4R,6R,6aR)-6-(6-Amino-9H-purin-9-yl)-2,2-dimethyl-tetr-
ahydro-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}(methyl)amino)cyclobutyl]prop-
anoic acid (0.620 g, 1.39 mmol),
4-(1-methoxy-2-methylpropan-2-yl)benzene-1,2-diamine (0.466 g, 2.08
mmol, 87% pure), EDC.HCl (0.532 g, 2.78 mmol) and OXYMA
(ethyl-cyano(hydroxyimino)acetate) (0.395 g, 2.78 mmol) were added
to a flask with a stirrer then purged with N2. Dry DCM (22 ml) and
dry Et.sub.3N (1.2 ml, 8.33 mmol) were added at r.t. and the
reaction left overnight. The reaction was quenched by the addition
of sat. NaHCO.sub.3 solution (25 ml) and the organic layer
separated. The aqueous layer was extracted with DCM (2.times.25 ml)
and the combined organic layers were dried over MgSO.sub.4,
filtered and concentrated in vacuo to give the crude product. The
product was purified by silica flash column chromatography using
first 100% EtOAc to elute the dianiline then between 100% DCM to
20% 2M NH.sub.3 in MeOH: 80% DCM as eluent to give the product as a
brown oil (1.081 g, quant.): MS (ESI.sup.+) for
C.sub.32H.sub.46N.sub.8O.sub.5 m/z 623.4 [M+H].sup.+; LC purity 98%
(ret. time, 1.36 min); .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta..sub.H 8.39 (d, J=7.8 Hz, 1H), 7.99 (s, 1H), 7.42-7.30 (m,
1H), 7.24-7.00 (m, 1H), 6.96-6.81 (m, 1H), 6.12 (d, J=12.6 Hz, 1H),
5.77 (d, J=6.7 Hz, 1H), 5.63 (d, J=5.0 Hz, 1H), 5.14-4.93 (m, 1H),
4.60-4.31 (m, 1H), 4.28-3.69 (m, 2H), 3.48-3.03 (m, 5H), 2.65-2.48
(m, 1H), 2.46-2.33 (m, 1H), 2.34-2.10 (m, 7H), 2.10-1.98 (m, 1H),
2.00-1.66 (m, 4H), 1.62 (d, J=15.7 Hz, 13H), 1.44 (d, J=6.5 Hz,
4H), 1.30 (t, J=3.3 Hz, 7H), 1.12 (d, J=52.3 Hz, 1H).
Step 11:
9-[(3aR,4R,6R,6aR)-6-{[(3-{2-[5-(1-Methoxy-2-methylpropan-2-yl)-1-
H-1,3-benzodiazol-2-yl]ethyl}cyclobutyl)(methyl)amino]methyl}-2,2-dimethyl-
-tetrahydro-2H-furo[3,4-d][1,3]dioxol-4-yl]-9H-purin-6-amine
##STR00412##
[1083]
3-[3-({[(3aR,4R,6R,6aR)-6-(6-Amino-9H-purin-9-yl)-2,2-dimethyl-tetr-
ahydro-2H-furo[3,4-d][1,3]dioxol-4-yl]methyl}(methyl)amino)cyclobutyl]-N-[-
2-amino-4-(1-methoxy-2-methylpropan-2-yl)phenyl]propanamide (1.036
g, 1.66 mmol) was dissolved in AcOH (17 ml) and heated to
50.degree. C. for 5 h. The reaction was concentrated in vacuo and
the residue was dissolved in DCM (75 ml) and sat. NaHCO.sub.3
solution (75 ml) was added. The organic layer was separated and the
aqueous layer was extracted with DCM (2.times.75 ml). The combined
organic layers were dried over MgSO.sub.4, filtered and
concentrated in vacuo to give the crude product as a dark orange
oil (0.850 g, 94%). The product was purified using neutral
prep-HPLC to give the product as a pale yellow oil (0.485 g, 47%,
88% pure): MS (ESI.sup.+) for C.sub.32H.sub.44N.sub.8O.sub.4 m/z
605.4 [M+H].sup.+; LC purity 88% (ret. time, 1.25 min); .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta..sub.H 10.44 (dd, J=207.7, 22.9 Hz,
1H), 8.41 (d, J=17.1 Hz, 1H), 8.16 (d, J=13.8 Hz, 1H), 7.89-7.58
(m, 1H), 7.54-7.29 (m, 2H), 6.15 (d, J=10.9 Hz, 1H), 5.92-5.56 (m,
3H), 4.98 (d, J=12.5 Hz, 1H), 4.76-4.39 (m, 1H), 3.47 (d, J=6.2 Hz,
2H), 3.33 (d, J=4.8 Hz, 3H), 2.98-2.43 (m, 4H), 2.42-2.00 (m, 6H),
1.99-1.88 (m, 2H), 1.77-1.66 (m, 1H), 1.64 (s, 3H), 1.61-1.48 (m,
1H), 1.44 (d, J=8.1 Hz, 3H), 1.40 (d, J=7.9 Hz, 6H), 1.28 (dd,
J=16.0, 9.0 Hz, 1H).
Step 12:
(2R,3R,4S,5R)-2-(6-Amino-9H-purin-9-yl)-5-{[(3-{2-[5-(1-methoxy-2-
-ethylpropan-2-yl)-1H-1,3-benzodiazol-2-yl]ethyl}cyclobutyl)(methyl)amino]-
methyl}oxolane-3,4-diol
##STR00413##
[1085]
9-[(3aR,4R,6R,6aR)-6-{[(3-{2-[5-(1-Methoxy-2-methylpropan-2-yl)-1H--
1,3-benzodiazol-2-yl]ethyl}cyclobutyl)(methyl)amino]methyl}-2,2-dimethyl-t-
etrahydro-2H-furo[3,4-d][1,3]dioxol-4-yl]-9H-purin-6-amine (0.485
g, 0.706 mmol, 88% pure) was dissolved in MeOH (20 ml) and conc.
HCl (4.9 ml, 10 vol) was added at r.t. and the reaction left for
2.5 h. The reaction was concentrated in vacuo and the residue
dissolved in the minimum amount of MeOH (.about.2 ml). The reaction
was quenched by addition of sat. NaHCO.sub.3 solution (10 ml) and
EtOAc (30 ml). The layers were separated and the aqueous layer was
extracted with EtOAc (2.times.30 ml). The combined organic layers
were dried over MgSO.sub.4, filtered and concentrated in vacuo to
give the crude product. The product was purified by silica flash
column chromatography using between 100% DCM and 20% 2M NH.sub.3 in
MeOH:80% DCM as eluent to give the product as a white foam (0.213
g, 53%): MS (ESI.sup.+) for C.sub.29H.sub.40N.sub.8O.sub.4 m/z
565.4 [M+H].sup.+; LC purity 100% (ret. time, 2.11 min) (7 min); 1H
NMR (500 MHz, d4-MeOD) .delta..sub.H 8.28 (d, J=4.1 Hz, 1H), 8.22
(d, J=3.2 Hz, 1H), 7.51 (s, 1H), 7.42 (dd, J=8.5, 2.1 Hz, 1H), 7.29
(dd, J=8.5, 1.5 Hz, 1H), 6.01 (t, J=3.9 Hz, 1H), 4.84-4.74 (m, 1H),
4.41-4.29 (m, 1H), 4.29-4.19 (m, 1H), 3.48 (s, 2H), 3.30 (s, 3H),
3.14-2.98 (m, 1H), 2.99-2.89 (m, 1H), 2.90-2.73 (m, 2H), 2.39 (s,
3H), 2.37-2.30 (m, 1H), 2.28-2.10 (m, 2H), 2.08-1.86 (m, 4H),
1.70-1.51 (m, 1H), 1.38 (s, 6H).
Compound 69:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino-
)methyl)cyclopentane-1,2-diol
##STR00414## ##STR00415##
[1086]
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,-
3S)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl-
)amino)methyl)cyclopentane-1,2-diol
[1087] .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.06 (s, 1H),
7.47 (brs, 1H), 7.38 (d, J=8.0 HZ, 1H), 7.27 (d, J=8.0 Hz, 1H),
7.19 (s, 1H), 6.60 (s, 1H), 4.32 (s, 1H), 3.88 (s, 1H), 2.80 (brs,
2H), 2.68 (brs, 1H), 2.49-2.10 (m, 9H), 1.90 (brs, 2H), 1.62-1.49
(m, 3H), 1.35 (s, 9H) ppm; ESI-MS (m/z): 532.3 [M+1].sup.+.
benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrol-
o[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-
methyl)amino)cyclobutyl)propanoate
[1088] A solution of
7-((3aR,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]-
dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
(20 g, 43.91 mmol), benzyl 3-(3-oxocyclobutyl)propanoate (12.2 g,
52.69 mmol) and HOAc (15 mL) in DCE (200 mL) was stirred at
30.degree. C. for 3 h. NaBH(OAc)3 (18.6 g, 87.81 mmol) was added
and the reaction mixture was stirred at 30.degree. C. for another 1
h. The mixture was washed with water (100 mL.times.2) and brine
(100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated.
The residue was purified by silica gel column chromatography using
EA:DCM:MeOH=10:10:1 as eluent to afford the desired compound (21 g,
yield: 65%, cis/trans=52/47) as a yellow solid. .sup.1H NMR (500
MHz, MeOD): & 8.16 (brs, 1H), 7.36-7.29 (m, 5H), 7.22 (d, J=3.5
Hz, 1H), 7.15 (d, J=8.5 Hz, 1H), 6.66 (brs, 1H), 6.54 (d, J=2.0 Hz,
1H), 6.43 (dd, J=8.5 and 2.0 Hz, 1H), 6.20 (brs, 1H), 5.41-5.39 (m,
1H), 5.08 (d, J=3.0 Hz, 1H), 4.99-4.95 (m, 1H), 4.66 (s, 2H),
4.30-4.25 (m, 1H), 3.83 (s, 3H), 3.76 (s, 3H), 3.38-3.35 (m, 0.5H),
3.11-3.06 (m, 0.5H), 2.92-2.82 (m, 2H), 2.29-2.18 (m, 3H),
2.12-2.05 (m, 0.5H), 1.90-1.68 (m, 4H), 1.63-1.61 (m, 1H), 1.60 (s,
3H), 1.38 (s, 3H), 1.35-1.28 (m, 0.5H), 1.25 (t, J=6.5 Hz, 2H),
1.15-1.12 (m, 0.5H) ppm; ESI-MS (negative mode, m/z): 670.3
[M-1].sup.+.
[1089] benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(isopropyl)amino)cyclobutyl)propanoate
[1090] A mixture of benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)amino)cyclobutyl)propanoate (4 g, 5.95 mmol), 2-iodopropane (6 g,
35.72 mmol) and K.sub.2CO.sub.3 (2.5 g, 17.86 mmol) in CH.sub.3CN
(50 mL) was stirred at reflux for 2 days. The mixture was cooled to
rt, filtered and the filtered cake was washed with CH.sub.3CN (20
mL). The filtrate was concentrated and the residue was purified by
Combi-Flash (80 g silica gel, start EA:DCM:MeOH=10:10:0 to 10:10:1
by gradient, 60 mL/min, 40 min, 2.4 L total solvent volume) to
afford the desired compound (3 g, yield: 71%) as a yellow solid.
.sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.15 (s, 1H), 7.38-7.29
(m, 5H), 7.19 (d, J=4.0 Hz, 1H), 7.14 (d, J=8.0 Hz, 1H), 6.65 (d,
J=3.0 Hz, 1H), 6.53 (d, J=2.0 Hz, 1H), 6.41 (dd, J=8.5 and 2.0 Hz,
1H), 6.20 (d, J=2.5 Hz, 1H), 5.36-5.32 (m, 1H), 5.09 (s, 1H), 5.07
(s, 1H), 4.94-4.89 (m, 3H), 4.65 (s, 2H), 4.17-4.14 (m, 1H), 3.82
(s, 3H), 3.75 (s, 3H), 3.41-3.35 (m, 0.5H), 3.04-2.95 (m, 0.5H),
2.94-2.86 (m, 1H), 2.72-2.52 (m, 2H), 2.25 (t, J=7.5 Hz, 1H), 2.21
(t, J=8.0 Hz, 1H), 2.05-1.90 (m, 2H), 1.90-1.82 (m, 0.5H),
1.76-1.70 (m, 0.5H), 1.65-1.55 (m, 5H), 1.42-1.34 (m, 4H), 0.95 (d,
J=6.5 Hz, 3H), 0.83-0.80 (m, 3H) ppm; ESI-MS (m/z): 714.4
[M+1].sup.+.
3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d-
]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)-
(isopropyl)amino)cyclobutyl)propanoic acid
[1091] To a solution of benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(isopropyl)amino)cyclobutyl)propanoate (2.7 g, 3.78 mmol) in
THF/MeOH (15 mL/15 mL) was added a solution of LiOH.H.sub.2O (1.6
g, 37.82 mmol) in water (5 mL). The mixture was stirred at
30.degree. C. for 2 h. The volatiles were removed under reduced
pressure. To the residue was diluted with water (10 mL) and
extracted with EA (15 mL.times.2). The suspension water layers were
adjusted to pH=3-4 with 1 N HCl solution and extracted with EA (30
mL.times.3). The combined organic layers were washed with brine (50
mL). The organic phase was dried over Na.sub.2SO.sub.4, filtered
and concentrated to afford the desired compound as a yellow solid
(2.9 g). .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.20 (s, 1H),
7.92 (s, 0.5H), 7.38-7.32 (m, 3H), 7.28-7.23 (m, 1.5H), 7.14 (d,
J=8.5 Hz, 1H), 6.68 (brs, 1H), 6.56 (d, J=2.0 Hz, 1H), 6.44 (d,
J=8.5 Hz, 1H), 6.25 (s, 1H), 5.51-5.47 (m, 1H), 5.18-5.13 (m, 1H),
4.66 (s, 2H), 4.61 (s, 1H), 4.43-4.40 (m, 1H), 3.96-3.90 (m, 0.5H),
3.85 (s, 3H), 3.77 (s, 3H), 3.65-3.58 (m, 0.5H), 3.50-3.40 (m, 2H),
2.46-2.36 (m, 1H), 2.20-2.00 (m, 4H), 1.98-1.70 (m, 2.5H),
1.70-1.58 (m, 4.5H), 1.40 (s, 3H), 1.18 (d, J=5.0 Hz, 3H), 0.90 (t,
J=6.0 Hz, 3H) ppm; ESI-MS (m/z): 624.3 [M+1].sup.+.
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimeth-
oxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofu-
ro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)propanamide
[1092] To a solution of
3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(isopropyl)amino)cyclobutyl)propanoic acid (2.7 g, 4.33 mmol),
4-(trifluoromethoxy)benzene-1,2-diamine (1.23 g, 6.49 mmol), HATU
(2.5 g, 6.49 mmol) and HOAT (0.88 g, 6.49 mmol) in DCM (30 mL) was
added TEA (1.8 mL, 12.99 mmol). The mixture was stirred at rt for 2
h. The mixture was added DCM (70 mL) and washed with water (20
mL.times.2) and brine (50 mL). The organic phase was dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by Combi-Flash (80 g silica gel, start EA:DCM:MeOH=10:10:0
to 10:10:2 by gradient, 60 mL/min, 40 min, 2.4 L total solvent
volume) to afford to afford the desired compound (2.2 g, yield: 67%
(two steps) as a brown solid. .sup.1H NMR (500 MHz, MeOD):
.delta..sub.H 8.21 (s, 1H), 7.28-7.12 (m, 3H), 6.73 (brs, 1H), 6.69
(brs, 1H), 6.56-6.52 (m, 2H), 6.42 (dd, J=8.0 and 2.5 Hz, 1H), 6.26
(d, J=1.5 Hz, 1H), 5.48 (d, J=6.0 Hz, 1H), 5.20-5.16 (m, 1H), 4.66
(s, 2H), 4.46-4.42 (m, 1H), 4.08-3.98 (m, 0.5H), 3.84 (s, 3H), 3.76
(s, 3H), 3.75-3.69 (m, 0.5H), 3.60-3.38 (m, 2H), 2.60-2.30 (m, 3H),
1.92-1.86 (m, 1H), 1.80-1.70 (m, 1H), 1.59 (d, J=3.5 Hz, 3H), 1.40
(s, 3H), 1.25 (t, J=7.5 Hz, 3H),), 1.00-0.80 (m, 2H) ppm; ESI-MS
(m/z): 798.3 [M+1].sup.+.
N-(2,4-dimethoxybenzyl)-7-((3aR,4R,6R,6aR)-6-((isopropyl(3-(2-(5-(trifluor-
omethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)-2,2-dime-
thyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-am-
ine
[1093] A solution of
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimet-
hoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)propanamide
[1094] (2.2 g, 2.5 mmol) in HOAc (20 mL) was stirred at 65.degree.
C. for 5 h. The mixture was cooled to rt and concentrated. The
residue was dissolved in DCM (50 mL), washed with 15%
Na.sub.2CO.sub.3 solution (20 mL.times.2), water (20 mL) and brine
(30 mL). The organic phase was dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was purified by Combi-Flash
(80 g silica gel, start EA:DCM:MeOH=10:10:0 to 10:10:2 by gradient,
60 mL/min, 35 min, 2.1 L total solvent volume) to afford to afford
the desired compound (1.4 g) as a brown solid, which was separated
by chiral HPLC to afford cis-isomer (600 mg, yield: 28%) and the
trans isomer (480 mg, yield: 22%).
[1095] cis-isomer: .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.17
(s, 1H), 7.52 (d, J=9.0 Hz, 1H), 7.39 (s, 1H), 7.20 (d, J=4.0 Hz,
1H), 7.12 (d, J=8.0 Hz, 1H), 6.68 (brs, 1H), 6.49 (d, J=2.5 Hz,
1H), 6.39 (dd, J=8.5 and 2.0 Hz, 1H), 6.19 (d, J=2.0 Hz, 1H), 5.35
(dd, J=6.0 and 2.0 Hz, 1H), 4.68-4.60 (m, 2H), 4.18-4.13 (m, 1H),
3.78 (s, 3H), 3.73 (s, 3H), 3.10-3.02 (m, 1H), 2.95-2.88 (m, 1H),
2.78-2.72 (m, 2H), 2.69-2.57 (m, 2H), 2.12-2.02 (m, 2H), 1.86-1.76
(m, 2H), 1.57 (s, 3H), 1.52-1.40 (m, 2H), 1.37 (s, 3H), 0.96 (d,
J=6.5 Hz, 3H), 0.82 (d, J=6.5 Hz, 3H) ppm; ESI-MS (m/z): 780.4
[M+1].sup.+.
[1096] trans-isomer: .sup.1H NMR (500 MHz, MeOD): .delta..sub.H
8.15 (s, 1H), 7.52 (d, J=9.0 Hz, 1H), 7.40 (s, 1H), 7.20 (d, J=3.5
Hz, 1H), 7.12 (d, J=8.0 Hz, 1H), 6.68 (brs, 1H), 6.51 (d, J=2.0 Hz,
1H), 6.40 (dd, J=8.0 and 2.5 Hz, 1H), 6.20 (d, J=2.5 Hz, 1H), 5.35
(dd, J=6.0 and 2.0 Hz, 1H), 4.64 (s, 2H), 4.20-4.16 (m, 1H), 3.82
(s, 3H), 3.73 (s, 3H), 3.50-3.42 (m, 1H), 2.95-2.90 (m, 1H), 2.80
(t, J=6.0 Hz, 2H), 2.75-2.58 (m, 2H), 2.12-1.95 (m, 4H), 1.75-1.65
(m, 2H), 1.58 (s, 3H), 1.38 (s, 3H), 0.97 (d, J=6.5 Hz, 3H), 0.83
(d, J=6.5 Hz, 3H) ppm; ESI-MS (m/z): 780.4 [M+1].sup.+.
Compound 70:
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino-
)methyl)cyclopentane-1,2-diol
##STR00416##
[1097] methyl
3-((1R,3s)-3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrr-
olo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dio-
xol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoate
[1098] A solution of methyl
3-((1R,3s)-3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrr-
olo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dio-
xol-4-yl)methyl)amino)cyclobutyl)propanoate (1.85 g, 3.12 mmol) and
NaBH.sub.3CN (590 mg, 9.36 mmol) in MeOH (25 mL) was adjusted pH=6
with AcOH, then formaldehyde (936 mg, 31.2 mmol) added. The
reaction was stirred at 25.degree. C. overnight. The reaction was
quenched with sat. NaHCO.sub.3 (5 mL), evaporated, added water (10
mL), extracted with DCM (150 mL.times.3), washed with brine (80
mL), dried and concentrated. The residue was purified by SGC to
obtain the desired compound (1.85 g, Yield 97%) as white solid.
.sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.12 (s, 1H), 7.22 (d,
J=3.5 Hz, 1H), 7.14 (d, J=8.5 Hz, 1H), 6.64 (d, J=3.0 Hz, 1H), 6.55
(d, J=2.5 Hz, 1H), 6.44 (dd, J=8.5 and 2.5 Hz, 1H), 5.01-4.98 (m,
2H), 4.65 (s, 2H), 4.60-4.58 (m, 1H), 4.30 (s, 1H), 3.84 (s, 3H),
3.76 (s, 3H), 3.66 (s, 3H), 3.44-3.38 (m, 1H), 2.80-2.73 (m, 2H),
2.48-2.43 (m, 4H), 2.32 (t, J=7.5 Hz, 2H), 2.20-2.16 (m, 4H),
1.95-1.94 (m, 2H), 1.82-1.81 (m, 2H), 1.56 (s, 3H), 1.31 (s, 3H)
ppm; ESI-MS (m/z): 608.3 [M+1].sup.+.
3-((1R,3s)-3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrro-
lo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]diox-
ol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoic acid
[1099] A solution of methyl
3-((1R,3s)-3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrr-
olo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dio-
xol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoate
[1100] (1.85 g, 3.04 mmol) and LiOH (382 mg, 15.24 mmol) in
THF/MeOH/H.sub.2O (1:1:1, 30 mL) were stirred at 50.degree. C. for
2 h. The reaction was concentrated to obtain the desired compound
(2.25 g, salt, purity 85%) as white solid. The crude was used
directly next step without further purification. .sup.1H NMR (500
MHz, MeOD): .delta..sub.H 8.11 (s, 1H), 7.92 (s, 1H), 7.30 (d,
J=3.0 Hz, 1H), 7.14 (d, J=8.0 Hz, 1H), 6.67 (d, J=3.0 Hz, 1H), 6.54
(d, J=2.0 Hz, 1H), 6.44 (dd, J=10.0 and 2.5 Hz, 1H), 5.05-5.03 (m,
2H), 4.73 (d, J=6.0 Hz, 1H), 4.65 (s, 2H), 3.90-3.85 (m, 1H), 3.85
(s, 3H), 3.77 (s, 3H), 3.30-3.18 (m, 2H), 2.82 (s, 3H), 2.65-2.55
(m, 1H), 2.53-2.46 (m, 3H), 2.27-2.20 (m, 3H), 2.12-2.11 (m, 2H),
1.83-1.82 (m, 2H), 1.57 (s, 3H), 1.32 (s, 3H) ppm; ESI-MS (m/z):
594.3 [M+1].sup.+.
N-(2-amino-4-(tert-butyl)phenyl)-3-((1R,3s)-3-((((3aR,4R,6R,6aS)-6-(4-((2,-
4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetr-
ahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)p-
ropanamide
[1101] A solution of
3-((1R,3s)-3-((((3aR,4R,6R,6aS)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrr-
olo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-3aH-cyclopenta[d][1,3]dio-
xol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoic acid (2.25 g,
3.8 mmol) HOAt (680 mg, 5 mmol) and HATU (1.9 g, 5 mmol) in DCM (60
mL) was stirred at room temperature for 1 h, then
4-tert-butylbenzendiamine (656 mg, 4 mmol) and TEA (1.21 g, 12
mmol) in DCM (3 mL) were added dropwise. The reaction was stirred
at rt overnight. The reaction was added water (20 mL) and DCM (60
mL), extracted with DCM (60 mL.times.2), washed with brine (10 mL),
dried and concentrated. The residue was purified by SGC to obtain
the desired compound (1.1 g, Yield 46%) as yellowish solid. .sup.1H
NMR (500 MHz, MeOD): .delta..sub.H 8.11 (s, 1H), 7.21 (d, J=4.0 Hz,
1H), 7.13 (d, J=8.5 Hz, 1H), 6.98 (d, J=8.5 Hz, 1H), 6.92 (d, J=2.0
Hz, 1H), 6.77 (dd, J=8.0 and 1.5 Hz, 1H), 6.64 (d, J=3.5 Hz, 1H),
6.54 (d, J=2.0 Hz, 1H), 6.43 (dd, J=8.0 and 1.5 Hz, 1H), 5.02-4.99
(m, 2H), 4.65-4.61 (m, 3H), 3.83 (s, 3H), 3.75 (s, 3H), 3.60-3.52
(m, 1H), 2.97-2.83 (m, 2H), 2.58 (s, 3H), 2.54-2.38 (m, 4H),
2.33-2.29 (m, 2H), 2.18-2.04 (m, 3H), 1.94-1.91 (m, 2H), 1.55 (s,
3H), 1.30 (s, 3H), 1.25 (s, 9H) ppm; ESI-MS (m/z): 740.5
[M+1].sup.+.
7-((3aS,4R,6R,6aR)-6-((((1s,3R)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-
-yl)ethyl)cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cycl-
openta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimi-
din-4-amine
[1102] A solution of
N-(2-amino-4-(tert-butyl)phenyl)-3-((1R,3s)-3-((((3aR,4R,6R,6aS)-6-(4-((2-
,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltet-
rahydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)-
propanamide (1.1 g, 1.49 mmol) in AcOH (8 mL) were heated to
65.degree. C. for 3 h. The reaction was evaporated, dissolved in
MeOH (5 mL) adjusted pH=8 with saturated NaHCO.sub.3 solution,
concentrated and purified by Prep-TLC to obtain the desired
compound (620 mg, 58%) as white solid. .sup.1H NMR (500 MHz, MeOD):
.delta..sub.H 8.10 (s, 1H), 7.48 (brs, 1H), 7.38 (brs, 1H), 7.28
(d, J=8.5 Hz, 1H), 7.21 (d, J=3.0 Hz, 1H), 7.13 (d, J=8.5 Hz, 1H),
6.62 (brs, 1H), 6.53 (s, 1H), 6.42 (d, J=8.5 Hz, 1H), 4.98-4.90 (m,
2H), 4.64 (s, 2H), 4.50 (brs, 1H), 3.83 (s, 3H), 3.75 (s, 3H),
3.01-2.98 (m, 1H), 2.83 (d, J=7.5 Hz, 1H), 2.44-2.34 (m, 4H), 2.16
(s, 3H), 2.12-1.96 (m, 6H), 1.84 (t, J=7.5 Hz, 2H), 1.53 (s, 3H),
1.36 (s, 9H), 1.28 (s, 3H) ppm; ESI-MS (m/z): 722.4
[M+1].sup.+.
(1R,2S,3R,5R)-3-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3-(-
2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)-
methyl)cyclopentane-1,2-diol
[1103] A solution of
7-((3aS,4R,6R,6aR)-6-((((1s,3R)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol--
2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydro-3aH-cyc-
lopenta[d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrim-
idin-4-amine (620 mg, 0.86 mmol) in TFA (5 mL, 90%) was stirred at
25.degree. C. for 1 hour. The reaction was concentrated to dryness,
dissolved in MeOH (5 mL) and adjusted pH=8 with saturated
K.sub.2CO.sub.3 solution. The mixture was stirred at rt for 0.5 h.
Then the reaction was concentrated to obtain the residue. The
residue was purified by Prep-HPLC to obtain the desired compound
(330 mg, Yield 73%) as white solid. .sup.1H NMR (500 MHz, MeOD):
.delta..sub.H 8.07 (s, 1H), 7.49 (d, J=1.5 Hz, 1H), 7.40 (d, J=11.0
Hz, 1H), 7.29 (dd, J=11.0 and 2.5 Hz, 1H), 7.21 (d, J=5.0 Hz, 1H),
6.60 (d, J=4.0 Hz, 1H), 4.92-4.89 (m, 1H), 4.33 (dd, J=9.5 and 8.5
Hz, 1H), 3.90 (d, J=5.5 Hz, 1H), 3.03-2.99 (m, 1H), 2.85 (d, J=9.5
Hz, 1H), 2.50-2.22 (m, 4H), 2.16 (s, 3H), 2.15-1.98 (m, 5H), 1.88
(t, J=10.0 Hz, 2H), 1.68-1.59 (m, 1H), 1.37 (s, 9H) ppm; ESI-MS
(m/z): 532.3 [M+1].sup.+.
Compound 71:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((((1r,3S)-3-(2-(5-(tert-butyl)-
-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)tetrahydro-
furan-3,4-diol
##STR00417##
[1105] A mixture of
cis-9-((3aR,4R,6R,6aR)-6-((((1r,3S)-3-(2-(5-(tert-butyl)-1H-benzo[d]imida-
zol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydrofuro-
[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine (920 mg, 1.60 mmol) in 3
M HCl/MeOH (20 mL) was stirred at 35.degree. C. for 2 h and
evaporated to dryness. The residue was dissolved in MeOH (15 mL)
and saturated K.sub.2CO.sub.3 solution was added to adjust the
solution to pH 8. Then, the mixture was stirred for 5 min and
filtered. The filtrate was concentrated and the crude was purified
by Prep-HPLC to obtain the target (400 mg, yield: 47%) as a white
solid. .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.27 (s, 1H),
8.20 (s, 1H), 7.47 (s, 1H), 7.47 (s, 1H), 7.40-7.37 (m, 1H),
7.29-7.26 (m, 1H), 5.98 (d, J=4.5 Hz, 1H), 4.69 (t, J=4.5 Hz, 1H),
4.24-4.20 (m, 1H), 4.18-4.15 (m, 1H), 2.81-2.76 (m, 2H), 2.75-2.69
(m, 1H), 2.67-2.62 (m, 2H), 2.25-2.18 (m, 1H), 2.14 (s, 3H),
1.90-1.85 (m, 3H), 1.49-1.41 (m, 2H), 1.36 (s, 9H) ppm; ESI-MS
(m/z): 535.3 [M+1].sup.+.
Compound 72:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((((1s,3R)-3-(2-(5-(tert-butyl)-
-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)tetrahydro-
furan-3,4-diol
##STR00418##
[1107] A mixture of
trans-9-((3aR,4R,6R,6aR)-6-((((1r,3S)-3-(2-(5-(tert-butyl)-1H-benzo[d]imi-
dazol-2-yl)ethyl)cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydrofu-
ro[3,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine (420 mg, 0.73 mmol) in
3 M HCl/MeOH (20 mL) was stirred at 35.degree. C. for 2 h and
evaporated to dryness. The residue was dissolved in MeOH (15 mL)
and saturated K.sub.2CO.sub.3 solution was added to adjust the
solution to pH 8. Then, the mixture was stirred for 5 min and
filtered. The filtrate was concentrated and the crude was purified
by Prep-HPLC to obtain the target (198 mg, yield: 51%) as a white
solid. .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.28 (s, 1H),
8.19 (s, 1H), 7.48 (s, 1H), 7.38 (d, J=8.0 Hz, 1H), 7.28-7.26 (m,
1H), 5.98 (d, J=4.0 Hz, 1H), 4.69 (t, J=5.5 Hz, 1H), 4.23 (t, J=5.0
Hz, 1H), 4.19-4.16 (m, 1H), 3.06-3.03 (m, 1H), 2.80 (t, J=7.5 Hz,
2H), 2.68-2.64 (m, 2H), 2.16 (s, 3H), 2.09-1.95 (m, 5H), 1.85-1.80
(m, 2H), 1.36 (s, 9H) ppm; ESI-MS (m/z): 535.3 [M+1].sup.+.
Benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetra-
hydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(ethyl)amino)cyclobutyl)propanoate
[1108] To a mixture of benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoate (3.76
g, 7.2 mmol) and NaBH.sub.3CN (5.9 g, 93.6 mmol) in MeOH (40 mL)
was added AcOH to adjust to pH=6. Then, 40% MeCHO (8.7 mL, 122.5
mmol) was added and the mixture was stirred at 30.degree. C. for
1.5 h. Water (15 mL) was added and the mixture was concentrated in
vacuo. Then, the mixture was extracted with DCM (30 mL.times.3).
The combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. The crude was purified
by SGC (DCM:MeOH=100: 1-20:1) to obtain the target (2.6 g, yield:
66%) as a white solid. .sup.1H NMR (500 MHz, MeOD): & 8.26 (s,
1H), 8.23 (m, 1H), 7.37-7.28 (m, 5H), 6.24 (s, 1H), 5.52-5.49 (m,
1H), 5.10-5.06 (m, 3H), 4.41-4.40 (m, 1H), 3.20-2.90 (m, 2H),
2.80-2.60 (m, 2H), 2.12-1.77 (m, 4H), 1.74-1.60 (m, 3H), 1.39 (s,
3H), 1.26-1.22 (m, 3H), 0.94-0.89 (m, 3H) ppm; ESI-MS (m/z): 551.3
[M+1].sup.+.
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofu-
ro[3,4-d][1,3]dioxol-4-yl)methyl)(ethyl)amino)cyclobutyl)propanoic
acid
[1109] To a solution of Benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(ethyl)amino)cyclobutyl)propanoate
(2.6 g, 4.73 mmol) in MeOH (40 mL) was added 10% Pd/C (2.3 g) and
the mixture was stirred under H2 atmosphere at 50.degree. C. for 20
h. The mixture was filtered and the filtrate was concentrated to
obtain the target (2 g, yield: 92%) as a white solid. .sup.1H NMR
(500 MHz, MeOD): .delta..sub.H 8.27 (s, 1H), 8.25 (s, 1H), 6.30 (s,
1H), 5.52-5.49 (m, 1H), 5.14-5.12 (m, 1H), 4.51-4.47 (m, 1H),
3.43-3.36 (m, 2H), 3.22-3.15 (m, 1H), 2.94-2.89 (m, 2H), 2.28-2.05
(m, 4H), 1.96-1.80 (m, 2H), 1.74-1.69 (m, 1H), 1.65-1.59 (m, 5H),
1.41-1.36 (m, 4H), 1.03-0.99 (m, 3H) ppm; ESI-MS (m/z): 461.3
[M+1].sup.+.
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-pur-
in-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(ethyl)a-
mino)cyclobutyl)propanamide
[1110] To a solution of
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(ethyl)amino)cyclobutyl)propanoic
acid (2 g, 4.35 mmol), HOAT (768 mg, 5.65 mmol), HATU (2.2 g, 5.65
mmol) and TEA (3 mL, 21.3 mmol) in DCM (40 mL) was added
4-tert-butylbenzene-1, 2-diamine (785 mg, 4.79 mmol) and the
mixture was stirred at rt for 2 h. Water (15 mL) was added and the
mixture was extracted with DCM (30 mL.times.2). The combined
organic phase was washed with H.sub.2O (20 mL.times.2). The
combined organic layers were dried over Na.sub.2SO.sub.4, filtered
and concentrated. The crude was purified by SGC
(DCM:MeOH=70:1-20:1) to obtain the target (1.6 g, yield: 61%) as a
white solid. .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.28-8.27
(m, 1H), 8.24-8.23 (m, 1H), 7.10-6.92 (m, 2H), 6.81-6.76 (m, 1H),
6.22 (s, 1H), 5.54-5.52 (m, 1H), 5.03 (s, 1H), 4.36 (s, 1H),
3.03-2.50 (m, 5H), 2.32-2.26 (m, 2H), 2.16-1.80 (m, 4H), 1.73-1.69
(m, 2H), 1.59 (s, 3H), 1.39 (s, 3H), 1.28-1.24 (m, 9H), 0.94-0.85
(m, 3H) ppm; ESI-MS (m/z): 607.3 [M+1].sup.+.
9-((3aR,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethy-
l)cyclobutyl)(ethyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]di-
oxol-4-yl)-9H-purin-6-amine
[1111] A solution of
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-pu-
rin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(ethyl)-
amino)cyclobutyl)propanamide
[1112] (1.6 g, 2.64 mmol) in AcOH (20 mL) was stirred at 65.degree.
C. for 15 h. The solution was concentrated in vacuo and diluted
with DCM (30 mL). The mixture was washed with saturated NaHCO.sub.3
solution (20 mL.times.2) and brine (20 mL.times.1). The combined
organic phase was dried over Na.sub.2SO.sub.4, filtered and
concentrated to obtain the target 1.5 g (yield: 97%) as a white
solid. .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.28-8.26 (m,
1H), 8.22 (s, 1H), 7.48 (s, 1H), 7.40-7.38 (m, 1H), 7.30-7.27 (m,
1H), 6.20-6.18 (m, 1H), 5.52-5.49 (m, 1H), 5.02-4.98 (m, 1H),
4.34-4.31 (m, 1H), 2.95-2.92 (m, 1H), 2.79-2.68 (m, 4H), 2.56-2.50
(m, 2H), 2.09-1.81 (m, 5H), 1.71-1.63 (m, 1H), 1.58 (s, 3H), 1.38
(s, 3H), 1.36 (s, 9H), 1.35-1.28 (m, 1H), 0.89-0.85 (m, 3H) ppm;
ESI-MS (m/z): 589.3 [M+1].sup.+.
[1113] A mixture of
9-((3aR,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)eth-
yl)cyclobutyl)(ethyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]d-
ioxol-4-yl)-9H-purin-6-amine (1.35 g, 2.30 mmol) in 3 M HCl/MeOH
(20 mL) was stirred at 35.degree. C. for 2 h and evaporated to
dryness. The residue was dissolved in MeOH (15 mL) and saturated
K.sub.2CO.sub.3 solution was added to adjust the solution to pH 8.
Then, the mixture was stirred for 5 min and filtered. The filtrate
was concentrated and the crude was separated by chiral HPLC and
purified by Prep-HPLC to obtain the cis product (280 mg, total
yield: 22%) and trans product (150 mg, total yield: 12%) as a white
solids.
Compound 73:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((((1r,3S)-3-(2-(5-(tert-butyl)-
-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(ethyl)amino)methyl)tetrahydrof-
uran-3,4-diol
##STR00419##
[1115] Cis-isomer: .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.27
(s, 1H), 8.20 (s, 1H), 7.47 (s, 1H), 7.39-7.37 (m, 1H), 7.29-7.26
(m, 1H), 5.97 (d, J=4.5 Hz, 1H), 4.67 (t, J=5.0 Hz, 1H), 4.24 (t,
J=5.5 Hz, 1H), 4.18-4.14 (m, 1H), 3.06-3.03 (m, 1H), 2.91-2.81 (m,
2H), 2.79-2.75 (m, 2H), 2.64-2.58 (m, 2H), 2.25-2.19 (m, 1H),
1.89-1.86 (m, 3H), 1.52-1.47 (m, 2H), 1.36 (s, 9H), 0.98 (t, J=7.0
Hz, 3H) ppm; ESI-MS (m/z): 549.3 [M+1].sup.+.
Compound 74:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((((1s,3R)-3-(2-(5-(tert-butyl)-
-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(ethyl)amino)methyl)tetrahydrof-
uran-3,4-diol
##STR00420##
[1117] Trans-isomer: .sup.1H NMR (500 MHz, MeOD): .delta..sub.H
8.28 (s, 1H), 8.20 (s, 1H), 7.48 (s, 1H), 7.39-7.37 (m, 1H),
7.29-7.26 (m, 1H), 5.97 (d, J=4.0 Hz, 1H), 4.67 (t, J=5.0 Hz, 1H),
4.24 (t, J=6.0 Hz, 1H), 4.18-4.14 (m, 1H), 3.42-3.35 (m, 1H),
2.91-2.78 (m, 4H), 2.64-2.58 (m, 2H), 2.10-2.07 (m, 3H), 1.99-1.96
(m, 2H), 1.85-1.79 (m, 2H), 1.35 (m, 9H), 0.98 (t, J=6.5 Hz, 3H)
ppm; ESI-MS (m/z): 549.3 [M+1].sup.+.
Compound 76:
(2R,3R,4S,5R)-2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((isopropyl((-
1s,3R)-3-(2-(5-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobuty-
l)amino)methyl)tetrahydrofuran-3,4-diol
##STR00421##
[1119] A solution of
trans-N-(2,4-dimethoxybenzyl)-7-((3aR,4R,6R,6aR)-6-((isopropyl(3-(2-(5-(t-
rifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)-2-
,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimid-
in-4-amine (480 mg, 0.62 mmol) in 90% TFA (5 mL) was stirred at
30.degree. C. for 2 h. The volatiles were removed under reduced
pressure. To the residue was added MeOH (6 mL) and adjusted to
pH=9-10 with NH.sub.3.H.sub.2O. The mixture was stirred at rt for
30 min and concentrated. The residue was purified by Prep-HPLC to
afford the desired compound (182 mg, yield: 50%) as a white solid.
.sup.1H NMR (400 MHz, MeOD): .delta..sub.H 8.09 (s, 1H), 7.52 (d,
J=8.8 Hz, 1H), 7.39 (s, 1H), 7.27 (d, J=3.6 Hz, 1H), 7.12 (d, J=8.8
Hz, 1H), 6.64 (d, J=3.2 Hz, 1H), 6.11 (d, J=4.0 Hz, 1H), 4.44 (t,
J=4.8 Hz, 1H), 4.12 (t, J=6.0 Hz, 1H), 4.06-4.01 (m, 1H), 3.62-3.53
(m, 1H), 3.10-3.00 (m, 1H), 2.92-2.65 (m, 4H), 2.25-2.15 (m, 2H),
2.10-1.98 (m, 3H), 1.85-1.76 (m, 2H), 1.03 (d, J=6.4 Hz, 3H), 0.98
(d, J=6.4 Hz, 3H) ppm; .sup.19F NMR (400 MHz, MeOD): .delta.-59.80
ppm; ESI-MS (m/z): 590.3 [M+1].sup.+.
Compounds 77 and 78
##STR00422## ##STR00423##
[1120] Compound 78:
(2R,3R,4S,5R)-2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(2,2,2-triflu-
oroethyl)amino)methyl)tetrahydrofuran-3,4-diol
##STR00424##
[1122] .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.09 (s, 1H),
7.48 (brs, 1H), 7.40-7.37 (m, 1H), 7.28 (dd, J=10.5 and 1.5 Hz,
1H), 7.21 (d, J=4.5 Hz, 1H), 6.65 (d, J=4.5 Hz, 1H), 6.11 (d, J=5.0
Hz, 1H), 4.44 (t, J=6.0 Hz, 1H), 4.13-4.08 (m, 2H), 3.30-3.15 (m,
3H), 3.09-3.03 (m, 1H), 2.97-2.90 (m, 1H), 2.78 (t, J=9.0 Hz, 2H),
2.28-2.20 (m, 2H), 1.92-1.78 (m, 3H), 1.55-1.45 (m, 2H), 1.37 (s,
9H) ppm; LC-MS (m/z): 602.3 [M+1].sup.+.
Compound 77:
(2R,3R,4S,5R)-2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(2,2,2-triflu-
oroethyl)amino)methyl)tetrahydrofuran-3,4-diol
##STR00425##
[1124] .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.09 (s, 1H),
7.48 (brs, 1H), 7.39 (d, J=10.5 Hz, 1H), 7.28 (dd, J=10.5 and 2.0
Hz, 1H), 7.21 (d, J=5.0 Hz, 1H), 6.64 (d, J=4.5 Hz, 1H), 6.12 (d,
J=6.0 Hz, 1H), 4.45 (t, J=6.0 Hz, 1H), 4.14-4.09 (m, 2H), 3.68-3.60
(m, 1H), 3.30-3.15 (m, 2H), 3.11-3.04 (m, 1H), 2.97-2.90 (m, 1H),
2.80 (t, J=9.5 Hz, 2H), 2.12-2.03 (m, 3H), 2.00-1.90 (m, 2H),
1.90-1.80 (m, 2H), 1.36 (s, 9H) ppm; LC-MS (m/z): 602.3
[M+1].sup.+.
Benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetra-
hydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoat-
e
[1125] To a mixture of benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)amino)cyclobutyl)propanoate (3.76
g, 7.2 mmol) and NaBH.sub.3CN (5.9 g, 93.6 mmol) in MeOH (40 mL)
was added AcOH to adjust to pH=6. Then, 37% HCHO (8.7 mL, 122.4
mmol) was added and the mixture was stirred at 30.degree. C. for
1.5 h. Water (15 mL) was added and the mixture was concentrated in
vacuo. Then, the mixture was extracted with DCM (30 mL.times.3).
The combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. The crude was purified
by SGC (DCM:MeOH=100: 1-20:1) to obtain the target (2.4 g, yield:
67%) as a white solid. .sup.1H NMR (500 MHz, MeOD): .delta..sub.H
8.27 (s, 1H), 8.22 (d, J=2.5 Hz, 1H), 7.35-7.30 (m, 5H), 6.22 (s,
1H), 5.55-5.52 (m, 1H), 5.09 (s, 2H), 5.04-5.01 (m, 1H), 4.40-4.38
(m, 1H), 2.76-2.65 (m, 3H), 2.29-2.22 (m, 2H), 2.18 (s, 3H),
2.11-1.95 (m, 2H), 1.78-1.71 (m, 2H), 1.64-1.61 (m, 2H), 1.59 (s,
3H), 1.41-1.39 (m, 1H), 1.38 (s, 3H) ppm; ESI-MS (m/z): 537.3
[M+1].sup.+.
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofu-
ro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoic
acid
[1126] To a solution of benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoate
[1127] (2.4 g, 4.48 mmol) in MeOH (40 mL) was added 10% Pd/C (2.3
g) and the mixture was stirred under H2 atmosphere at 50.degree. C.
for 15 h. The mixture was filtered and the filtrate was
concentrated to obtain the target (1.9 g, yield: 95%) as a white
solid. .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.27 (s, 1H),
8.24 (s, 1H), 6.27 (s, 1H), 5.54-5.52 (m, 1H), 5.09-5.07 (m, 1H),
4.50-4.47 (m, 1H), 3.17-3.07 (m, 2H), 3.02-2.90 (m, 1H), 2.39-2.35
(m, 3H), 2.31-2.05 (m, 4H), 1.91-1.70 (m, 2H), 1.64-1.51 (m, 5H),
1.39 (s, 3H), 1.23-1.15 (m, 1H) ppm; ESI-MS (m/z): 447.2
[M+1].sup.+.
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-pur-
in-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)-
amino)cyclobutyl)propanamide
[1128] To a solution of
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoic
acid
[1129] (1.9 g, 4.26 mmol), HOAT (753 mg, 5.54 mmol), HATU (2.1 g,
5.54 mmol) and TEA (3 mL, 21.3 mmol) in DCM (40 mL) was added
4-tert-butylbenzene-1, 2-diamine (769 mg, 4.69 mmol) and the
mixture was stirred at rt for 2 h. Water (15 mL) was added and the
mixture was extracted with DCM (30 mL.times.2). The combined
organic phase was washed with H.sub.2O (20 mL.times.2). The
combined organic layers were dried over Na.sub.2SO.sub.4, filtered
and concentrated. The crude was purified by SGC (DCM:MeOH=70:
1-20:1) to obtain the target (1.8 g, yield: 72%) as a white solid.
.sup.1H NMR (500 MHz, MeOD): .delta. 8.28 (s, 1H), 8.23 (d, J=3.0
Hz, 1H), 7.10-6.92 (m, 2H), 6.81-6.76 (m, 1H), 6.21 (s, 1H), 5.56
(s, 1H), 5.49 (d, J=3.0 Hz, 1H), 5.01 (s, 1H), 4.36 (s, 1H),
2.69-2.49 (m, 3H), 2.31-2.27 (m, 2H), 2.13-2.00 (m, 5H), 1.88-1.81
(m, 2H), 1.75-1.65 (m, 2H), 1.60 (s, 3H), 1.41-1.35 (m, 4H),
1.28-1.25 (m, 9H) ppm; ESI-MS (m/z): 593.4 [M+1].sup.+.
9-((3aR,4R,6R,6aR)-6-((((1r,3S)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-
-yl)ethyl)cyclobutyl)(methyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3,4--
d][1,3]dioxol-4-yl)-9H-purin-6-amine
[1130] A solution of
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-pu-
rin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl-
)amino)cyclobutyl)propanamide (1.8 g, 3.04 mmol) in AcOH (20 mL)
was stirred at 65.degree. C. for 15 h. The solution was
concentrated in vacuo and diluted with DCM (30 mL). The mixture was
washed with saturated NaHCO.sub.3 solution (20 mL.times.2) and
brine (20 mL.times.1). The combined organic phase was dried over
Na.sub.2SO.sub.4, filtered and concentrated to obtain 1.7 g (yield:
97%) of the desired product.
[1131] The product was separated by chiral HPLC to obtain 920 mg of
cis-isomer and 420 mg of trans-isomer.
[1132] Cis-Isomer: .sup.1H NMR (500 MHz, MeOD): .delta. 8.27 (s,
1H), 8.21 (s, 1H), 7.48 (s, 1H), 7.40-7.38 (m, 1H), 7.29-7.26 (m,
1H), 6.19 (d, J=2.5 Hz, 1H), 5.55-5.52 (m, 1H), 4.99-4.97 (m, 1H),
4.35-4.31 (m, 1H), 2.77-2.73 (m, 2H), 2.62-2.46 (m, 3H), 2.10-2.01
(m, 4H), 1.84-1.81 (m, 3H), 1.58 (s, 3H), 1.38-1.36 (m, 12H),
1.19-1.14 (m, 3H) ppm; ESI-MS (m/z): 575.3 [M+1].sup.+.
[1133] Trans-isomer: .sup.1H NMR (500 MHz, MeOD): .delta..sub.H
8.28 (s, 1H), 8.21 (s, 1H), 7.48 (s, 1H), 7.40-7.38 (m, 1H),
7.30-7.27 (m, 1H), 6.19 (d, J=1.5 Hz, 1H), 5.55-5.52 (m, 1H),
5.01-4.98 (m, 1H), 4.36-4.34 (m, 1H), 2.96-2.92 (m, 2H), 2.77 (t,
J=7.5 Hz, 2H), 2.58-2.50 (m, 2H), 2.09 (s, 3H), 2.04-1.90 (m, 4H),
1.82-1.79 (m, 1H), 1.70-1.66 (m, 2H), 1.59 (s, 3H), 1.38 (s, 3H),
1.36 (s, 9H) ppm; ESI-MS (m/z): 575.3 [M+1].sup.+.
Compound 87:
(2R,3R,4S,5R)-2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((isopropyl((-
1r,3S)-3-(2-(5-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobuty-
l)amino)methyl)tetrahydrofuran-3,4-diol
##STR00426##
[1135] A solution of
cis-N-(2,4-dimethoxybenzyl)-7-((3aR,4R,6R,6aR)-6-((isopropyl(3-(2-(5-(tri-
fluoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)-2,2-
-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-
-4-amine (600 mg, 0.77 mmol) in 90% TFA (5 mL) was stirred at
30.degree. C. for 2 h. The volatiles were removed under reduced
pressure. To the residue was added MeOH (6 mL) and adjusted to
pH=9-10 with NH.sub.3.H.sub.2O. The mixture was stirred at rt for
30 min and concentrated. The residue was purified by Prep-HPLC to
afford the desired compound (260 mg, yield: 57%) as a white solid.
.sup.1H NMR (400 MHz, MeOD): .delta..sub.H 8.09 (s, 1H), 7.52 (d,
J=8.8 Hz, 1H), 7.39 (s, 1H), 7.27 (d, J=4.0 Hz, 1H), 7.12 (dd,
J=8.4 and 0.8 Hz, 1H), 6.64 (d, J=4.0 Hz, 1H), 6.12 (d, J=4.4 Hz,
1H), 4.43 (t, J=5.2 Hz, 1H), 4.11 (t, J=5.2 Hz, 1H), 4.05-4.01 (m,
1H), 3.20-3.10 (m, 1H), 3.08-3.00 (m, 1H), 2.88-2.65 (m, 4H),
2.25-2.15 (m, 2H), 1.92-1.82 (m, 3H), 1.65-1.55 (m, 2H), 1.02 (d,
J=6.4 Hz, 3H), 0.98 (d, J=6.4 Hz, 3H) ppm; .sup.19F NMR (400 MHz,
MeOD): .delta. -59.80 ppm; ESI-MS (m/z): 590.3 [M+1].sup.+.
Compounds 90 and 75
benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrol-
o[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-
methyl)amino)cyclobutyl)propanoate
[1136] To a solution of
7-((3aR,4R,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]-
dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
(2.5 g, 5.49 mmol), benzyl 3-(3-oxocyclobutyl)propanoate (1.66 g,
7.14 mmol) and HOAc (329 mg, 5.49 mmol) in DCE (40 mL) was added
NaB(OAc).sub.3H (2.33 g, 11 mmol) in one portion. Then the
resulting reaction mixture was stirred at rt overnight. Saturated
aqueous NaHCO.sub.3 (40 mL) was added to quench the reaction, then
was extracted with DCM (50 mL.times.3), dried over anhydrous
Na.sub.2SO.sub.4 and concentrated. The crude was purified by SGC
(DCM:MeOH=100:1 to 50:1) to afford the desired compound (2.3 g,
yield: 64%) as a white solid. .sup.1H NMR (500 MHz, MeOD):
.delta..sub.H 8.14 (d, J=2.5 Hz, 1H), 7.35-7.28 (m, 5H), 7.20 (d,
J=4.0 Hz, 1H), 7.13 (d, J=8.0 Hz, 1H), 6.64 (d, J=3.0 Hz, 1H), 6.53
(d, J=2.0 Hz, 1H), 6.42 (d, J=8.5 Hz, 1H), 6.18 (d, J=2.5 Hz, 1H),
5.40-5.39 (m, 1H), 5.08-5.07 (m, 2H), 4.96-4.94 (m, 1H), 4.64 (s,
2H), 4.26-4.24 (m, 1H), 3.82 (s, 3H), 3.75 (s, 3H), 3.10-3.05 (m,
0.55H), 2.86-2.82 (m, 2H), 2.26-2.20 (m, 3H), 2.10-1.59 (m, 5H),
1.58 (s, 3H), 1.37 (s, 3H), 1.35-1.25 (m, 0.6H), 1.15-1.08 (m,
0.5H) ppm; LC-MS (m/z): 672.4 [M+1].sup.+.
benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrol-
o[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-
methyl)(isopropyl)amino)cyclobutyl)propanoate
[1137] Benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)amino)cyclobutyl)propanoate (2.3 g, 3.43 mmol) was mixed with
K.sub.2CO.sub.3 (3.3 g, 24 mmol) and 2-iodopropane (5.8 g, 34.3
mmol) in MeCN (25 mL) in a sealed tube, then heated to 95.degree.
C. with stirring for 20 h. The reaction mixture was filtered and
rinsed with MeCN (30 mL), the filtrate was evaporated in vacuo to
afford the desired compound (2.1 g, yield: 88%) as a white solid,
which was used for next step without further purification. .sup.1H
NMR (500 MHz, MeOD): .delta..sub.H 8.13 (s, 1H), 7.35-7.29 (m, 5H),
7.18 (d, J=3.0 Hz, 1H), 7.13 (d, J=8.5 Hz, 1H), 6.64 (d, J=3.0 Hz,
1H), 6.53 (d, J=2.5 Hz, 1H), 6.41 (dd, J=8.5 and 2.5 Hz, 1H), 6.18
(d, J=2.5 Hz, 1H), 5.33-5.32 (m, 1H), 5.08-5.06 (m, 2H), 4.90-4.89
(m, 1H), 4.64 (s, 2H), 4.15-4.14 (m, 1H), 3.83 (s, 3H), 3.75 (s,
3H), 3.37-3.36 (m, 0.43H), 3.01-2.98 (m, 0.59H), 2.92-2.88 (m, 1H),
2.70-2.40 (m, 3H), 2.24-2.18 (m, 2H), 2.10-1.80 (m, 3H), 1.75-1.69
(m, 2H), 1.61-1.52 (m, 5H), 1.38 (s, 3H), 0.94 (d, J=6.5 Hz, 3H),
0.81-0.79 (m, 3H) ppm; LC-MS (m/z): 714.0 [M+1].sup.+.
3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d-
]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)-
(isopropyl)amino)cyclobutyl)propanoic acid
[1138] Benzyl
3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(isopropyl)amino)cyclobutyl)propanoate
[1139] (1.5 g, 2.1 mmol) was dissolved in MeOH (25 mL), Pd/C (10%
on carbon, 70% water, 742 mg) was added and the resultant mixture
was stirred at 35.degree. C. under 1 atm H2 overnight. The mixture
was then filtered and rinsed with MeOH (15 mL.times.3), the
filtrate was evaporated in vacuo to afford the desired compound
(1.12 g, yield: 85%) as a white solid, which was used for next step
without further purification. .sup.1H NMR (500 MHz, MeOD):
.delta..sub.H 8.18 (s, 1H), 7.90 (s, 1H), 7.35-7.32 (m, 1H),
7.23-7.22 (m, 1H), 7.12 (d, J=8.5 Hz, 1H), 6.66 (brs, 1H), 6.54 (d,
J=2.0 Hz, 1H), 6.43-6.41 (m, 1H), 6.24-6.23 (m, 1H), 5.47-5.46 (m,
1H), 5.13-5.12 (m, 2H), 4.64 (s, 2H), 4.41-4.37 (m, 1H), 3.84 (s,
3H), 3.75 (s, 3H), 3.64-3.58 (0.6H), 3.46-3.40 (m, 1.7H), 2.45-1.60
(m, 9H), 1.57 (s, 3H), 1.38 (s, 3H), 1.11 (d, J=7.0 Hz, 3H), 0.87
(d, J=6.5 Hz, 3H) ppm; LC-MS (m/z): 624.0 [M+1].sup.+.
N-(2-amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimeth-
oxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofu-
ro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)propanamide
[1140] To a solution of
3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-dimethoxybenzyl)amino)-7H-pyrrolo[2,3--
d]pyrimidin-7-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl-
)(isopropyl)amino)cyclobutyl)propanoic acid (1.1 g, 1.76 mmol),
HATU (1 g, 2.64 mmol), HOAT (359 mg, 2.64 mmol) in DCM (30 mL) was
added a solution of 4-tert-butylbenzene-1,2-diamine (433 mg, 2.64
mmol) and TEA (533 mg, 5.28 mmol) in DCM (10 mL) dropwise, then the
resultant reaction mixture was stirred at rt overnight. After
diluted with DCM (50 mL), the mixture was washed water (30
mL.times.3), dried and concentrated. The crude was purified by SGC
(DCM:MeOH=100:1 to 40:1) to afford the desired compound (680 mg,
yield: 50%) as a white solid. .sup.1H NMR (500 MHz, MeOD):
.delta..sub.H 8.16 (s, 1H), 7.19 (d, J=3.5 Hz, 1H), 7.14-7.10 (m,
1.7H), 6.99-6.97 (m, 0.6H), 6.92 (s, 0.6H), 6.77 (dd, J=8.0, 2.0
Hz, 1H), 6.66-6.65 (m, 1H), 6.54-6.53 (m, 1H), 6.42 (d, J=8.0 Hz,
1H), 6.20 (d, J=2.0 Hz, 1H), 5.36-5.35 (m, 1H), 4.96-4.95 (m, 1H),
4.65 (s, 2H), 3.83 (s, 3H), 3.75 (s, 3H), 3.17-2.73 (m, 4H),
2.33-1.71 (m, 8H), 1.58 (s, 3H), 1.53-1.50 (m, 1H), 1.38 (s, 3H),
1.28 (s, 9H), 1.0 (d, J=5.5 Hz, 3H), 0.84 (d, J=5.0 Hz, 3H) ppm;
LC-MS (m/z): 770.0 [M+1].sup.+.
7-((3aR,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethy-
l)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,-
3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
[1141]
N-(2-Amino-4-(tert-butyl)phenyl)-3-(3-((((3aR,4R,6R,6aR)-6-(4-((2,4-
-dimethoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetra-
hydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(isopropyl)amino)cyclobutyl)propan-
amide (670 mg, 0.87 mmol) was dissolved in HOAc (8 mL) and then
heated to 65.degree. C. with stirring overnight. Solvent was
removed in vacuo. The residue was dissolved in DCM (60 mL), then
washed with NaHCO.sub.3 (sat. 20 mL) and water (20 mL), the organic
phase was dried and concentrated. The crude was purified by
Prep-TLC (DCM:MeOH=10:1) to afford the desired compound (470 mg,
yield: 73%) as a white solid, which was then separated by Chiral
HPLC to afford cis (243 mg) and trans isomers (180 mg) as a white
solids.
[1142] Cis-isomer: .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.15
(s, 1H), 7.47 (s, 1H), 7.38-7.37 (m, 1H), 7.27 (dd, J=8.5 and 1.5
Hz, 1H), 7.17 (d, J=4.0 Hz, 1H), 7.10 (d, J=8.5 Hz, 1H), 6.65 (d,
J=3.0 Hz, 1H), 6.49 (d, J=2.5 Hz, 1H), 6.38 (dd, J=8.0 and 2.5 Hz,
1H), 6.18 (d, J=2.5 Hz, 1H), 5.33 (dd, J=6.5 and 2.5 Hz, 1H),
4.92-4.91 (m, 1H), 4.63 (s, 2H), 4.16-4.15 (m, 1H), 3.78 (s, 3H),
3.72 (s, 3H), 3.08-3.07 (m, 1H), 2.97-2.96 (m, 1H), 2.72-2.66 (m,
4H), 2.09-2.03 (m, 2H), 1.82-1.78 (m, 3H), 1.56 (s, 3H), 1.48-1.40
(m, 2H), 1.38 (s, 3H), 1.36 (s, 9H), 0.95 (d, J=7.0 Hz, 3H), 0.81
(d, J=6.5 Hz, 3H) ppm; LC-MS (m/z): 752.0 [M+1].sup.+.
[1143] Trans-isomer: .sup.1H NMR (500 MHz, MeOD): .delta..sub.H
8.14 (s, 1H), 7.47 (s, 1H), 7.38-7.37 (m, 1H), 7.27 (dd, J=8.5 and
1.5 Hz, 1H), 7.18 (d, J=3.5 Hz, 1H), 7.10 (d, J=8.0 Hz, 1H), 6.64
(d, J=4.0 Hz, 1H), 6.50 (d, J=2.0 Hz, 1H), 6.39-6.37 (m, 1H), 6.18
(d, J=2.0 Hz, 1H), 5.33 (dd, J=6.0 and 2.5 Hz, 1H), 4.90 (dd, J=6.5
and 3.5 Hz, 1H), 4.62 (s, 2H), 4.17-4.15 (m, 1H), 3.80 (s, 3H),
3.71 (s, 3H), 3.44-3.43 (m, 1H), 2.93-2.90 (m, 1H), 2.77-2.68 (m,
3H), 2.62-2.60 (m, 1H), 2.05-1.93 (m, 5H), 1.68-1.67 (m, 2H), 1.56
(s, 3H), 1.36 (s, 12H), 0.95 (d, J=7.0 Hz, 3H), 0.81 (d, J=6.5 Hz,
3H) ppm; LC-MS (m/z): 752.0 [M+1].sup.+.
Compound 90:
(2R,3R,4S,5R)-2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1r,3S)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)am-
ino)methyl)tetrahydrofuran-3,4-diol
##STR00427##
[1145] To a mixture of TFA (2.7 mL) and water (0.3 mL) was added
cis
isomer-7-((3aR,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-
-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3-
,4-d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin--
4-amine (235 mg, 0.31 mmol). The solution was allowed to stand at
35.degree. C. for 2 h and evaporated to dryness. The residue was
co-evaporated with methanol twice. Then the residue was dissolved
in MeOH (20 mL). The solution was neutralized by K.sub.2CO.sub.3
(124 mg, dissolved in 1 mL of H2O) with stirring at rt for 1 h.
Solvent was removed in vacuo, then the crude was purified by
Prep-HPLC to afford the desired compound (90 mg, yield: 51%) as a
white solid. .sup.1H NMR (500 MHz, MeOD): .delta. 8.08 (s, 1H),
7.47 (s, 1H), 7.39-7.37 (m, 1H), 7.28 (d, J=2.5 Hz, 1H), 7.26 (d,
J=4.5 Hz, 1H), 6.62 (d, J=4.5 Hz, 1H), 6.10 (d, J=5.5 Hz, 1H),
4.43-4.41 (m, 1H), 4.12-4.09 (m, 1H), 4.04-4.01 (m, 1H), 3.15-3.13
(m, 1H), 3.04-3.01 (m, 1H), 2.86-2.68 (m, 4H), 2.21-2.17 (m, 2H),
1.88-1.85 (m, 3H), 1.60-1.57 (m, 2H), 1.36 (s, 9H), 1.01 (d, J=8.0
Hz, 3H), 0.97 (d, J=8.0 Hz, 3H) ppm; LC-MS (m/z): 562.5
[M+1].sup.+.
Compound 75:
(2R,3R,4S,5R)-2-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((((1s,3R)-3--
(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)am-
ino)methyl)tetrahydrofuran-3,4-diol
##STR00428##
[1147] To a mixture of TFA (2.7 mL) and water (0.3 mL) was added
trans isomer
7-((3aR,4R,6R,6aR)-6-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-
-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3-
,4-d][1,3]dioxol-4-yl)-N-(2,4-dimethoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin--
4-amine (175 mg, 0.23 mmol). The solution was allowed to stand at
35.degree. C. for 2 h and evaporated to dryness. The residue was
co-evaporated with methanol twice. Then the residue was dissolved
in MeOH (20 mL). The solution was neutralized by K.sub.2CO.sub.3
(97 mg, dissolved in 1 mL of H2O) with stirring at rt for 1 h.
Solvent was removed in vacuo, then the crude was purified by
Prep-HPLC to afford the desired compound (95 mg, yield: 71%) as a
white solid. .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.09 (s,
1H), 7.49 (s, 1H), 7.41-7.39 (m, 1H), 7.30-7.27 (m, 2H), 6.64 (d,
J=4.0 Hz, 1H), 6.12 (d, J=5.5 Hz, 1H), 4.45-4.42 (m, 1H), 4.13-4.11
(m, 1H), 4.05-4.03 (m, 1H), 3.58-3.54 (m, 1H), 3.06-3.02 (m, 1H),
2.89-2.80 (m, 3H), 2.74-2.70 (m, 1H), 2.20-2.17 (m, 2H), 2.03-1.99
(m, 3H), 1.84-1.81 (m, 3H), 1.38 (s, 9H), 1.03 (d, J=8.5 Hz, 3H),
0.98 (d, J=8.0 Hz, 3H) ppm; LC-MS (m/z): 562.5 [M+1].sup.+.
Compound 96:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((methyl((1r,3S)-3-(2-(5-(trifl-
uoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrah-
ydrofuran-3,4-diol
##STR00429##
[1148]
N-(2-amino-4-(trifluoromethoxy)phenyl)-3-(3-((((3aR,4R,6R,6aR)-6-(6-
-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)me-
thyl)(methyl)amino)cyclobutyl)propanamide
[1149] To a solution of
3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyltetrahydrof-
uro[3,4-d][1,3]dioxol-4-yl)methyl)(methyl)amino)cyclobutyl)propanoic
acid (1.2 g, 2.91 mmol), HATU (2.17 g, 5.83 mmol) and HOAT (0.91 g,
5.83 mmol) in DCM (17 mL) were added 4-(trifluoromethoxy)
benzene-1,2-diamine (1.1 g, 5.83 mmol) and TEA (2.05 mL, 14.56
mmol). The mixture was stirred at rt overnight. The mixture was
diluted with DCM (50 mL) and washed with water (15 mL.times.3) and
brine (30 mL). The organic phase was dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was purified by Combi-Flash
(40 g silica gel, start EA:DCM:MeOH=10:10:0 to 10:10:8 by gradient,
40 mL/min, 50 min, 2.0 L total solvent volume) to afford the
desired compound (1.0 g, yield: 60%) as a yellow solid. .sup.1H NMR
(500 MHz, MeOD): .delta..sub.H 8.29 (s, 1H), 8.25-8.24 (m, 1H),
7.20-7.13 (m, 1H), 6.95-6.85 (m, 0.4H), 6.73 (brs, 0.8H), 6.54 (d,
J=7.5 Hz, 0.8H), 6.22 (d, J=2.0 Hz, 1H), 5.60-5.55 (m, 1H),
5.03-5.00 (m, 1H), 4.40-4.35 (m, 1H), 3.40-3.35 (m, 0.3H),
3.00-2.92 (m, 0.7H), 2.70-2.47 (m, 3H), 2.36-2.28 (m, 2H),
2.20-1.80 (m, 6H), 1.76-1.66 (m, 2H), 1.60 (s, 3H), 1.40 (s, 3H),
1.26-1.16 (m, 1H) ppm; ESI-MS (m/z): 621.3 [M+1].sup.+.
9-((3aR,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(5-(trifluoromethoxy)-1H-b-
enzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydrofuro[3,4-d][1-
,3]dioxol-4-yl)-9H-purin-6-amine
[1150] A solution of
N-(2-amino-4-(trifluoromethoxy)phenyl)-3-(3-((((3aR,4R,6R,6aR)-6-(6-amino-
-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)(-
methyl)amino)cyclobutyl)propanamide (1.0 g, 1.61 mmol) in HOAc (10
mL) was stirred at 65.degree. C. overnight. The mixture was cooled
to rt and concentrated. The residue was dissolved in DCM (50 mL),
washed with saturation NaHCO.sub.3 solution (10 mL.times.2), water
(20 mL) and brine (30 mL). The residue was separated by chiral HPLC
to afford the cis isomer (460 mg, yield: 47%) and the trans isomer
(220 mg, yield: 23%).
[1151] cis-isomer: .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.28
(s, 1H), 8.23 (s, 1H), 7.53 (d, J=9.0 Hz, 1H), 7.40 (s, 1H), 7.13
(dd, J=8.5 and 1.0 Hz, 1H), 6.21 (d, J=2.0 Hz, 1H), 5.55 (dd,
J=6.5, 2.5 Hz, 1H), 5.01 (q, J=3.5 Hz, 1H), 4.33-4.37 (m, 1H),
2.81-2.78 (m, 2H), 2.66-2.58 (m, 2H), 2.53-2.49 (m, 1H), 2.13-2.03
(m, 5H), 1.86-1.84 (m, 3H), 1.59 (s, 3H), 1.43-1.39 (m, 4H),
1.22-1.17 (m, 1H) ppm; LC-MS (m/z): 603.3 [M+1].sup.+.
[1152] trans-isomer: .sup.1H NMR (500 MHz, MeOD): .delta..sub.H
8.29 (s, 1H), 8.22 (s, 1H), 7.52 (brs, 1H), 7.40 (s, 1H), 7.13 (d,
J=9.0 Hz, 1H), 6.21 (d, J=2.0 Hz, 1H), 5.55 (dd, J=6.5 and 2.0 Hz,
1H), 5.01 (q, J=3.0 Hz, 1H), 4.38-4.34 (m, 1H), 2.96-2.92 (m, 1H),
2.84-2.81 (m, 2H), 2.59-2.49 (m, 2H), 2.10 (s, 3H), 2.05-1.91 (m,
4H), 1.85-1.79 (m, 1H), 1.73-1.66 (m, 2H), 1.60 (s, 3H), 1.39 (s,
3H) ppm; LC-MS (m/z): 603.3 [M+1].sup.+.
Compound 96
[1153] A solution of
cis-9-((3aR,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(5-(trifluoromethoxy)-
-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydrofuro[3,4-
-d][1,3]dioxol-4-yl)-9H-purin-6-amine (460 mg, 0.77 mmol) in 1 N
HCl/MeOH (10 mL) was stirred at 30.degree. C. for 4 h. The
volatiles were removed under reduced pressure. To the residue was
added MeOH (10 mL) and adjusted to pH=10.about.11 with
NH.sub.3.H.sub.2O. The mixture was stirred at rt for 30 min and
concentrated. The residue was purified by Prep-HPLC to afford the
desired compound (215 mg, yield: 43%) as a white solid. .sup.1H NMR
(500 MHz, M eOD): .delta..sub.H 8.28 (s, 1H), 8.21 (s, 1H), 7.52
(d, J=8.0 Hz, 1H), 7.40 (s, 1H), 7.12 (dd, J=8.5 and 0.5 Hz, 1H),
6.00 (d, J=4.0 Hz, 1H), 4.71 (t, J=4.5 Hz, 1H), 4.24 (t, J=5.5 Hz,
1H), 4.18 (t, J=6.0 Hz, 1H), 2.84-2.81 (m, 2H), 2.77-2.68 (m, 3H),
2.25-2.22 (m, 2H), 2.17 (s, 3H), 1.91-1.89 (m, 3H), 1.51-1.45 (m,
2H) ppm; LC-MS (m/z): 563.3 [M+1].sup.+.
Compound 97:
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((methyl((1s,3R)-3-(2-(5-(trifl-
uoromethoxy)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrah-
ydrofuran-3,4-diol
##STR00430##
[1155] A solution of
trans-9-((3aR,4R,6R,6aR)-2,2-dimethyl-6-((methyl(3-(2-(5-(trifluoromethox-
y)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)amino)methyl)tetrahydrofuro[3-
,4-d][1,3]dioxol-4-yl)-9H-purin-6-amine (220 mg, 0.37 mmol) in 1 N
HCl/MeOH (5 mL) was stirred at 30.degree. C. for 4 h. The volatiles
were removed under reduced pressure. To the residue was added MeOH
(10 mL) and adjusted to pH=10-11 with NH.sub.3.H.sub.2O. The
mixture was stirred at rt for 30 min and concentrated. The residue
was purified by Prep-HPLC to afford the desired compound (80 mg,
yield: 39%) as a white solid. .sup.1H NMR (500 MHz, MeOD):
&8.29 (s, 1H), 8.20 (s, 1H), 7.52 (d, J=8.5 Hz, 1H), 7.40 (s,
1H), 7.11 (d, J=8.5 Hz, 1H), 6.00 (d, J=4.0 Hz, 1H), 4.72 (t, J=4.5
Hz, 1H), 4.25 (t, J=5.5 Hz, 1H), 4.18-4.20 (m, 1H), 3.04-3.08 (m,
1H), 2.83-2.86 (m, 2H), 2.64-2.72 (m, 2H), 2.17 (s, 3H), 1.97-2.10
(m, 5H), 1.82-1.85 (m, 2H) ppm; LC-MS (m/z): 563.3 [M+1].sup.+.
Compound 106
##STR00431##
[1157] To a solution of
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((((1r,3
S)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopro-
pyl)amino)methyl)tetrahydrofuran-3,4-diol (300 mg, 0.53 mmol) in
30% aqueous dioxane (20 mL) was added MCPBA (91 mg, 0.53 mmol). The
mixture was stirred at rt for 3 h and was then concentrated. The
crude was purified by Prep-HPLC to obtain the desired product (160
mg, Yield: 45%) as a white solid. .sup.1H NMR (500 MHz, MeOD):
&8.24 (s, 1H), 8.22 (s, 1H), 7.49 (brs, 1H), 7.42-7.38 (m, 1H),
7.31-7.29 (m, 1H), 6.00-5.96 (m, 1H), 4.68-4.65 (m, 2H), 4.43-4.36
(m, 1H), 4.05-4.00 (m, 1H), 3.88-3.76 (m, 1H), 3.68-3.49 (m, 1H),
3.46-3.37 (m, 1H), 2.84-2.81 (m, 2H), 2.42-2.15 (m, 4H), 1.95-1.89
(m, 3H), 1.39 (s, 1H), 1.35-1.27 (m, 3H), 1.25-1.21 (m, 3H) ppm;
ESI-MS (m/z): 579.4 [M+1].sup.+.
Compound 107
##STR00432##
[1159] To a solution of
(2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((((1s,3R)-3-(2-(5-(tert-butyl)-
-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)tetrahy-
drofuran-3,4-diol (300 mg, 0.53 mmol) in 30% aqueous dioxane (20
mL) was added MCPBA (91 mg, 0.53 mmol). The mixture was stirred at
rt for 3 h and was then concentrated. The crude was purified by
Prep-HPLC to obtain the desired product (80 mg, Yield: 26%) as a
white solid. .sup.1H NMR (500 MHz, MeOD): .delta..sub.H 8.25-8.22
(m, 2H), 7.49 (brs, 1H), 7.42-7.38 (m, 1H), 7.31-7.28 (m, 1H),
6.02-5.97 (m, 1H), 4.69-4.60 (m, 2H), 4.47-4.32 min, 2H), 3.86-3.74
(m, 1H), 3.70-3.54 (m, 1H), 3.45-3.35 (m, 1H), 2.97-2.72 (m, 4H),
2.15-1.75 (m, 5H), 1.39 (s, 1H), 1.34-1.28(3H), 1.26-1.22 (3H) ppm;
ES-MS (m/z): 579.7 [M+1].sup.+.
Example 9: Supercritical Fluid Chromatography
[1160] Compounds were purified by Supercritical Fluid
Chromatography (SFC) using known techniques. Such as methods
employed by Lotus Separations, LLC, Princeton, N.J. See, e.g.
http://www.lotussep.com. See also,
http://www.greenchemistrygroup.org/Program2009.html.
[1161] SFC separation conditions for certain examples are listed
below. Other compounds described herein can be separated by similar
methods.
TABLE-US-00003 Cmpd Lotus prep separation conditions Lotus
analytical separation conditions 28 AD-H (2 .times. 15 cm) 40%
AD-H(15 .times. 0.46 cm) 40% isopropanol(0.1% DEA))/CO2, 100
isopropanol(DEA)/CO2, 100 bar 5 bar 70 mL/min, 220 nm. inj vol.: 1
mL/min, 220 and 254 nm mL, 2.5 mg/mL methanol 30 AD-H (2 .times. 15
cm) 35% AD-H(15 .times. 0.46 cm) 40% isopropanol(0.1% DEA))/CO2,
100 isopropanol(DEA)/CO2, 100 bar 3 bar 70 mL/min, 220 nm. inj
vol.: mL/min, 220 and 254 nm 0.5-2 mL, 13 mg/mL ethanol 31 AD-H (2
.times. 15 cm) 40% AD-H(15 .times. 0.46 cm) 40% isopropanol(0.1%
DEA))/CO2, 100 isopropanol(DEA)/CO2, 100 bar 5 bar 70 mL/min, 220
nm. inj vol.: 1 mL/min, 220 and 254 nm mL, 2.5 mg/mL methanol 34
AD-H (2 .times. 15 cm) 35% AD-H(15 .times. 0.46 cm) 40%
isopropanol(0.1% DEA))/CO2, 100 isopropanol(DEA)/CO2, 100 bar 3 bar
70 mL/min, 220 nm. inj vol.: mL/min, 220 and 254 nm 0.5-2 mL, 13
mg/mL ethanol 35 IC (2 .times. 15 cm) 35% IC (15 .times. 0.46 cm)
40% isopropanol(0.2% DEA))/CO2, 100 isopropanol(DEA)/CO2, 100 bar 3
bar 60 mL/min, 220 nm. inj vol.: mL/min, 220 nm 0.75 mL, 4 mg/mL
methanol 36 AD-H (2 .times. 15 cm) 35% AD-H(15 .times. 0.46 cm) 40%
isopropanol(0.1% DEA))/CO2, 100 isopropanol(DEA)/CO2, 100 bar 3 bar
70 mL/min, 220 nm. inj vol.: mL/min, 220 nm 0.5 mL, 6.7 mg/mL 9:1
methanol:DCM 37 IC (2 .times. 15 cm) 30% IC(15 .times. 0.46 cm) 30%
isopropanol(0.2% DEA))/CO2, 100 isopropanol(DEA)/CO2, 100 bar 3 bar
65 mL/min, 220 nm. inj vol.: 1 mL/min, 280 nm mL, 2.6 mg/mL
methanol 38 Lux-3 (2 .times. 15 cm) 30% Lux-3 (15 .times. 0.46 cm)
25% ethanol(0.2% DEA))/CO2, 100 bar ethanol(NH4OH)/CO2, 100 bar 3
65 mL/min, 220 nm. inj vol.: 0.4 mL/min, 220 nm mL, 6.2 mg/mL
methanol 39 AD-H (2 .times. 15 cm) 35% AD-H(15 .times. 0.46 cm) 40%
isopropanol(0.1% DEA))/CO2, 100 isopropanol(DEA)/CO2, 100 bar 3 bar
70 mL/min, 220 nm. inj vol.: mL/min, 220 nm 0.5 mL, 6.7 mg/mL 9:1
methanol:DCM 40 AD-H (2 .times. 15 cm) 40% AD-H (15 .times. 0.46
cm) 40% isopropanol(0.1% DEA))/CO2, 100 isopropanol(DEA)/CO2, 100
bar 3 bar 65 mL/min, 220 nm. inj vol.: mL/min, 280 nm 0.5-1 mL, 8.1
mg/mL methanol 41 Premier (2 .times. 25 cm) 30% Premier (25 .times.
0.46 cm) 25% methanol(0.1% DEA))CO2, 100 bar methanol(DEA)/CO2, 100
bar 3 60 ml/min, 220 nm. Inj vol.: 1 mL, mL/min, 220 nm 20 mg/mL
methanol 46 Lux-3 (2 .times. 15 cm) 25% Lux-3 (15 .times. 0.46 cm)
30% ethanol(0.1% NH4OH))/CO2, 100 ethanol(NH4OH)/CO2, 100 bar 3 bar
65 mL/min, 220 nm. inj vol.: mL/min, 220 nm 0.3 mL, 2 mg/mL
methanol 47 AD-H (2 .times. 15 cm) 30% AD-H(15 .times. 0.46 cm) 40%
ethanol(0.1% DEA))/CO2, 100 bar ethanol(DEA)/CO2, 100 bar 3 65
mL/min, 220 nm. inj vol.: 0.5 mL/min, 280 nm mL, 12 mg/mL methanol
48 AD-H (2 .times. 15 cm) 40% 1:1 AD-H(15 .times. 0.46 cm) 50% 1:1
heptane:ethanol(0.1% DEA))/CO2, heptane:ethanol(DEA)/CO2, 100 bar 3
100 bar 65 mL/min, 220 nm. inj mL/min, 280 nm vol.: 0.5 mL, 25
mg/mL ethanol 49 Lux-3 (2 .times. 15 cm) 30% Lux-3 (15 .times. 0.46
cm) 25% ethanol(0.2% DEA))/CO2, 100 bar ethanol(NH4OH)/CO2, 100 bar
3 65 mL/min, 220 nm. inj vol.: 0.4 mL/min, 220 nm mL, 6.2 mg/mL
methanol 51 Lux-3 (2 .times. 15 cm) 25% Lux-3 (15 .times. 0.46 cm)
30% ethanol(0.1% NH4OH))/CO2, 100 ethanol(NH4OH)/CO2, 100 bar 3 bar
65 mL/min, 220 nm. inj vol.: mL/min, 220 nm 0.3 mL, 2 mg/mL
methanol 52 AD-H (2 .times. 15 cm) 40% AD-H (15 .times. 0.46 cm)
40% ethanol(0.2% DEA))/CO2, 100 bar ethanol(DEA)/CO2, 100 bar 3 60
mL/min, 220 nm. inj vol.: 0.5 mL/min, 280 nm mL, 10 mg/mL methanol
53 AD-H (2 .times. 15 cm) 40% 1:1 AD-H(15 .times. 0.46 cm) 50% 1:1
heptane:ethanol(0.1% DEA))/CO2, heptane:ethanol(DEA)/CO2, 100 bar 3
100 bar 65 mL/min, 220 nm. inj mL/min, 280 nm vol.: 0.5 mL, 25
mg/mL ethanol 54 AD-H (2 .times. 15 cm) 40% AD-H (15 .times. 0.46
cm) 40% isopropanol(0.1% DEA))/CO2, 100 isopropanol(DEA)/CO2, 100
bar 3 bar 65 mL/min, 220 nm. inj vol.: mL/min, 280 nm 0.5-1 mL, 8.1
mg/mL methanol 55 AD-H (2 .times. 15 cm) 30% AD-H(15 .times. 0.46
cm) 40% ethanol(0.1% DEA))/CO2, 100 bar ethanol(DEA)/CO2, 100 bar 3
65 mL/min, 220 nm. inj vol.: 0.5 mL/min, 280 nm mL, 12 mg/mL
methanol 57 IC (2 .times. 15 cm) 30% IC(15 .times. 0.46 cm) 30%
isopropanol(0.2% DEA))/CO2, 100 isopropanol(DEA)/CO2, 100 bar 3 bar
65 mL/min, 220 nm. inj vol.: 1 mL/min, 280 nm mL, 2.6 mg/mL
methanol 58 AD-H (2 .times. 15 cm) 40% AD-H (15 .times. 0.46 cm)
40% ethanol(0.2% DEA))/CO2, 100 bar ethanol(DEA)/CO2, 100 bar 3 60
mL/min, 220 nm. inj vol.: 0.5 mL/min, 280 nm mL, 10 mg/mL methanol
60 AD-H (2 .times. 15 cm) 30% AD-H(15 .times. 0.46 cm) 25%
ethanol(0.1% DEA))/CO2, 100 bar ethanol(DEA)/CO2, 100 bar 3 60
mL/min, 220 nm, inj vol.: 0.5 mL/min, 280 nm mL, 10 mg/mL methanol
61 AD-H (2 .times. 15 cm) 30% AD-H(15 .times. 0.46 cm) 25%
ethanol(0.1% DEA))/CO2, 100 bar ethanol(DEA)/CO2, 100 bar 3 60
mL/min, 220 nm. inj vol.: 0.5 mL/min, 280 nm mL, 10 mg/mL methanol
63 AD-H (2 .times. 15 cm) 38% AD-H(15 .times. 0.46 cm) 40%
methanol(0.1% DEA))/CO2, 100 methanol(DEA)/CO2, 100 bar 3 bar 70
mL/min, 220 nm. inj vol.: mL/min, 220 and 254 nm 1.5 mL, 11.8 mg/mL
methanol 65 AD-H (2 .times. 15 cm) 38% AD-H(15 .times. 0.46 cm) 40%
methanol(0.1% DEA))/CO2, 100 methanol(DEA)/CO2, 100 bar 3 bar 70
mL/min, 220 nm. inj vol.: mL/min, 220 and 254 nm 1.5 mL, 11.8 mg/mL
methanol 69 IC (2 .times. 15 cm) 35% IC (15 .times. 0.46 cm) 40%
isopropanol(0.2% DEA))/CO2, 100 isopropanol(DEA)/CO2, 100 bar 3 bar
60 mL/min, 220 nm. inj vol.: mL/min, 220 nm 0.75 mL, 4 mg/mL
methanol 114 LUX2 (2 .times. 15 cm) 45% LUX2 (15 .times. 0.46 cm)
45% methanol(0.1% DEA))/CO2, 100 methanol(DEA)/CO2, 100 bar 5 bar
60 mL/min, 220 nm. inj vol.: mL/min, 220 and 254 nm 0.75 mL, 5.6
mg/mL methanol:DCM 114 OZ-H (3 .times. 25 cm) 40% OZ-H(15 .times.
0.46 cm) 40% methanol(0.1% DEA))/CO2, 100 methanol(DEA)/CO2, 100
bar 3 bar 65 mL/min, 220 nm. inj vol.: mL/min, 220 and 254 nm 1.75
mL, 10 mg/mL methanol 115 LUX2 (2 .times. 15 cm) 45% LUX2 (15
.times. 0.46 cm) 45% methanol(0.1% DEA))/CO2, 100
methanol(DEA)/CO2, 100 bar 5 bar 60 mL/min, 220 nm. inj vol.:
mL/min, 220 and 254 nm 0.75 mL, 5.6 mg/mL methanol:DCM 115 OZ-H (3
.times. 25 cm) 40% OZ-H(15 .times. 0.46 cm) 40% methanol(0.1%
DEA))/CO2, 100 methanol(DEA)/CO2, 100 bar 3 bar 65 mL/min, 220 nm.
inj vol.: mL/min, 220 and 254 nm 1.75 mL, 10 mg/mL methanol 116
LUX-2 (2 .times. 15 cm) 35% LUX-2(15 .times. 0.46 cm) 45%
methanol(0.1% DEA))/CO2, 100 methanol(DEA)/CO2, 100 bar 3 bar 70
mL/min, 220 nm. inj vol.: 1 mL/min, 220 nm mL, 8.5 mg/mL methanol
119 LUX-2 (2 .times. 15 cm) 35% LUX-2(15 .times. 0.46 cm) 45%
methanol(0.1% DEA))/CO2, 100 methanol(DEA)/CO2, 100 bar 3 bar 70
mL/min, 220 nm. inj vol.: 1 mL/min, 220 nm mL, 8.5 mg/mL methanol
121 OZ-H (3 .times. 25 cm) 40% OZ-H(15 .times. 0.46 cm) 40%
methanol(0.1% DEA))/CO2, 100 methanol(DEA)/CO2, 100 bar 3 bar 65
mL/min, 220 nm. inj vol.: 2 mL/min, 220 and 254 nm mL, 14 mg/mL
methanol 122 OZ-H (3 .times. 25 cm) 40% OZ-H(15 .times. 0.46 cm)
40% methanol(0.1% DEA))/CO2, 100 methanol(DEA)/CO2, 100 bar 3 bar
65 mL/min, 220 nm. inj vol.: 2 mL/min, 220 and 254 nm mL, 14 mg/mL
methanol 123 OZ-H (3 .times. 25 cm) 40% OZ-H(15 .times. 0.46 cm)
40% methanol(0.1% DEA))/CO2, 100 methanol(DEA)/CO2, 100 bar 3 bar
65 mL/min, 220 nm. inj vol.: mL/min, 220 and 254 nm 0.8 mL, 19
mg/mL methanol 124 OZ-H (3 .times. 25 cm) 40% OZ-H(15 .times. 0.46
cm) 40% methanol(0.1% DEA))/CO2, 100 methanol(DEA)/CO2, 100 bar 3
bar 65 mL/min, 220 nm. inj vol.: mL/min, 220 and 254 nm 0.8 mL, 19
mg/mL methanol 128 AD-H (2 .times. 15 cm) 30% AD-H(15 .times. 0.46
cm) 40% isopropanol(0.1% DEA))/CO2, 100 isopropanol(DEA)/CO2, 100
bar 3 bar 65 mL/min, 220 nm. inj vol.: mL/min, 220 and 254 nm 0.5-2
mL, 20 mg/mL methanol 129 AD-H (2 .times. 15 cm) 30% AD-H(15
.times. 0.46 cm) 40% isopropanol(0.1% DEA))/CO2, 100
isopropanol(DEA)/CO2, 100 bar 3 bar 65 mL/min, 220 nm, inj vol.:
mL/min, 220 and 254 nm 0.5-2 mL, 20 mg/mL methanol 131 AD-H (2
.times. 15 cm) 30% AD-H(15 .times. 0.46 cm) 30% methanol(0.1%
DEA))/CO2, 100 isopropanol(DEA)/CO2, 100 bar 3 bar 70 mL/min, 220
nm. inj vol.: 1 mL/min, 220 and 254 nm mL, 15 mg/mL methanol 132
AD-H (2 .times. 15 cm) 30% AD-H(15 .times. 0.46 cm) 30%
methanol(0.1% DEA))/CO2, 100 isopropanol(DEA)/CO2, 100 bar 3 bar 70
mL/min, 220 nm. inj vol.: 1 mL/min, 220 and 254 nm mL, 15 mg/mL
methanol 133 AD-H (2 .times. 15 cm) 30% AD-H(15 .times. 0.46 cm)
30% methanol(0.1% DEA))/CO2, 100 methanol(DEA)/CO2, 100 bar 4 bar
70 mL/min, 220 nm. inj vol.: mL/min, 220 and 254 nm 0.5 mL, 7.7
mg/mL methanol 134 AD-H (2 .times. 15 cm) 30% AD-H(15 .times. 0.46
cm) 30% methanol(0.1% DEA))/CO2, 100 methanol(DEA)/CO2, 100 bar 4
bar 70 mL/min, 220 nm. inj vol.: mL/min, 220 and 254 nm 0.5 mL, 7.7
mg/mL methanol 135 AD-H (2 .times. 15 cm) 30% AD-H(15 .times. 0.46
cm) 30% isopropanol(0.1% DEA))/CO2, 100 isopropanol(DEA)/CO2, 100
bar 3 bar 70 mL/min, 220 nm. inj vol.: mL/min, 220 and 254 nm 0.75
mL, 15 mg/mL methanol 136 AD-H (2 .times. 15 cm) 30% AD-H(15
.times. 0.46 cm) 30% isopropanol(0.1% DEA))/CO2, 100
isopropanol(DEA)/CO2, 100 bar 3 bar 70 mL/min, 220 nm. inj vol.:
mL/min, 220 and 254 nm 0.75 mL, 15 mg/mL methanol 137 IC (2 .times.
15 cm) 35% IC(15 .times. 0.46 cm) 35% isopropanol(0.1% DEA))/CO2,
100 isopropanol(DEA)/CO2, 100 bar 3 bar 60 mL/min, 220 nm. inj
vol.: mL/min, 220 and 254 nm 0.5 mL, 10 mg/mL methanol 138 IC (2
.times. 15 cm) 35% IC(15 .times. 0.46 cm) 35% isopropanol(0.1%
DEA))/CO2, 100 isopropanol(DEA)/CO2, 100 bar 3 bar 60 mL/min, 220
nm. inj vol.: mL/min, 220 and 254 nm 0.5 mL, 10 mg/mL methanol
Example 10: Bioassay protocol and General Methods
[1162] Cell Culture. Human hematological tumor cell lines THP-1,
RS4;11, and MV4-11 were obtained from ATCC, MOLM-13 cells were
obtained from DSMZ. All lines were grown in RPMI 1640 containing
10% FBS and maintained using the vendors recommended cell densities
and environmental conditions. Media was supplemented with non
essential amino acids and L-Glutamine. THP-1 cells were also
supplemented with 0.05 mM P-Mercaptoethanol.
[1163] Methylation Analysis. Cells were seeded at 5.times.10.sup.5
cells/mL in a 12 well plate at a final volume of 2 mLs. Cells were
dosed with compounds to the appropriate concentration from a 50 mM
DMSO stock solution. Compound and media were refreshed every two
days over the course of seven day incubation by counting cells
using trypan blue exclusion (Vicell), pelleting at 200 g for 5
minutes and resuspending in fresh media containing compound at a
final cell concentration of 5.times.10.sup.5 cells/mL. Following
compound incubation, histones were extracted from 1.times.10.sup.6
cells using a commercial histone extraction kit (Active Motif).
Purified histones were quantitated using the BCA protein assay
(Pierce) with a BSA standard curve. 400 ng of isolated histones
were fractionated by SDS-PAGE on a 4-20% gel and transferred to
nitrocellulose membranes. Membranes were incubated with various
primary and secondary antibodies and imaged on the Licor imaging
system (Odyssey). The H3K79-Me2 rabbit polyclonal was purchased
from Abcam. Other rabbit polyclonal antibodies including H3K4-Me3,
H3K9-Me3, H3K27-Me2, and H3K27-Me3 were purchased from Cell
Signaling Technologies (CST). A mouse monoclonal total H3 antibody
was used as a loading control (CST). Fluorescently labeled
secondary antibodies were purchased from Odyssey.
[1164] Cell Growth and Viability Analysis. Cells were harvested
from exponentially growing cell cultures and seeded at
3.times.10.sup.4 cells per well. Samples were maintained in a 96
well black walled clear bottom plate (Corning). A final
concentration of 50 uM compound in 0.2% DMSO was added to the
appropriate wells on Day 0. Treatment of MV4-11 and MOLM-13 lasted
14 days, while THP-1 cells were treated for 18 days. Compound and
media were replaced every two days during incubation by
transferring samples to a V-bottom plate (Corning), spinning at 200
g for 5 minutes in a room temperature rotor, resuspending in fresh
media containing compound and transferring back to the assay plate.
Cells were counted periodically using the Guava Viacount assay and
read on the EasyCyte Plus instrument (Millipore). Assay plates were
split when necessary to within recommended cell densities. Final
cell counts were adjusted to take cell splits into account and
reported as total viable cells/well.
[1165] HOXA9 (qPCR). Cells were treated with compound for 7 days
similar to methylation assay. Cell were pelleted at 200 g in a room
temperature rotor and total RNA isolated using the Qiagen RNeasy
kit. RNA concentration and quality was determined by using the
Nanovue (GE Healthcare). Total RNA was reverse transcribed using a
high capacity cDNA reverse transcription kit (Applied Biosystems).
A predesigned labeled primer set for HOXA9 was purchased from
Applied Biosystems. qPCR reactions contained 50 ng cDNA, 1.times.
labeled primer and 1.times. Taqman universal PCR master mix
(Applied Biosystems). Samples were run on a 7900 HT Fast Real Time
PCR machine (Applied Biosystems) with PCR conditions of 2 min
50.degree. C., 10 min 95.degree. C., 40 cycles at 15 sec 95.degree.
C. and 1 min 60.degree. C. HOXA9 cycle numbers were normalized to
the house keeping gene B2 microglobulin (B2M predesigned control
from Applied Biosystems). Percent of DMSO control was calculated
with the equation, percent control=(2{circumflex over (
)}-.sup..DELTA..DELTA.CT)*100 where the .DELTA..DELTA.CT is the
difference between normalized HOXA9 sample and control (ACT sample
-ACT control=.DELTA..DELTA.CT).
[1166] Determination of IC.sub.50. Test compounds were serially
diluted 3 fold in DMSO for 10 points and 1 .mu.l was plated in a
384 well microtiter plate. Positive control (100% inhibition
standard) was 2.5 uM final concentration of
S-adenosyl-L-homocysteine and negative control (0% inhibition
standard) contained 1 .mu.l of DMSO. Compound was then incubated
for 30 minutes with 40 .mu.l per well of DOT1L(1-416) (0.25 nM
final concentration in assay buffer: 20 mM TRIS, pH 8.0, 10 mM
NaCl, 0.002% Tween20, 0.005% Bovine Skin Gelatin, 100 mM KCl, and
0.5 mM DTT). 10 .mu.l per well of substrate mix (same assay buffer
with 200 nM S-[methyl-.sup.3H]-adenosyl-L methionine, 600 nM of
unlabeled S-[methyl-.sup.3H]-adenosyl-L methionine, and 20 nM
oligonucleosome) was added to initiate the reaction. Reaction was
incubated for 120 minutes at room temperature and quenched with 10
.mu.l per well of 100 .mu.M S-methyl-adenosyl-L methionine. For
detection, substrate from 50 .mu.l of reaction was immobilized on a
384 well Streptavidin coated Flashplate (Perkin Elmer) (also coated
with 0.2% polyethyleneimine) and read on a Top Count scintillation
counter (Perkin Elmer). IC.sub.50 values are presented in the table
below. In this table, "A" indicates IC.sub.50 values of <0.1
.mu.M; "B" indicates IC.sub.50 values of >0.1 .mu.M and <1
.mu.M; "C" indicates IC.sub.50 values of >1 .mu.M and <10
.mu.M; and "D" indicates IC.sub.50 values of >10 .mu.M and
<50 .mu.M
TABLE-US-00004 DOT1L Compound# IC.sub.50 2 A 3 A 4 A 5 A 6 A 7 A 8
D 9 A 10 A 11 A 12 A 13 A 14 A 15 C 16 C 17 B 18 B 19 A 20 A 21 C
22 B 23 A 24 A 25 A 26 A 27 A 28 A 29 A 30 A 31 A 32 A 33 A 34 A 35
A 36 A 37 A 38 A 39 A 40 A 41 A 42 A 43 A 44 A 45 A 46 A 47 A 48 A
49 A 50 A 51 A 52 A 53 A 54 A 55 A 56 A 57 A 58 A 59 A 60 A 61 B 62
B 63 B 64 B 65 B 66 B 67 A 68 A 69 A 70 A 71 A 72 A 73 A 74 A 75 A
76 A 77 B 78 C 79 A 80 B 81 B 82 B 83 B 84 C 85 D 86 A 87 A 88 A 89
A 90 A 91 A 92 A 93 A 94 A 95 A 96 A 97 A 98 A 99 A 100 A 101 B 102
B 103 C 104 C 105 C 106 A 107 A 110 A 111 A 112 A 114 A 115 A 116 A
117 A 118 A 119 A 120 A 121 A 122 A 123 A 124 A 128 A 129 A 130 A
131 A 132 A 133 A 134 A 135 A 136 A 137 A 138 A 139 A 140 A
Example 11: Tumor Anti-Proliferation Assays
[1167] In Vitro Anti-Proliferative Assay. The potency and
selectivity of the anti-proliferative activity of the compounds of
the present invention were assessed using a panel of MLL-rearranged
and non-MLL-rearranged human leukemia cell lines. The cell lines
used in the study are listed in FIG. 1A. The MLL-rearranged panel
included cell lines derived from ALL, AML and biphenotypic
leukemias harboring MLL-AF4, MLL-AF9 or MLL-ENL fusions. These cell
lines recruit DOT1L. The panel also included five cell lines that
do not possess an MLL-rearrangement, and one cell line that bears a
partial tandem duplication of the MLL gene (MLL-PTD).
[1168] Exponentially growing cells were plated, in triplicate, in
96-well plates at a density of 3.times.10.sup.4 cells/well in a
final volume of 150 .mu.l. Cells were incubated in the presence of
increasing concentrations of Compound 2. Anti-proliferative
activity was determined by cell viability measurements every 3-4
days for up to 14 days. On days of cell counts, growth media and
Compound 2 were replaced and cells split back to a density of
5.times.10.sup.4 cells/well.
[1169] The half maximal inhibitory concentration (IC.sub.50)
results in FIG. 1 show that Compound 2 demonstrates potent
nanomolar anti-proliferative activity against three of four
MLL-rearranged cell lines tested (MV4;11 (MLL-AF4), MOLM-13
(MLL-AF9), and KOPN-8 (MLL-ENL). EOL-1 cells which express an
MLL-PTD were also highly sensitive to Compound 2 (IC.sub.50=11 nM).
RS4;11 cells and two non MLL-rearranged cells (Reh and Kasumi-1)
were 1-3 log orders less sensitive, and two non-MLL-rearranged
cells (Jurkat and HL-60) showed no activity. Overall, the results
indicate that Compound 2 potently and selectively inhibits the
proliferation of MLL-rearranged leukemia cell lines and a subset of
non-MLL-rearranged leukemia cell lines.
[1170] In Vivo Anti-Proliferative Assay. The in vivo anti-tumor
activity of the compounds of the present invention were assessed in
a mouse xenograft model of MLL-rearranged leukemia.
[1171] Four groups of 20 (Groups 1, 3, 4 and 5), and one group of 8
(Group 2) female nude mice (average weight of 0.023 kg) bearing
MV4-11 xenograft tumors of sizes ranging from 80-120 mm.sup.3 were
implanted subcutaneously with minipumps (Alzet Model 2001). Group 1
received vehicle only from the pump. Group 2 received vehicle only
from the pump plus ip injections tid (8 hours apart) of vehicle.
Group 3 received 112 mg/kg/day from the pump plus ip injections tid
(8 hours apart) of 20 mg/kg of Compound 2 for a total daily dose of
172 mg/kg/day. Group 4 and 5 received 112 and 56 mg/kg/day of
Compound 2 from the pumps, respectively. Pumps were designed to
last for 7 days and were exchanged twice to give total infusion
duration of 21 days exposure.
[1172] A single blood sample was taken from all animals in Groups 4
and 5 on days 7, 14, and 21 and assayed for plasma levels of
Compound 2. Blood samples were taken from Group 3 on days 7 and 14
at the following time points (3 animals per time point): 5 minutes
pre-ip dose, and 15 min, 30 min, 1, 2, and 4 hours post ip dose. On
day 21 three hours after the last ip injection, a single blood
sample was taken from Group 3. Tumor size was measured every 4
days. After 21 days the study was terminated, and mean TGI
calculated.
[1173] FIG. 2 shows the tumor growth over the 21 days of dosing.
There was no difference in tumor size between the two vehicle
control groups. The high dose minipump group supplemented with ip
dosing showed a statistically significant TGI of >70% compared
to the controls. The 56 and 112 mg/kg/day groups showed
non-statistically significant TGI values of 43 and 38%,
respectively, compared to controls. Compound 2 is referred to as
Ex. 2 in FIG. 2.
[1174] FIG. 3A shows the estimated steady state plasma
concentrations of Compound 2 in Groups 4 and 5 as determined by the
averaged blood samples taken on days 7, 14, and 21. The data
suggest that the average steady state Compound 2 plasma levels
ranged from 99 to 152 ng/ml for Group 4, and 52 to 238 ng/ml for
Group 5. The average plasma level from the last sampling on day 21
was 99 ng/ml for Group 4 and 52 ng/ml for Group 5.
[1175] FIG. 3B shows the Compound 2 plasma concentrations plotted
against time after ip injection. The ip injections produced a
significant increase in plasma exposure to Compound 2 in terms of
both the C.sub.max (4200 to 5000 ng/ml) after each of the tid
injections and the daily AUCs over those produced by the steady
state plasma level resulting from the continuous infusion. Overall,
the results indicate that Compound 2 demonstrated significant
anti-tumor activity in a mouse xenograft model of MLL-rearranged
leukemia.
[1176] The entire disclosure of each of the patent documents and
scientific articles referred to herein is incorporated by reference
for all purposes.
[1177] The invention can be embodied in other specific forms
without departing from the spirit or essential characteristics
thereof. The foregoing embodiments are therefore to be considered
in all respects illustrative rather than limiting on the invention
described herein. Scope of the invention is thus indicated by the
appended claims rather than by the foregoing description, and all
changes that come within the meaning and range of equivalency of
the claims are intended to be embraced therein.
* * * * *
References