U.S. patent application number 15/262230 was filed with the patent office on 2016-12-29 for substituted dihydroisoquinolinone compounds.
This patent application is currently assigned to Pfizer Inc.. The applicant listed for this patent is Pfizer Inc.. Invention is credited to Michael Raymond COLLINS, Robert Steven Kania, Robert Arnold Kumpf, Pei-Pei Kung, Daniel Tyler Richter, Scott Channing Sutton, Martin James Wythes.
Application Number | 20160376254 15/262230 |
Document ID | / |
Family ID | 53434412 |
Filed Date | 2016-12-29 |
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United States Patent
Application |
20160376254 |
Kind Code |
A1 |
COLLINS; Michael Raymond ;
et al. |
December 29, 2016 |
SUBSTITUTED DIHYDROISOQUINOLINONE COMPOUNDS
Abstract
This invention relates to compounds of general formula (I)
##STR00001## in which R.sup.1, R.sup.2, R.sup.3, R.sup.4, L, X and
Z are as defined herein, and the pharmaceutically acceptable salts
thereof, to pharmaceutical compositions comprising such compounds
and salts, and to methods of using such compounds, salts and
compositions for the treatment of abnormal cell growth, including
cancer.
Inventors: |
COLLINS; Michael Raymond;
(San Diego, CA) ; Kania; Robert Steven; (San
Diego, CA) ; Kumpf; Robert Arnold; (Carlsbad, CA)
; Kung; Pei-Pei; (San Diego, CA) ; Richter; Daniel
Tyler; (San Diego, CA) ; Sutton; Scott Channing;
(San Diego, CA) ; Wythes; Martin James; (Solana
Beach, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Pfizer Inc. |
New York |
NY |
US |
|
|
Assignee: |
Pfizer Inc.
New York
NY
|
Family ID: |
53434412 |
Appl. No.: |
15/262230 |
Filed: |
September 12, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14740439 |
Jun 16, 2015 |
9481666 |
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15262230 |
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62156533 |
May 4, 2015 |
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62013410 |
Jun 17, 2014 |
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Current U.S.
Class: |
514/210.18 |
Current CPC
Class: |
C07B 59/002 20130101;
C07D 401/14 20130101; C07B 2200/05 20130101; A61P 35/00 20180101;
C07D 413/14 20130101; C07D 401/06 20130101; C07D 405/14 20130101;
C07D 409/14 20130101; A61P 43/00 20180101 |
International
Class: |
C07D 401/06 20060101
C07D401/06; C07D 409/14 20060101 C07D409/14; C07D 413/14 20060101
C07D413/14; C07B 59/00 20060101 C07B059/00; C07D 405/14 20060101
C07D405/14; C07D 401/14 20060101 C07D401/14 |
Claims
1. A compound of formula (I): ##STR00258## or a pharmaceutically
acceptable salt thereof, wherein: R.sup.1 is selected from the
group consisting of H, F, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4
alkoxy, C(O)R.sup.5, C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl and 5-12 membered heteroaryl, where each said
C.sub.1-C.sub.4 alkyl or C.sub.1-C.sub.4 alkoxy is optionally
substituted by one or more R.sup.6, and each said C.sub.3-C.sub.8
cycloalkyl, 3-12 membered heterocyclyl or 5-12 membered heteroaryl
is optionally substituted by one or more R.sup.7; R.sup.2 is H, F
or C.sub.1-C.sub.4 alkyl; L is a bond or a C.sub.1-C.sub.4
alkylene; R.sup.3 is selected from the group consisting of
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy, OH, CN, C(O)R.sup.8,
COOR.sup.9, NR.sup.10R.sup.11, OR.sup.12, C.sub.3-C.sub.8
cycloalkyl, 3-12 membered heterocyclyl and 5-12 membered
heteroaryl, where each said C.sub.1-C.sub.4 alkyl or
C.sub.1-C.sub.4 alkoxy is optionally substituted by one or more
R.sup.6, and each said C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl or 5-12 membered heteroaryl is optionally substituted
by one or more R.sup.7; R.sup.4 is H, halo or C.sub.1-C.sub.4
alkyl, where each said C.sub.1-C.sub.4 alkyl is optionally
substituted by one or more R.sup.6; R.sup.5 is C.sub.1-C.sub.4
alkyl, where each said C.sub.1-C.sub.4 alkyl is optionally
substituted by one or more R.sup.14; each R.sup.6 is independently
OH, F, CN or C.sub.1-C.sub.4 alkoxy; each R.sup.7 is independently
C.sub.1-C.sub.4 alkyl, OH, F, CN, C.sub.1-C.sub.4 alkoxy, .dbd.O,
CHO, C(O)R.sup.13, SO.sub.2R.sup.13 or 3-6 membered heterocyclyl;
R.sup.8 is C.sub.1-C.sub.4 alkyl, where each said C.sub.1-C.sub.4
alkyl is optionally substituted by one or more R.sup.14; R.sup.9 is
H or C.sub.1-C.sub.4 alkyl, where each said C.sub.1-C.sub.4 alkyl
is optionally substituted by one or more R.sup.14; R.sup.10 and
R.sup.11 are independently H or C.sub.1-C.sub.4 alkyl, where each
said C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14; R.sup.12 is selected from the group consisting of
C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl and 5-12
membered heteroaryl, where each said C.sub.3-C.sub.8 cycloalkyl,
3-12 membered heterocyclyl or 5-12 membered heteroaryl is
optionally substituted by one or more R.sup.7; each R.sup.13 is
independently C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.15; each R.sup.14 and R.sup.15 is independently OH, F, CN or
C.sub.1-C.sub.4 alkoxy; and X and Z are independently
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4
alkoxy or C.sub.1-C.sub.4 fluoroalkoxy.
2. The compound or salt of claim 1, wherein R.sup.2 is H.
3. The compound or salt of claim 1, wherein R.sup.4 is Cl, F, Br or
CH.sub.3.
4. The compound or salt of claim 1, wherein X is CH.sub.3,
OCH.sub.3 or OCHF.sub.2 and Z is CH.sub.3.
5. The compound or salt of claim 1, wherein R.sup.1 is
C.sub.1-C.sub.4 alkoxy optionally substituted by one or more
R.sup.6.
6. The compound or salt of claim 5, wherein said C.sub.1-C.sub.4
alkoxy is OCH.sub.3.
7. The compound or salt of claim 1, wherein R.sup.1 is
C.sub.1-C.sub.4 alkyl optionally substituted by one or more
R.sup.6.
8. The compound or salt of claim 1, wherein L is a bond and R.sup.3
is 3-12 membered heterocyclyl optionally substituted by one or more
R.sup.7.
9. The compound or salt of claim 8, wherein said 3-12 membered
heterocyclyl is selected from the group consisting of oxetanyl,
tetrahydrofuranyl and tetrahydropyranyl, each optionally
substituted by one or more R.sup.7.
10. A compound of Formula (I-A): ##STR00259## or a pharmaceutically
acceptable salt thereof, wherein: R.sup.1 is C.sub.1-C.sub.4
alkoxy; R.sup.2 is H; L is a bond; R.sup.3 is 3-12 membered
heterocyclyl, optionally substituted by one or more R.sup.7;
R.sup.4 is H or Cl; each R.sup.7 is independently C.sub.1-C.sub.4
alkyl, OH, F, CN, C.sub.1-C.sub.4 alkoxy, .dbd.O, CHO,
C(O)R.sup.13, SO.sub.2R.sup.13 or 3-6 membered heterocyclyl; each
R.sup.13 is independently C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.15; each R.sup.15 is independently OH, F, CN or
C.sub.1-C.sub.4 alkoxy; and X and Z are independently
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4
alkoxy or C.sub.1-C.sub.4 fluoroalkoxy.
11. The compound or salt of claim 10, wherein R.sup.3 is 3-12
membered heterocyclyl selected from the group consisting of
oxetanyl, tetrahydrofuranyl and tetrahydropyranyl, each optionally
substituted by one or more R.sup.7.
12. A compound that is
5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydro-pyridin-3-yl)methyl-
]-7-[(R)-methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-(2H)-one.
13. A compound of formula (III): ##STR00260## or a pharmaceutically
acceptable salt thereof, wherein: R.sup.1 and R.sup.3 are taken
together to form a 3-12 membered heterocyclyl optionally
substituted by one or more R.sup.7; R.sup.2 is H, F or
C.sub.1-C.sub.4 alkyl; R.sup.4 is H, halo or C.sub.1-C.sub.4 alkyl,
where each said C.sub.1-C.sub.4 alkyl is optionally substituted by
one or more R.sup.6; each R.sup.6 is independently OH, F, CN or
C.sub.1-C.sub.4 alkoxy; each R.sup.7 is independently
C.sub.1-C.sub.4 alkyl, OH, F, CN, C.sub.1-C.sub.4 alkoxy, .dbd.O,
CHO, C(O)R.sup.13, SO.sub.2R.sup.13 or 3-6 membered heterocyclyl;
each R.sup.13 is independently C.sub.1-C.sub.4 alkyl, where each
said C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.15; each R.sup.15 is independently OH, F, CN or
C.sub.1-C.sub.4 alkoxy; and X and Z are independently
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4
alkoxy or C.sub.1-C.sub.4 fluoroalkoxy.
14. The compound or salt of claim 13, wherein R.sup.1 and R.sup.3
are taken together to form a 3-12 membered heterocyclyl selected
from the group consisting of azetidinyl, pyrrolidinyl, piperidinyl
and homopiperidinyl, each optionally substituted by one or more
R.sup.7.
15. The compound or salt of claim 14, wherein R.sup.7 is CHO,
C(O)R.sup.13 or SO.sub.2R.sup.13 and each R.sup.13 is independently
C.sub.1-C.sub.4 alkyl optionally substituted by one or more
R.sup.15.
16. A pharmaceutical composition comprising a compound of claim 1,
or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier or excipient.
17. A pharmaceutical composition comprising the compound of claim
12 and a pharmaceutically acceptable carrier or excipient.
18. A pharmaceutical composition comprising a compound of claim 13,
or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier or excipient.
19. A method for the treatment of abnormal cell growth in a
subject, comprising administering to the subject a therapeutically
effective amount of a compound of claim 1, or a pharmaceutically
acceptable salt thereof.
20. A method for the treatment of abnormal cell growth in a
subject, comprising administering to the subject a therapeutically
effective amount of a compound of claim 13, or a pharmaceutically
acceptable salt thereof.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This is a continuation from U.S. application Ser. No.
14/740,439, filed on Jun. 16 2015, which application claims the
benefit of priority to U.S. Provisional Application No. 62/013,410,
filed on Jun. 17, 2014, and to U.S. Provisional Application No.
62/156,533, filed on May 4, 2015, each of which is incorporated by
reference herein in its entirety.
BACKGROUND OF THE INVENTION
[0002] Field of the Invention
[0003] The present invention relates to compounds of Formulae I,
I', II, II' and Ill, and their pharmaceutically acceptable salts,
to pharmaceutical compositions comprising such compounds and salts,
and to the uses thereof. The compounds, salts and compositions of
the present invention may be useful for treating or ameliorating
abnormal cell proliferative disorders, such as cancer.
[0004] Description of Related Art
[0005] Epigenetic alterations play an important role in the
regulation of cellular processes, including cell proliferation,
cell differentiation and cell survival. The epigenetic silencing of
tumor suppressor genes and activation of oncogenes may occur
through alteration of CpG island methylation patterns, histone
modification, and dysregulation of DNA binding protein. Polycomb
genes are a set of epigenetic effectors. EZH2 (enhancer of zeste
homolog 2) is the catalytic component of the Polycomb Repressor
Complex 2 (PRC2), a conserved multi-subunit complex that represses
gene transcription by methylating lysine 27 on Histone H3 (H3K27).
EZH2 plans a key role in regulating gene expression patterns that
regulate cell fate decisions, such as differentiation and
self-renewal. EZH2 is overexpressed in certain cancer cells, where
it has been linked to cell proliferation, cell invasion,
chemoresistance and metastasis.
[0006] High EZH2 expression has been correlated with poor
prognosis, high grade, and high stage in several cancer types,
including breast, colorectal, endometrial, gastric, liver, kidney,
lung, melanoma, ovarian, pancreatic, prostate, and bladder cancers.
See Crea et al., Crit. Rev. Oncol. Hematol. 2012, 83:184-193, and
references cited therein; see also Kleer et al., Proc. Natl. Acad.
Sci. USA 2003, 100:11606-11; Mimori et al., Eur. J. Surg. Oncol.
2005, 31:376-80; Bachmann et al., J. Clin. Oncol. 2006, 24:268-273;
Matsukawa et al., Cancer Sci. 2006, 97:484-491; Sasaki et al. Lab.
Invest. 2008, 88:873-882; Sudo et al., Br. J. Cancer 2005,
92(9):1754-1758; Breuer et al., Neoplasia 2004, 6:736-43; Lu et
al., Cancer Res. 2007, 67:1757-1768; Ougolkov et al., Clin. Cancer
Res. 2008, 14:6790-6796; Varambally et al., Nature 2002,
419:624-629; Wagener et al., Int. J. Cancer 2008, 123:1545-1550;
and Weikert et al., Int. J. Mol. Med. 2005, 16:349-353.
[0007] Recurring somatic mutations in EZH2 have been identified in
diffuse large B-cell lymphoma (DLBCL) and follicular lymphomas
(FL). Mutations altering EZH2 tyrosine 641 (e.g., Y641C, Y641F,
Y641N, Y641S, and Y641H) were reportedly observed in up to 22% of
germinal center B-cell DLBCL and 7% of F L. Morin et al. Nat.
Genetics 2010 February; 42(2):181-185. Mutations of alanine 677
(A677) and alanine 687 (A687) have also been reported. McCabe et
al., Proc. Natl. Acad. Sci. USA 2012, 109:2989-2994; Majer et al.
FEBS Letters 2012, 586:3448-3451. EZH2 activating mutations have
been suggested to alter substrate specificity resulting in elevated
levels of trimethylated H3K27 (H3K27me3).
[0008] Accordingly, compounds that inhibit the activity of wild
type and/or mutant forms of EZH2 may be of interest for the
treatment of cancer.
BRIEF SUMMARY OF THE INVENTION
[0009] The present invention provides, in part, novel compounds and
pharmaceutically acceptable salts. Such compounds may modulate the
activity of EZH2, thereby effecting biological functions, for
example by inhibiting cell proliferation and cell invasiveness,
inhibiting metastasis, inducing apoptosis or inhibiting
angiogenesis. Also provided are pharmaceutical compositions and
medicaments, comprising the compounds or salts of the invention,
alone or in combination with other therapeutic agents or palliative
agents. The present invention also provides, in part, methods for
preparing the novel compounds, salts and compositions thereof, and
methods of using the foregoing.
[0010] In one aspect, the invention provides a compound of formula
(I):
##STR00002##
[0011] or a pharmaceutically acceptable salt thereof,
[0012] wherein:
[0013] R.sup.1 is selected from the group consisting of H, F,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy, C(O)R.sup.5,
C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl and 5-12
membered heteroaryl, where each said C.sub.1-C.sub.4 alkyl or
C.sub.1-C.sub.4 alkoxy is optionally substituted by one or more
R.sup.6, and each said C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl or 5-12 membered heteroaryl is optionally substituted
by one or more R.sup.7;
[0014] R.sup.2 is H, F or C.sub.1-C.sub.4 alkyl;
[0015] L is a bond or a C.sub.1-C.sub.4 alkylene;
[0016] R.sup.3 is selected from the group consisting of
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy, OH, CN, C(O)R.sup.8,
COOR.sup.9, NR.sup.10R.sup.11, OR.sup.12, C.sub.3-C.sub.8
cycloalkyl, 3-12 membered heterocyclyl and 5-12 membered
heteroaryl, where each said C.sub.1-C.sub.4 alkyl or
C.sub.1-C.sub.4 alkoxy is optionally substituted by one or more
R.sup.6, and each said C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl or 5-12 membered heteroaryl is optionally substituted
by one or more R.sup.7;
[0017] R.sup.4 is H, halo or C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.6;
[0018] R.sup.5 is C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14;
[0019] each R.sup.6 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy;
[0020] each R.sup.7 is independently C.sub.1-C.sub.4 alkyl, OH, F,
CN, C.sub.1-C.sub.4 alkoxy, .dbd.O, CHO, C(O)R.sup.13,
SO.sub.2R.sup.13 or 3-6 membered heterocyclyl;
[0021] R.sup.8 is C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14;
[0022] R.sup.9 is H or C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14;
[0023] R.sup.10 and R.sup.11 are independently H or C.sub.1-C.sub.4
alkyl, where each said C.sub.1-C.sub.4 alkyl is optionally
substituted by one or more R.sup.14;
[0024] R.sup.12 is selected from the group consisting of
C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl and 5-12
membered heteroaryl, where each said C.sub.3-C.sub.8 cycloalkyl,
3-12 membered heterocyclyl or 5-12 membered heteroaryl is
optionally substituted by one or more R.sup.7;
[0025] each R.sup.13 is independently C.sub.1-C.sub.4 alkyl, where
each said C.sub.1-C.sub.4 alkyl is optionally substituted by one or
more R.sup.15;
[0026] each R.sup.14 and R.sup.15 is independently OH, F, CN or
C.sub.1-C.sub.4 alkoxy; and
[0027] X and Z are independently C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4 alkoxy or
C.sub.1-C.sub.4 fluoroalkoxy.
[0028] In some embodiments, the compound of Formula (I) has the
absolute stereochemistry at the carbon atom bearing the R.sup.1 and
R.sup.2 substituents as shown in Formula (I-A) or (I-B):
##STR00003##
[0029] or a pharmaceutically acceptable salt thereof,
[0030] wherein:
[0031] R.sup.1, R.sup.2, L, R.sup.3, R.sup.4, X and Z are defined
as for Formula (I).
[0032] In another aspect, the invention provides a compound of
Formula (II), (II-A) or (II-B):
##STR00004##
[0033] or a pharmaceutically acceptable salt thereof,
[0034] wherein:
[0035] R.sup.1, L, R.sup.3 and X are defined as for Formula (I);
and
[0036] R.sup.4 is H, Cl, Br, F or CH.sub.3.
[0037] In another aspect, the invention provides a compound of
formula (III):
##STR00005##
[0038] or a pharmaceutically acceptable salt thereof,
[0039] wherein:
[0040] R.sup.1 and R.sup.3 are taken together to form a 3-12
membered heterocyclyl optionally substituted by one or more
R.sup.7;
[0041] R.sup.2 is H, F or C.sub.1-C.sub.4 alkyl;
[0042] R.sup.4 is H, halo or C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.6;
[0043] each R.sup.6 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy;
[0044] each R.sup.7 is independently C.sub.1-C.sub.4 alkyl, OH, F,
CN, C.sub.1-C.sub.4 alkoxy, .dbd.O, CHO, C(O)R.sup.13,
SO.sub.2R.sup.13 or 3-6 membered heterocyclyl;
[0045] each R.sup.13 is independently C.sub.1-C.sub.4 alkyl, where
each said C.sub.1-C.sub.4 alkyl is optionally substituted by one or
more R.sup.15;
[0046] each R.sup.15 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy; and
[0047] X and Z are independently C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4 alkoxy or
C.sub.1-C.sub.4 fluoroalkoxy.
[0048] In another aspect, the invention provides a pharmaceutical
composition comprising a compound of one of the formulae described
herein, or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier or excipient. In some
embodiments, the pharmaceutical composition comprises two or more
pharmaceutically acceptable carriers and/or excipients.
[0049] The invention also provides therapeutic methods and uses
comprising administering a compound of the invention, or a
pharmaceutically acceptable salt thereof.
[0050] In one aspect, the invention provides a method for the
treatment of abnormal cell growth in a subject comprising
administering to the subject a therapeutically effective amount of
a compound of the invention, or a pharmaceutically acceptable salt
thereof.
[0051] In frequent embodiments of the methods provided herein, the
abnormal cell growth is cancer. In some embodiments, the methods
provided result in one or more of the following effects: (1)
inhibiting cancer cell proliferation; (2) inhibiting cancer cell
invasiveness; (3) inducing apoptosis of cancer cells; (4)
inhibiting cancer cell metastasis; or (5) inhibiting
angiogenesis.
[0052] In another aspect, the invention provides a method for the
treatment of a disorder mediated by EZH2 in a subject comprising
administering to the subject a compound of the invention, or a
pharmaceutically acceptable salt thereof, in an amount that is
effective for treating said disorder. The compounds and salts of
the present invention may inhibit wild-type and certain mutant
forms of human histone methyltransferase EZH2.
[0053] In another aspect, the invention provides a compound of one
of the formulae described herein, or pharmaceutically acceptable
salt thereof, for use in the treatment of abnormal cell growth in a
subject.
[0054] In a further aspect, the invention provides the use of a
compound of one of the formulae described herein, or
pharmaceutically acceptable salt thereof, for the treatment of
abnormal cell growth in a subject.
[0055] In yet another aspect, the invention provides the use of a
compound of one of the formulae described herein, or a
pharmaceutically acceptable salt thereof, for the preparation of a
medicament for the treatment of abnormal cell growth.
[0056] In frequent embodiments, the abnormal cell growth is cancer
and the subject is a human.
[0057] In some embodiments, the methods described herein further
comprise administering to the subject an amount of an anti-cancer
therapeutic agent or a palliative agent, which amounts may be
together effective in treating said abnormal cell growth. In some
embodiments, the one or more anti-cancer therapeutic agent is
selected from anti-tumor agents, anti-angiogenesis agents, signal
transduction inhibitors and antiproliferative agents, which amounts
are together effective in treating said abnormal cell growth. In
some such embodiments, the anti-tumor agent is selected from the
group consisting of mitotic inhibitors, alkylating agents,
anti-metabolites, intercalating antibiotics, growth factor
inhibitors, radiation, cell cycle inhibitors, enzymes,
topoisomerase inhibitors, biological response modifiers,
antibodies, cytotoxics, anti-hormones, and anti-androgens.
[0058] In other embodiments, the uses described herein comprise the
use of a compound of one of the formulae described herein or
pharmaceutically acceptable salt thereof, in combination with one
or more substances selected from anti-tumor agents,
anti-angiogenesis agents, signal transduction inhibitors and
antiproliferative agents.
[0059] In some embodiments, the medicaments described herein may be
adapted for use in combination with one or more substances selected
from anti-tumor agents, anti-angiogenesis agents, signal
transduction inhibitors and antiproliferative agents.
[0060] Each of the embodiments of the compounds of the present
invention described below may be combined with one or more other
embodiments of the compounds of the present invention described
herein not inconsistent with the embodiment(s) with which it is
combined. In addition, each of the embodiments below describing the
invention envisions within its scope the pharmaceutically
acceptable salts of the compounds of the invention. Accordingly,
the phrase "or a pharmaceutically acceptable salt thereof" is
implicit in the description of all compounds described herein.
DETAILED DESCRIPTION OF THE INVENTION
[0061] The present invention may be understood more readily by
reference to the following detailed description of the preferred
embodiments of the invention and the Examples included herein. It
is to be understood that the terminology used herein is for the
purpose of describing specific embodiments only and is not intended
to be limiting. It is further to be understood that unless
specifically defined herein, the terminology used herein is to be
given its traditional meaning as known in the relevant art.
[0062] As used herein, the singular form "a", "an", and "the"
include plural references unless indicated otherwise. For example,
"a" substituent includes one or more substituents.
[0063] "Alkyl" refers to a saturated, monovalent aliphatic
hydrocarbon radical including straight chain and branched chain
groups having the specified number of carbon atoms. Alkyl
substituents typically contain 1 to 20 carbon atoms
("C.sub.1-C.sub.20 alkyl"), preferably 1 to 12 carbon atoms
("C.sub.1-C.sub.12 alkyl"), more preferably 1 to 8 carbon atoms
("C.sub.1-C.sub.8 alkyl"), or 1 to 6 carbon atoms ("C.sub.1-C.sub.6
alkyl"), or 1 to 4 carbon atoms ("C.sub.1-C.sub.4 alkyl"). Examples
of alkyl groups include methyl, ethyl, n-propyl, isopropyl,
n-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl,
n-hexyl, n-heptyl, n-octyl and the like. Alkyl groups may be
substituted or unsubstituted. In particular, unless otherwise
specified, alkyl groups may be substituted by one or more halo
groups, up to the total number of hydrogen atoms present on the
alkyl moiety. Thus, C.sub.1-C.sub.4 alkyl includes halogenated
alkyl groups, and in particular fluorinated alkyl groups, having 1
to 4 carbon atoms, e.g., trifluoromethyl or difluoroethyl (i.e.,
CF.sub.3 and --CH.sub.2CHF.sub.2).
[0064] Alkyl groups described herein as optionally substituted by
may be substituted by one or more substituent groups, which are
selected independently unless otherwise indicated. The total number
of substituent groups may equal the total number of hydrogen atoms
on the alkyl moiety, to the extent such substitution makes chemical
sense. Optionally substituted alkyl groups typically contain from 1
to 6 optional substituents, sometimes 1 to 5 optional substituents,
preferably from 1 to 4 optional substituents, or more preferably
from 1 to 3 optional substituents.
[0065] Optional substituent groups suitable for alkyl include, but
are not limited to C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl, C.sub.6-C.sub.12 aryl and 5-12 membered heteroaryl,
halo, .dbd.O (oxo), .dbd.S (thiono), .dbd.N--CN, .dbd.N--OR.sup.X,
.dbd.NR.sup.X, --CN, --C(O)R.sup.x, --CO.sub.2R.sup.x,
--C(O)NR.sup.xR.sup.y, --SR.sup.x, --SOR.sup.x, --SO.sub.2R.sup.x,
--SO.sub.2NR.sup.xR.sup.y, --NO.sub.2, --NR.sup.xR.sup.y,
--NR.sup.xC(O)R.sup.y, --NR.sup.xC(O)NR.sup.xR.sup.y,
--NR.sup.xC(O)OR.sup.x, --NR.sup.xSO.sub.2R.sup.y,
--NR.sup.xSO.sub.2NR.sup.xR.sup.y, --OR.sup.x, --OC(O)R.sup.x and
--OC(O)NR.sup.xR.sup.y; wherein each R.sup.x and R.sup.y is
independently H, C.sub.1-C.sub.8 alkyl, C.sub.1-C.sub.8 acyl,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8
cycloalkyl, 3-12 membered heterocyclyl, C.sub.6-C.sub.12 aryl, or
5-12 membered heteroaryl, or R.sup.x and R.sup.y may be taken
together with the N atom to which they are attached to form a 3-12
membered heterocyclyl or 5-12 membered heteroaryl, each optionally
containing 1, 2 or 3 additional heteroatoms selected from O, N and
S; each R.sup.x and R.sup.y is optionally substituted with 1 to 3
substituents independently selected from the group consisting of
halo, .dbd.O, .dbd.S, .dbd.N--CN, .dbd.N--OR', .dbd.NR', --CN,
--C(O)R', --CO.sub.2R', --C(O)NR'.sub.2, --SR', --SOR',
--SO.sub.2R', --SO.sub.2NR'.sub.2, --NO.sub.2, --NR'.sub.2,
--NR'C(O)R', --NR'C(O)NR'.sub.2, --NR'C(O)OR', --NR'SO.sub.2R',
--NR'SO.sub.2NR'.sub.2, --OR', --OC(O)R' and --OC(O)NR'.sub.2,
wherein each R' is independently H, C.sub.1-C.sub.8 alkyl,
C.sub.1-C.sub.8 acyl, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl,
C.sub.6-C.sub.12 aryl, or C.sub.5-C.sub.12 heteroaryl; and wherein
each said C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl,
C.sub.6-C.sub.12 aryl and 5-12 membered heteroaryl is optionally
substituted as further defined herein.
[0066] Typical substituent groups on alkyl include halo, --OH,
C.sub.1-C.sub.4 alkoxy, --O--C.sub.6-C.sub.12 aryl, --CN, .dbd.O,
--COOR.sup.x, --OC(O)R.sup.x, --C(O)NR.sup.xR.sup.y,
--NR.sup.xC(O)R.sup.y, --NR.sup.xR.sup.y, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.12 aryl, 5-12 membered heteroaryl and
3-12 membered heterocyclyl; where each R.sup.x and R.sup.y is
independently H or C.sub.1-C.sub.4 alkyl, or R.sup.x and R.sup.y
may be taken together with the N to which they are attached form a
3-12 membered heterocyclyl or 5-12 membered heteroaryl ring, each
optionally containing 1, 2 or 3 additional heteroatoms selected
from O, N and S; wherein each said C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.12 aryl, 5-12 membered heteroaryl and 3-12 membered
heterocyclyl is optionally substituted by 1 to 3 substituents
independently selected from the group consisting of halo, --OH,
.dbd.O, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 hydroxyalkyl,
C.sub.1-C.sub.4 alkoxy-C.sub.1-C.sub.6 alkyl, --CN, --NH.sub.2,
--NH(C.sub.1-C.sub.4 alkyl), and --N(C.sub.1-C.sub.4
alkyl).sub.2.
[0067] In some embodiments, alkyl is optionally substituted by one
or more substituents, and preferably by 1 to 3 substituents, which
are independently selected from the group consisting of halo, --OH,
C.sub.1-C.sub.4 alkoxy, --O--C.sub.6-C.sub.12 aryl, --CN, .dbd.O,
--COOR.sup.x, --OC(O)R.sup.x, --C(O)NR.sup.xR.sup.y,
--NR.sup.xC(O)R.sup.y, --NR.sup.xR.sup.y, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.12 aryl, 5-12 membered heteroaryl and
3-12 membered heterocyclyl; where each R.sup.x and R.sup.y is
independently H or C.sub.1-C.sub.4 alkyl, or R.sup.x and R.sup.y
may be taken together with the N to which they are attached form a
3-12 membered heterocyclyl or 5-12 membered heteroaryl ring, each
optionally containing 1, 2 or 3 additional heteroatoms selected
from O, N and S; and each said C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.12 aryl, 5-12 membered heteroaryl and 3-12 membered
heterocyclyl is optionally substituted by 1 to 3 substituents
independently selected from the group consisting of halo, --OH,
.dbd.O, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 hydroxyalkyl,
C.sub.1-C.sub.4 alkoxy-C.sub.1-C.sub.6 alkyl, --CN, --NH.sub.2,
--NH(C.sub.1-C.sub.4 alkyl) and --N(C.sub.1-C.sub.4
alkyl).sub.2.
[0068] In other embodiments, alkyl is optionally substituted by one
or more substituent, and preferably by 1 to 3 substituents,
independently selected from the group consisting of halo, --OH,
C.sub.1-C.sub.4 alkoxy, --CN, --NR.sup.xR.sup.y, C.sub.3-C.sub.8
cycloalkyl, 3-12 membered heterocyclyl, C.sub.6-C.sub.12 aryl and
5-12 membered heteroaryl; where each R.sup.x and R.sup.y is
independently H or C.sub.1-C.sub.4 alkyl, or R.sup.x and R.sup.y
may be taken together with the N to which they are attached form a
3-12 membered heterocyclyl or 5-12 membered heteroaryl ring, each
optionally containing 1, 2 or 3 additional heteroatoms selected
from O, N and S; and where each said cycloalkyl, heterocyclyl, aryl
or heteroaryl is optionally substituted by 1 to 3 substituents
independently selected from the group consisting of halo, --OH,
.dbd.O, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 hydroxyalkyl,
C.sub.1-C.sub.4 alkoxy-C.sub.1-C.sub.6 alkyl, --CN, --NH.sub.2,
--NH(C.sub.1-C.sub.4 alkyl) and --N(C.sub.1-C.sub.4
alkyl).sub.2.
[0069] In some instances, substituted alkyl groups may be
specifically named with reference to the substituent group. For
example, "haloalkyl" refers to an alkyl group having the specified
number of carbon atoms that is substituted by one or more halo
substituents, and typically contain 1-6 carbon atoms and 1, 2 or 3
halo atoms (i.e., "C.sub.1-C.sub.6 haloalkyl") or sometimes 1-4
carbon atoms and 1, 2 or 3 halo atoms (i.e., "C.sub.1-C.sub.4
haloalkyl"). Thus, a C.sub.1-C.sub.4 haloalkyl group includes
trifluoromethyl (--CF.sub.3) and difluoromethyl (--CF.sub.2H). More
specifically, fluorinated alkyl groups may be specifically referred
to as fluoroalkyl groups, e.g., C.sub.1-C.sub.6 or C.sub.1-C.sub.4
fluoroalkyl groups.
[0070] Similarly, "hydroxyalkyl" refers to an alkyl group having
the specified number of carbon atoms that is substituted by one or
more hydroxy substituents, and typically contain 1-6 carbon atoms
and 1, 2 or 3 hydroxy (i.e., "C.sub.1-C.sub.6 hydroxyalkyl"). Thus,
C.sub.1-C.sub.6 hydroxyalkyl includes hydroxymethyl (--CH.sub.2OH)
and 2-hydroxyethyl (--CH.sub.2CH.sub.2OH).
[0071] "Alkoxyalkyl" refers to an alkyl group having the specified
number of carbon atoms that is substituted by one or more alkoxy
substituents. Alkoxyalkyl groups typically contain 1-6 carbon atoms
in the alkyl portion and are substituted by 1, 2 or 3
C.sub.1-C.sub.4 alkoxy substituents. Such groups are sometimes
described herein as C.sub.1-C.sub.4 alkyoxy-C.sub.1-C.sub.6
alkyl.
[0072] "Aminoalkyl" refers to alkyl group having the specified
number of carbon atoms that is substituted by one or more
substituted or unsubstituted amino groups, as such groups are
further defined herein. Aminoalkyl groups typically contain 1-6
carbon atoms in the alkyl portion and are substituted by 1, 2 or 3
amino substituents. Thus, a C.sub.1-C.sub.6 aminoalkyl group
includes, for example, aminomethyl (--CH.sub.2NH.sub.2),
N,N-dimethylamino-ethyl (--CH.sub.2CH.sub.2N(CH.sub.3).sub.2),
3-(N-cyclopropylamino)propyl
(--CH.sub.2CH.sub.2CH.sub.2NH--.sup.cPr) and N-pyrrolidinylethyl
(--CH.sub.2CH.sub.2-N-pyrrolidinyl).
[0073] "Alkenyl" refers to an alkyl group, as defined herein,
consisting of at least two carbon atoms and at least one
carbon-carbon double bond. Typically, alkenyl groups have 2 to 20
carbon atoms ("C.sub.2-C.sub.20 alkenyl"), preferably 2 to 12
carbon atoms ("C.sub.2-C.sub.12 alkenyl"), more preferably 2 to 8
carbon atoms ("C.sub.2-C.sub.8 alkenyl"), or 2 to 6 carbon atoms
("C.sub.2-C.sub.6 alkenyl"), or 2 to 4 carbon atoms
("C.sub.2-C.sub.4 alkenyl"). Representative examples include, but
are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-, 2-, or
3-butenyl, and the like. Alkenyl groups may be unsubstituted or
substituted by the same groups that are described herein as
suitable for alkyl.
[0074] "Alkynyl" refers to an alkyl group, as defined herein,
consisting of at least two carbon atoms and at least one
carbon-carbon triple bond. Alkynyl groups have 2 to 20 carbon atoms
("C.sub.2-C.sub.20 alkynyl"), preferably 2 to 12 carbon atoms
("C.sub.2-C.sub.12 alkynyl"), more preferably 2 to 8 carbon atoms
("C.sub.2-C.sub.8 alkynyl"), or 2 to 6 carbon atoms
("C.sub.2-C.sub.6 alkynyl"), or 2 to 4 carbon atoms
("C.sub.2-C.sub.4 alkynyl"). Representative examples include, but
are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-, 2-, or
3-butynyl, and the like. Alkynyl groups may be unsubstituted or
substituted by the same groups that are described herein as
suitable for alkyl.
[0075] "Alkylene" as used herein refers to a divalent hydrocarbyl
group having the specified number of carbon atoms which can link
two other groups together. Sometimes it refers to a group
--(CH.sub.2).sub.n-- where n is 1-8, and preferably n is 1-4. Where
specified, an alkylene may also be substituted by other groups and
may include one or more degrees of unsaturation (i.e., an
alkenylene or alkynlene moiety) or rings. The open valences of an
alkylene need not be at opposite ends of the chain. Thus branched
alkylene groups such as --CH(Me)- and --C(Me).sub.2- are also
included within the scope of the term `alkylenes`, as are cyclic
groups such as cyclopropan-1,1-diyl and unsaturated groups such as
ethylene (--CH.dbd.CH--) or propylene (--CH.sub.2--CH.dbd.CH--).
Where an alkylene group is described as optionally substituted, the
substituents include those typically present on alkyl groups as
described herein.
[0076] "Heteroalkylene" refers to an alkylene group as described
above, wherein one or more non-contiguous carbon atoms of the
alkylene chain are replaced by --N(R)--, --O-- or --S(O).sub.q--,
where R is H or C.sub.1-C.sub.4 alkyl and q is 0-2. For example,
the group --O--(CH.sub.2).sub.1-4-- is a
`C.sub.2-C.sub.5`-heteroalkylene group, where one of the carbon
atoms of the corresponding alkylene is replaced by O.
[0077] "Alkoxy" refers to a monovalent --O-alkyl group, wherein the
alkyl portion has the specified number of carbon atoms. Alkoxy
groups typically contain 1 to 8 carbon atoms ("C.sub.1-C.sub.8
alkoxy"), or 1 to 6 carbon atoms ("C.sub.1-C.sub.6 alkoxy"), or 1
to 4 carbon atoms ("C.sub.1-C.sub.4 alkoxy"). For example,
C.sub.1-C.sub.4 alkoxy includes --OCH.sub.3, --OCH.sub.2CH.sub.3,
--OCH(CH.sub.3).sub.2, --OC(CH.sub.3).sub.3, and the like. Such
groups may also be referred to herein as methoxy, ethoxy,
isopropoxy, tert-butyloxy, etc. Alkoxy groups may be unsubstituted
or substituted on the alkyl portion by the same groups that are
described herein as suitable for alkyl. In particular, alkoxy
groups may be substituted by one or more halo groups, up to the
total number of hydrogen atoms present on the alkyl portion. Thus,
C.sub.1-C.sub.4 alkoxy includes halogenated alkoxy groups, e.g.,
trifluoromethoxy and 2,2-difluoroethoxy (i.e., --OCF.sub.3 and
--OCH.sub.2CHF.sub.2). In some instances, such groups may be
referred to as "haloalkoxy" (or, where fluorinated, more
specifically as "fluoroalkoxy") groups having the specified number
of carbon atoms and substituted by one or more halo substituents,
and typically contain 1-6 carbon atoms and 1, 2 or 3 halo atoms
(i.e., "C.sub.1-C.sub.6 haloalkoxy") or sometimes 1-4 carbon atoms
and 1, 2 or 3 halo atoms (i.e., "C.sub.1-C.sub.4 haloalkoxy").
Thus, a C.sub.1-C.sub.4 haloalkoxy group includes trifluoromethoxy
(--OCF.sub.3) and difluoromethoxy (--OCF.sub.2H). More
specifically, fluorinated alkyl groups may be specifically referred
to as fluoroalkoxy groups, e.g., C.sub.1-C.sub.6 or C.sub.1-C.sub.4
fluoroalkoxy groups.
[0078] Similarly, "thioalkoxy" refers to a monovalent --S-alkyl
group, wherein the alkyl portion has the specified number of carbon
atoms, and may be optionally substituted on the alkyl portion by
the same groups that are described herein as suitable for alkyl.
For example, a C.sub.1-C.sub.4 thioalkoxy includes --SCH.sub.3 and
--SCH.sub.2CH.sub.3.
[0079] "Cycloalkyl" refers to a non-aromatic, saturated or
partially unsaturated carbocyclic ring system containing the
specified number of carbon atoms, which may be a monocyclic,
bridged or fused bicyclic or polycyclic ring system that is
connected to the base molecule through a carbon atom of the
cycloalkyl ring. Typically, the cycloalkyl groups of the invention
contain 3 to 12 carbon atoms ("C.sub.3-C.sub.12 cycloalkyl"),
preferably 3 to 8 carbon atoms ("C.sub.3-C.sub.8 cycloalkyl").
Representative examples include, e.g., cyclopropane, cyclobutane,
cyclopentane, cyclopentene, cyclohexane, cyclohexene,
cyclohexadiene, cycloheptane, cycloheptatriene, adamantane, and the
like. Cycloalkyl groups may be unsubstituted or substituted by the
same groups that are described herein as suitable for alkyl.
[0080] Illustrative examples of cycloalkyl rings include, but are
not limited to, the following:
##STR00006##
[0081] "Cycloalkylalkyl" may be used to describe a cycloalkyl ring,
typically a C.sub.3-C.sub.8 cycloalkyl, which is connected to the
base molecule through an alkylene linker, typically a
C.sub.1-C.sub.4 alkylene. Cycloalkylalkyl groups are described by
the total number of carbon atoms in the carbocyclic ring and
linker, and typically contain from 4-12 carbon atoms
("C.sub.4-C.sub.12 cycloalkylalkyl"). Thus a cyclopropylmethyl
group is a C.sub.4-cycloalkylalkyl group and a cyclohexylethyl is a
C.sub.8-cycloalkylalkyl. Cycloalkylalkyl groups may be
unsubstituted or substituted on the cycloalkyl and/or alkylene
portions by the same groups that are described herein as suitable
for alkyl groups.
[0082] The terms "heterocyclyl", "heterocyclic" or
"heteroalicyclic" may be used interchangeably herein to refer to a
non-aromatic, saturated or partially unsaturated ring system
containing the specified number of ring atoms, including at least
one heteroatom selected from N, O and S as a ring member, wherein
the heterocyclic ring is connected to the base molecule via a ring
atom, which may be C or N. Heterocyclic rings may be fused to one
or more other heterocyclic or carbocyclic rings, which fused rings
may be saturated, partially unsaturated or aromatic. Preferably,
heterocyclic rings contain 1 to 4 heteroatoms selected from N, O,
and S as ring members, and more preferably 1 to 2 ring heteroatoms,
provided that such heterocyclic rings do not contain two contiguous
oxygen atoms. Heterocyclyl groups may be unsubstituted or
substituted by the same groups that are described herein as
suitable for alkyl, aryl or heteroaryl. In addition, ring N atoms
may be optionally substituted by groups suitable for an amine,
e.g., alkyl, acyl, carbamoyl, sulfonyl substituents, etc., and ring
S atoms may be optionally substituted by one or two oxo groups
(i.e., S(O).sub.q, where q is 0, 1 or 2).
[0083] Preferred heterocycles include 3-12 membered heterocyclyl
groups in accordance with the definition herein.
[0084] Illustrative examples of partially unsaturated heterocyclic
groups include, but are not limited to:
##STR00007##
[0085] Illustrative examples of bridged and fused heterocyclic
groups include, but are not limited to:
##STR00008##
[0086] Illustrative examples of saturated heterocyclic groups
include, but are not limited to:
##STR00009##
[0087] In frequent embodiments, heterocyclic groups contain 3-12
ring members, including both carbon and non-carbon heteroatoms, and
preferably 4-6 ring members. In certain embodiments, substituent
groups comprising 3-12 membered heterocycles are selected from the
group consisting of azetidinyl, pyrrolidinyl, piperidinyl,
piperazinyl, morpholinyl and thiomorpholinyl, each of which may be
optionally substituted to the extent such substitution makes
chemical sense. In other embodiments, substituent groups comprising
3-12 membered heterocycles are selected from the group consisting
of oxetanyl, tetrahydrofuranyl and tetrahydropyranyl, each of which
may be optionally substituted to the extent such substitution makes
chemical sense. In particular embodiments, said 3-12 membered
heterocycle is oxetanyl, optionally substituted to the extent such
substitution makes chemical sense.
[0088] It is understood that no more than two N, O or S atoms are
ordinarily connected sequentially except where an oxo group is
attached to N or S to form a nitro or sulfonyl group, or in the
case of certain heteroaromatic rings, such as triazine, triazole,
tetrazole, oxadiazole, thiadiazole, and the like.
[0089] The term "heterocyclylalkyl" may be used to describe a
heterocyclic group of the specified size that is connected to the
base molecule through an alkylene linker of the specified length.
Typically, such groups contain an optionally substituted 3-12
membered heterocycle attached to the base molecule through a
C.sub.1-C.sub.4 alkylene linker. Where so indicated, such groups
may be optionally substituted on the alkylene portion by the same
groups that are described herein as suitable for alkyl groups and
on the heterocyclic portion by groups described as suitable for
heterocyclic rings.
[0090] "Aryl" or "aromatic" refer to an optionally substituted
monocyclic, biaryl or fused bicyclic or polycyclic ring systems,
having the well-known characteristics of aromaticity, wherein at
least one ring contains a completely conjugated pi-electron system.
Typically, aryl groups contain 6 to 20 carbon atoms
("C.sub.6-C.sub.20 aryl") as ring members, preferably 6 to 14
carbon atoms ("C.sub.6-C.sub.14 aryl") or more preferably, 6 to 12
carbon atoms ("C.sub.6-C.sub.12 aryl"). Fused aryl groups may
include an aryl ring (e.g., a phenyl ring) fused to another aryl
ring, or fused to a saturated or partially unsaturated carbocyclic
or heterocyclic ring. The point of attachment to the base molecule
on such fused aryl ring systems may be a C atom the aromatic
portion or a C or N atom of the non-aromatic portion of the ring
system. Examples, without limitation, of aryl groups include
phenyl, biphenyl, naphthyl, anthracenyl, phenanthrenyl, indanyl,
indenyl, and tetrahydronaphthyl. The aryl group may be
unsubstituted or substituted as further described herein.
[0091] Similarly, "heteroaryl" or "heteroaromatic" refer to
monocyclic, heterobiaryl or fused bicyclic or polycyclic ring
systems having the well-known characteristics of aromaticity that
contain the specified number of ring atoms and include at least one
heteroatom selected from N, O and S as a ring member in an aromatic
ring. The inclusion of a heteroatom permits aromaticity in
5-membered rings as well as 6-membered rings. Typically, heteroaryl
groups contain 5 to 20 ring atoms ("5-20 membered heteroaryl"),
preferably 5 to 14 ring atoms ("5-14 membered heteroaryl"), and
more preferably 5 to 12 ring atoms ("5-12 membered heteroaryl").
Heteroaryl rings are attached to the base molecule via a ring atom
of the heteroaromatic ring, such that aromaticity is maintained.
Thus, 6-membered heteroaryl rings may be attached to the base
molecule via a ring C atom, while 5-membered heteroaryl rings may
be attached to the base molecule via a ring C or N atom. Examples
of unsubstituted heteroaryl groups often include, but are not
limited to, pyrrole, furan, thiophene, pyrazole, imidazole,
isoxazole, oxazole, isothiazole, thiazole, triazole, oxadiazole,
thiadiazole, tetrazole, pyridine, pyridazine, pyrimidine, pyrazine,
benzofuran, benzothiophene, indole, benzimidazole, indazole,
quinoline, isoquinoline, purine, triazine, naphthryidine and
carbazole. In frequent preferred embodiments, 5- or 6-membered
heteroaryl groups are selected from the group consisting of
pyrrolyl, furanyl, thiophenyl, pyrazolyl, imidazolyl, isoxazolyl,
oxazolyl, isothiazolyl, thiazolyl, triazolyl, pyridinyl,
pyrimidinyl, pyrazinyl and pyridazinyl rings. The heteroaryl group
may be unsubstituted or substituted as further described
herein.
[0092] Aryl, heteroaryl and heterocyclyl moieties described herein
as optionally substituted by may be substituted by one or more
substituent groups, which are selected independently unless
otherwise indicated. The total number of substituent groups may
equal the total number of hydrogen atoms on the aryl, heteroaryl or
heterocyclyl moiety, to the extent such substitution makes chemical
sense and aromaticity is maintained in the case of aryl and
heteroaryl rings. Optionally substituted aryl, heteroaryl or
heterocyclyl groups typically contain from 1 to 5 optional
substituents, sometimes 1 to 4 optional substituents, preferably 1
to 3 optional substituents, or more preferably 1-2 optional
substituents.
[0093] Optional substituent groups suitable for aryl, heteroaryl
and heterocyclyl rings include, but are not limited to:
C.sub.1-C.sub.8 alkyl, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl,
C.sub.6-C.sub.12 aryl and 5-12 membered heteroaryl; and halo,
.dbd.O, --CN, --C(O)R.sup.x, --CO.sub.2R.sup.x,
--C(O)NR.sup.xR.sup.y, --SR.sup.x, --SOR.sup.x, --SO.sub.2R.sup.x,
--SO.sub.2NR.sup.xR.sup.y, --NO.sub.2, --NR.sup.xR.sup.y,
--NR.sup.xC(O)R.sup.y, --NR.sup.xC(O)NR.sup.xR.sup.y,
--NR.sup.xC(O)OR.sup.x, --NR.sup.xSO.sub.2R.sup.y,
--NR.sup.xSO.sub.2NR.sup.xR.sup.y, --OR.sup.x, --OC(O)R.sup.x and
--OC(O)NR.sup.xR.sup.y; where each R.sup.x and R.sup.y is
independently H, C.sub.1-C.sub.8 alkyl, C.sub.1-C.sub.8 acyl,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8
cycloalkyl, 3-12 membered heterocyclyl, C.sub.6-C.sub.12 aryl, or
5-12 membered heteroaryl, or R.sup.x and R.sup.y may be taken
together with the N atom to which they are attached to form a 3-12
membered heterocyclyl or 5-12 membered heteroaryl, each optionally
containing 1, 2 or 3 additional heteroatoms selected from O, N and
S; each R.sup.x and R.sup.y is optionally substituted with 1 to 3
substituents independently selected from the group consisting of
halo, .dbd.O, .dbd.S, .dbd.N--CN, .dbd.N--OR', .dbd.NR', --CN,
--C(O)R', --CO.sub.2R', --C(O)NR'.sub.2, --SR', --SOR',
--SO.sub.2R', --SO.sub.2NR'.sub.2, --NO.sub.2, --NR'.sub.2,
--NR'C(O)R', --NR'C(O)NR'.sub.2, --NR'C(O)OR', --NR'SO.sub.2R',
--NR'SO.sub.2NR'.sub.2, --OR', --OC(O)R' and --OC(O)NR'.sub.2,
wherein each R' is independently H, C.sub.1-C.sub.8 alkyl,
C.sub.1-C.sub.8 acyl, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl,
C.sub.6-C.sub.12 aryl, or 5-12 membered heteroaryl; and each said
C.sub.1-C.sub.8 alkyl, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl,
C.sub.6-C.sub.12 aryl and 5-12 membered heteroaryl is optionally
substituted as further defined herein.
[0094] In typical embodiments, optional substitution on aryl,
heteroaryl and heterocyclyl rings includes one or more
substituents, and preferably 1 to 3 substituents, independently
selected from the group consisting of halo, C.sub.1-C.sub.8 alkyl,
--OH, C.sub.1-C.sub.8 alkoxy, CN, .dbd.O, --C(O)R.sup.x,
--COOR.sup.x, --OC(O)R.sup.x, --C(O)NR.sup.xR.sup.y,
--NR.sup.xC(O)R.sup.y, --SR.sup.x, --SOR.sup.x, --SO.sub.2R.sup.x,
--SO.sub.2NR.sup.xR.sup.y, --NO.sub.2, --NR.sup.xR.sup.y,
--NR.sup.xC(O)R.sup.y, --NR.sup.xC(O)NR.sup.xR.sup.y,
--NR.sup.xC(O)OR.sup.y --NR.sup.xSO.sub.2R.sup.y,
--NR.sup.xSO.sub.2NR.sup.xR.sup.y, --OC(O)R.sup.x,
--OC(O)NR.sup.xR.sup.y, C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl, C.sub.6-C.sub.12 aryl, 5-12 membered heteroaryl,
--O--(C.sub.3-C.sub.8 cycloalkyl), --O-(3-12 membered
heterocyclyl), --O--(C.sub.6-C.sub.12 aryl) and --O-(5-12 membered
heteroaryl); where each R.sup.x and R.sup.y is independently H or
C.sub.1-C.sub.4 alkyl, or R.sup.x and R.sup.y may be taken together
with the N to which they are attached form a 3-12 membered
heterocyclyl or 5-12 membered heteroaryl ring, each optionally
containing 1, 2 or 3 additional heteroatoms selected from O, N and
S; and wherein each said C.sub.1-C.sub.8 alkyl, C.sub.1-C.sub.8
alkoxy, C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl,
C.sub.6-C.sub.12 aryl, 5-12 membered heteroaryl,
--O--(C.sub.3-C.sub.8 cycloalkyl), --O-(3-12 membered
heterocyclyl), --O--(C.sub.6-C.sub.12 aryl) and --O-(5-12 membered
heteroaryl) that is described as an optional substituent or is part
of R.sup.x or R.sup.y is optionally substituted by 1 to 3
substituents independently selected from the group consisting of
halo, --OH, .dbd.O, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 hydroxyalkyl,
C.sub.1-C.sub.4 alkoxy-C.sub.1-C.sub.6 alkyl, --CN, --NH.sub.2,
--NH(C.sub.1-C.sub.4 alkyl), --N(C.sub.1-C.sub.4 alkyl).sub.2 and
N-pyrrolidinyl.
[0095] Illustrative examples of monocyclic heteroaryl groups
include, but are not limited to:
##STR00010##
[0096] Illustrative examples of fused ring heteroaryl groups
include, but are not limited to:
##STR00011## ##STR00012## ##STR00013##
[0097] An "arylalkyl" group refers to an aryl group as described
herein which is linked to the base molecule through an alkylene or
similar linker. Arylalkyl groups are described by the total number
of carbon atoms in the ring and linker. Thus a benzyl group is a
C.sub.7-arylalkyl group and a phenylethyl is a C.sub.8-arylalkyl.
Typically, arylalkyl groups contain 7-16 carbon atoms
("C.sub.7-C.sub.16 arylalkyl"), wherein the aryl portion contains
6-12 carbon atoms and the alkylene portion contains 1-4 carbon
atoms. Such groups may also be represented as --C.sub.1-C.sub.4
alkylene-C.sub.6-C.sub.12 aryl.
[0098] "Heteroarylalkyl" refers to a heteroaryl group as described
above that is attached to the base molecule through an alkylene
linker, and differs from "arylalkyl" in that at least one ring atom
of the aromatic moiety is a heteroatom selected from N, O and S.
Heteroarylalkyl groups are sometimes described herein according to
the total number of non-hydrogen atoms (i.e., C, N, S and O atoms)
in the ring and linker combined, excluding substituent groups.
Thus, for example, pyridinylmethyl may be referred to as a
"C.sub.7"-heteroarylalkyl. Typically, unsubstituted heteroarylalkyl
groups contain 6-20 non-hydrogen atoms (including C, N, S and O
atoms), wherein the heteroaryl portion typically contains 5-12
atoms and the alkylene portion typically contains 1-4 carbon atoms.
Such groups may also be represented as --C.sub.1-C.sub.4
alkylene-5-12 membered heteroaryl.
[0099] Similarly, "arylalkoxy" and "heteroarylalkoxy" refer to aryl
and heteroaryl groups, attached to the base molecule through a
heteroalkylene linker (i.e., --O-alkylene-), wherein the groups are
described according to the total number of non-hydrogen atoms
(i.e., C, N, S and O atoms) in the ring and linker combined. Thus,
--O--CH.sub.2-phenyl and --O--CH.sub.2-pyridinyl groups would be
referred to as C.sub.8-arylalkoxy and C.sub.8-heteroarylalkoxy
groups, respectively.
[0100] Where an arylalkyl, arylalkoxy, heteroarylalkyl or
heteroarylalkoxy group is described as optionally substituted, the
substituents may be on either the divalent linker portion or on the
aryl or heteroaryl portion of the group. The substituents
optionally present on the alkylene or heteroalkylene portion are
the same as those described above for alkyl or alkoxy groups
generally, while the substituents optionally present on the aryl or
heteroaryl portion are the same as those described above for aryl
or heteroaryl groups generally.
[0101] "Hydroxy" refers to an --OH group.
[0102] "Acyloxy" refers to a monovalent group --OC(O)alkyl, wherein
the alkyl portion has the specified number of carbon atoms
(typically C.sub.1-C.sub.8, preferably C.sub.1-C.sub.6 or
C.sub.1-C.sub.4) and may be optionally substituted by groups
suitable for alkyl. Thus, C.sub.1-C.sub.4 acyloxy includes an
--OC(O)C.sub.1-C.sub.4 alkyl substituent, e.g.,
--OC(O)CH.sub.3.
[0103] "Acylamino" refers to a monovalent group, --NHC(O)alkyl or
--NRC(O)alkyl, wherein the alkyl portion has the specified number
of carbon atoms (typically C.sub.1-C.sub.8, preferably
C.sub.1-C.sub.6 or C.sub.1-C.sub.4) and may be optionally
substituted by groups suitable for alkyl. Thus, C.sub.1-C.sub.4
acylamino includes an --NHC(O)C.sub.1-C.sub.4 alkyl substituent,
e.g., --NHC(O)CH.sub.3.
[0104] "Aryloxy" or "heteroaryloxy" refer to optionally substituted
--O-aryl or --O-heteroaryl, in each case where aryl and heteroaryl
are as further defined herein.
[0105] "Arylamino" or "heteroarylamino" refer to optionally
substituted --NH-aryl, --NR-aryl, --NH-heteroaryl or
--NR-heteroaryl, in each case where aryl and heteroaryl are as
further defined herein and R represents a substituent suitable for
an amine, e.g., an alkyl, acyl, carbamoyl or sulfonyl group, or the
like.
[0106] "Cyano" refers to a --C.ident.N group.
[0107] "Unsubstituted amino" refers to a group --NH.sub.2. Where
the amino is described as substituted or optionally substituted,
the term includes groups of the form --NR.sup.xR.sup.y, where each
or R.sup.x and R.sup.y is independently H, alkyl, alkenyl, alkynyl,
cycloalkyl, heterocyclyl, acyl, thioacyl, aryl, heteroaryl,
cycloalkylalkyl, arylalkyl or heteroarylalkyl, in each case having
the specified number of atoms and optionally substituted as
described herein. For example, "alkylamino" refers to a group
--NR.sup.xR.sup.y, wherein one of R.sup.x and R.sup.y is an alkyl
moiety and the other is H, and "dialkylamino" refers to
--NR.sup.xR.sup.y wherein both of R.sup.x and R.sup.y are alkyl
moieties, where the alkyl moieties having the specified number of
carbon atoms (e.g., --NH--C.sub.1-C.sub.4 alkyl or
--N(C.sub.1-C.sub.4 alkyl).sub.2). Typically, alkyl substituents on
amines contain 1 to 8 carbon atoms, preferably 1 to 6 carbon atoms,
or more preferably 1 to 4 carbon atoms. The term also includes
forms wherein R.sup.x and R.sup.y are taken together with the N
atom to which they are attached to form a 3-12 membered
heterocyclyl or 5-12 membered heteroaryl ring, each of which may
itself be optionally substituted as described herein for
heterocyclyl or heteroaryl rings, and which may contain 1 to 3
additional heteroatoms selected from N, O and S as ring members,
provided that such rings do not contain two contiguous oxygen
atoms.
[0108] "Halogen" or "halo" refers to fluoro, chloro, bromo and iodo
(F, Cl, Br, I). Preferably, halo refers to fluoro or chloro (F or
Cl).
[0109] "Heteroform" is sometimes used herein to refer to a
derivative of a group such as, e.g., an alkyl, aryl, or acyl,
wherein at least one carbon atom of the designated carbocyclic
group has been replaced by a heteroatom selected from N, O and S.
Thus the heteroforms of alkyl, alkenyl, alkynyl, acyl, aryl, and
arylalkyl are heteroalkyl, heteroalkenyl, heteroalkynyl,
heteroacyl, heteroaryl, and heteroarylalkyl, respectively. It is
understood that no more than two N, O or S atoms are ordinarily
connected sequentially, except where an oxo group is attached to N
or S to form a nitro or sulfonyl group.
[0110] "Optional" or "optionally" means that the subsequently
described event or circumstance may but need not occur, and the
description includes instances where the event or circumstance
occurs and instances in which it does not.
[0111] The terms "optionally substituted" and "substituted or
unsubstituted" may be used interchangeably to indicate that the
particular group being described may have no non-hydrogen
substituents (i.e., unsubstituted), or the group may have one or
more non-hydrogen substituents (i.e., substituted). If not
otherwise specified, the total number of substituents that may be
present is equal to the number of H atoms present on the
unsubstituted form of the group being described, to the extent that
such substitution makes chemical sense. Where an optional
substituent is attached via a double bond, such as an oxo (.dbd.O)
substituent, the group occupies two available valences, so the
total number of other substituents that may be included is reduced
by two. In the case where optional substituents are selected
independently from a list of alternatives, the selected groups may
be the same or different.
[0112] In one aspect, the invention provides a compound of formula
(I):
##STR00014##
[0113] or a pharmaceutically acceptable salt thereof,
[0114] wherein:
[0115] R.sup.1 is selected from the group consisting of H, F,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy, C(O)R.sup.5,
C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl and 5-12
membered heteroaryl, where each said C.sub.1-C.sub.4 alkyl or
C.sub.1-C.sub.4 alkoxy is optionally substituted by one or more
R.sup.6, and each said C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl or 5-12 membered heteroaryl is optionally substituted
by one or more R.sup.7;
[0116] R.sup.2 is H, F or C.sub.1-C.sub.4 alkyl;
[0117] L is a bond or a C.sub.1-C.sub.4 alkylene;
[0118] R.sup.3 is selected from the group consisting of
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy, OH, CN, C(O)R.sup.8,
COOR.sup.9, NR.sup.10R.sup.11, OR.sup.12, C.sub.3-C.sub.8
cycloalkyl, 3-12 membered heterocyclyl and 5-12 membered
heteroaryl, where each said C.sub.1-C.sub.4 alkyl or
C.sub.1-C.sub.4 alkoxy is optionally substituted by one or more
R.sup.6, and each said C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl or 5-12 membered heteroaryl is optionally substituted
by one or more R.sup.7;
[0119] R.sup.4 is H, halo or C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.6;
[0120] R.sup.5 is C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14;
[0121] each R.sup.6 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy;
[0122] each R.sup.7 is independently C.sub.1-C.sub.4 alkyl, OH, F,
CN, C.sub.1-C.sub.4 alkoxy, .dbd.O, CHO, C(O)R.sup.13,
SO.sub.2R.sup.13 or 3-6 membered heterocyclyl;
[0123] R.sup.8 is C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14;
[0124] R.sup.9 is H or C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14;
[0125] R.sup.10 and R.sup.11 are independently H or C.sub.1-C.sub.4
alkyl, where each said C.sub.1-C.sub.4 alkyl is optionally
substituted by one or more R.sup.14;
[0126] R.sup.12 is selected from the group consisting of
C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl and 5-12
membered heteroaryl, where each said C.sub.3-C.sub.8 cycloalkyl,
3-12 membered heterocyclyl or 5-12 membered heteroaryl is
optionally substituted by one or more R.sup.7;
[0127] each R.sup.13 is independently C.sub.1-C.sub.4 alkyl, where
each said C.sub.1-C.sub.4 alkyl is optionally substituted by one or
more R.sup.15;
[0128] each R.sup.14 and R.sup.15 is independently OH, F, CN or
C.sub.1-C.sub.4 alkoxy; and
[0129] X and Z are independently C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4 alkoxy or
C.sub.1-C.sub.4 fluoroalkoxy.
[0130] In some embodiments, the compound of Formula (I) has the
absolute stereochemistry at the carbon atom bearing the R.sup.1 and
R.sup.2 substituents as shown in Formula (I-A) or (I-B):
##STR00015##
[0131] or a pharmaceutically acceptable salt thereof,
[0132] wherein:
[0133] R.sup.1, R.sup.2, L, R.sup.3, R.sup.4, X and Z are defined
as for Formula (I).
[0134] Each of the aspects and embodiments described herein with
respect to Formula (I) is also applicable to compounds of Formula
(I-A) or (I-B).
[0135] In frequent embodiments of Formula (I), R.sup.2 is H.
[0136] In frequent embodiments of Formula (I), R.sup.4 is H, Cl,
Br, F or CH.sub.3. In some such embodiments, R.sup.4 is H or Cl. In
some embodiments, R.sup.4 is H. In other embodiments, R.sup.4 is
Cl. In further embodiments, R.sup.4 is Cl or Br.
[0137] In compounds of Formula (I), X and Z are independently
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4
alkoxy or C.sub.1-C.sub.4 fluoroalkoxy. In some embodiments, Z is
C.sub.1-C.sub.4 alkyl, for example CH.sub.3 or C.sub.2H.sub.5
(i.e., methyl or ethyl). In some embodiments, X is C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4 alkoxy. In
specific embodiments, X is CH.sub.3, OCH.sub.3 or OCHF.sub.2 (i.e.,
methyl, methoxy or difluoromethoxy). In further embodiments, X is
CH.sub.3, OCH.sub.3 or OCHF.sub.2, and Z is CH.sub.3.
[0138] In some embodiments of Formula (I), R.sup.1 is H or F.
[0139] In other embodiments of Formula (I), R.sup.1 is
C.sub.1-C.sub.4 alkyl or C.sub.1-C.sub.4 alkoxy, each optionally
substituted by one or more R.sup.6. In some such embodiments, said
alkyl or alkoxy is substituted by at least one OH or CN. In
specific embodiments, R.sup.1 is CH.sub.3, C.sub.2H.sub.5,
CH.sub.2OH, CH.sub.2CH.sub.2OH, CH(OH)CH.sub.3 or CH.sub.2CN (i.e.,
methyl, ethyl, hydroxymethyl, 2-hydroxyethyl, 1-hydroxyethyl or
cyanomethyl). In other specific embodiments, R.sup.1 is OCH.sub.3
(i.e., methoxy).
[0140] In other embodiments of Formula (I), R.sup.1 is C(O)R.sup.5,
where R.sup.5 is C.sub.1-C.sub.4 alkyl optionally substituted by
one or more R.sup.14. In some such embodiments, R.sup.5 is
C.sub.1-C.sub.4 alkyl optionally substituted by OH. In specific
embodiments, R.sup.5 is CH.sub.3, CH.sub.2OH, CH.sub.2CH.sub.2OH or
CH(CH.sub.3)OH such that R.sup.1 is C(O)CH.sub.3, C(O)CH.sub.2OH,
C(O)CH.sub.2CH.sub.2OH or C(O)CH(CH.sub.3)OH (i.e., acetyl,
.alpha.-hydroxyacetyl, 3-hydroxypropionyl or
2-hydroxypropionyl).
[0141] In still other embodiments of Formula (I), R.sup.1 is
C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl or 5-12
membered heteroaryl, where each said C.sub.3-C.sub.8 cycloalkyl,
3-12 membered heterocyclyl or 5-12 membered heteroaryl is
optionally substituted by one or more R.sup.7.
[0142] In some such embodiments, R.sup.1 is C.sub.3-C.sub.8
cycloalkyl optionally substituted by one or more R.sup.7. In some
such embodiments, R.sup.1 is cyclopropyl, cyclobutyl, cyclopentyl
or cyclohexyl, each optionally substituted by one or more
R.sup.7.
[0143] In other embodiments, R.sup.1 is 3-12 membered heterocyclyl
optionally substituted by one or more R.sup.7. In some such
embodiments, said 3-12 membered heterocyclyl is selected from the
group consisting of tetrahydrofuranyl, tetrahydropyranyl,
tetrahydrothiophenyl, tetrahydrothiopyranyl, pyrrolidinyl,
piperidinyl and morpholinyl, each optionally substituted by one or
more R.sup.7. In other such embodiments, said 3-12 membered
heterocyclyl is selected from the group consisting of oxetanyl,
tetrahydrofuranyl and tetrahydropyranyl, each optionally
substituted by one or more R.sup.7. In specific embodiments, said
3-12 membered heterocyclyl is oxetanyl optionally substituted by
one or more R.sup.7. In some such embodiments, said oxetanyl is
unsubstituted.
[0144] In still other such embodiments, R.sup.1 is 5-12 membered
heteroaryl, where each said 5-12 membered heteroaryl is optionally
substituted by one or more R.sup.7. In some such embodiments,
R.sup.1 is a 5- or 6-membered heteroaryl. In specific embodiments,
said 5- or 6-membered heteroaryl is selected from the group
consisting of pyrazolyl, imidazolyl, isoxazolyl, oxazolyl,
isothiazolyl, thiazolyl, and triazolyl groups, each optionally
substituted by one or more R.sup.7.
[0145] In certain embodiments, when R.sup.1 is C.sub.3-C.sub.8
cycloalkyl, 3-12 membered heterocyclyl or 5-12 membered heteroaryl,
each R.sup.7 is independently CH.sub.3, OH, F, CN, OCH.sub.3,
.dbd.O, CHO, C(O)R.sup.13, SO.sub.2R.sup.13 or 3-6 membered
heterocyclyl, where R.sup.13 is CH.sub.3 or C.sub.2H.sub.5 each
optionally substituted by OH (e.g., R.sup.13 is CH.sub.3,
CH.sub.2OH, CH.sub.2CH.sub.2OH or CH(CH.sub.3)OH.
[0146] In compounds of Formula (I), L is a bond or a
C.sub.1-C.sub.4 alkylene. In some embodiments of Formula (I), L is
a bond. In other embodiments of Formula (I), L is a C.sub.1-C.sub.4
alkylene. In specific embodiments, L is a methylene or
ethylene.
[0147] In compounds of Formula (I), R.sup.3 is selected from the
group consisting of C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy,
OH, CN, C(O)R.sup.8, COOR.sup.9, NR.sup.10R.sup.11, OR.sup.12,
C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl and 5-12
membered heteroaryl, where each said C.sub.1-C.sub.4 alkyl or
C.sub.1-C.sub.4 alkoxy is optionally substituted by one or more
R.sup.6, and each said C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl or 5-12 membered heteroaryl is optionally substituted
by one or more R.sup.7.
[0148] In some embodiments, R.sup.3 is C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 alkoxy or 3-12 membered heterocyclyl, each
optionally substituted as described above. In some embodiments,
R.sup.3 is C.sub.1-C.sub.4 alkyl or C.sub.1-C.sub.4 alkoxy, in
particular CH.sub.3 or OCH.sub.3 (i.e., methyl or methoxy).
[0149] In further embodiments, R.sup.3 is 3-12 membered
heterocyclyl optionally substituted by one or more R.sup.7. In some
such embodiments, said 3-12 membered heterocyclyl is selected from
the group consisting of tetrahydrofuranyl, tetrahydropyranyl,
tetrahydrothiophenyl, tetrahydrothiopyranyl, pyrrolidinyl,
piperidinyl and morpholinyl, each optionally substituted by one or
more R.sup.7. In other such embodiments, said 3-12 membered
heterocyclyl is selected from the group consisting of oxetanyl,
tetrahydrofuranyl and tetrahydropyranyl, each optionally
substituted by one or more R.sup.7. In specific embodiments, said
3-12 membered heterocyclyl is oxetanyl, optionally substituted by
one or more R.sup.7. In some such embodiments, said oxetanyl is
unsubstituted.
[0150] In further embodiments, L is a bond and R.sup.3 is
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy or 3-12 membered
heterocyclyl, each optionally substituted as described above. In
specific embodiments, L is a bond and R.sup.3 is C.sub.1-C.sub.4
alkyl or C.sub.1-C.sub.4 alkoxy, in particular CH.sub.3 or
OCH.sub.3 (i.e., methyl or methoxy).
[0151] In still further embodiments, L is a bond and R.sup.3 is
3-12 membered heterocyclyl optionally substituted by one or more
R.sup.7. In some such embodiments, L is a bond and said 3-12
membered heterocyclyl is selected from the group consisting of
tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl,
tetrahydrothiopyranyl, pyrrolidinyl, piperidinyl and morpholinyl,
each optionally substituted by one or more R.sup.7. In other such
embodiments, L is a bond and said 3-12 membered heterocyclyl is
selected from the group consisting of oxetanyl, tetrahydrofuranyl
and tetrahydropyranyl, each optionally substituted by one or more
R.sup.7. In specific embodiments, L is a bond and said 3-12
membered heterocyclyl is oxetanyl, optionally substituted by one or
more R.sup.7. In some such embodiments, said oxetanyl is
unsubstituted.
[0152] In other such embodiments, L is a C.sub.1-C.sub.4 alkylene
and R.sup.3 is C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy or
3-12 membered heterocyclyl, each optionally substituted as
described above.
[0153] In still other embodiments, R.sup.3 is OH, CN, C(O)R.sup.8
or COOR.sup.9, where R.sup.8 is C.sub.1-C.sub.4 alkyl optionally
substituted by one or more R.sup.14, and R.sup.9 is H or
C.sub.1-C.sub.4 alkyl optionally substituted by one or more
R.sup.14.
[0154] In some such embodiments, L is a bond and R.sup.3 is OH, CN,
C(O)R.sup.8 or COOR.sup.9, where R.sup.8 and R.sup.9 are described
as above.
[0155] In other embodiments, L is a C.sub.1-C.sub.4 alkylene and
R.sup.3 is OH, CN, C(O)R.sup.8 or COOR.sup.9 where R.sup.8 and
R.sup.9 are described as above. In specific embodiments, L is a
C.sub.1-C.sub.4 alkylene, for example methylene or ethylene, and
R.sup.3 is OH or CN.
[0156] In further embodiments, R.sup.3 is OR.sup.12,
C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl or 5-12
membered heteroaryl, where each said C.sub.3-C.sub.8 cycloalkyl,
3-12 membered heterocyclyl or 5-12 membered heteroaryl is
optionally substituted by one or more R.sup.7, and where R.sup.12
is C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl or 5-12
membered heteroaryl, each optionally substituted by one or more
R.sup.7.
[0157] In some such embodiments, L is a bond and R.sup.3 is
OR.sup.12, C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl
or 5-12 membered heteroaryl, as described above. In other such
embodiments, L is a C.sub.1-C.sub.4 alkylene and R.sup.3 is
OR.sup.12, C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl
or 5-12 membered heteroaryl, as described above.
[0158] In compounds of Formula (I), each R.sup.6 is independently
OH, F, CN or C.sub.1-C.sub.4 alkoxy. In frequent embodiments, at
least one R.sup.6 is OH or F.
[0159] In compounds of Formula (I), each R.sup.7 is independently
C.sub.1-C.sub.4 alkyl, OH, F, CN, C.sub.1-C.sub.4 alkoxy, .dbd.O,
CHO, C(O)R.sup.13, SO.sub.2R.sup.13 or 3-6 membered heterocyclyl.
In some embodiments, at least one R.sup.7 is C(O)R.sup.13, where
R.sup.13 is C.sub.1-C.sub.4 alkyl and said C.sub.1-C.sub.4 alkyl is
optionally further substituted by one or more R.sup.15. In some
embodiments, at least one R.sup.7 is SO.sub.2R.sup.13, where
R.sup.13 is C.sub.1-C.sub.4 alkyl and said C.sub.1-C.sub.4 alkyl is
optionally further substituted by one or more R.sup.15. In other
specific embodiments, at least one R.sup.7 is OH, F or
C.sub.1-C.sub.4 alkyl, e.g., CH.sub.3.
[0160] In specific embodiments, R.sup.1 and/or R.sup.3 is a 3-12
membered heterocyclyl substituted by one or more R.sup.7, where at
least one R.sup.7 is CHO or C(O)R.sup.13, and where R.sup.13 is
CH.sub.3, CH.sub.2OH or CH.sub.2CN, such that R.sup.7 is CHO,
C(O)CH.sub.3, C(O)CH.sub.2OH or C(O)CH.sub.2CN (i.e., formyl,
acetyl, hydroxyacetyl or cyanoacetyl, respectively).
[0161] In specific embodiments, R.sup.1 and/or R.sup.3 is a 3-12
membered heterocyclyl substituted by one or more R.sup.7, where at
least one R.sup.7 is SO.sub.2R.sup.13, and where R.sup.13 is
CH.sub.3, CH.sub.2OH or CH.sub.2CN, such that R.sup.7 is
SO.sub.2CH.sub.3, SO.sub.2CH.sub.2OH or SO.sub.2CH.sub.2CN.
[0162] In further specific embodiments, R.sup.1 and/or R.sup.3 is a
3-12 membered heterocyclyl substituted by one or more R.sup.7,
where at least one R.sup.7 is OH.
[0163] In compounds of Formula (I), R.sup.8 is C.sub.1-C.sub.4
alkyl, where each said C.sub.1-C.sub.4 alkyl is optionally
substituted by one or more R.sup.14.
[0164] In compounds of Formula (I), R.sup.9 is H or C.sub.1-C.sub.4
alkyl, where each said C.sub.1-C.sub.4 alkyl is optionally
substituted by one or more R.sup.14. In some such embodiments,
R.sup.9 is H. In other such embodiments, R.sup.9 is C.sub.1-C.sub.4
alkyl, optionally substituted as described above.
[0165] In compounds of Formula (I), R.sup.10 and R.sup.11 are
independently H or C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14.
[0166] Each R.sup.14 and R.sup.15 is independently OH, F, CN or
C.sub.1-C.sub.4 alkoxy.
[0167] In one preferred embodiment, the invention provides a
compound of Formula (I), (I-A) or (I-B), or a pharmaceutically
acceptable salt thereof, wherein:
[0168] R.sup.1 is 3-12 membered heterocyclyl or 5-12 membered
heteroaryl, where each said 3-12 membered heterocyclyl or 5-12
membered heteroaryl is optionally substituted by one or more
R.sup.7;
[0169] R.sup.2 is H;
[0170] L is C.sub.1-C.sub.4 alkylene;
[0171] R.sup.3 is OH or CN;
[0172] R.sup.4 is H or Cl;
[0173] each R.sup.7 is independently C.sub.1-C.sub.4 alkyl, OH, F,
CN, C.sub.1-C.sub.4 alkoxy, .dbd.O, CHO, C(O)R.sup.13,
SO.sub.2R.sup.13 or 3-6 membered heterocyclyl;
[0174] each R.sup.13 is independently C.sub.1-C.sub.4 alkyl, where
each said C.sub.1-C.sub.4 alkyl is optionally substituted by one or
more R.sup.15;
[0175] each R.sup.15 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy; and
[0176] X and Z are independently C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4 alkoxy or
C.sub.1-C.sub.4 fluoroalkoxy.
[0177] In another preferred embodiment, the invention provides a
compound of Formula (I), (I-A) or (I-B), or a pharmaceutically
acceptable salt thereof, wherein:
[0178] R.sup.1 is H or C.sub.1-C.sub.4 alkyl, where said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.6;
[0179] R.sup.2 is H;
[0180] L is a bond or C.sub.1-C.sub.4 alkylene;
[0181] R.sup.3 is selected from the group consisting of
C.sub.1-C.sub.4 alkyl, OH, CN, C(O)R.sup.8, COOR.sup.9,
NR.sup.10R.sup.11, C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl and 5-12 membered heteroaryl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.6, and each said C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl or 5-12 membered heteroaryl is optionally substituted
by one or more R.sup.7;
[0182] R.sup.4 is H or Cl;
[0183] each R.sup.6 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy;
[0184] each R.sup.7 is independently C.sub.1-C.sub.4 alkyl, OH, F,
CN, C.sub.1-C.sub.4 alkoxy, .dbd.O, CHO, C(O)R.sup.13,
SO.sub.2R.sup.13 or 3-6 membered heterocyclyl;
[0185] R.sup.8 is C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14;
[0186] R.sup.9 is H or C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14;
[0187] R.sup.10 and R.sup.11 are independently H or C.sub.1-C.sub.4
alkyl, where each said C.sub.1-C.sub.4 alkyl is optionally
substituted by one or more R.sup.14;
[0188] each R.sup.13 is independently C.sub.1-C.sub.4 alkyl, where
each said C.sub.1-C.sub.4 alkyl is optionally substituted by one or
more R.sup.15;
[0189] each R.sup.14 and R.sup.15 is independently OH, F, CN or
C.sub.1-C.sub.4 alkoxy; and
[0190] X and Z are independently C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4 alkoxy or
C.sub.1-C.sub.4 fluoroalkoxy.
[0191] In another preferred embodiment, the invention provides a
compound of Formula (I), (I-A) or (I-B), or a pharmaceutically
acceptable salt thereof, wherein:
[0192] R.sup.1 is C.sub.1-C.sub.4 alkoxy, where said
C.sub.1-C.sub.4 alkoxy is optionally substituted by one or more
R.sup.6;
[0193] R.sup.2 is H;
[0194] L is a bond or a C.sub.1-C.sub.4 alkylene;
[0195] R.sup.3 is selected from the group consisting of
C.sub.1-C.sub.4 alkyl, OH, C(O)R.sup.8 and 3-12 membered
heterocyclyl, where each said C.sub.1-C.sub.4 alkyl is optionally
substituted by one or more R.sup.6, and each said 3-12 membered
heterocyclyl is optionally substituted by one or more R.sup.7;
[0196] R.sup.4 is H or Cl;
[0197] each R.sup.6 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy;
[0198] each R.sup.7 is independently C.sub.1-C.sub.4 alkyl, OH, F,
CN, C.sub.1-C.sub.4 alkoxy, .dbd.O, CHO, C(O)R.sup.13,
SO.sub.2R.sup.13 or 3-6 membered heterocyclyl;
[0199] R.sup.8 is C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14;
[0200] each R.sup.13 is independently C.sub.1-C.sub.4 alkyl, where
each said C.sub.1-C.sub.4 alkyl is optionally substituted by one or
more R.sup.15;
[0201] each R.sup.14 and R.sup.15 is independently OH, F, CN or
C.sub.1-C.sub.4 alkoxy; and
[0202] X and Z are independently C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4 alkoxy or
C.sub.1-C.sub.4 fluoroalkoxy.
[0203] In another preferred embodiment, the invention provides a
compound of Formula (I), (I-A) or (I-B), or a pharmaceutically
acceptable salt thereof, wherein:
[0204] R.sup.1 is C.sub.1-C.sub.4 alkyl, where said C.sub.1-C.sub.4
alkyl is optionally substituted by one or more R.sup.6;
[0205] R.sup.2 is H;
[0206] L is a bond or a C.sub.1-C.sub.4 alkylene;
[0207] R.sup.3 is OR.sup.12;
[0208] R.sup.4 is H or Cl;
[0209] each R.sup.6 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy;
[0210] each R.sup.7 is independently C.sub.1-C.sub.4 alkyl, OH, F,
CN, C.sub.1-C.sub.4 alkoxy, .dbd.O, CHO, C(O)R.sup.13,
SO.sub.2R.sup.13 or 3-6 membered heterocyclyl;
[0211] R.sup.12 is selected from the group consisting of
C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl and 5-12
membered heteroaryl, where each said C.sub.3-C.sub.8 cycloalkyl,
3-12 membered heterocyclyl or 5-12 membered heteroaryl is
optionally substituted by one or more R.sup.7;
[0212] each R.sup.13 is independently C.sub.1-C.sub.4 alkyl, where
each said C.sub.1-C.sub.4 alkyl is optionally substituted by one or
more R.sup.15;
[0213] each R.sup.15 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy; and
[0214] X and Z are independently C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4 alkoxy or
C.sub.1-C.sub.4 fluoroalkoxy.
[0215] In another preferred embodiment, the invention provides a
compound of Formula (I), (I-A) or (I-B), or a pharmaceutically
acceptable salt thereof, wherein:
[0216] R.sup.1 is C.sub.1-C.sub.4 alkoxy;
[0217] R.sup.2 is H;
[0218] L is a bond;
[0219] R.sup.3 is 3-12 membered heterocyclyl, preferably selected
from the group consisting of oxetanyl, tetrahydrofuranyl and
tetrahydropyranyl, each optionally substituted by one or more
R.sup.7;
[0220] R.sup.4 is H or Cl;
[0221] each R.sup.7 is independently C.sub.1-C.sub.4 alkyl, OH, F,
CN, C.sub.1-C.sub.4 alkoxy, .dbd.O, CHO, C(O)R.sup.13,
SO.sub.2R.sup.13 or 3-6 membered heterocyclyl;
[0222] each R.sup.13 is independently C.sub.1-C.sub.4 alkyl, where
each said C.sub.1-C.sub.4 alkyl is optionally substituted by one or
more R.sup.15;
[0223] each R.sup.15 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy; and
[0224] X and Z are independently C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4 alkoxy or
C.sub.1-C.sub.4 fluoroalkoxy.
[0225] In another preferred embodiment, the invention provides a
compound of Formula (I), (I-A) or (I-B), or a pharmaceutically
acceptable salt thereof, wherein:
[0226] R.sup.1 is 3-12 membered heterocyclyl, preferably selected
from the group consisting of oxetanyl, tetrahydrofuranyl and
tetrahydropyranyl, each optionally substituted by one or more
R.sup.7;
[0227] R.sup.2 is H;
[0228] L is a bond;
[0229] R.sup.3 is C.sub.1-C.sub.4 alkoxy, optionally substituted by
one or more R.sup.6,
[0230] R.sup.4 is H or Cl;
[0231] each R.sup.6 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy;
[0232] each R.sup.7 is independently C.sub.1-C.sub.4 alkyl, OH, F,
CN, C.sub.1-C.sub.4 alkoxy, .dbd.O, CHO, C(O)R.sup.13,
SO.sub.2R.sup.13 or 3-6 membered heterocyclyl;
[0233] each R.sup.13 is independently C.sub.1-C.sub.4 alkyl, where
each said C.sub.1-C.sub.4 alkyl is optionally substituted by one or
more R.sup.15;
[0234] each R.sup.15 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy; and
[0235] X and Z are independently C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4 alkoxy or
C.sub.1-C.sub.4 fluoroalkoxy.
[0236] In another aspect, the invention provides a compound of
formula (II), (II-A) or (II-B):
##STR00016##
[0237] or a pharmaceutically acceptable salt thereof,
[0238] wherein:
[0239] R.sup.1, L, R.sup.3 and X are defined as for Formula (I);
and
[0240] R.sup.4 is H, Cl, Br, F or CH.sub.3.
[0241] The embodiments described herein for Formula (I), (I-A) and
(I-B) are also applicable to compounds of Formulae (II), (II-A) and
(II-B) to the extent they are not inconsistent.
[0242] In a further aspect, the invention provides a compound of
formula (III):
##STR00017##
[0243] or a pharmaceutically acceptable salt thereof,
[0244] wherein:
[0245] R.sup.1 and R.sup.3 are taken together to form a 3-12
membered heterocyclyl optionally substituted by one or more
R.sup.7;
[0246] R.sup.2 is H, F or C.sub.1-C.sub.4 alkyl;
[0247] R.sup.4 is H, halo or C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.6;
[0248] each R.sup.6 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy;
[0249] each R.sup.7 is independently C.sub.1-C.sub.4 alkyl, OH, F,
CN, C.sub.1-C.sub.4 alkoxy, .dbd.O, CHO, C(O)R.sup.13,
SO.sub.2R.sup.13 or 3-6 membered heterocyclyl;
[0250] each R.sup.13 is independently C.sub.1-C.sub.4 alkyl, where
each said C.sub.1-C.sub.4 alkyl is optionally substituted by one or
more R.sup.15;
[0251] each R.sup.15 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy; and
[0252] X and Z are independently C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4 alkoxy or
C.sub.1-C.sub.4 fluoroalkoxy.
[0253] In some embodiments of Formula (III), R.sup.2 is F or
CH.sub.3.
[0254] In compounds of Formula (III), R.sup.1 and R.sup.3 are taken
together to form a 3-12 membered heterocyclyl optionally
substituted by one or more R.sup.7. In some such embodiments, said
3-12 membered heterocyclyl is selected from the group consisting of
azetidinyl, pyrrolidinyl, piperidinyl and homopiperidinyl, each
optionally substituted by one or more R.sup.7. In other such
embodiments, said 3-12 membered heterocyclyl is selected from the
group consisting of oxetanyl, tetrahydrofuranyl and
tetrahydropyranyl, each optionally substituted by one or more
R.sup.7.
[0255] In compounds of Formula (III), each R.sup.7 is independently
C.sub.1-C.sub.4 alkyl, OH, F, CN, C.sub.1-C.sub.4 alkoxy, .dbd.O,
CHO, C(O)R.sup.13, SO.sub.2R.sup.13 or 3-6 membered heterocyclyl.
In some embodiments, R.sup.7 is CHO, C(O)R.sup.13 or
SO.sub.2R.sup.13, where each R.sup.13 is independently
C.sub.1-C.sub.4 alkyl optionally substituted by one or more
R.sup.15. In some such embodiments, R.sup.13 is C.sub.1-C.sub.4
alkyl optionally substituted by one or more R.sup.15 and each
R.sup.15 is independently OH, F, CN or C.sub.1-C.sub.4 alkoxy. In
particular embodiments, R.sup.13 is C.sub.1-C.sub.4 alkyl
optionally substituted by OH. In specific embodiments, R.sup.13 is
CH.sub.3 or CH.sub.2OH, such that R.sup.7 is C(O)CH.sub.3,
C(O)CH.sub.2OH, SO.sub.2CH.sub.3 or SO.sub.2CH.sub.2OH (i.e.,
acetyl, .alpha.-hydroxyacetyl, methylsulfonyl or
.alpha.-hydroxymethylsulfonyl).
[0256] In some embodiments, R.sup.4 is H, CH.sub.3 or Cl.
[0257] In some embodiments, Z is CH.sub.3.
[0258] In some embodiments, X is CH.sub.3 or OCH.sub.3.
[0259] In a preferred embodiment, the invention provides a compound
of Formula (III), or a pharmaceutically acceptable salt thereof,
wherein:
[0260] R.sup.2 is F or CH.sub.3;
[0261] R.sup.1 and R.sup.3 are taken together to form a 3-12
membered heterocyclyl, each optionally substituted by one or more
R.sup.7;
[0262] R.sup.4 is H, CH.sub.3 or Cl;
[0263] Z is CH.sub.3; and
[0264] X is CH.sub.3 or OCH.sub.3.
[0265] In another preferred embodiment, the invention provides a
compound of Formula (III), or a pharmaceutically acceptable salt
thereof, wherein:
[0266] R.sup.2 is F or CH.sub.3;
[0267] R.sup.1 and R.sup.3 are taken together to form a 3-12
membered heterocyclyl selected from the group consisting of
azetidinyl, pyrrolidinyl, piperidinyl and homopiperidinyl, each
optionally substituted by one or more R.sup.7;
[0268] R.sup.4 is H, CH.sub.3 or Cl;
[0269] R.sup.7 is C(O)R.sup.13 or SO.sub.2R.sup.13;
[0270] each R.sup.13 is independently C.sub.1-C.sub.4 alkyl
optionally substituted by one or more R.sup.15;
[0271] each R.sup.15 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy;
[0272] Z is CH.sub.3; and
[0273] X is CH.sub.3 or OCH.sub.3.
[0274] In another aspect, the invention provides a compound of
formula (I'):
##STR00018##
[0275] or a pharmaceutically acceptable salt thereof,
[0276] wherein:
[0277] R.sup.1 is selected from the group consisting of H, F,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy, C(O)R.sup.5,
C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl and 5-12
membered heteroaryl, where each said C.sub.1-C.sub.4 alkyl or
C.sub.1-C.sub.4 alkoxy is optionally substituted by one or more
R.sup.6, and each said C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl or 5-12 membered heteroaryl is optionally substituted
by one or more R.sup.7;
[0278] R.sup.2 is H, F or C.sub.1-C.sub.4 alkyl;
[0279] L is a bond or a C.sub.1-C.sub.4 alkylene;
[0280] R.sup.3 is selected from the group consisting of
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy, OH, CN, C(O)R.sup.8,
COOR.sup.9, NR.sup.10R.sup.11, OR.sup.12, C.sub.3-C.sub.8
cycloalkyl, 3-12 membered heterocyclyl and 5-12 membered
heteroaryl, where each said C.sub.1-C.sub.4 alkyl or
C.sub.1-C.sub.4 alkoxy is optionally substituted by one or more
R.sup.6, and each said C.sub.3-C.sub.8 cycloalkyl, 3-12 membered
heterocyclyl or 5-12 membered heteroaryl is optionally substituted
by one or more R.sup.7;
[0281] R.sup.4 is H, halo or C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.6;
[0282] R.sup.5 is C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14;
[0283] each R.sup.6 is independently OH, F, CN or C.sub.1-C.sub.4
alkoxy;
[0284] each R.sup.7 is independently C.sub.1-C.sub.4 alkyl, OH, F,
CN, C.sub.1-C.sub.4 alkoxy, .dbd.O or C(O)R.sup.13;
[0285] R.sup.8 is C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14;
[0286] R.sup.9 is H or C.sub.1-C.sub.4 alkyl, where each said
C.sub.1-C.sub.4 alkyl is optionally substituted by one or more
R.sup.14;
[0287] R.sup.10 and R.sup.11 are independently H or C.sub.1-C.sub.4
alkyl, where each said C.sub.1-C.sub.4 alkyl is optionally
substituted by one or more R.sup.14;
[0288] R.sup.12 is selected from the group consisting of
C.sub.3-C.sub.8 cycloalkyl, 3-12 membered heterocyclyl and 5-12
membered heteroaryl, where each said C.sub.3-C.sub.8 cycloalkyl,
3-12 membered heterocyclyl or 5-12 membered heteroaryl is
optionally substituted by one or more R.sup.7;
[0289] each R.sup.13 is independently H or C.sub.1-C.sub.4 alkyl,
where each said C.sub.1-C.sub.4 alkyl is optionally substituted by
one or more R.sup.15;
[0290] each R.sup.14 and R.sup.15 is independently OH, F, CN or
C.sub.1-C.sub.4 alkoxy; and
[0291] X and Z are independently C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 fluoroalkyl, C.sub.1-C.sub.4 alkoxy or
C.sub.1-C.sub.4 fluoroalkoxy.
[0292] In some embodiments, the compound of Formula (I') has the
absolute stereochemistry at the carbon atom bearing the R.sup.1 and
R.sup.2 substituents as shown in Formula (I-A') or (I-B'):
##STR00019##
[0293] or a pharmaceutically acceptable salt thereof,
[0294] wherein:
[0295] R.sup.1, R.sup.2, L, R.sup.3, R.sup.4, X and Z are defined
as for Formula (I).
[0296] The embodiments described herein for Formula (I), (I-A) and
(I-B) are also applicable to compounds of Formulae (I'), (I-A') and
(I-B') to the extent they are not inconsistent.
[0297] In another aspect, the invention provides a compound of
Formula (II'), (II-A') or (II-B'):
##STR00020##
[0298] or a pharmaceutically acceptable salt thereof,
[0299] wherein:
[0300] R.sup.1, L, R.sup.3 and X are defined as for Formula (I');
and
[0301] R.sup.4 is H, Cl, Br, F or CH.sub.3.
[0302] The embodiments described herein for Formula (I), (I-A) and
(I-B) are also applicable to compounds of Formulae (II'), (II-A')
and (II-B') to the extent they are not inconsistent.
[0303] A "pharmaceutical composition" refers to a mixture of one or
more of the compounds described herein, or a pharmaceutically
acceptable salt, solvate, hydrate or prodrug thereof as an active
ingredient, and at least one pharmaceutically acceptable carrier or
excipient.
[0304] In another aspect the invention provides a pharmaceutical
composition comprising a compound of one of the formulae described
herein, or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier or excipient. In some
embodiments, the pharmaceutical composition comprises two or more
pharmaceutically acceptable carriers and/or excipients.
[0305] In some embodiments, the pharmaceutical composition may
further comprise at least one additional an anti-cancer therapeutic
agent or a palliative agent. In some such embodiments, the at least
one additional medicinal or pharmaceutical agent is an anti-cancer
agent as described below. In some such embodiments, the combination
provides an additive, greater than additive, or synergistic
anti-cancer effect. In some such embodiments, the one or more
anti-cancer therapeutic agent is selected from the group consisting
of anti-tumor agents, anti-angiogenesis agents, signal transduction
inhibitors and antiproliferative agents.
[0306] In one aspect, the invention provides a method for the
treatment of abnormal cell growth in a subject comprising
administering to the subject a therapeutically effective amount of
a compound of the invention, or a pharmaceutically acceptable salt
thereof.
[0307] In some embodiments, the methods described herein further
comprise administering to the subject an amount of an anti-cancer
therapeutic agent or a palliative agent, which amounts are together
effective in treating said abnormal cell growth. In some
embodiments, the one or more anti-cancer therapeutic agent are
selected from anti-tumor agents, anti-angiogenesis agents, signal
transduction inhibitors and antiproliferative agents, which amounts
are together effective in treating said abnormal cell growth. In
some such embodiments, the anti-tumor agent is selected from the
group consisting of mitotic inhibitors, alkylating agents,
anti-metabolites, intercalating antibiotics, growth factor
inhibitors, radiation, cell cycle inhibitors, enzymes,
topoisomerase inhibitors, biological response modifiers,
antibodies, cytotoxics, anti-hormones, and anti-androgens.
[0308] In frequent embodiments of the methods provided herein, the
abnormal cell growth is cancer. In some embodiments, the methods
provided result in one or more of the following effects: (1)
inhibiting cancer cell proliferation; (2) inhibiting cancer cell
invasiveness; (3) inducing apoptosis of cancer cells; (4)
inhibiting cancer cell metastasis; or (5) inhibiting
angiogenesis.
[0309] In another aspect, the invention provides a method for the
treatment of a disorder mediated by EZH2 in a subject comprising
administering to the subject a compound of the invention, or a
pharmaceutically acceptable salt thereof, in an amount that is
effective for treating said disorder.
[0310] Unless indicated otherwise, all references herein to the
inventive compounds include references to salts, solvates, hydrates
and complexes thereof, and to solvates, hydrates and complexes of
salts thereof, including polymorphs, stereoisomers, and
isotopically labeled versions thereof.
[0311] Compounds of the invention may exist in the form of
pharmaceutically acceptable salts such as, e.g., acid addition
salts and base addition salts of the compounds of one of the
formulae provided herein. As used herein, the term
"pharmaceutically acceptable salt" refers to those salts which
retain the biological effectiveness and properties of the parent
compound. The phrase "pharmaceutically acceptable salt(s)", as used
herein, unless otherwise indicated, includes salts of acidic or
basic groups which may be present in the compounds of the formulae
disclosed herein.
[0312] For example, the compounds of the invention that are basic
in nature may be capable of forming a wide variety of salts with
various inorganic and organic acids. Although such salts must be
pharmaceutically acceptable for administration to animals, it is
often desirable in practice to initially isolate the compound of
the present invention from the reaction mixture as a
pharmaceutically unacceptable salt and then simply convert the
latter back to the free base compound by treatment with an alkaline
reagent and subsequently convert the latter free base to a
pharmaceutically acceptable acid addition salt. The acid addition
salts of the base compounds of this invention may be prepared by
treating the base compound with a substantially equivalent amount
of the selected mineral or organic acid in an aqueous solvent
medium or in a suitable organic solvent, such as methanol or
ethanol. Upon evaporation of the solvent, the desired solid salt is
obtained. The desired acid salt may also be precipitated from a
solution of the free base in an organic solvent by adding an
appropriate mineral or organic acid to the solution.
[0313] The acids that may be used to prepare pharmaceutically
acceptable acid addition salts of such basic compounds of those
that form non-toxic acid addition salts, i.e., salts containing
pharmacologically acceptable anions, such as the hydrochloride,
hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate,
acid phosphate, isonicotinate, acetate, lactate, salicylate,
citrate, acid citrate, tartrate, pantothenate, bitartrate,
ascorbate, succinate, maleate, gentisinate, fumarate, gluconate,
glucuronate, saccharate, formate, benzoate, glutamate,
methanesulfonate, ethanesulfonate, benzenesulfonate, p
toluenesulfonate and pamoate [i.e.,
1,1'-methylene-bis-(2-hydroxy-3-naphthoate)] salts.
[0314] Examples of salts include, but are not limited to, acetate,
acrylate, benzenesulfonate, benzoate (such as chlorobenzoate,
methylbenzoate, dinitrobenzoate, hydroxybenzoate, and
methoxybenzoate), bicarbonate, bisulfate, bisulfite, bitartrate,
borate, bromide, butyne-1,4-dioate, calcium edetate, camsylate,
carbonate, chloride, caproate, caprylate, clavulanate, citrate,
decanoate, dihydrochloride, dihydrogenphosphate, edetate,
edislyate, estolate, esylate, ethylsuccinate, formate, fumarate,
gluceptate, gluconate, glutamate, glycollate, glycollylarsanilate,
heptanoate, hexyne-1,6-dioate, hexylresorcinate, hydrabamine,
hydrobromide, hydrochloride, .gamma.-hydroxybutyrate, iodide,
isobutyrate, isothionate, lactate, lactobionate, laurate, malate,
maleate, malonate, mandelate, mesylate, metaphosphate,
methane-sulfonate, methylsulfate, monohydrogenphosphate, mucate,
napsylate, naphthalene-1-sulfonate, naphthalene-2-sulfonate,
nitrate, oleate, oxalate, pamoate (embonate), palmitate,
pantothenate, phenylacetates, phenylbutyrate, phenylpropionate,
phthalate, phospate/diphosphate, polygalacturonate,
propanesulfonate, propionate, propiolate, pyrophosphate,
pyrosulfate, salicylate, stearate, subacetate, suberate, succinate,
sulfate, sulfonate, sulfite, tannate, tartrate, teoclate, tosylate,
triethiodode, and valerate salts.
[0315] Illustrative examples of suitable salts include organic
salts derived from amino acids, such as glycine and arginine,
ammonia, primary, secondary, and tertiary amines, and cyclic
amines, such as piperidine, morpholine and piperazine, and
inorganic salts derived from sodium, calcium, potassium, magnesium,
manganese, iron, copper, zinc, aluminum and lithium.
[0316] The compounds of the invention that include a basic moiety,
such as an amino group, may form pharmaceutically acceptable salts
with various amino acids, in addition to the acids mentioned
above.
[0317] Those compounds of the invention that are acidic in nature
may be capable of forming base salts with various pharmacologically
acceptable cations. Examples of such salts include the alkali metal
or alkaline-earth metal salts and particularly, the sodium and
potassium salts. These salts may be prepared by conventional
techniques. The chemical bases which may be used as reagents to
prepare the pharmaceutically acceptable base salts of this
invention include those which form non-toxic base salts with the
acidic compounds herein. These salts may be prepared by any
suitable method, for example, treatment of the free acid with an
inorganic or organic base, such as an amine (primary, secondary or
tertiary), an alkali metal hydroxide or alkaline earth metal
hydroxide, or the like. These salts may also be prepared by
treating the corresponding acidic compounds with an aqueous
solution containing the desired pharmacologically acceptable
cations, and then evaporating the resulting solution to dryness,
preferably under reduced pressure. Alternatively, they may also be
prepared by mixing lower alkanolic solutions of the acidic
compounds and the desired alkali metal alkoxide together, and then
evaporating the resulting solution to dryness in the same manner as
before. In either case, stoichiometric quantities of reagents may
be employed in order to ensure completeness of reaction and maximum
yields of the desired final product.
[0318] The chemical bases that may be used as reagents to prepare
pharmaceutically acceptable base salts of the compounds of the
invention that are acidic in nature are those that form non-toxic
base salts with such compounds. Such non-toxic base salts include,
but are not limited to, those derived from such pharmacologically
acceptable cations such as alkali metal cations (e.g., potassium
and sodium) and alkaline earth metal cations (e.g., calcium and
magnesium), ammonium or water-soluble amine addition salts such as
N-methylglucamine-(meglumine), and the lower alkanolammonium and
other base salts of pharmaceutically acceptable organic amines.
[0319] Hemisalts of acids and bases may also be formed, for
example, hemisulphate and hemicalcium salts.
[0320] For a review on suitable salts, see Handbook of
Pharmaceutical Salts: Properties, Selection, and Use by Stahl and
Wermuth (Wiley-VCH, 2002). Methods for making pharmaceutically
acceptable salts are known to those of skill in the art.
[0321] Salts of the present invention may be prepared according to
methods known to those of skill in the art. A pharmaceutically
acceptable salt of the inventive compounds may be readily prepared
by mixing together solutions of the compound and the desired acid
or base, as appropriate. The salt may precipitate from solution and
be collected by filtration or may be recovered by evaporation of
the solvent. The degree of ionization in the salt may vary from
completely ionized to almost non-ionized.
[0322] It will be understood by those of skill in the art that the
compounds of the invention in free base form having a basic
functionality may be converted to the acid addition salts by
treating with a stoichiometric excess of the appropriate acid. The
acid addition salts of the compounds of the invention may be
reconverted to the corresponding free base by treating with a
stoichiometric excess of a suitable base, such as potassium
carbonate or sodium hydroxide, typically in the presence of aqueous
solvent, and at a temperature of between about 0.degree. C. and
100.degree. C. The free base form may be isolated by conventional
means, such as extraction with an organic solvent. In addition,
acid addition salts of the compounds of the invention may be
interchanged by taking advantage of differential solubilities of
the salts, volatilities or acidities of the acids, or by treating
with the appropriately loaded ion exchange resin. For example, the
interchange may be affected by the reaction of a salt of the
compounds of the invention with a slight stoichiometric excess of
an acid of a lower pK than the acid component of the starting salt.
This conversion is typically carried out at a temperature between
about 0.degree. C. and the boiling point of the solvent being used
as the medium for the procedure. Similar exchanges are possible
with base addition salts, typically via the intermediacy of the
free base form.
[0323] The compounds of the invention may exist in both unsolvated
and solvated forms. When the solvent or water is tightly bound, the
complex will have a well-defined stoichiometry independent of
humidity. When, however, the solvent or water is weakly bound, as
in channel solvates and hygroscopic compounds, the water/solvent
content will be dependent on humidity and drying conditions. In
such cases, non-stoichiometry will be the norm. The term `solvate`
is used herein to describe a molecular complex comprising the
compound of the invention and one or more pharmaceutically
acceptable solvent molecules, for example, ethanol. The term
`hydrate` is employed when the solvent is water. Pharmaceutically
acceptable solvates in accordance with the invention include
hydrates and solvates wherein the solvent of crystallization may be
isotopically substituted, e.g. D.sub.2O, d.sub.6-acetone and
d.sub.6-DMSO.
[0324] Also included within the scope of the invention are
complexes such as clathrates, drug-host inclusion complexes
wherein, in contrast to the aforementioned solvates, the drug and
host are present in stoichiometric or non-stoichiometric amounts.
Also included are complexes of the drug containing two or more
organic and/or inorganic components which may be in stoichiometric
or non-stoichiometric amounts. The resulting complexes may be
ionized, partially ionized, or non-ionized. For a review of such
complexes, see J Pharm Sci, 64 (8), 1269-1288 by Haleblian (August
1975), the disclosure of which is incorporated herein by reference
in its entirety.
[0325] The invention also relates to prodrugs of the compounds of
the formulae provided herein. Thus, certain derivatives of
compounds of the invention which may have little or no
pharmacological activity themselves may, when administered to a
subject, be converted into the inventive compounds, for example, by
hydrolytic cleavage. Such derivatives are referred to as
`prodrugs`. Further information on the use of prodrugs may be found
in `Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium
Series (T Higuchi and W Stella) and `Bioreversible Carriers in Drug
Design`, Pergamon Press, 1987 (ed. E B Roche, American
Pharmaceutical Association), the disclosures of which are
incorporated herein by reference in their entireties. As used
herein, "subject" may refer to a human or animal subject.
[0326] Prodrugs in accordance with the invention may, for example,
be produced by replacing appropriate functionalities present in the
inventive compounds with certain moieties known to those skilled in
the art as `pro-moieties` as described, for example, in "Design of
Prodrugs" by H Bundgaard (Elsevier, 1985), the disclosure of which
is incorporated herein by reference in its entirety.
[0327] Some non-limiting examples of potential prodrugs in
accordance with the invention include:
[0328] (i) where the compound contains a carboxylic acid
functionality (--COOH), an ester thereof, for example, replacement
of the hydrogen with (C.sub.1-C.sub.8)alkyl;
[0329] (ii) where the compound contains an alcohol functionality
(--OH), an ether thereof, for example, replacement of the hydrogen
with (C.sub.1-C.sub.6)alkanoyloxymethyl; and
[0330] (iii) where the compound contains a primary or secondary
amino functionality (--NH.sub.2 or --NHR where R.noteq.H), an amide
thereof, for example, replacement of one or both hydrogens with a
suitably metabolically labile group, such as an amide, carbamate,
urea, phosphonate, sulfonate, etc.
[0331] Further examples of replacement groups in accordance with
the foregoing examples and examples of other potential prodrug
types may be found in the aforementioned references.
[0332] Finally, certain inventive compounds may themselves act as
potential prodrugs of other of the inventive compounds.
[0333] Also included within the scope of the invention are
metabolites of compounds of the formulae described herein, i.e.,
compounds formed in vivo upon administration of a drug.
[0334] The compounds of the formulae provided herein may have
asymmetric carbon atoms. The carbon-carbon bonds of the compounds
of the invention may be depicted herein using a solid line (), a
solid wedge (), or a dotted wedge (). The use of a solid line to
depict bonds to asymmetric carbon atoms is meant to indicate that
all possible stereoisomers (e.g. specific enantiomers, racemic
mixtures, etc.) at that carbon atom are included. The use of either
a solid or dotted wedge to depict bonds to asymmetric carbon atoms
is meant to indicate that only the stereoisomer shown is meant to
be included. It is possible that compounds of the invention may
contain more than one asymmetric carbon atom. In those compounds,
the use of a solid line to depict bonds to asymmetric carbon atoms
is meant to indicate that all possible stereoisomers are meant to
be included. For example, unless stated otherwise, it is intended
that the compounds of the invention may exist as enantiomers and
diastereomers or as racemates and mixtures thereof. The use of a
solid line to depict bonds to one or more asymmetric carbon atoms
in a compound of the invention and the use of a solid or dotted
wedge to depict bonds to other asymmetric carbon atoms in the same
compound is meant to indicate that a mixture of diastereomers is
present.
[0335] Compounds of the invention that have chiral centers may
exist as stereoisomers, such as racemates, enantiomers, or
diastereomers.
[0336] Stereoisomers of the compounds of the formulae herein may
include cis and trans isomers, optical isomers such as (R) and (S)
enantiomers, diastereomers, geometric isomers, rotational isomers,
atropisomers, conformational isomers, and tautomers of the
compounds of the invention, including compounds exhibiting more
than one type of isomerism; and mixtures thereof (such as racemates
and diastereomeric pairs). Also included are acid addition or base
addition salts wherein the counterion is optically active, for
example, d-lactate or I-lysine, or racemic, for example,
dl-tartrate or dl-arginine.
[0337] When a racemate crystallizes, crystals of two different
types may be possible. The first type is the racemic compound (true
racemate) referred to above wherein one homogeneous form of crystal
is produced containing both enantiomers in equimolar amounts. The
second type is the racemic mixture or conglomerate wherein two
forms of crystal are produced in equimolar amounts each comprising
a single enantiomer.
[0338] The compounds of the invention may exhibit the phenomena of
tautomerism and structural isomerism. For example, the compounds
may exist in several tautomeric forms, including the enol and imine
form, and the keto and enamine form and geometric isomers and
mixtures thereof. All such tautomeric forms are included within the
scope of compounds of the invention. Tautomers exist as mixtures of
a tautomeric set in solution. In solid form, usually one tautomer
predominates. Even though one tautomer may be described, the
present invention includes all tautomers of the compounds of the
formulae provided.
[0339] In addition, some of the compounds of the invention may form
atropisomers (e.g., substituted biaryls). Atropisomers are
conformational stereoisomers which occur when rotation about a
single bond in the molecule is prevented, or greatly slowed, as a
result of steric interactions with other parts of the molecule and
the substituents at both ends of the single bond are unsymmetrical.
The interconversion of atropisomers is slow enough to allow
separation and isolation under predetermined conditions. The energy
barrier to thermal racemization may be determined by the steric
hindrance to free rotation of one or more bonds forming a chiral
axis.
[0340] Where a compound of the invention contains an alkenyl or
alkenylene group, geometric cis/trans (or Z/E) isomers may be
possible. Cis/trans isomers may be separated by conventional
techniques well known to those skilled in the art, for example,
chromatography or fractional crystallization.
[0341] Conventional techniques for the preparation/isolation of
individual enantiomers include chiral synthesis from a suitable
optically pure precursor or resolution of the racemate (or the
racemate of a salt or derivative) using, for example, chiral high
pressure liquid chromatography (HPLC).
[0342] Alternatively, the racemate (or a racemic precursor) may be
reacted with a suitable optically active compound, for example, an
alcohol, or, in the case where the compound contains an acidic or
basic moiety, an acid or base such as tartaric acid or
1-phenylethylamine. The resulting diastereomeric mixture may be
separated by chromatography and/or fractional crystallization and
one or both of the diastereoisomers converted to the corresponding
pure enantiomer(s) by means well known to one skilled in the
art.
[0343] Chiral compounds of the invention (and chiral precursors
thereof) may be obtained in enantiomerically-enriched form using
chromatography, typically HPLC, on an asymmetric resin with a
mobile phase consisting of a hydrocarbon, typically heptane or
hexane, containing from 0 to 50% isopropanol, typically from 2 to
20%, and from 0 to 5% of an alkylamine, typically 0.1%
diethylamine. Concentration of the eluate affords the enriched
mixture.
[0344] Stereoisomeric conglomerates may be separated by
conventional techniques known to those skilled in the art; see, for
example, "Stereochemistry of Organic Compounds" by E L Eliel
(Wiley, New York, 1994), the disclosure of which is incorporated
herein by reference in its entirety.
[0345] "Enantiomerically pure" as used herein, describes a compound
that is present as a single enantiomer and which is described in
terms of enantiomeric excess (e.e.). Preferably, wherein the
compound is present as an enantiomer, the enantiomer is present at
an enantiomeric excess of greater than or equal to about 80%, more
preferably, at an enantiomeric excess of greater than or equal to
about 90%, more preferably still, at an enantiomeric excess of
greater than or equal to about 95%, more preferably still, at an
enantiomeric excess of greater than or equal to about 98%, most
preferably, at an enantiomeric excess of greater than or equal to
about 99%. Similarly, "diastereomerically pure" as used herein,
describes a compound that is present as a diastereomer and which is
described in terms of diasteriomeric excess (d.e.). Preferably,
wherein the compound is present as a diastereomer, the diastereomer
is present at an diastereomeric excess of greater than or equal to
about 80%, more preferably, at an diastereomeric excess of greater
than or equal to about 90%, more preferably still, at an
diastereomeric excess of greater than or equal to about 95%, more
preferably still, at an diastereomeric excess of greater than or
equal to about 98%, most preferably, at an diastereomeric excess of
greater than or equal to about 99%.
[0346] The present invention also includes isotopically-labeled
compounds, which are identical to those recited in one of the
formulae provided, but for the fact that one or more atoms are
replaced by an atom having an atomic mass or mass number different
from the atomic mass or mass number usually found in nature.
[0347] Isotopically-labeled compounds of the invention may
generally be prepared by conventional techniques known to those
skilled in the art or by processes analogous to those described
herein, using an appropriate isotopically-labeled reagent in place
of the non-labeled reagent otherwise employed.
[0348] Examples of isotopes that may be incorporated into compounds
of the invention include isotopes of hydrogen, carbon, nitrogen,
oxygen, phosphorus, fluorine and chlorine, such as, but not limited
to, .sup.2H, .sup.3H, .sup.13C, .sup.14C, .sup.15N, .sup.18O,
.sup.17O, .sup.31P, .sup.32P, .sup.35S, .sup.18F, and .sup.36Cl.
Certain isotopically-labeled compounds of the invention, for
example those into which radioactive isotopes such as .sup.3H and
.sup.14C are incorporated, may be useful in drug and/or substrate
tissue distribution assays. Tritiated, i.e., .sup.3H, and
carbon-14, i.e., .sup.14C, isotopes may be particularly preferred
for their ease of preparation and detectability. Further,
substitution with heavier isotopes such as deuterium, i.e.,
.sup.2H, may afford certain potential therapeutic advantages
resulting from potentially greater metabolic stability, for example
potentially increased in vivo half-life or potentially reduced
dosage requirements and, hence, may be preferred in some
circumstances. Isotopically-labeled compounds of the invention may
generally be prepared by carrying out the procedures disclosed in
the Schemes and/or in the Examples and Preparations below, by
substituting an isotopically-labeled reagent for a
non-isotopically-labeled reagent.
[0349] Compounds of the invention may be used as crystalline or
amorphous products, or mixtures thereof. They may be obtained, for
example, as solid plugs, powders, or films by methods such as
precipitation, crystallization, freeze drying, spray drying, or
evaporative drying. Microwave or radio frequency drying may be used
for this purpose.
Therapeutic Methods and Uses
[0350] The invention further provides therapeutic methods and uses
comprising administering a compound of the invention, or
pharmaceutically acceptable salt thereof, alone or in combination
with one or more other therapeutic agents or palliative agents.
[0351] In one aspect, the invention provides a method for the
treatment of abnormal cell growth in a subject comprising
administering to the subject a therapeutically effective amount of
a compound of the invention, or a pharmaceutically acceptable salt
thereof.
[0352] In another aspect, the invention provides a method for the
treatment of abnormal cell growth in a subject comprising
administering to the subject an amount of a compound of the
invention, or a pharmaceutically acceptable salt thereof, in
combination with an amount of an anti-tumor agent, which amounts
are together effective in treating said abnormal cell growth. In
some such embodiments, the anti-tumor agent is selected from the
group consisting of mitotic inhibitors, alkylating agents,
anti-metabolites, intercalating antibiotics, growth factor
inhibitors, radiation, cell cycle inhibitors, enzymes,
topoisomerase inhibitors, biological response modifiers,
antibodies, cytotoxics, anti-hormones, and anti-androgens.
[0353] Compounds of the invention include compounds of any of the
formulae described herein, namely compounds of formulae I, I', II,
II', I-A, I-A', I-B, I-B', II-A, II-A', II-B, II-B' and III as
provided and defined herein, or a pharmaceutically acceptable salt
thereof.
[0354] In another aspect, the invention provides a method for the
treatment of abnormal cell growth in a subject comprising
administering to the subject an amount of a compound of the
invention, or a pharmaceutically acceptable salt thereof, that is
effective in treating abnormal cell growth.
[0355] In still another aspect, the invention provides a method of
inhibiting cancer cell proliferation in a subject, comprising
administering to the subject a compound of the invention, or
pharmaceutically acceptable salt thereof, in an amount effective to
inhibit cell proliferation.
[0356] In another aspect, the invention provides a method of
inhibiting cancer cell invasiveness in a subject, comprising
administering to the subject a compound of the invention, or
pharmaceutically acceptable salt thereof, in an amount effective to
inhibit cell invasiveness.
[0357] In another aspect, the invention provides a method of
inducing apoptosis in cancer cells in a subject, comprising
administering to the subject a compound of the invention, or
pharmaceutically acceptable salt thereof, in an amount effective to
induce apoptosis.
[0358] In a further aspect, the invention provides a method of
inducing apoptosis in a subject, comprising administering to the
subject a therapeutically effective amount of a compound of one of
the formulae described herein, or pharmaceutically acceptable salt
thereof.
[0359] In frequent embodiments of the methods provided herein, the
abnormal cell growth is cancer, wherein said cancer is selected
from the group consisting of basal cell cancer, medulloblastoma
cancer, liver cancer, rhabdomyosarcoma, lung cancer, bone cancer,
pancreatic cancer, skin cancer, cancer of the head or neck,
cutaneous or intraocular melanoma, uterine cancer, ovarian cancer,
rectal cancer, cancer of the anal region, stomach cancer, colon
cancer, breast cancer, uterine cancer, carcinoma of the fallopian
tubes, carcinoma of the endometrium, carcinoma of the cervix,
carcinoma of the vagina, carcinoma of the vulva, Hodgkin's disease,
cancer of the esophagus, cancer of the small intestine, cancer of
the endocrine system, cancer of the thyroid gland, cancer of the
parathyroid gland, cancer of the adrenal gland, sarcoma of soft
tissue, cancer of the urethra, cancer of the penis, prostate
cancer, chronic or acute leukemia, lymphocytic lymphomas, cancer of
the bladder, cancer of the kidney or ureter, renal cell carcinoma,
carcinoma of the renal pelvis, neoplasms of the central nervous
system (CNS), primary CNS lymphoma, spinal axis tumors, brain stem
glioma, pituitary adenoma, or a combination of one or more of the
foregoing cancers.
[0360] In some embodiments, a compound of the invention is
selective for the mutant form of the EZH2, such that trimethylation
of H3K27, which is associated with certain cancers, is inhibited.
The methods and uses provided herein may be used to treat cancers
including follicular lymphoma and diffuse large B-cell lymphoma
(DLBCL).
[0361] Compounds of the invention may be useful for the treatment
of cancers, such as tumors such as brain, breast, cervical,
colorectal, endometrial, esophageal, gastric/stomach, head and
neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate,
testicular and thyroid carcinomas and sarcomas.
[0362] The term "therapeutically effective amount" as used herein
refers to that amount of a compound being administered which will
relieve to some extent one or more of the symptoms of the disorder
being treated. In reference to the treatment of cancer, a
therapeutically effective amount refers to that amount which has
the effect of (1) reducing the size of the tumor, (2) inhibiting
(that is, slowing to some extent, preferably stopping) tumor
metastasis, (3) inhibiting to some extent (that is, slowing to some
extent, preferably stopping) tumor growth or tumor invasiveness,
and/or (4) relieving to some extent (or, preferably, eliminating)
one or more signs or symptoms associated with the cancer.
[0363] As used herein, "subject" refers to a human or animal
subject. In certain preferred embodiments, the subject is a
human.
[0364] The term "treating", as used herein, unless otherwise
indicated, means reversing, alleviating, inhibiting the progress
of, or preventing the disorder or condition to which such term
applies, or one or more symptoms of such disorder or condition. The
term "treatment", as used herein, unless otherwise indicated,
refers to the act of treating as "treating" is defined immediately
above. The term "treating" also includes adjuvant and neo-adjuvant
treatment of a subject.
[0365] The terms "abnormal cell growth" and "hyperproliferative
disorder" are used interchangeably in this application.
[0366] "Abnormal cell growth", as used herein, unless otherwise
indicated, refers to cell growth that is independent of normal
regulatory mechanisms (e.g., loss of contact inhibition). Abnormal
cell growth may be benign (not cancerous), or malignant
(cancerous). This includes the abnormal growth of: (1) tumor cells
(tumors) that show increased expression of EZH2; (2) benign and
malignant cells of other proliferative diseases in which EZH2 is
over-expressed; (3) tumors that proliferate by aberrant EZH2
activation; and (4) benign and malignant cells of other
proliferative diseases in which aberrant EZH2 activation
occurs.
[0367] As used herein "cancer" refers to any malignant and/or
invasive growth or tumor caused by abnormal cell growth. As used
herein "cancer" refers to solid tumors named for the type of cells
that form them, cancer of blood, bone marrow, or the lymphatic
system. Examples of solid tumors include but not limited to
sarcomas and carcinomas. Examples of cancers of the blood include
but not limited to leukemias, lymphomas and myeloma. The term
"cancer" includes but is not limited to a primary cancer that
originates at a specific site in the body, a metastatic cancer that
has spread from the place in which it started to other parts of the
body, a recurrence from the original primary cancer after
remission, and a second primary cancer that is a new primary cancer
in a person with a history of previous cancer of different type
from latter one. The compounds of the invention may inhibit EZH2,
and thus may be useful as antiproliferative agents (e.g., cancer)
or antitumor agent (e.g., effect against solid tumors) in mammals,
particularly in humans. In particular, the compounds of the
invention may be useful in the prevention and treatment of a
variety of human hyperproliferative disorders, such as malignant or
benign abnormal cell growth.
[0368] The compounds, compositions and methods provided herein may
be useful for the treatment of cancers including but not limited to
cancers of the:
[0369] circulatory system, for example, heart (sarcoma
[angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma],
myxoma, rhabdomyoma, fibroma, lipoma and teratoma), mediastinum and
pleura, and other intrathoracic organs, vascular tumors and
tumor-associated vascular tissue;
[0370] respiratory tract, for example, nasal cavity and middle ear,
accessory sinuses, larynx, trachea, bronchus and lung such as small
cell lung cancer (SCLC), non-small cell lung cancer (NSCLC),
bronchogenic carcinoma (squamous cell, undifferentiated small cell,
undifferentiated large cell, adenocarcinoma), alveolar
(bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma,
chondromatous hamartoma, mesothelioma;
[0371] gastrointestinal system, for example, esophagus (squamous
cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach
(carcinoma, lymphoma, leiomyosarcoma), gastric, pancreas (ductal
adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid
tumors, vipoma), small bowel (adenocarcinoma, lymphoma, carcinoid
tumors, Karposi's sarcoma, leiomyoma, hemangioma, lipoma,
neurofibroma, fibroma), large bowel (adenocarcinoma, tubular
adenoma, villous adenoma, hamartoma, leiomyoma);
[0372] genitourinary tract, for example, kidney (adenocarcinoma,
Wilm's tumor [nephroblastoma], lymphoma, leukemia), bladder and/or
urethra (squamous cell carcinoma, transitional cell carcinoma,
adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis
(seminoma, teratoma, embryonal carcinoma, teratocarcinoma,
choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma,
fibroadenoma, adenomatoid tumors, lipoma);
[0373] liver, for example, hepatoma (hepatocellular carcinoma),
cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular
adenoma, hemangioma, pancreatic endocrine tumors (such as
pheochromocytoma, insulinoma, vasoactive intestinal peptide tumor,
islet cell tumor and glucagonoma);
[0374] bone, for example, osteogenic sarcoma (osteosarcoma),
fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma,
Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma),
multiple myeloma, malignant giant cell tumor chordoma,
osteochronfroma (osteocartilaginous exostoses), benign chondroma,
chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell
tumors;
[0375] nervous system, for example, neoplasms of the central
nervous system (CNS), primary CNS lymphoma, skull cancer (osteoma,
hemangioma, granuloma, xanthoma, osteitis deformans), meninges
(meningioma, meningiosarcoma, gliomatosis), brain cancer
(astrocytoma, medulloblastoma, glioma, ependymoma, germinoma
[pinealoma], glioblastoma multiform, oligodendroglioma, schwannoma,
retinoblastoma, congenital tumors), spinal cord neurofibroma,
meningioma, glioma, sarcoma);
[0376] reproductive system, for example, gynecological, uterus
(endometrial carcinoma), cervix (cervical carcinoma, pre-tumor
cervical dysplasia), ovaries (ovarian carcinoma [serous
cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified
carcinoma], granulosa-thecal cell tumors, Sertoli-Leydig cell
tumors, dysgerminoma, malignant teratoma), vulva (squamous cell
carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma,
melanoma), vagina (clear cell carcinoma, squamous cell carcinoma,
botryoid sarcoma (embryonal rhabdomyosarcoma), fallopian tubes
(carcinoma) and other sites associated with female genital organs;
placenta, penis, prostate, testis, and other sites associated with
male genital organs;
[0377] hematologic system, for example, blood (myeloid leukemia
[acute and chronic], acute lymphoblastic leukemia, chronic
lymphocytic leukemia, myeloproliferative diseases, multiple
myeloma, myelodysplastic syndrome), Hodgkin's disease,
non-Hodgkin's lymphoma [malignant lymphoma];
[0378] oral cavity, for example, lip, tongue, gum, floor of mouth,
palate, and other parts of mouth, parotid gland, and other parts of
the salivary glands, tonsil, oropharynx, nasopharynx, pyriform
sinus, hypopharynx, and other sites in the lip, oral cavity and
pharynx;
[0379] skin, for example, malignant melanoma, cutaneous melanoma,
basal cell carcinoma, squamous cell carcinoma, Karposi's sarcoma,
moles dysplastic nevi, lipoma, angioma, dermatofibroma, and
keloids;
[0380] adrenal glands: neuroblastoma; and
[0381] other tissues including connective and soft tissue,
retroperitoneum and peritoneum, eye, intraocular melanoma, and
adnexa, breast, head or/and neck, anal region, thyroid,
parathyroid, adrenal gland and other endocrine glands and related
structures, secondary and unspecified malignant neoplasm of lymph
nodes, secondary malignant neoplasm of respiratory and digestive
systems and secondary malignant neoplasm of other sites.
[0382] More specifically, examples of "cancer" when used herein in
connection with the present invention include cancer selected from
lung cancer (NSCLC and SCLC), cancer of the head or neck, ovarian
cancer, colon cancer, rectal cancer, cancer of the anal region,
stomach cancer, breast cancer, cancer of the kidney or ureter,
renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of
the central nervous system (CNS), primary CNS lymphoma,
non-Hodgkin's lymphoma, spinal axis tumors, or a combination of one
or more of the foregoing cancers.
[0383] Still more specifically, examples of "cancer" when used
herein in connection with the present invention include cancer
selected from lung cancer (NSCLC and SCLC), breast cancer, ovarian
cancer, colon cancer, rectal cancer, cancer of the anal region, or
a combination of one or more of the foregoing cancers.
[0384] In one embodiment of the present invention the non-cancerous
conditions include such hyperplastic conditions such as benign
hyperplasia of the skin (e.g., psoriasis) and benign hyperplasia of
the prostate (e.g., BPH).
[0385] In another aspect, the invention provides a method for
inhibiting cell proliferation, comprising contacting cells with a
compound of the invention or a pharmaceutically acceptable salt
thereof in an amount effective to inhibit proliferation of the
cells.
[0386] In another aspect, the invention provides methods for
inducing cell apoptosis, comprising contacting cells with a
compound described herein in an amount effective to induce
apoptosis of the cells.
[0387] "Contacting" refers to bringing a compound or
pharmaceutically acceptable salt of the invention and a cell
expressing EZH2 together in such a manner that the compound may
affect the activity of EZH2, either directly or indirectly.
Contacting may be accomplished in vitro (i.e., in an artificial
environment such as, e.g., without limitation, in a test tube or
culture medium) or in vivo (i.e., within a living organism such as,
without limitation, a mouse, rat or rabbit.)
[0388] In some embodiments, the cells are in a cell line, such as a
cancer cell line. In other embodiments, the cells are in a tissue
or tumor, and the tissue or tumor may be in a subject, including a
human.
Dosage Forms and Regimens
[0389] Administration of compounds of the invention may be effected
by any method that enables delivery of the compounds to the site of
action. These methods include oral routes, intraduodenal routes,
parenteral injection (including intravenous, subcutaneous,
intramuscular, intravascular or infusion), topical, and rectal
administration.
[0390] Dosage regimens may be adjusted to provide the optimum
desired response. For example, a single bolus may be administered,
several divided doses may be administered over time or the dose may
be proportionally reduced or increased as indicated by the
exigencies of the therapeutic situation. It is especially
advantageous to formulate 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 mammalian subjects 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 may be dictated by and directly
dependent on (a) the unique characteristics of the chemotherapeutic
agent and the particular therapeutic or prophylactic effect to be
achieved, and (b) the limitations inherent in the art of
compounding such an active compound for the treatment of
sensitivity in individuals.
[0391] Thus, the skilled artisan would appreciate, based upon the
disclosure provided herein, that the dose and dosing regimen is
adjusted in accordance with methods well-known in the therapeutic
arts. That is, the maximum tolerable dose may be readily
established, and the effective amount providing a detectable
therapeutic benefit to a patient may also be determined, as can the
temporal requirements for administering each agent to provide a
detectable therapeutic benefit to the patient. Accordingly, while
certain dose and administration regimens are exemplified herein,
these examples in no way limit the dose and administration regimen
that may be provided to a patient in practicing the present
invention.
[0392] It is to be noted that dosage values may vary with the type
and severity of the condition to be alleviated, and may include
single or multiple doses. It is to be further understood that for
any particular subject, specific dosage regimens should be adjusted
over time according to the individual need and the professional
judgment of the person administering or supervising the
administration of the compositions, and that dosage ranges set
forth herein are exemplary only and are not intended to limit the
scope or practice of the claimed composition. For example, doses
may be adjusted based on pharmacokinetic or pharmacodynamic
parameters, which may include clinical effects such as toxic
effects and/or laboratory values. Thus, the present invention
encompasses intra-patient dose-escalation as determined by the
skilled artisan. Determining appropriate dosages and regimens for
administration of the chemotherapeutic agent are well-known in the
relevant art and would be understood to be encompassed by the
skilled artisan once provided the teachings disclosed herein.
[0393] The amount of the compound of the invention administered
will be dependent on the subject being treated, the severity of the
disorder or condition, the rate of administration, the disposition
of the compound and the discretion of the prescribing physician.
However, an effective dosage is in the range of about 0.001 to
about 100 mg per kg body weight per day, preferably about 1 to
about 35 mg/kg/day, in single or divided doses. For a 70 kg human,
this would amount to about 0.05 to about 7 g/day, preferably about
0.1 to about 2.5 g/day. In some instances, dosage levels below the
lower limit of the aforesaid range may be more than adequate, while
in other cases still larger doses may be employed without causing
any harmful side effect, provided that such larger doses are first
divided into several small doses for administration throughout the
day.
Formulations and Routes of Administration
[0394] As used herein, a "pharmaceutically acceptable carrier"
refers to a carrier or diluent that does not cause significant
irritation to an organism and does not abrogate the biological
activity and properties of the active compound.
[0395] The pharmaceutical acceptable carrier may comprise any
conventional pharmaceutical carrier or excipient. The choice of
carrier and/or excipient will to a large extent depend on factors
such as the particular mode of administration, the effect of the
excipient on solubility and stability, and the nature of the dosage
form.
[0396] Suitable pharmaceutical carriers include inert diluents or
fillers, water and various organic solvents (such as hydrates and
solvates). The pharmaceutical compositions may, if desired, contain
additional ingredients such as flavorings, binders, excipients and
the like. Thus for oral administration, tablets containing various
excipients, such as citric acid may be employed together with
various disintegrants such as starch, alginic acid and certain
complex silicates and with binding agents such as sucrose, gelatin
and acacia. Examples, without limitation, of excipients include
calcium carbonate, calcium phosphate, various sugars and types of
starch, cellulose derivatives, gelatin, vegetable oils and
polyethylene glycols. Additionally, lubricating agents such as
magnesium stearate, sodium lauryl sulfate and talc are often useful
for tableting purposes. Solid compositions of a similar type may
also be employed in soft and hard filled gelatin capsules.
Non-limiting examples of materials, therefore, include lactose or
milk sugar and high molecular weight polyethylene glycols. When
aqueous suspensions or elixirs are desired for oral administration
the active compound therein may be combined with various sweetening
or flavoring agents, coloring matters or dyes and, if desired,
emulsifying agents or suspending agents, together with diluents
such as water, ethanol, propylene glycol, glycerin, or combinations
thereof.
[0397] The pharmaceutical composition may, for example, be in a
form suitable for oral administration as a tablet, capsule, pill,
powder, sustained release formulation, solution or suspension, for
parenteral injection as a sterile solution, suspension or emulsion,
for topical administration as an ointment or cream, or for rectal
administration as a suppository.
[0398] Exemplary parenteral administration forms include solutions
or suspensions of an active compound in a sterile aqueous solution,
for example, aqueous propylene glycol or dextrose solutions. Such
dosage forms may be suitably buffered, if desired.
[0399] The pharmaceutical composition may be in unit dosage forms
suitable for single administration of precise amounts.
[0400] Pharmaceutical compositions suitable for the delivery of
active agents and methods for their preparation will be readily
apparent to those skilled in the art. Such compositions and methods
for their preparation may be found, for example, in `Remington's
Pharmaceutical Sciences`, 19th Edition (Mack Publishing Company,
1995), the disclosure of which is incorporated herein by reference
in its entirety.
[0401] Compounds of the invention may be administered orally. Oral
administration may involve swallowing, so that the compound enters
the gastrointestinal tract, or buccal or sublingual administration
may be employed by which the compound enters the blood stream
directly from the mouth.
[0402] Formulations suitable for oral administration include solid
formulations such as tablets, capsules containing particulates,
liquids, or powders, lozenges (including liquid-filled), chews,
multi- and nano-particulates, gels, solid solution, liposome, films
(including muco-adhesive), ovules, sprays and liquid
formulations.
[0403] Liquid formulations include suspensions, solutions, syrups
and elixirs. Such formulations may be used as fillers in soft or
hard capsules and typically include a carrier, for example, water,
ethanol, polyethylene glycol, propylene glycol, methylcellulose, or
a suitable oil, and one or more emulsifying agents and/or
suspending agents. Liquid formulations may also be prepared by the
reconstitution of a solid, for example, from a sachet.
[0404] Compounds of the invention may also be used in
fast-dissolving, fast-disintegrating dosage forms such as those
described in Expert Opinion in Therapeutic Patents, 11 (6), 981-986
by Liang and Chen (2001), the disclosure of which is incorporated
herein by reference in its entirety.
[0405] For tablet dosage forms, the active agent may make up from 1
wt % to 80 wt % of the dosage form, more typically from 5 wt % to
60 wt % of the dosage form. In addition to the active agent,
tablets generally contain a disintegrant. Examples of disintegrants
include sodium starch glycolate, sodium carboxymethyl cellulose,
calcium carboxymethyl cellulose, croscarmellose sodium,
crospovidone, polyvinylpyrrolidone, methyl cellulose,
microcrystalline cellulose, lower alkyl-substituted hydroxypropyl
cellulose, starch, pregelatinized starch and sodium alginate.
Generally, the disintegrant may comprise from 1 wt % to 25 wt %,
preferably from 5 wt % to 20 wt % of the dosage form.
[0406] Binders are generally used to impart cohesive qualities to a
tablet formulation. Suitable binders include microcrystalline
cellulose, gelatin, sugars, polyethylene glycol, natural and
synthetic gums, polyvinylpyrrolidone, pregelatinized starch,
hydroxypropyl cellulose and hydroxypropyl methylcellulose. Tablets
may also contain diluents, such as lactose (monohydrate,
spray-dried monohydrate, anhydrous and the like), mannitol,
xylitol, dextrose, sucrose, sorbitol, microcrystalline cellulose,
starch and dibasic calcium phosphate dihydrate.
[0407] Tablets may also optionally include surface active agents,
such as sodium lauryl sulfate and polysorbate 80, and glidants such
as silicon dioxide and talc. When present, surface active agents
are typically in amounts of from 0.2 wt % to 5 wt % of the tablet,
and glidants typically from 0.2 wt % to 1 wt % of the tablet.
[0408] Tablets also generally contain lubricants such as magnesium
stearate, calcium stearate, zinc stearate, sodium stearyl fumarate,
and mixtures of magnesium stearate with sodium lauryl sulphate.
Lubricants generally are present in amounts from 0.25 wt % to 10 wt
%, preferably from 0.5 wt % to 3 wt % of the tablet.
[0409] Other conventional ingredients include anti-oxidants,
colorants, flavoring agents, preservatives and taste-masking
agents.
[0410] Exemplary tablets may contain up to about 80 wt % active
agent, from about 10 wt % to about 90 wt % binder, from about 0 wt
% to about 85 wt % diluent, from about 2 wt % to about 10 wt %
disintegrant, and from about 0.25 wt % to about 10 wt %
lubricant.
[0411] Tablet blends may be compressed directly or by roller to
form tablets. Tablet blends or portions of blends may alternatively
be wet-, dry-, or melt-granulated, melt congealed, or extruded
before tableting. The final formulation may include one or more
layers and may be coated or uncoated; or encapsulated.
[0412] The formulation of tablets is discussed in detail in
"Pharmaceutical Dosage Forms: Tablets, Vol. 1", by H. Lieberman and
L. Lachman, Marcel Dekker, N.Y., N.Y., 1980 (ISBN 0-8247-6918-X),
the disclosure of which is incorporated herein by reference in its
entirety.
[0413] Solid formulations for oral administration may be formulated
to be immediate and/or modified release. Modified release
formulations include delayed-, sustained-, pulsed-, controlled-,
targeted and programmed release.
[0414] Suitable modified release formulations are described in U.S.
Pat. No. 6,106,864. Details of other suitable release technologies
such as high energy dispersions and osmotic and coated particles
may be found in Verma et al, Pharmaceutical Technology On-line,
25(2), 1-14 (2001). The use of chewing gum to achieve controlled
release is described in WO 00/35298. The disclosures of these
references are incorporated herein by reference in their
entireties.
Parenteral Administration
[0415] Compounds of the invention may also be administered directly
into the blood stream, into muscle, or into an internal organ.
Suitable means for parenteral administration include intravenous,
intraarterial, intraperitoneal, intrathecal, intraventricular,
intraurethral, intrasternal, intracranial, intramuscular and
subcutaneous. Suitable devices for parenteral administration
include needle (including micro needle) injectors, needle-free
injectors and infusion techniques.
[0416] Parenteral formulations are typically aqueous solutions
which may contain excipients such as salts, carbohydrates and
buffering agents (preferably to a pH of from 3 to 9), but, for some
applications, they may be more suitably formulated as a sterile
non-aqueous solution or as a dried form to be used in conjunction
with a suitable vehicle such as sterile, pyrogen-free water.
[0417] The preparation of parenteral formulations under sterile
conditions, for example, by lyophilization, may readily be
accomplished using standard pharmaceutical techniques well known to
those skilled in the art.
[0418] The solubility of compounds of the invention used in the
preparation of parenteral solutions may potentially be increased by
the use of appropriate formulation techniques, such as the
incorporation of solubility-enhancing agents.
[0419] Formulations for parenteral administration may be formulated
to be immediate and/or modified release. Modified release
formulations include delayed-, sustained-, pulsed-, controlled-,
targeted and programmed release. Thus compounds of the invention
may potentially be formulated as a solid, semi-solid, or
thixotropic liquid for administration as an implanted depot
providing modified release of the active compound. Examples of such
formulations include drug-coated stents and PGLA microspheres.
[0420] The compounds of the invention may also potentially be
administered topically to the skin or mucosa, that is, dermally or
transdermally. Typical formulations for this purpose include gels,
hydrogels, lotions, solutions, creams, ointments, dusting powders,
dressings, foams, films, skin patches, wafers, implants, sponges,
fibers, bandages and microemulsions. Liposomes may also be used.
Typical carriers include alcohol, water, mineral oil, liquid
petrolatum, white petrolatum, glycerin, polyethylene glycol and
propylene glycol. Penetration enhancers may be incorporated; see,
for example, J Pharm Sci, 88 (10), 955-958 by Finnin and Morgan
(October 1999). Other means of topical administration include
delivery by electroporation, iontophoresis, phonophoresis,
sonophoresis and micro needle or needle-free (e.g. Powderject.TM.,
Bioject.TM., etc.) injection. The disclosures of these references
are incorporated herein by reference in their entireties.
[0421] Formulations for topical administration may be formulated to
be immediate and/or modified release. Modified release formulations
include delayed-, sustained-, pulsed-, controlled-, targeted and
programmed release.
[0422] Compounds of the invention may also potentially be
administered intranasally or by inhalation, typically in the form
of a dry powder (either alone, as a mixture, for example, in a dry
blend with lactose, or as a mixed component particle, for example,
mixed with phospholipids, such as phosphatidylcholine) from a dry
powder inhaler or as an aerosol spray from a pressurized container,
pump, spray, atomizer (preferably an atomizer using
electrohydrodynamics to produce a fine mist), or nebulizer, with or
without the use of a suitable propellant, such as
1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane. For
intranasal use, the powder may include a bioadhesive agent, for
example, chitosan or cyclodextrin.
[0423] The pressurized container, pump, spray, atomizer, or
nebulizer may contain a solution or suspension of the compound(s)
of the invention comprising, for example, ethanol, aqueous ethanol,
or a suitable alternative agent for dispersing, solubilizing, or
extending release of the active, a propellant(s) as solvent and an
optional surfactant, such as sorbitan trioleate, oleic acid, or an
oligolactic acid.
[0424] Prior to use in a dry powder or suspension formulation, the
compound may be micronized to a size suitable for delivery by
inhalation (typically less than 5 microns). This may be achieved by
any appropriate comminuting method, such as spiral jet milling,
fluid bed jet milling, supercritical fluid processing to form
nanoparticles, high pressure homogenization, or spray drying.
[0425] Capsules (made, for example, from gelatin or HPMC), blisters
and cartridges for use in an inhaler or insufflator may be
formulated to contain a powder mix of the compound of the
invention, a suitable powder base such as lactose or starch and a
performance modifier such as I-leucine, mannitol, or magnesium
stearate. The lactose may be anhydrous or in the form of the
monohydrate, preferably the latter. Other suitable excipients
include dextran, glucose, maltose, sorbitol, xylitol, fructose,
sucrose and trehalose.
[0426] A suitable solution formulation for use in an atomizer using
electrohydrodynamics to produce a fine mist may contain from 1
.mu.g to 20 mg of the compound of the invention per actuation and
the actuation volume may vary from 1 .mu.L to 100 .mu.L. A typical
formulation includes a compound of the invention, propylene glycol,
sterile water, ethanol and sodium chloride. Alternative solvents
which may be used instead of propylene glycol include glycerol and
polyethylene glycol.
[0427] Suitable flavors, such as menthol and levomenthol, or
sweeteners, such as saccharin or saccharin sodium, may be added to
those formulations of the invention intended for inhaled/intranasal
administration.
[0428] Formulations for inhaled/intranasal administration may be
formulated to be immediate and/or modified release using, for
example, poly(DL-lactic-coglycolic acid (PGLA). Modified release
formulations include delayed-, sustained-, pulsed-, controlled-,
targeted and programmed release.
[0429] In the case of dry powder inhalers and aerosols, the dosage
unit is determined by means of a valve which delivers a metered
amount. Units in accordance with the invention are typically
arranged to administer a metered dose or "puff" containing a
desired mount of the compound of the invention. The overall daily
dose may be administered in a single dose or, more usually, as
divided doses throughout the day.
[0430] Compounds of the invention may potentially be administered
rectally or vaginally, for example, in the form of a suppository,
pessary, or enema. Cocoa butter is a traditional suppository base,
but various alternatives may be used as appropriate.
[0431] Formulations for rectal/vaginal administration may be
formulated to be immediate and/or modified release. Modified
release formulations include delayed-, sustained-, pulsed-,
controlled-, targeted and programmed release.
[0432] Compounds of the invention may also potentially be
administered directly to the eye or ear, typically in the form of
drops of a micronized suspension or solution in isotonic,
pH-adjusted, sterile saline. Other formulations suitable for ocular
and aural administration may include ointments, biodegradable (e.g.
absorbable gel sponges, collagen) and non-biodegradable (e.g.
silicone) implants, wafers, lenses and particulate or vesicular
systems, such as niosomes or liposomes. A polymer such as
crossed-linked polyacrylic acid, polyvinylalcohol, hyaluronic acid,
a cellulosic polymer, for example, hydroxypropylmethylcellulose,
hydroxyethylcellulose, or methyl cellulose, or a
heteropolysaccharide polymer, for example, gelan gum, may be
incorporated together with a preservative, such as benzalkonium
chloride. Such formulations may also be delivered by
iontophoresis.
[0433] Formulations for ocular/aural administration may be
formulated to be immediate and/or modified release. Modified
release formulations include delayed-, sustained-, pulsed-,
controlled-, targeted, or programmed release.
Other Technologies
[0434] Compounds of the invention may be combined with soluble
macromolecular entities, such as cyclodextrin and suitable
derivatives thereof or polyethylene glycol-containing polymers, in
order to improve their solubility, dissolution rate, taste-masking,
bioavailability and/or stability for use in any of the
aforementioned modes of administration.
[0435] Drug-cyclodextrin complexes, for example, may be useful for
different dosage forms and administration routes. Both inclusion
and non-inclusion complexes may potentially be used. As an
alternative to direct complexation with the drug, the cyclodextrin
may be used as an auxiliary additive, i.e. as a carrier, diluent,
or solubilizer. Most commonly used for these purposes are alpha-,
beta- and gamma-cyclodextrins, examples of which may be found in
PCT Publication Nos. WO 91/11172, WO 94/02518 and WO 98/55148, the
disclosures of which are incorporated herein by reference in their
entireties.
Dosage
[0436] The amount of the active compound administered will be
dependent on the subject being treated, the severity of the
disorder or condition, the rate of administration, the disposition
of the compound and the discretion of the prescribing physician.
However, an effective dosage is typically in the range of about
0.001 to about 100 mg per kg body weight per day, preferably about
0.01 to about 35 mg/kg/day, in single or divided doses. For a 70 kg
human, this would amount to about 0.07 to about 7000 mg/day,
preferably about 0.7 to about 2500 mg/day. In some instances,
dosage levels below the lower limit of the aforesaid range may be
more than adequate, while in other cases still larger doses may be
used without causing any harmful side effect, with such larger
doses typically divided into several smaller doses for
administration throughout the day.
Kit-of-Parts
[0437] Inasmuch as it may desirable to administer a combination of
active compounds, for example, for the purpose of treating a
particular disease or condition, it is within the scope of the
present invention that two or more pharmaceutical compositions, at
least one of which contains a compound in accordance with the
invention, may conveniently be combined in the form of a kit
suitable for coadministration of the compositions. Thus the kit of
the invention includes two or more separate pharmaceutical
compositions, at least one of which contains a compound of the
invention, and means for separately retaining said compositions,
such as a container, divided bottle, or divided foil packet. An
example of such a kit is the familiar blister pack used for the
packaging of tablets, capsules and the like.
[0438] The kit of the invention may be particularly suitable for
administering different dosage forms, for example, oral and
parenteral, for administering the separate compositions at
different dosage intervals, or for titrating the separate
compositions against one another. To assist compliance, the kit
typically includes directions for administration and may be
provided with a memory aid.
Combination Therapy
[0439] As used herein, the term "combination therapy" refers to the
administration of a compound of the invention together with an at
least one additional pharmaceutical or medicinal agent (e.g., an
anti-cancer agent), either sequentially or simultaneously.
[0440] As noted above, the compounds of the invention may
potentially be used in combination with one or more additional
anti-cancer agents which are described below. When a combination
therapy is used, the one or more additional anti-cancer agents may
be administered sequentially or simultaneously with the compound of
the invention. In one embodiment, the additional anti-cancer agent
is administered to a mammal (e.g., a human) prior to administration
of the compound of the invention. In another embodiment, the
additional anti-cancer agent is administered to the mammal after
administration of the compound of the invention. In another
embodiment, the additional anti-cancer agent is administered to the
mammal (e.g., a human) simultaneously with the administration of
the compound of the invention.
[0441] The invention also relates to a pharmaceutical composition
for the treatment of abnormal cell growth in a mammal, including a
human, which comprises an amount of a compound of the invention, as
defined above (including hydrates, solvates and polymorphs of said
compound or pharmaceutically acceptable salts thereof), in
combination with one or more (preferably one to three) anti-cancer
agents selected from the group consisting of anti-angiogenesis
agents and signal transduction inhibitors and a pharmaceutically
acceptable carrier, wherein the amounts of the active agent and the
combination anti-cancer agents when taken as a whole is
therapeutically effective for treating said abnormal cell
growth.
[0442] In one embodiment of the present invention the anti-cancer
agent used in conjunction with a compound of the invention and
pharmaceutical compositions described herein is an
anti-angiogenesis agent (e.g., an agent that stops tumors from
developing new blood vessels). Examples of anti-angiogenesis agents
include for example VEGF inhibitors, VEGFR inhibitors, TIE-2
inhibitors, PDGFR inhibitors, angiopoetin inhibitors, PKC.beta.
inhibitors, COX-2 (cyclooxygenase II) inhibitors, integrins
(alpha-v/beta-3), MMP-2 (matrix-metalloprotienase 2) inhibitors,
and MMP-9 (matrix-metalloprotienase 9) inhibitors.
[0443] Preferred anti-angiogenesis agents include sunitinib
(Sutent.TM.), bevacizumab (Avastin.TM.), axitinib (AG 13736), SU
14813 (Pfizer), and AG 13958 (Pfizer).
[0444] Additional anti-angiogenesis agents include vatalanib (CGP
79787), Sorafenib (Nexavar.TM.), pegaptanib octasodium
(Macugen.TM.), vandetanib (Zactima.TM.), PF-0337210 (Pfizer), SU
14843 (Pfizer), AZD 2171 (AstraZeneca), ranibizumab (Lucentis.TM.),
Neovastat.TM. (AE 941), tetrathiomolybdata (Coprexa.TM.), AMG 706
(Amgen), VEGF Trap (AVE 0005), CEP 7055 (Sanofi-Aventis), XL 880
(Exelixis), telatinib (BAY 57-9352), and CP-868,596 (Pfizer).
[0445] Other anti-angiogenesis agents include enzastaurin (LY
317615), midostaurin (CGP 41251), perifosine (KRX 0401), teprenone
(Selbex.TM.) and UCN 01 (Kyowa Hakko).
[0446] Other examples of anti-angiogenesis agents which may be used
in conjunction with a compound of the invention and pharmaceutical
compositions described herein include celecoxib (Celebrex.TM.),
parecoxib (Dynastat.TM.), deracoxib (SC 59046), lumiracoxib
(Preige.TM.), valdecoxib (Bextra.TM.), rofecoxib (Vioxx.TM.),
iguratimod (Careram.TM.), IP 751 (Invedus), SC-58125 (Pharmacia)
and etoricoxib (Arcoxia.TM.).
[0447] Other anti-angiogenesis agents include exisulind
(Aptosyn.TM.), salsalate (Amigesic.TM.), diflunisal (Dolobid.TM.),
ibuprofen (Motrin.TM.), ketoprofen (Orudis.TM.), nabumetone
(Relafen.TM.), piroxicam (Feldene.TM.), naproxen (Aleve.TM.,
Naprosyn.TM.), diclofenac (Voltaren.TM.), indomethacin
(Indocin.TM.), sulindac (Clinoril.TM.), tolmetin (Tolectin.TM.),
etodolac (Lodine.TM.), ketorolac (Toradol.TM.), and oxaprozin
(Daypro.TM.).
[0448] Other anti-angiogenesis agents include ABT 510 (Abbott),
apratastat (TMI 005), AZD 8955 (AstraZeneca), incyclinide
(Metastat.TM.), and PCK 3145 (Procyon).
[0449] Other anti-angiogenesis agents include acitretin
(Neotigason.TM.), plitidepsin (Aplidine.TM.), cilengtide (EMD
121974), combretastatin A4 (CA4P), fenretinide (4 HPR),
halofuginone (Tempostatin.TM.), Panzem.TM. (2-methoxyestradiol),
PF-03446962 (Pfizer), rebimastat (BMS 275291), catumaxomab
(Removab.TM.), lenalidomide (Revlimid.TM.), squalamine
(EVIZON.TM.), thalidomide (Thalomid.TM.), Ukrain.TM. (NSC 631570),
Vitaxin.TM. (MEDI 522), and zoledronic acid (Zometa.TM.).
[0450] In another embodiment the anti-cancer agent is a so called
signal transduction inhibitor (e.g., inhibiting the means by which
regulatory molecules that govern the fundamental processes of cell
growth, differentiation, and survival communicated within the
cell). Signal transduction inhibitors include small molecules,
antibodies, and antisense molecules. Signal transduction inhibitors
include for example kinase inhibitors (e.g., tyrosine kinase
inhibitors or serine/threonine kinase inhibitors) and cell cycle
inhibitors. More specifically signal transduction inhibitors
include, for example, farnesyl protein transferase inhibitors, EGF
inhibitor, ErbB-1 (EGFR), ErbB-2, pan erb, IGF1R inhibitors, MEK,
c-Kit inhibitors, FLT-3 inhibitors, K-Ras inhibitors, PI3 kinase
inhibitors, JAK inhibitors, STAT inhibitors, Raf kinase inhibitors,
Akt inhibitors, mTOR inhibitor, P70S6 kinase inhibitors, inhibitors
of the WNT pathway and so called multi-targeted kinase
inhibitors.
[0451] Preferred signal transduction inhibitors include gefitinib
(Iressa.TM.), cetuximab (Erbitux.TM.), erlotinib (Tarceva.TM.),
trastuzumab (Herceptin.TM.), sunitinib (Sutent.TM.), imatinib
(Gleevec.TM.), and PD325901 (Pfizer).
[0452] Additional examples of signal transduction inhibitors which
may be used in conjunction with a compound of the invention and
pharmaceutical compositions described herein include BMS 214662
(Bristol-Myers Squibb), lonafarnib (Sarasar.TM.), pelitrexol (AG
2037), matuzumab (EMD 7200), nimotuzumab (TheraCIM h-R3.TM.),
panitumumab (Vectibix.TM.), Vandetanib (Zactima.TM.), pazopanib (SB
786034), ALT 110 (Alteris Therapeutics), BIBW 2992 (Boehringer
Ingelheim), and Cervene.TM. (TP 38).
[0453] Other examples of signal transduction inhibitor include
PF-2341066 (Pfizer), PF-299804 (Pfizer), canertinib (CI 1033),
pertuzumab (Omnitarg.TM.), Lapatinib (Tycerb.TM.), pelitinib (EKB
569), miltefosine (Miltefosin.TM.), BMS 599626 (Bristol-Myers
Squibb), Lapuleucel-T (Neuvenge.TM.), NeuVax.TM. (E75 cancer
vaccine), Osidem.TM. (IDM 1), mubritinib (TAK-165), CP-724,714
(Pfizer), panitumumab (Vectibix.TM.), lapatinib (Tycerb.TM.),
PF-299804 (Pfizer), pelitinib (EKB 569), and pertuzumab
(Omnitarg.TM.).
[0454] Other examples of signal transduction inhibitors include
ARRY 142886 (Array Biopharm), everolimus (Certican.TM.),
zotarolimus (Endeavor.TM.), temsirolimus (Torisel.TM.), AP 23573
(ARIAD), and VX 680 (Vertex).
[0455] Additionally, other signal transduction inhibitors include
XL 647 (Exelixis), sorafenib (Nexavar.TM.), LE-AON (Georgetown
University), and GI-4000 (Globelmmune).
[0456] Other signal transduction inhibitors include ABT 751
(Abbott), alvocidib (flavopiridol), BMS 387032 (Bristol Myers), EM
1421 (Erimos), indisulam (E 7070), seliciclib (CYC 200), BIO 112
(Onc Bio), BMS 387032 (Bristol-Myers Squibb), PD 0332991 (Pfizer),
and AG 024322 (Pfizer).
[0457] This invention contemplates the use of a compound of the
invention together with classical antineoplastic agents. Classical
antineoplastic agents include but are not limited to hormonal
modulators such as hormonal, anti-hormonal, androgen agonist,
androgen antagonist and anti-estrogen therapeutic agents, histone
deacetylase (HDAC) inhibitors, gene silencing agents or gene
activating agents, ribonucleases, proteosomics, Topoisomerase I
inhibitors, Camptothecin derivatives, Topoisomerase II inhibitors,
alkylating agents, antimetabolites, poly(ADP-ribose) polymerase-1
(PARP-1) inhibitor, microtubulin inhibitors, antibiotics, plant
derived spindle inhibitors, platinum-coordinated compounds, gene
therapeutic agents, antisense oligonucleotides, vascular targeting
agents (VTAs), and statins
[0458] Examples of classical antineoplastic agents used in
combination therapy with a compound of the invention, optionally
with one or more other agents include, but are not limited to,
glucocorticoids, such as dexamethasone, prednisone, prednisolone,
methylprednisolone, hydrocortisone, and progestins such as
medroxyprogesterone, megestrol acetate (Megace), mifepristone
(RU-486), Selective Estrogen Receptor Modulators (SERMs; such as
tamoxifen, raloxifene, lasofoxifene, afimoxifene, arzoxifene,
bazedoxifene, fispemifene, ormeloxifene, ospemifene, tesmilifene,
toremifene, trilostane and CHF 4227 (Cheisi), Selective
Estrogen-Receptor Downregulators (SERD's; such as fulvestrant),
exemestane (Aromasin), anastrozole (Arimidex), atamestane,
fadrozole, letrozole (Femara), gonadotropin-releasing hormone
(GnRH; also commonly referred to as luteinizing hormone-releasing
hormone [LHRH]) agonists such as buserelin (Suprefact), goserelin
(Zoladex), leuprorelin (Lupron), and triptorelin (Trelstar),
abarelix (Plenaxis), bicalutamide (Casodex), cyproterone, flutamide
(Eulexin), megestrol, nilutamide (Nilandron), and osaterone,
dutasteride, epristeride, finasteride, Serenoa repens, PHL 00801,
abarelix, goserelin, leuprorelin, triptorelin, bicalutamide,
tamoxifen, exemestane, anastrozole, fadrozole, formestane,
letrozole, and combinations thereof.
[0459] Other examples of classical antineoplastic agents used in
combination with a compound of the invention include but are not
limited to suberolanilide hydroxamic acid (SAHA, Merck Inc./Aton
Pharmaceuticals), depsipeptide (FR901228 or FK228), G2M-777,
MS-275, pivaloyloxymethyl butyrate and PXD-101; Onconase
(ranpirnase), PS-341 (MLN-341), Velcade (bortezomib),
9-aminocamptothecin, belotecan, BN-80915 (Roche), camptothecin,
diflomotecan, edotecarin, exatecan (Daiichi), gimatecan,
10-hydroxycamptothecin, irinotecan HCl (Camptosar), lurtotecan,
Orathecin (rubitecan, Supergen), SN-38, topotecan, camptothecin,
10-hydroxycamptothecin, 9-aminocamptothecin, irinotecan, SN-38,
edotecarin, topotecan, aclarubicin, adriamycin, amonafide,
amrubicin, annamycin, daunorubicin, doxorubicin, elsamitrucin,
epirubicin, etoposide, idarubicin, galarubicin, hydroxycarbamide,
nemorubicin, novantrone (mitoxantrone), pirarubicin, pixantrone,
procarbazine, rebeccamycin, sobuzoxane, tafluposide, valrubicin,
Zinecard (dexrazoxane), nitrogen mustard N-oxide, cyclophosphamide,
AMD-473, altretamine, AP-5280, apaziquone, brostallicin,
bendamustine, busulfan, carboquone, carmustine, chlorambucil,
dacarbazine, estramustine, fotemustine, glufosfamide, ifosfamide,
KW-2170, lomustine, mafosfamide, mechlorethamine, melphalan,
mitobronitol, mitolactol, mitomycin C, mitoxatrone, nimustine,
ranimustine, temozolomide, thiotepa, and platinum-coordinated
alkylating compounds such as cisplatin, Paraplatin (carboplatin),
eptaplatin, lobaplatin, nedaplatin, Eloxatin (oxaliplatin, Sanofi),
streptozocin, satrplatin, and combinations thereof.
[0460] The invention also contemplates the use of the a compound of
the invention together with dihydrofolate reductase inhibitors
(such as methotrexate and NeuTrexin (trimetresate glucuronate)),
purine antagonists (such as 6-mercaptopurine riboside,
mercaptopurine, 6-thioguanine, cladribine, clofarabine (Clolar),
fludarabine, nelarabine, and raltitrexed), pyrimidine antagonists
(such as 5-fluorouracil (5-FU), Alimta (premetrexed disodium,
LY231514, MTA), capecitabine (Xeloda.TM.), cytosine arabinoside,
Gemzar.TM. (gemcitabine, Eli Lilly), Tegafur (UFT Orzel or Uforal
and including TS-1 combination of tegafur, gimestat and otostat),
doxifluridine, carmofur, cytarabine (including ocfosfate, phosphate
stearate, sustained release and liposomal forms), enocitabine,
5-azacitidine (Vidaza), decitabine, and ethynylcytidine) and other
antimetabolites such as eflornithine, hydroxyurea, leucovorin,
nolatrexed (Thymitaq), triapine, trimetrexate,
N-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]--
2-thenoyl)-L-glutamic acid, AG-014699 (Pfizer Inc.), ABT-472
(Abbott Laboratories), INO-1001 (Inotek Pharmaceuticals), KU-0687
(KuDOS Pharmaceuticals) and GPI 18180 (Guilford Pharm Inc) and
combinations thereof.
[0461] Other examples of classical antineoplastic cytotoxic agents
used in combination therapy with a compound of the invention,
optionally with one or more other agents include, but are not
limited to, Abraxane (Abraxis BioScience, Inc.), Batabulin (Amgen),
EPO 906 (Novartis), Vinflunine (Bristol-Myers Squibb Company),
actinomycin D, bleomycin, mitomycin C, neocarzinostatin
(Zinostatin), vinblastine, vincristine, vindesine, vinorelbine
(Navelbine), docetaxel (Taxotere), Ortataxel, paclitaxel (including
Taxoprexin a DHA/paciltaxel conjugate), cisplatin, carboplatin,
Nedaplatin, oxaliplatin (Eloxatin), Satraplatin, Camptosar,
capecitabine (Xeloda), oxaliplatin (Eloxatin), Taxotere
alitretinoin, Canfosfamide (Telcyta.TM.), DMXAA (Antisoma),
ibandronic acid, L-asparaginase, pegaspargase (Oncaspar.TM.),
Efaproxiral (Efaproxyn.TM.--radiation therapy), bexarotene
(Targretin.TM.), Tesmilifene (DPPE--enhances efficacy of
cytotoxics), Theratope.TM. (Biomira), Tretinoin (Vesanoid.TM.),
tirapazamine (Trizaone.TM.), motexafin gadolinium (Xcytrin.TM.)
Cotara.TM. (mAb), and NBI-3001 (Protox Therapeutics),
polyglutamate-paclitaxel (Xyotax.TM.) and combinations thereof.
[0462] Further examples of classical antineoplastic agents used in
combination therapy with a compound of the invention, optionally
with one or more other agents include, but are not limited to, as
Advexin (ING 201), TNFerade (GeneVec, a compound which express
TNFalpha in response to radiotherapy), RB94 (Baylor College of
Medicine), Genasense (Oblimersen, Genta), Combretastatin A4P
(CA4P), Oxi-4503, AVE-8062, ZD-6126, TZT-1027, Atorvastatin
(Lipitor, Pfizer Inc.), Provastatin (Pravachol, Bristol-Myers
Squibb), Lovastatin (Mevacor, Merck Inc.), Simvastatin (Zocor,
Merck Inc.), Fluvastatin (Lescol, Novartis), Cerivastatin (Baycol,
Bayer), Rosuvastatin (Crestor, AstraZeneca), Lovostatin, Niacin
(Advicor, Kos Pharmaceuticals), Caduet, Lipitor, torcetrapib, and
combinations thereof.
[0463] Another embodiment of the present invention of particular
interest relates to a method for the treatment of breast cancer in
a human in need of such treatment, comprising administering to said
human an amount of a compound of the invention, in combination with
one or more (preferably one to three) anti-cancer agents selected
from the group consisting of trastuzumab, tamoxifen, docetaxel,
paclitaxel, capecitabine, gemcitabine, vinorelbine, exemestane,
letrozole and anastrozole.
[0464] In one embodiment the invention provides a method of
treating colorectal cancer in a mammal, such as a human, in need of
such treatment, by administering an amount of a compound of the
invention, in combination with one or more (preferably one to
three) anti-cancer agents. Examples of particular anti-cancer
agents include those typically used in adjuvant chemotherapy, such
as FOLFOX, a combination of 5-fluorouracil (5-FU) or capecitabine
(Xeloda), leucovorin and oxaliplatin (Eloxatin). Further examples
of particular anti-cancer agents include those typically used in
chemotherapy for metastatic disease, such as FOLFOX or FOLFOX in
combination with bevacizumab (Avastin); and FOLFIRI, a combination
of 5-FU or capecitabine, leucovorin and irinotecan (Camptosar).
Further examples include 17-DMAG, ABX-EFR, AMG-706, AMT-2003,
ANX-510 (CoFactor), aplidine (plitidepsin, Aplidin), Aroplatin,
axitinib (AG-13736), AZD-0530, AZD-2171, bacillus Calmette-Guerin
(BCG), bevacizumab (Avastin), BIO-117, BIO-145, BMS-184476,
BMS-275183, BMS-528664, bortezomib (Velcade), C-1311 (Symadex),
cantuzumab mertansine, capecitabine (Xeloda), cetuximab (Erbitux),
clofarabine (Clofarex), CMD-193, combretastatin, Cotara, CT-2106,
CV-247, decitabine (Dacogen), E-7070, E-7820, edotecarin,
EMD-273066, enzastaurin (LY-317615)epothilone B (EPO-906),
erlotinib (Tarceva), flavopyridol, GCAN-101, gefitinib (Iressa),
huA33, huC242-DM4, imatinib (Gleevec), indisulam, ING-1, irinotecan
(CPT-11, Camptosar) ISIS 2503, ixabepilone, lapatinib (Tykerb),
mapatumumab (HGS-ETR1), MBT-0206, MEDI-522 (Abregrin), Mitomycin,
MK-0457 (VX-680), MLN-8054, NB-1011, NGR-TNF, NV-1020, oblimersen
(Genasense, G3139), OncoVex, ONYX 015 (CI-1042), oxaliplatin
(Eloxatin), panitumumab (ABX-EGF, Vectibix), pelitinib (EKB-569),
pemetrexed (Alimta), PD-325901, PF-0337210, PF-2341066, RAD-001
(Everolimus), RAV-12, Resveratrol, Rexin-G, S-1 (TS-1), seliciclib,
SN-38 liposome, Sodium stibogluconate (SSG), sorafenib (Nexavar),
SU-14813, sunitinib (Sutent), temsirolimus (CCI 779),
tetrathiomolybdate, thalomide, TLK-286 (Telcyta), topotecan
(Hycamtin), trabectedin (Yondelis), vatalanib (PTK-787), vorinostat
(SAHA, Zolinza), WX-UK1, and ZYC300, wherein the amounts of the
active agent together with the amounts of the combination
anticancer agents are effective in treating colorectal cancer.
[0465] Another embodiment of the present invention of particular
interest relates to a method for the treatment of renal cell
carcinoma in a human in need of such treatment, comprising
administering to said human an amount of a compound of the
invention, in combination with one or more (preferably one to
three) anti-cancer agents selected from the group consisting of
axitinib (AG 13736), capecitabine (Xeloda), interferon alpha,
interleukin-2, bevacizumab (Avastin), gemcitabine (Gemzar),
thalidomide, cetuximab (Erbitux), vatalanib (PTK-787), sunitinib
(Sutent.TM.), AG-13736, SU-11248, Tarceva, Iressa, Lapatinib and
Gleevec, wherein the amounts of the active agent together with the
amounts of the combination anticancer agents is effective in
treating renal cell carcinoma.
[0466] Another embodiment of the present invention of particular
interest relates to a method for the treatment of melanoma in a
human in need of such treatment, comprising administering to said
human an amount of a compound of the invention, in combination with
one or more (preferably one to three) anti-cancer agents selected
from the group consisting of interferon alpha, interleukin-2,
temozolomide (Temodar), docetaxel (Taxotere), paclitaxel,
Dacarbazine (DTIC), carmustine (also known as BCNU), Cisplatin,
vinblastine, tamoxifen, PD-325,901, axitinib (AG 13736),
bevacizumab (Avastin), thalidomide, sorafanib, vatalanib (PTK-787),
sunitinib (Sutent.TM.), CpG-7909, AG-13736, Iressa, Lapatinib and
Gleevec, wherein the amounts of the active agent together with the
amounts of the combination anticancer agents is effective in
treating melanoma.
[0467] Another embodiment of the present invention of particular
interest relates to a method for the treatment of lung cancer in a
human in need of such treatment, comprising administering to said
human an amount of a compound of the invention, in combination with
one or more (preferably one to three) anti-cancer agents selected
from the group consisting of capecitabine (Xeloda), axitinib (AG
13736), bevacizumab (Avastin), gemcitabine (Gemzar), docetaxel
(Taxotere), paclitaxel, premetrexed disodium (Alimta), Tarceva,
Iressa, Vinorelbine, Irinotecan, Etoposide, Vinblastine, sunitinib
(Sutent.TM.), and Paraplatin (carboplatin), wherein the amounts of
the active agent together with the amounts of the combination
anticancer agents is effective in treating lung cancer.
Synthetic Methods
[0468] Compounds of the invention may be prepared according to the
exemplary procedures provided herein and modifications thereof
known to those of skill in the art. In addition, synthetic routes
for the formation of various compounds useful as staring materials
for the preparation of the compounds claimed herein are described
in International Application No. PCT/IB2013/060682, the content of
which is incorporated by reference herein in its entirely.
[0469] These and other methods are exemplified in the preparation
of the examples provided herein. It will be understood by those of
skill in the art that the selection of starting materials and the
particular order of the steps, including, e.g., formation of the
lactam ring, installation or manipulation of various substituent
groups on the fused lactam or its precursors, and installation of
the pyridinone moiety, may be varied by selection of a suitable
synthetic strategy.
[0470] Synthetic examples are provided throughout the Examples and
in Table 1 below. EZH2 IC.sub.50 values (.mu.M) for WT EZH2 and
Mutant Y641N EZH2 are provided in Table 2 for exemplary compounds
of the invention.
[0471] The following abbreviations are used throughout the
Examples: "Ac" means acetyl, "AcO" or "OAc" means acetoxy,
"Ac.sub.2O" means acetic anhydride, "ACN" or "MeCN" means
acetonitrile, "AIBN" means azobisisobutyronitrile, "BOC", "Boc" or
"boc" means N-tert-butoxycarbonyl, "Bn" means benzyl, "BPO" means
dibenzoyl peroxide, "Bu" means butyl, "iBu" means isobutyl, "sBu"
means sec-butyl, "tBu" means tert-butyl, "tBuOK" or "KOtBu" means
potassium tert-butoxide, "CDI" means carbonyldiimidazole, "DCE"
means 1,2-dichloroethane, "DCM" (CH.sub.2Cl.sub.2) means methylene
chloride, "DEAD" means diethyl azodicarboxylate, "DIAD" means
diisopropyl azodicarboxylate, "DIPEA" or "DIEA" means diisopropyl
ethyl amine, "DBU" means 1,8-diazabicyclo[5.4.0]undec-7-ene,
"DIBAL-H" means diisobutylaluminum hydride, "DMA" means
N,N-dimethylacetamide, "DMAP" means 4-dimethylaminopyridine, "DME"
means dimethoxyethane, "DMF" means N--N-dimethyl formamide, "DMS"
means dimethylsulfide, "DMSO" means dimethylsulfoxide, "dppf" means
(diphenylphosphino)ferrocene, "DPPP" means
1,3-bis(diphenylphosphino)propane, "Et" means ethyl, "EtOAc" means
ethyl acetate, "EtOH" means ethanol, "HATU" means
2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate, "HOAc" or "AcOH" means acetic acid, "i-Pr" or
".sup.iPr" means isopropyl, "IPA" means isopropyl alcohol, "KHMDS"
means potassium hexamethyldisilazide (potassium
bis(trimethylsilyl)amide), "LiHMDS" means lithium
hexamethyldisilazide (lithium bis(trimethylsilyl)amide), "mCPBA"
means meta-chloroperoxy-benzoic acid, "Me" means methyl, "MeOH"
means methanol, "Ms" means methanesulfonate (commonly called
`mesylate`), "MTBE" means methyl t-butyl ether, "NBS" means
N-bromosuccinimide, "NCS" means N-chlorosuccinimide, "NIS" means
N-iodosuccinimide, "NMM" means N-methylmorpholine, "NMP" means
1-methyl 2-pyrrolidinone, "Ph" means phenyl, "RuPhos" means
2-Dicyclohexylphosphino-2',6'-diisopropoxybiphenyl, "Selectfluor"
means Chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane
bis(tetrafluoroborate), "TEA" means triethylamine, "TFA" means
trifluoroacetic acid, "Tf" means trifluoromethanesulfonate
(commonly called `triflate`), "THF" means tetrahydrofuran, "TMS"
means trimethylsilyl, "TMSA" means trimethylsilylazide, "TsCl"
means toluenesulfonyl chloride (commonly called `tosylate`), "SFC"
means supercritical fluid chromatography, "TLC" means thin layer
chromatography, "Rf" means retention fraction, ".about." means
approximately, "rt" means room temperature, "h" means hours, "min"
means minutes, "eq." means equivalents.
Preparation of Synthetic Intermediates
Compound D: 2-(benzyloxy)-3-(chloromethyl)-4,6-dimethylpyridine
##STR00021##
[0473] To a solution of
2-hydroxy-4,6-dimethylpyridine-3-carbonitrile (85.0 g, 0.574 mol)
and benzyl chloride (87.0 g, 0.688 mol) in toluene (800 mL) was
added Ag.sub.2O (146 g, 0.631 mol). The reaction mixture was
stirred at 110.degree. C. overnight. The reaction mixture was
filtered through CELITE.RTM. and the solids washed with
dichloromethane. The filtrate was concentrated under vacuum and
purified by column chromatography (petroleum ether/ethyl acetate)
to give 2-(benzyloxy)-4,6-dimethylpyridine-3-carbonitrile (Cpd A,
89 g, 65%) as a white solid.
[0474] 44.5 g.times.2 batches: To a stirred solution of
2-(benzyloxy)-4,6-dimethylpyridine-3-carbonitrile (Cpd A, 44.5 g,
187 mmol) in dichloromethane (500 mL) was added drop wise DIBAL-H
(224 mL, 224 mmol, 1M in toluene) at 0.about.5.degree. C. The
reaction mixture was allowed to warm to room temperature and
stirred for an additional 3 hours. The mixture was quenched with 1N
HCl (200 mL) and was stirred vigorously for 30 minutes. The
reaction mixture was neutralized with 4N NaOH (20 mL) and the
biphasic mixture was filtered and washed with dichloromethane (500
mL). The aqueous layer was extracted with dichloromethane (200 mL),
the combined organic layers were dried over sodium sulfate, and
concentrated under vacuum. The residue was purified by column
chromatography (petroleum ether/EtOAc) to give
2-(benzyloxy)-4,6-dimethylpyridine-3-carbaldehyde (Cpd B, 70 g,
78%) as a yellow solid.
[0475] 35 g.times.2 batches: To a 0.degree. C. solution of
2-(benzyloxy)-4,6-dimethylpyridine-3-carbaldehyde (Cpd B, 35.0 g,
145 mmol) in methanol (1000 mL) was added sodium borohydride (6.60
g, 174 mmol) in portions. The reaction mixture was stirred at room
temperature for 2 hours. The reaction mixture was concentrated
under vacuum and the residue was diluted with NaHCO.sub.3 (sat.,
aq.). After the bubbling had stopped, the aqueous solution was
extracted with ethyl acetate (2.times.500 mL). The combined organic
layers were dried over sodium sulfate, concentrated under vacuum,
and purified by column chromatography (petroleum ether/ethyl
acetate) to give [2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methanol
(Cpd C, 43 g, 61%) as a colorless oil.
[0476] 21.5 g.times.2 batches: To a solution of
[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methanol (Cpd C, 21.5 g,
88.5 mmol) in anhydrous dichloromethane (400 mL) was added thionyl
chloride (16.0 g, 133 mmol) at -40.degree. C. under N.sub.2. The
mixture was stirred at -40.degree. C. for 30 minutes. The reaction
mixture was poured into ice-water (300 mL) and adjusted to pH
7.about.8 with NaHCO.sub.3 (solid). The mixture was separated and
the aqueous layer was extracted with dichloromethane (300 mL). The
combined organic layers were washed with brine (300 mL), dried over
sodium sulfate, and concentrated under vacuum. The residue was
purified by column chromatography (petroleum ether/ethyl acetate,
100:1) to give 2-(benzyloxy)-3-(chloromethyl)-4,6-dimethylpyridine
(Cpd D, 27.5 g, 60%) as a white solid. .sup.1H NMR (400 MHz, CDCl3)
.delta. 7.51-7.49 (d, 2H), 7.41-7.37 (t, 2H), 7.34-7.30 (t, 1H),
6.62 (s, 1H), 5.45 (s, 2H), 4.73 (s, 2H), 2.42 (s, 3H), 2.37 (s,
3H). MS: 261.9 [M+H].sup.+.
Compound L:
2-(benzyloxy)-3-(chloromethyl)-4-(difluoromethoxy)-6-methylpyridine
##STR00022##
[0478] To a cooled (-10.degree. C.) suspension of sodium hydride
(60 wt % dispersion in mineral oil, 59.9 g, 1500 mmol) in dry
tetrahydrofuran (1200 mL) was added solution of malononitrile (100
g, 1190 mmol) in dry tetrahydrofuran (30 mL) dropwise, slowly
enough to maintain the internal temperature below 5.degree. C.
After the addition was complete, the mixture was stirred at
0.degree. C. for 1.5 hours, then diketene (80.1 g, 1190 mmol) was
added dropwise, slowly enough to maintain the internal temperature
below 0.degree. C. The mixture was stirred at -10.degree. C. for
1.5 hours, then neutralized with 4N aq. HCl, and concentrated to
remove volatiles. The remaining suspension in 4N aq. HCl (2000 mL)
was stirred at reflux for 5 hours, then stirred at room temperature
overnight. The resulting white precipitate was collected by suction
filtration. The filter cake was washed sequentially with water (500
mL), ethanol (500 mL) and MTBE (300 mL). The solid was dried to
obtain 4-hydroxy-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile
(Cpd E, 108 g, 60.3%) as a yellow powder. .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 12.40 (br. s., 1H), 11.72 (br. s., 1H), 5.82 (s,
1H), 2.17 (s, 3H).
[0479] A suspension of
4-hydroxy-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile (Cpd E,
91 g, 610 mmol), phosphorus oxychloride (195 g, 1270 mmol) and
phosphorus pentachloride (265 g, 1270 mmol) in chloroform (1200 mL)
was heated at reflux for 5 hours, resulting in a red homogeneous
mixture. The mixture was poured into water (2000 mL) carefully with
stirring, then neutralized by ammonium hydroxide (28% aqueous). The
resulting solid precipitate was filtered off, washed sequential
with dichloromethane (400 mL) and ethanol (500 mL), and dried to
give 4-chloro-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile
(Cpd F, 78 g, 76%) as a yellow solid. .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 12.43 (br. s., 1H), 6.53 (s, 1H), 2.28 (s,
3H).
[0480] A suspension of
4-chloro-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile (Cpd F,
90 g, 530 mmol), silver(I) oxide (136 g, 587 mmol) and benzyl
chloride (81.1 g, 641 mmol) in anhydrous toluene (1500 mL) was
heated at reflux for 12 hours. The mixture was filtered through a
CELITE.RTM. pad and the filter cake washed with dichloromethane
(500 mL). The filtrate was concentrated to give a residue
(.about.100 g), which was purified by column chromatography (silica
gel, petroleum ether/EtOAc=50:1.about.30:1), affording
2-(benzyloxy)-4-chloro-6-methylnicotinonitrile (Cpd G, 70 g, 51%)
as a light yellow solid. .sup.1H NMR (400 MHz, CDCl3) .delta.
7.49-7.47 (m, 2H), 7.40-7.33 (m, 3H), 6.91 (s, 1H), 5.05 (s, 2H),
2.50 (s, 3H).
[0481] To a stirred solution of
2-(benzyloxy)-4-chloro-6-methylnicotinonitrile (Cpd G, 70 g, 270.58
mmol) in N,N-dimethylformamide (300 mL) was added cesium acetate
(156.0 g, 812 mmol) at room temperature. The resulting mixture was
stirred at 80.degree. C. for 40 hours. The mixture was diluted with
ethyl acetate (500 mL) and washed with brine (3.times.400 mL). The
organic phase was dried over sodium sulfate, filtered, and
concentrated to give a residue (.about.50 g), which was purified by
column chromatography (silica gel, petroleum
ether/EtOAc=10:1.about.3:1) to give
2-(benzyloxy)-4-hydroxy-6-methylnicotinonitrile (Cpd H, 31 g, 48%)
as a light yellow solid. .sup.1H NMR (400 MHz, DMSO-d6) .delta.
12.28 (br. s., 1H), 7.51-6.98 (m, 5H), 6.50 (s, 1H), 5.41 (s, 2H),
2.34 (s, 3H). MS 226.8 [M+Na].sup.+.
[0482] To a suspension of
2-(benzyloxy)-4-hydroxy-6-methylnicotinonitrile (Cpd H, 20.0 g, 83
mmol) and sodium chlorodifluoroacetate (25.4 g, 166 mmol) in
N,N-dimethylformamide (200 mL) was added potassium carbonate (34.5
g, 250 mmol) at room temperature. The resulting mixture was heated
to 100.degree. C. for 10 minutes. The reaction mixture was diluted
with ethyl acetate (300 mL) and washed with sat. aq. NH4Cl
(3.times.400 mL), and brine (3.times.400 mL). The aqueous layer was
back-extracted with ethyl acetate (400 mL). The combined organic
layers were dried over sodium sulfate, filtered, and concentrated
to give a residue (18 g), which was purified by column
chromatography (silica gel, petroleum ether/EtOAc=50:1.about.20:1)
to give 2-(benzyloxy)-4-(difluoromethoxy)-6-methylnicotinonitrile
(Cpd I, 16.3 g, 67%) as light yellow solid. .sup.1H NMR (400 MHz,
CDCl3) .delta. 7.49-7.46 (m, 2H), 7.40-7.33 (m, 3H), 6.69 (t, J=71
Hz, 1H), 6.67 (s, 1H), 5.51 (s, 2H), 2.52 (s, 3H).
[0483] To a solution of
2-(benzyloxy)-4-(difluoromethoxy)-6-methylnicotinonitrile (Cpd I,
11 g, 38 mmol) in dry dichloromethane (250 mL) under nitrogen was
added diisobutylaluminium hydride (1.0 M in toluene, 72 mL, 72
mmol) dropwise at 0.degree. C. After the addition was complete, the
mixture was stirred at room temperature for 2.5 hours. The mixture
was acidified to pH.about.5 with 1M aq. HCl. After stirring at room
temperature for 2 hours, the mixture was neutralized with 4.0 M aq.
NaOH. The mixture was filtered off through a CELITE.RTM. pad and
the filter cake was washed with dichloromethane (300 mL). The
filtrate was extracted with dichloromethane (2.times.500 mL). The
combined organic layers were washed with brine (800 mL), dried over
sodium sulfate, and concentrated to give a residue (13.4 g), which
was purified by column chromatography (silica gel, petroleum
ether/EtOAc=30:1.about.10:1) to give
2-(benzyloxy)-4-(difluoromethoxy)-6-methylnicotinaldehyde (Cpd J, 6
g, 50%) as a light yellow solid. .sup.1H NMR (400 MHz, CDCl3)
.delta. 10.40 (s, 1H), 7.49-7.48 (m, 2H), 7.40-7.31 (m, 3H), 6.68
(t, J=72 Hz, 1H), 6.62 (s, 1H), 5.53 (s, 2H), 2.50 (s, 3H).
[0484] To a solution of
2-(benzyloxy)-4-(difluoromethoxy)-6-methylnicotinaldehyde (Cpd J,
12 g, 41 mmol) in methanol (120 mL) was added sodium borohydride
(1.86 g, 49.16 mmol) portion-wise at 0.degree. C. After the
addition was complete, the mixture was stirred at room temperature
for 2 hours. The reaction was quenched with sat. aq. NH4Cl (50 mL),
then diluted with ethyl acetate (500 mL) and water (100 mL) and
extracted with ethyl acetate (2.times.100 mL). The combined organic
layers were washed with brine (300 mL), dried over sodium sulfate
and concentrated to give a residue (.about.13.1 g), which was
purified by column chromatography (silica gel, petroleum ether:
EtOAc=6:1) to give
(2-(benzyloxy)-4-(difluoromethoxy)-6-methylpyridin-3-yl)methanol
(Cpd K, 11.7 g, 97%) as a white solid. .sup.1H NMR (400 MHz, CDCl3)
.delta. 7.52-7.46 (m, 2H), 7.44-7.33 (m, 3H), 6.60 (t, J=73 Hz,
1H), 6.55 (s, 1H), 5.46 (s, 2H), 2.46 (s, 3H).
[0485] To a solution of
(2-(benzyloxy)-4-(difluoromethoxy)-6-methylpyridin-3-yl) methanol
(Cpd K, 7.6 g, 26 mmol) in anhydrous dichloromethane (120 mL) was
added thionyl chloride (3.67 g, 30.9 mmol) dropwise at -20.degree.
C. The mixture was stirred at -20.degree. C. for 1 hour, then
poured into water (50 mL), and neutralized with saturated aq.
NaHCO3. The aqueous phase was extracted with dichloromethane
(2.times.90 mL). The combined organic phases were dried over sodium
sulfate, filtered and concentrated to give a residue (.about.6.1
g), which was purified by silica gel chromatography (petroleum
ether/EtOAc=6:1) to give the title compound,
2-(benzyloxy)-3-(chloromethyl)-4-(difluoromethoxy)-6-methylpyridine
(Cpd L, 5.7 g, 71%) as a white solid. .sup.1H NMR (400 MHz, CDCl3)
.delta. 7.50 (d, J=7.2, 2H), 7.41-7.33 (m, 3H), 6.64 (t, J=73 Hz,
1H), 6.56 (s, 1H), 5.48 (s, 2H), 4.69 (s, 2H), 2.47 (s, 3H). MS:
314 [M+H].sup.+.
Compound S:
2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3-
,4-dihydroisoquinolin-1(2H)-one
##STR00023##
[0487] A mixture of 3-chloro-2-methylbenzoic acid (100 g, 0.58
mol), N-chlorosuccinimide (90 g, 0.67 mol) and palladium (II)
acetate (14.7 g, 65.7 mmol) in N,N-dimethylformamide (1 L) was
stirred at 110.degree. C. under a nitrogen atmosphere overnight.
After cooling to room temperature, cesium carbonate (378 g, 1.16
mol) and iodoethane (317 g, 2.03 mol) were added and stirring
continued at room temperature for 1.5 hours. The reaction mixture
was poured into a mixture of water (1 L) and methyl tert-butyl
ether (800 mL). Solids were removed by filtration, and the filtrate
layers separated. The aqueous layer was extracted with more methyl
tert-butyl ether (600 mL). The combined organic extracts were
washed with saturated aqueous sodium chloride solution (1.2 L),
dried over sodium sulfate, and concentrated in vacuo. The residue
was purified by silica gel chromatography (eluting with 50:1
petroleum ether/ethyl acetate), affording ethyl
3,6-dichloro-2-methylbenzoate (Cpd N, 110 g, .about.80% pure, 80%
yield) as a yellow oil.
[0488] A solution of ethyl 3,6-dichloro-2-methylbenzoate (Cpd N,
120 g, 0.52 mol) and N-bromosuccinimide (147 g, 0.82 mol) in
chloroform (1 L) was treated with azobisisobutyronitrile (25.3 g,
0.15 mol) and the mixture refluxed overnight. After cooling to room
temperature, the mixture was diluted with dichloromethane (800 mL)
and washed with water (1.2 L). The aqueous layer was extracted with
dichloromethane (800 mL). The combined organic extracts were washed
with saturated aqueous sodium chloride solution (1.5 L), dried over
sodium sulfate, and concentrated in vacuo to give ethyl
2-(bromomethyl)-3,6-dichlorobenzoate (Cpd O, 160 g, 100% yield)
which was used without further purification.
[0489] A solution of sodium cyanide (75.12 g, 1.53 mol) in water
(300 mL) was added dropwise to a solution of ethyl
2-(bromomethyl)-3,6-dichlorobenzoate (Cpd O, 320 g, 1.03 mol) in
dimethysulfoxide (2.4 L) at room temperature. The mixture was
stirred at room temperature for 1.5 hours. The reaction mixture was
poured into a mixture of water (4 L) and methyl tert-butyl ether (2
L), and the layers separated. The organic layer was washed with
water (2 L) and with saturated aqueous sodium chloride solution (2
L), dried over sodium sulfate, and concentrated in vacuo. The
residue was purified by silica gel chromatography (eluting with
30:1 petroleum ether/ethyl acetate), affording ethyl
3,6-dichloro-2-(cyanomethyl)benzoate (Cpd P, 150 g, .about.75%
pure, 47% yield) as a yellow oil.
[0490] Cobalt (II) chloride hexahydrate (166 g, 0.70 mol) was added
to a room temperature solution of ethyl
3,6-dichloro-2-(cyanomethyl)benzoate (Cpd P, 90 g, 0.35 mol) in
ethanol (1.5 L), and the resulting mixture cooled to 0.degree. C.
Sodium borohydride (66.3 g, 1.74 mol) was added in portions. The
mixture was stirred at room temperature for 1 hour, and then
refluxed overnight. The resulting suspension was filtered and the
filtrate concentrated in vacuo. The solids in the filter cake were
stirred in ethyl acetate (600 mL), and then filtered again. This
procedure was repeated a second time. The combined filtrates were
added to the original filtrate residue, and this organic solution
washed with water (800 mL) and saturated aqueous sodium chloride
solution (800 mL), dried over sodium sulfate, and concentrated in
vacuo to give 5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd Q,
29.3 g, 39% yield) as an off-white solid.
[0491] To a solution of
5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd Q, 40 g, 0.186
mol) in concentrated sulfuric acid (200 mL) at 60.degree. C. was
added N-bromosuccinimide (49.7 g, 0.279 mol) in portions. Stirring
was continued at 60.degree. C. for 2 hours, then more
N-bromosuccinimide (5 g. 28 mmol) was added. After stirring at
60.degree. C. for 1 hour more, the mixture was poured onto ice
water (500 mL), then extracted with dichloromethane (3.times.500
mL). The combined organic extracts were washed with saturated
aqueous sodium chloride solution (800 mL), dried over sodium
sulfate, and concentrated in vacuo. The residue was stirred in
ethyl acetate (40 mL) and petroleum ether (20 mL), and the
resulting solids collected by filtration and dried under vacuum to
give 7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd R,
41 g, 75% yield) as an off-white solid.
[0492] Potassium tert-butoxide solution in tetrahydrofuran (1.0 M,
190 mL, 0.19 mol) was added dropwise to a cooled (0.degree. C.)
solution of 7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one
(Cpd R, 47 g, 0.16 mol) in anhydrous N,N-dimethylformamide (500 mL)
under a nitrogen atmosphere. Stirring was continued at 0.degree. C.
for 5 minutes, then
2-(benzyloxy)-3-(chloromethyl)-4,6-dimethylpyridine (Cpd D, 40.2 g,
0.15 mol) was added in one portion. After stirring for 10 minutes
at 0.degree. C., the mixture was treated with concentrated acetic
acid (2 mL) and poured into methyl tert-butyl ether (600 mL). The
organic solution was washed with water (800 mL) and saturated
aqueous sodium chloride solution (800 mL), dried over sodium
sulfate, and concentrated in vacuo. The residue was purified by
silica gel chromatography (eluting with 30:1 to 20:1 petroleum
ether/ethyl acetate), affording 2-{[2-(benzyloxy)-4,6-dimethyl
pyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-on-
e (Cpd S, 50 g, 64% yield) as an off-white solid. .sup.1H NMR (400
MHz, DMSO-d6) .delta. 8.08 (s, 1H), 7.45-7.43 (m, 2H), 7.32-7.29
(m, 3H), 6.76 (s, 1H), 5.38 (s, 2H), 4.71 (s, 2H), 3.24 (t, J=6 Hz,
2H), 2.72 (t, J=6 Hz, 2H), 2.36 (s, 3H), 2.31 (s, 3H). MS: 521
[M+H].sup.+.
Compound T: Methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)acetate
##STR00024##
[0494] A mixture of
2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3-
,4-dihydroisoquinolin-1(2H)-one (Cpd S, 1.0 g, 1.9222 mmol),
1-(tert-butyldimethylsilyloxy)-1-methoxyethene (1.09 g, 5.77 mmol),
bis(tri-tert-butylphosphine)palladium(0) (98.2 mg, 0.192 mmol) and
lithium fluoride (299 mg, 11.5 mmol) in dry N,N-dimethylformamide
(18 mL) was degassed with nitrogen for 10 minutes. Then the mixture
was heated in a microwave reactor at 100.degree. C. for 3 hours.
Water (20 mL) was added to the reaction mixture, which was then
extracted with ethyl acetate (2.times.40 mL). The combined organic
layers were washed with brine (4.times.50 mL), dried over sodium
sulfate, and concentrated under vacuum. The crude product was
purified by silica gel chromatography (petroleum ether/EtOAc=3:1,
Rf.about.0.45) to give methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)acetate (Cpd T, 600 mg, 60.8%)
as a light yellow solid. .sup.1H NMR (400 MHz, CDCl3) .delta. 7.45
(d, J=6.8 Hz, 2H), 7.37-7.30 (m, 4H), 6.62 (s, 1H), 5.42 (s, 2H),
4.87 (s, 2H), 3.80 (s, 2H), 3.72 (s, 3H), 3.28 (t, J=6.4 Hz, 2H),
2.73 (t, J=6.4 Hz, 2H), 2.42 (s, 3H), 2.32 (s, 3H). MS: 535.0
[M+Na].sup.+
Compound U: Methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)-2-diazoacetate
##STR00025##
[0496] To a solution of methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)acetate (500 mg, 0.974 mmol) and
4-acetyl aminobenzenesulfonyl azide (281 mg, 1.17 mmol) in
anhydrous acetonitrile (8 mL) was added
1,8-diazabicyclo[5.4.0]undec-7-ene (0.22 mL, 1.47 mmol). The
resulting reaction mixture was stirred at room temperature for 3 h.
After removing solvent, the resulting residue was purified by a
silica gel column with a gradient elution of 0.fwdarw.40%
EtOAc/heptane to afford methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)-2-diazoacetate as a foam like
solid (Cpd U, 454 mg, 86% yield). LCMS: 511.10/512.10 (M-N.sub.2).
.sup.1H NMR (400 MHz, CDCl3) .delta. 7.64 (s, 1H), 7.44 (d, J=6.60
Hz, 2H), 7.29-7.39 (m, 3H), 6.63 (s, 1H), 5.47 (s, 2H), 4.89 (s,
2H), 3.86 (s, 3H), 3.30 (t, J=5.99 Hz, 2H), 2.74-2.86 (m, 2H), 2.43
(s, 3H), 2.36 (s, 3H).
Compound W: Methyl
2-(2-((2-(benzyloxy)-4-(difluoromethoxy)-6-methylpyridin-3-yl)methyl)-5,8-
-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)acetate
##STR00026##
[0498] Potassium tert-butoxide solution in tetrahydrofuran (1.0 M,
3.2 mL, 3.2 mmol) was added dropwise to a cooled (0.degree. C.)
solution of 7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one
(Cpd R, 750 mg, 2.54 mmol) in dry N,N-dimethylformamide (15 mL).
The mixture was stirred at 0.degree. C. for 15 minutes, and then a
solution of
2-(benzyloxy)-3-(chloromethyl)-4-(difluoromethoxy)-6-methylpyridine
(Cpd L, 798 mg, 2.54 mmol) in dry N,N-dimethylformamide (5 mL) was
added dropwise. After stirring at 0.degree. C. for 30 minutes, the
solution was quenched with water (30 mL) and extracted with ethyl
acetate (3.times.15 mL). The combined organic layers were washed
sequentially with water (2.times.20 mL) and brine (20 mL), dried
over sodium sulfate, filtered, concentrated, and was purified by
column chromatography (silica gel, petroleum ether/EtOAc=7:1) to
give
2-((2-(benzyloxy)-4-(difluoromethoxy)-6-methylpyridin-3-yl)methyl)-7-brom-
o-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd V, 0.97 g,
67%) as a light-yellow solid.
[0499] A mixture of
2-((2-(benzyloxy)-4-(difluoromethoxy)-6-methylpyridin-3-yl)methyl)-7-brom-
o-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd V, 500 mg,
0.874 mmol), 1-(tert-butyldimethylsilyloxy)-1-methoxyethene (494
mg, 2.62 mmol), bis(tri-tert-butylphosphine)palladium(0) (67 mg,
0.313 mmol) and lithium fluoride (136 mg, 5.24 mmol) in dry
N,N-dimethylformamide (15 mL) was degassed with nitrogen for 10
minutes., then heated to 100.degree. C. in a microwave reactor for
3 hours. After cooling, the mixture was diluted with water (30 mL)
and extracted with ethyl acetate (4.times.20 mL). The combined
organic layers were washed with water (3.times.15 mL) and brine (15
mL), dried and concentrated. The residue was purified by prep. TLC
(silica gel, petroleum ether/EtOAc=2:1, Rf.about.0.35) to give
methyl
2-(2-((2-(benzyloxy)-4-(difluoromethoxy)-6-methylpyridin-3-yl)meth-
yl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)acetate
(Cpd W, 175 mg, 35.4%) as a white solid. .sup.1H NMR (400 MHz,
CDCl3) .delta. 7.42-7.41 (m, 2H), 7.37 (s, 1H), 7.29-7.27 (m, 3H),
6.66 (t, J=72 Hz, 1H), 6.62 (s, 1H), 5.45 (s, 2H), 4.82 (s, 2H),
3.80 (s, 2H), 3.72 (s, 3H), 3.26 (t, J=6.0 Hz, 2H), 2.73 (t, J=6.4
Hz, 2H), 2.46 (s, 3H).
Compound Z:
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-7-bromo-8-chloro-3,4-d-
ihydroisoquinolin-1(2H)-one
##STR00027##
[0501] A solution of 7-amino-3,4-dihydroisoquinolin-1(2H)-one (1.01
g, 6.23 mmol) and N-chlorosuccinimide (832 mg, 6.23 mmol) in
N,N-dimethylformamide (10 mL) was and heated to 55.degree. C. for 5
hours. The mixture was poured into water and extracted with ethyl
acetate (3.times.). The combined ethyl acetate layers were
concentrated, and residual DMF was removed on high vacuum
overnight. The resulting dark oil was purified on silica gel
(Biotage SNAP, 50 g, gradient of 50-100% ethyl acetate in heptanes)
to give 7-amino-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd X,
0.539 g, 44%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.87 (br. s., 1H), 6.96 (d, J=8.19 Hz, 1H), 6.87 (d, J=8.19
Hz, 1H), 5.32 (s, 2H), 3.20 (dt, J=3.79, 6.17 Hz, 2H), 2.69 (t,
J=6.24 Hz, 2H); MS 197 [M+H].sup.+.
[0502] A suspension of copper(I)bromide (1.04 g, 7.28 mmol) in
acetonitrile (20 mL) was stirred at 60.degree. C. for 10 minutes.
Isoamyl nitrite (0.348 mL, 2.91 mmol) was added, followed by
7-amino-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd X, 0.477 g,
2.43 mmol) in one portion. The reaction mixture was stirred at
60.degree. C. for 1 hour. After cooling to room temperature,
saturated aq. NH.sub.4Cl and EtOAc were added to the solution, and
the biphasic mixture stirred vigorously for 20 minutes. The layers
were separated, the organic layer concentrated, and the residue was
purified on silica gel (Biotage SNAP, 10 g, HP-Sil, gradient of
40-100% ethyl acetate in heptane) to give
7-bromo-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd Y, 0.287 g,
45%) as a yellow solid. .sup.1H NMR (400 MHz, CDCl3) .delta. 7.70
(d, J=8.07 Hz, 1H), 7.03 (d, J=8.07 Hz, 1H), 6.14 (br. s., 1H),
3.43-3.57 (m, 2H), 2.95 (t, J=6.36 Hz, 2H); MS 260, 262
[M+H].sup.+.
[0503] Potassium t-butoxide (1.3 mL, 1.3 mmol, 1.0 M in THF) was
added to a cooled (0.degree. C.) solution of
7-bromo-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd Y, 0.287 g,
1.10 mmol) in N,N-dimethylformamide (10 mL). After 5 minutes,
2-(benzyloxy)-3-(chloromethyl)-4,6-dimethylpyridine (Cpd D 0.311 g,
1.19 mmol) was added in one portion. The mixture was stirred for 30
minutes, then quenched with acetic acid (3 drops), diluted with
MTBE, and washed with water (2.times.). The organic layer was
concentrated, and the resulting oil purified on silica gel (Biotage
SNAP, 10 g, gradient of 0-25% ethyl acetate in heptane) to
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-7-bromo-8-chloro-3,4-d-
ihydroisoquinolin-1(2H)-one (Cpd Z, 0.387 g, 72%) as a clear gum.
.sup.1H NMR (400 MHz, CDCl3) .delta. 7.61 (d, J=8.07 Hz, 1H),
7.42-7.47 (m, 2H), 7.28-7.38 (m, 3H), 6.89 (d, J=8.07 Hz, 1H), 6.63
(s, 1H), 5.43 (s, 2H), 4.90 (s, 2H), 3.22-3.29 (m, 2H), 2.60-2.66
(m, 2H), 2.42 (s, 3H), 2.34 (s, 3H); MS 485, 487 [M+H].sup.+.
Compound FF:
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5-bromo-8-chloro-7-iod-
o-3,4-dihydroisoquinolin-1(2H)-one
##STR00028##
[0505] Two batches were run in parallel under the following
conditions, then combined for workup and purification: To a room
temperature (15-20.degree. C.) solution of
2-(2-bromo-5-chlorophenyl)acetic acid (25.0 g, 100.2 mmol) in
anhydrous THF (300 mL) was added oxalyl chloride (14.5 g, 9.97 mL,
114 mmol) and DMF (150 mg, 2.05 mmol), initiating gas evolution.
The mixture was stirred at room temperature for two hours, until
TLC showed the starting acid was completely consumed. The mixture
was cooled to 0.degree. C., and ammonium hydroxide (28 wt % in
water, 154 mL) was added in one portion, causing the internal
temperature to rise to 40.degree. C. The cooling bath was removed,
and the solution stirred vigorously at room temperature for one
hour. The two batches were combined, diluted with water (500 mL),
and extracted with ethyl acetate (2.times.1000 mL). The combined
organic extracts were washed with water (2.times.500 mL), 1N
aqueous HCl (500 mL), and brine (500 mL), then dried over anhydrous
sodium sulfate and concentrated to give crude product (.about.50 g)
as a yellow solid. The crude product was crystallized from 5/1
petroleum ether/ethyl acetate (200 mL.times.2) and dried to give
2-(2-bromo-5-chlorophenyl)acetamide (Cpd AA, 44.0 g, 88% combined
yield for the two batches) as a white solid. .sup.1H NMR (400 MHz,
CDCl3) .delta. 7.52 (d, J=8.8 Hz, 1H), 7.37 (d, J=2.8 Hz, 1H), 7.16
(dd, J=2.8, 8.8 Hz, 1H), 5.67 (br s, 1H), 5.50 (br s, 1H), 3.70 (s,
2H).
[0506] Two batches were run in parallel under the following
conditions, then combined for purification: Borane-THF complex (1.0
M in THF, 400 mL, 400 mmol) was added dropwise to a cooled
(0.degree. C.) suspension of 2-(2-bromo-5-chlorophenyl)acetamide
(Cpd AA, 22.0 g, 88.5 mmol) in anhydrous THF (300 mL). The
resulting clear solution was heated to 80.degree. C. for two hours,
then cooled again to 0.degree. C. The mixture was quenched by
sequential addition of water (45 mL) and conc. HCl (120 mL),
causing significant gas evolution. Stirring was continued at
10-15.degree. C. for 16 hours, after which the mixture was
concentrated to remove THF. The aqueous residue was cooled to
0.degree. C., then 12 N aqueous sodium hydroxide was added to bring
the pH to 11. The basified solution was extracted with ethyl
acetate (3.times.500 mL). The combined organic extracts were washed
with brine (500 mL), dried over anhydrous sodium sulfate, filtered,
and concentrated to give crude product (.about.25 g) as a yellow
oil. Two .about.25 g batches of this crude product were combined,
treated with 4N HCl/MeOH (500 mL), and stirred at 10-15.degree. C.
for 16 hours. The mixture was concentrated, and the residue stirred
in ethyl acetate (500 mL) for 30 minutes. The resulting white solid
was collected by filtration, and the filter cake washed with ethyl
acetate (3.times.100 mL). The solids were dissolved in water (500
mL), filtered to remove insolubles, and the filtrate extracted with
ethyl acetate (2.times.500 mL). The aqueous layer was basified with
solid NaOH to pH 10, then extracted with ethyl acetate (2.times.500
mL). The combined organic extracts were washed with brine (500 mL),
dried over anhydrous sodium sulfate, filtered, and concentrated to
give 2-(2-bromo-5-chlorophenyl)ethan-1-amine (Cpd BB, 30.0 g, 72%
combined yield for the two batches) as a colorless oil. .sup.1H NMR
(400 MHz, CDCl3) .delta. 7.47 (d, J=8.4 Hz, 1H), 7.23 (d, J=2.4 Hz,
1H), 7.07 (dd, J=2.4, 8.4 Hz, 1H), 2.98 (t, J=6.8 Hz, 2H), 2.86 (t,
J=6.8 Hz, 2H), 1.28 (m, 2H).
[0507] To a cooled (0.degree. C.) suspension of
2-(2-bromo-5-chlorophenyl)ethan-1-amine (Cpd BB, 28.0 g, 119 mmol)
and sodium carbonate (32.3 g, 304 mmol) in anhydrous
1,2-dichloroethane (600 mL) was added 4-nitrophenyl chloroformate
(25.5 g, 127 mmol). The mixture was stirred at 0.degree. C. for 30
minutes, then at 10-15.degree. C. for 16 hours. The solution was
diluted with water (1000 mL) and extracted with dichloromethane
(3.times.1000 mL). The combined organic extracts were washed with
water (1000 mL) and brine (1000 mL), dried over anhydrous sodium
sulfate, and concentrated. The crude product (.about.55 g, yellow
solid) was crystallized from 5/1 petroleum ether/EtOAc (100
mL.times.2) to give 4-nitrophenyl
(2-bromo-5-chlorophenethyl)carbamate (Cpd CC, 40.0 g, 84% yield) as
a white solid. .sup.1H NMR (400 MHz, CDCl3) .delta. 8.25 (d, J=9.2
Hz, 2H), 7.51 (d, J=8.4 Hz, 1H), 7.31 (m, 3H), 7.13 (dd, J=2.0, 8.4
Hz, 1H), 5.22 (br s, 1H), 3.57 (t, J=6.8 Hz, 2H), 3.05 (t, J=6.8
Hz, 2H).
[0508] Trifluoromethanesulfonic acid (150 g, 1000 mmol) was added
dropwise to a cooled (0.degree. C.) suspension of 4-nitrophenyl
(2-bromo-5-chlorophenethyl)carbamate (Cpd CC, 40.0 g, 100 mmol) in
anhydrous 1,2-dichloroethane (300 mL). Solids gradually dissolve
over the course of the addition, resulting in a clear yellow
solution. The mixture was stirred at 0.degree. C. for 10 minutes,
then heated at 60-70.degree. C. for 3 hours. The resulting brown
solution was poured into ice-water (1000 mL) and stirred until all
the ice had melted. The layers were separated, and the aqueous
layer extracted with dichloromethane (2.times.1000 mL). The
combined organic layers were washed with 2N aqueous sodium
hydroxide (3.times.500 mL), water (500 mL), and brine (500 mL),
then dried over anhydrous sodium sulfate and concentrated. The
crude product (.about.30 g brown solid) was crystalized from 2/1
petroleum ether/ethyl acetate (150 mL.times.2), to give
5-bromo-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd DD, 20.7 g,
80% yield) as a brown solid. .sup.1H NMR (400 MHz, DMSO-d6) .delta.
8.25 (br s, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.35 (d, J=8.4 Hz, 1H),
3.12 (t, J=4.4 Hz, 2H), 2.95 (t, J=6.2 Hz, 2H).
[0509] N-iodosuccinimide (53.7 g, 239 mmol) was added to a cooled
(0.degree. C.) solution of
5-bromo-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd DD, 20.7 g,
79.6 mmol) in conc. sulfuric acid (98% w/w, 300 mL). The resulting
brown suspension was stirred at 10-15.degree. C. for 16 hours.,
then poured into ice-water (1000 mL) and stirred until all the ice
had melted. The resulting aqueous suspension was extracted with
ethyl acetate (3.times.1000 mL). The combined organic extracts were
washed with saturated aqueous NaHSO3 (2.times.500 mL), 2N aqueous
sodium hydroxide (2.times.500 mL), and brine (500 mL), then dried
over anhydrous sodium sulfate and concentrated. The crude product
(.about.30 g yellow solid) was crystalized with 1/1 petroleum
ether/ethyl acetate (100 mL.times.2) to give
5-bromo-8-chloro-7-iodo-3,4-dihydroisoquinolin-1(2H)-one (Cpd EE,
23.0 g, 75% yield) as an off-white solid. .sup.1H NMR (400 MHz,
DMSO-d6) .delta. ppm 8.35 (br. s, 1H), 8.33 (s, 1H), 3.30-3.25 (2H,
m), 2.89 (t, J=6.0 Hz, 2H). MS: 386 [M+H].sup.+.
[0510] Potassium tert-butoxide (1.0M solution in THF, 7.30 mL, 7.30
mmol) was added dropwise to a cooled (0.degree. C.) suspension of
5-bromo-8-chloro-7-iodo-3,4-dihydroisoquinolin-1(2H)-one (Cpd EE,
2.35 g, 6.08 mmol) in anhydrous DMF (30 mL). The mixture was
stirred at 0.degree. C. for 30 minutes, then a solution of
2-(benzyloxy)-3-(chloromethyl)-4,6-dimethylpyridine (Cpd D, 1.75 g,
6.69 mmol) in anhydrous DMF (10 mL) was added, and stirring
continued at 0.degree. C. for 30 minutes. The reaction mixture was
partitioned between ethyl acetate (100 mL) and water (100 mL). The
organic phase was washed with water (1.times.100 mL) and brine
(1.times.100 mL), dried over sodium sulfate, concentrated to
dryness, and purified by silica gel chromatography, eluting with a
gradient of 0-40% ethyl acetate in heptane to afford
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5-bromo-8-ch-
loro-7-iodo-3,4-dihydroisoquinolin-1(2H)-one (Cpd FF, 2.95 g, 79%
yield) as a gum. .sup.1H NMR (400 MHz, CDCl3) .delta. ppm 8.11 (s,
1H), 7.40-7.47 (m, 2H), 7.27-7.37 (m, 3H), 6.62 (s, 1H), 5.42 (s,
2H), 4.85 (s, 2H), 3.25 (t, J=6.24 Hz, 2H), 2.68 (t, J=6.24 Hz,
2H), 2.41 (s, 3H), 2.32 (s, 3H). MS: 611, 613 [M+H].sup.+.
Compound KK:
2-(benzyloxy)-3-(chloromethyl)-4-methoxy-6-methylpyridine
##STR00029##
[0512] Benzyl bromide (19.1 g, 112 mmol) was added to a room
temperature solution of ethyl 2,4-dihydroxy-6-methylnicotinate
(20.0 g, 101.4 mmol) and silver carbonate (15.4 g, 55.8 mmol) in
THF (100 mL), then the mixture was heated to 60.degree. C. for 18
hours. After cooling to room temperature, the suspension was
filtered through a CELITE.RTM. pad, the filtrate concentrated and
purified by silica gel chromatography (eluting with 5% ethyl
acetate in heptane) to give ethyl
2-(benzyloxy)-4-hydroxy-6-methylnicotinate (Cpd GG, 18 g, 62%
yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d6) .delta.
11.14 (s, 1H), 7.40-7.44 (m, 2H), 7.36 (t, J=7.34 Hz, 2H),
7.27-7.33 (m, 1H), 6.44 (s, 1H), 5.35 (s, 2H), 4.23 (q, J=7.13 Hz,
2H), 2.30 (s, 3H), 1.22 (t, J=7.09 Hz, 3H). MS: 288
[M+H].sup.+.
[0513] A solution of ethyl
2-(benzyloxy)-4-hydroxy-6-methylnicotinate (Cpd GG, 18.0 g, 62.6
mmol) and potassium carbonate (9.52 g, 68.9 mmol) in DMF (50 mL)
was stirred at room temperature for 10 minutes, then iodomethane
(9.98 g, 68.9 mmol) was added and stirring continued at room
temperature for 18 hours. The mixture was partitioned between water
and ethyl acetate. The organic extracts were washed with sat. aq.
NaCl, dried over sodium sulfate, and concentrated. The residue was
purified by silica gel chromatography (eluting with 0-35% ethyl
acetate in heptane), affording ethyl
2-(benzyloxy)-4-methoxy-6-methylnicotinate (Cpd HH, 16.7 g, 89%
yield) as a colorless oil. .sup.1H NMR (400 MHz, DMSO-d6) .delta.
7.33-7.42 (m, 4H), 7.26-7.33 (m, 1H), 6.75 (s, 1H), 5.36 (s, 2H),
4.22 (q, J=7.13 Hz, 2H), 3.83 (s, 3H), 2.39 (s, 3H), 1.20 (t,
J=7.09 Hz, 3H).). MS: 302 [M+H].sup.+.
[0514] Lithium aluminium hydride solution (2.0M in THF) was added
dropwise to a cooled (0.degree. C.) solution of ethyl
2-(benzyloxy)-4-methoxy-6-methylnicotinate (Cpd HH, 16.7 g, 55.4
mmol) in THF (100 mL). After addition was complete, the solution
was allowed to gradually warm to room temperature with stirring for
18 hours. The mixture was diluted with THF (200 mL), cooled to
0.degree. C., and quenched by sequential dropwise addition of water
(3.4 mL), 15% aq. sodium hydroxide, and water (10.2 mL). The
resulting slurry was stirred at room temperature for 2 hours, then
filtered through a pad of CELITE.RTM.. Concentration of the
filtrate yielded
(2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methanol (Cpd JJ, 14
g, 97% yield) as colorless oil. .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 7.46 (d, J=7.09 Hz, 2H), 7.36 (t, J=7.40 Hz, 2H), 7.25-7.33
(m, 1H), 6.63 (s, 1H), 5.35 (s, 2H), 4.37-4.46 (m, 3H), 3.82 (s,
3H), 2.35 (s, 3H). MS: 260 [M+H].sup.+.
[0515] Thionyl chloride (6.57 g, 54.7 mmol) was added dropwise to a
cooled (0.degree. C.) solution of
(2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methanol (Cpd JJ,
13.5 g, 52.1 mmol) in ethyl acetate (300 mL), causing formation of
a solid precipitate. The slurry was stirred in the cooling bath for
30 minutes, then water was added to dissolve the solids. After
separation of the phases, the organic layer was washed with sat.
aq. NaCl, dried over sodium sulfate, and concentrated to dryness.
The residue was dissolved in heptane and again concentrated to
dryness, affording
2-(benzyloxy)-3-(chloromethyl)-4-methoxy-6-methylpyridine (Cpd KK,
13.9 g, 95% yield) as white solid. .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 7.47 (d, J=7.34 Hz, 2H), 7.38 (t, J=7.40 Hz, 2H), 7.27-7.34
(m, 1H), 6.71 (s, 1H), 5.40 (s, 2H), 4.66 (s, 2H), 3.89 (s, 3H),
2.38 (s, 3H). MS: 260 [M+H].sup.+.
Compound RR:
8-chloro-7-iodo-5-methyl-3,4-dihydroisoquinolin-1(2H)-one
##STR00030##
[0517] To a cooled (0.degree. C.) solution of
5-chloro-2-methylbenzoic acid (20.0 g, 117 mmol) in anhydrous
2-methyltetrahydrofuran (200 mL), borane-dimethylsulfide complex
(28.0 g, 35.0 mL, 369 mmol) was added dropwise over 1 hour, slowly
enough to maintain the internal temperature below 10.degree. C. Gas
evolution was observed, and some precipitate formed. After the
addition was complete, the cooling bath was removed and stirring
continued at room temperature overnight. Methanol (50 mL) was
carefully added to quench the mixture. The solution was
concentrated to dryness, and the residue partitioned between ether
(200 mL) and saturated aqueous sodium bicarbonate. The organic
layer was washed with brine (200 mL), dried over sodium sulfate,
filtered, and concentrated to give ethyl
2-(benzyloxy)-4-hydroxy-6-methylnicotinate (Cpd LL, 18.4 g, 100%
yield) as an oil. 1H NMR (400 MHz, CDCl3) .delta. 7.36 (d, J=2.08
Hz, 1H), 7.13-7.19 (m, 1H), 7.04-7.11 (m, 1H), 4.63 (s, 2H), 2.27
(s, 3H), 2.12 (s, 1H).
[0518] A solution of 2-(benzyloxy)-4-hydroxy-6-methylnicotinate
(Cpd LL, 18.0 g, 115 mmol) in anhydrous toluene (300 mL) was cooled
to below 10.degree. C. internal. Thionyl chloride (21.3 g, 179
mmol) was added dropwise, slowly enough to maintain the internal
temperature below 10.degree. C.
[0519] The mixture was stirred at this temperature for 30 minutes,
then the cooling bath was removed and stirring continued at room
temperature for 5 hours. The solution was concentrated to remove
volatiles, and the residue partitioned between ethyl acetate (200
mL) and sodium bicarbonate (200 mL). The organic phase was washed
with brine (200 mL), dried over sodium sulfate, and concentrated to
give 4-chloro-2-(chloromethyl)-1-methylbenzene (Cpd MM, 17.5 g, 87%
yield) as an oil. .sup.1H NMR (400 MHz, CDCl3) .delta. 7.33 (d,
J=2.20 Hz, 1H), 7.20-7.25 (m, 1H), 7.14 (d, J=8.07 Hz, 1H), 4.55
(s, 2H), 2.39 (s, 3H).
[0520] To a solution of 4-chloro-2-(chloromethyl)-1-methylbenzene
((Cpd MM, 17.5 g, 100 mmol) in DMSO (200.0 mL) and water (50.0 mL)
was added solid sodium cyanide (5.88 g, 120 mmol) in one portion.
The reaction was slightly exothermic, and the internal temperature
of the reaction mixture rose to 43.degree. C. Stirring was
continued for one hour. The reaction mixture was partitioned
between ethyl acetate (300 mL) and water (300 mL). The organic
phase was washed with sodium bicarbonate (300 mL) and brine (300
mL), dried over sodium sulfate, and concentrated to give
2-(5-chloro-2-methylphenyl)acetonitrile (Cpd NN, 16.1 g, 97% yield)
as an oil. .sup.1H NMR (400 MHz, CDCl3) .delta. 7.37 (d, J=1.96 Hz,
1H), 7.21-7.26 (m, 1H), 7.13-7.17 (m, 1H), 3.64 (s, 2H), 2.32 (s,
3H).
[0521] Borane dimethylsulfide complex (22.3 g, 293 mmol, 26.0 mL)
was added dropwise to a solution of
2-(5-chloro-2-methylphenyl)acetonitrile (Cpd NN, 16.0 g, 96.6 mmol)
in 2-methyl tetrahydrofuran (150 mL), causing gas evolution. After
the addition was complete, the mixture was heated to reflux for 5
hours. After cooling to room temperature, methanol was added to
quench the mixture until no more bubbles were generated. The
solution was concentrated to dryness. The residue was dissolved in
methanol and treated with 4M HCl/dioxane solution (100 mL) to break
up the boron complex. The solution was concentrated to dryness. The
white solid residue was dissolved in minimal methanol (.about.20
mL), ethyl acetate (.about.200 mL) was added, and the mixture
stirred vigorously until a thick paste formed. The solids were
collected by filtration, washed with ethyl acetate, and dried to
give 2-(5-chloro-2-methylphenyl)ethan-1-amine hydrochloride (Cpd
OO, 9.5 g, 48% yield) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 8.24 (br. s., 3H), 7.26 (s, 1H), 7.20 (d, J=1.34
Hz, 2H), 2.84-3.02 (m, 4H), 2.27 (s, 3H). MS: 170 [M+H].sup.+.
[0522] A cooled (0.degree. C.) solution of
2-(5-chloro-2-methylphenyl)ethanamine hydrochloride (Cpd OO, 8.41
g, 40.8 mmol) in DMF (200 mL) was stirred with triethylamine(10.3
g, 102 mmol) for 10 minutes, then solid 4-nitrophenyl chloroformate
(8.14 g, 38.8 mmol) was added in one portion. A thick paste formed,
and the reaction mixture became slightly yellow. Stirring was
continued at 0.degree. C. for 1 hour. The reaction mixture was
partitioned between ethyl acetate (300 mL) and water (300 mL). The
organic phase was washed with water (200 mL), 10% sodium carbonate
(200 mL), and brine (200 mL), then dried over sodium sulfate, and
concentrated to give 4-nitrophenyl
(5-chloro-2-methylphenethyl)carbamate (Cpd PP, 9.94 g, 73% yield)
as a solid. .sup.1H NMR (400 MHz, CDCl3) .delta. 8.18-8.29 (m, 2H),
7.24-7.32 (m, 2H), 7.11-7.17 (m, 3H), 5.28 (br. s., 1H), 3.45-3.53
(m, 2H), 2.85-2.92 (m, 2H), 2.32 (s, 3H).
[0523] A cooled (0.degree. C.) suspension of 4-nitrophenyl
(5-chloro-2-methylphenethyl)carbamate (Cpd PP, 9.94 g, 29.7 mmol)
in anhydrous 1,2-dichloroethane (120 mL) was treated with freshly
opened trifluoromethylsulfonic acid (45.8 g, 305 mmol, 27.0 mL),
and stirring continued at 0.degree. C. for 30 minutes. The reaction
mixture was then heated to 70.degree. C. for 3 hours. After cooling
to room temperature, the reaction mixture was carefully poured into
ice water (200 mL), and stirred until all the ice had melted. The
biphasic mixture was extracted with dichloromethane (2.times.200
mL). The combined organic extracts were washed with 2M sodium
carbonate (200 mL). The aqueous phase was back-extracted with
dichloromethane (200 mL). The dichloromethane extracts were
combined, dried over sodium sulfate, and concentrated to give
8-chloro-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd QQ, 4.17 g,
72% yield) as a solid. .sup.1H NMR (400 MHz, CDCl3) .delta.:
7.15-7.22 (m, 1H), 7.08 (d, J=13.69 Hz, 1H), 6.46 (br. s., 1H),
3.43-3.51 (m, 2H), 2.88 (t, J=6.17 Hz, 2H), 2.28 (s, 3H). MS: 196
[M+H].sup.+.
[0524] A flask containing
8-chloro-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd QQ, 6.0 g,
30.7 mmol) was cooled in an ice bath. Concentrated sulfuric acid
(125.0 mL) was added, and the mixture stirred at 0.degree. C. for
30 min. N-iodosuccinimide (20.7 g, 92.0 mmol) was added as a solid
in one portion, and the mixture stirred at 0.degree. C. for 3
hours. The solution was carefully poured into ice water (300 mL),
causing a precipitate to form. The suspension was extracted with
ethyl acetate (300 mL). Both organic and aqueous phases contained
precipitates. The organic phase was washed with 10%
Na.sub.2S.sub.2O.sub.3 (300 mL) to remove excess iodine, and the
aqueous layer was extracted with ethyl acetate (200 mL). The
combined organic phases were dried over sodium sulfate and
concentrated to dryness. The solid residue was stirred in methanol
(100 mL). Insolubles were collected by filtration, and the
precipitate (.about.14 g white solid) was slurried in carbon
disulfide (100 mL). Solids were collected by filtration, washed
with carbon disulfide, and dried under vacuum to yield
8-chloro-7-iodo-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd RR,
8.57 g, 87% yield) as a solid. .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 8.15 (br. s., 1H), 7.94 (s, 1H), 3.26 (td, J=6.17, 4.03 Hz,
2H), 2.75 (t, J=6.24 Hz, 2H), 2.22 (s, 3H). MS; 321
[M=H].sup.+.
Compound SS:
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-7-bromo-5,8-dich-
loro-3,4-dihydroisoquinolin-1(2H)-one
##STR00031##
[0526] A room-temperature suspension of
7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd R, 14.9
g, 50.4 mmol) in ethyl acetate (300 mL) was treated with potassium
tert-butoxide (1.0 M solution in THF, 65.5 mL, 65.5 mmol), causing
the solids to dissolve. After a few minutes, a precipitate begins
to form. To this was added
2-(benzyloxy)-3-(chloromethyl)-4-methoxy-6-methylpyridine (Cpd KK,
14.0 g, 50.4 mmol), and the resulting suspension heated at
75.degree. C. for 4 hours. After cooling to room temperature, the
mixture was washed with water (2.times.) and sat. aq. NaCl,
concentrated, and the residue crystalized from ethyl acetate,
affording
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-7-bromo-5,8-dich-
loro-3,4-dihydroisoquinolin-1(2H)-one (Cpd SS, 21.96 g, 81% yield)
as a white solid. .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.05 (s,
1H), 7.34-7.40 (m, 2H), 7.18-7.25 (m, 3H), 6.70 (s, 1H), 5.36 (s,
2H), 4.68 (s, 2H), 3.83 (s, 3H), 3.16 (t, J=6.17 Hz, 2H), 2.71 (t,
J=6.17 Hz, 2H), 2.38 (s, 3H). MS: 535, 537 [M+H].sup.+.
Compound TT:
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-8-chloro-7-iodo-5-meth-
yl-3,4-dihydroisoquinolin-1(2H)-one
##STR00032##
[0528] To a cooled (0.degree. C.) solution of
8-chloro-7-iodo-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd RR,
1.52 g, 4.73 mmol) in anhydrous DMF (20 mL) was added solid
potassium tert-butoxide (796 mg, 7.09 mmol) in portions. Stirring
was continued at 0.degree. C. for 30 minutes, then a solution of
2-(benzyloxy)-3-(chloromethyl)-4,6-dimethylpyridine (Cpd D, 1.18 g,
4.49 mmol) in anhydrous DMF (5 mL) was added dropwise. After
stirring for 20 more minutes at 0.degree. C., the mixture was
poured into ice-water (50 mL) and extracted with ethyl acetate
(4.times.50 mL). The combined organic layers were washed with brine
(4.times.50 mL), dried over sodium sulfate, filtered, and
concentrated. The crude product (.about.2.4 g yellow solid) was
purified by silica gel chromatography, eluting with 5/1 petroleum
ether/ethyl acetate, affording
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-8-chloro-7-iodo-5-meth-
yl-3,4-dihydroisoquinolin-1(2H)-one (Cpd TT, 1.7 g, 66% yield) as
an off-white solid. .sup.1H NMR (400 MHz, CDCl3) .delta. 7.76 (s,
1H), 7.45 (d, J=7.2 Hz, 2H), 7.36-7.30 (m, 3H), 6.62 (s, 1H), 5.42
(s, 2H), 4.88 (s, 2H), 3.24 (t, J=6.2 Hz, 2H), 2.50 (t, J=6 Hz,
2H), 2.42 (s, 3H), 2.32 (s, 3H), 2.13 (s, 3H). MS: 547
[M+H].sup.+.
Compound UU:
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-iodo--
5-methyl-3,4-dihydroisoquinolin-1(2H)-one
##STR00033##
[0530] To a cooled (0.degree. C.) solution of
8-chloro-7-iodo-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd RR,
3.2 g, 9.95 mmol) in anhydrous DMF (50 mL) was added solid
potassium tert-butoxide (1.68 g, 14.9 mmol) in portions. Stirring
was continued at 0.degree. C. for 30 minutes, then a solution of
2-(benzyloxy)-3-(chloromethyl)-4-methoxy-6-methylpyridine (Cpd KK,
2.63 g, 14.9 mmol) in anhydrous DMF (50 mL) was added dropwise.
After stirring for 30 more minutes at 0.degree. C., the mixture was
poured into ice-water (100 mL) and extracted with ethyl acetate
(3.times.100 mL). The combined organic layers were washed with
brine (4.times.100 mL), dried over sodium sulfate, filtered, and
concentrated. The crude product (.about.5 g yellow solid) was
purified by silica gel chromatography, eluting with 20-50% ethyl
acetate in petroleum ether. The resulting product was dissolved in
dichloromethane (10 mL), added to petroleum ether (30 mL) and
stirred at room temperature until a precipitate forms (30 minutes).
The precipitate was collected by filtration and dried to a white
solid. TLC of the precipitate still showed impurities, so it was
repurified by silica gel chromatography, eluting with 0-10%
methanol in dichloromethane, yielding
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-iodo--
5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd UU, 2.8 g, 50%
yield) as a white solid. .sup.1H NMR (400 MHz, CDCl3) .delta. 7.75
(s, 1H), 7.42-7.40 (m, 2H), 7.29-7.27 (m, 1H), 7.25-7.22 (m, 2H),
6.38 (s, 1H), 5.42 (s, 2H), 4.87 (s, 2H), 3.83 (s, 3H), 3.14 (t,
J=6.2 Hz, 2H), 2.50 (t, J=6.2 Hz, 2H), 2.44 (s, 3H), 2.13 (s, 3H).
MS: 563 [M+H].sup.+.
EXAMPLES
General Methods and Representative Examples
Method A
Example 1
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1R-
)-2-hydroxy-1-[(3R)-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquinolin-1(2-
H)-one
Example 2
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1R-
*)-2-hydroxy-1-[(3S*)-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquinolin-1-
(2H)-one
Example 3
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1S-
*)-2-hydroxy-1-[(3R*)-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquinolin-1-
(2H)-one
Example 4
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1S-
)-2-hydroxy-1-[(3S)-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquinolin-1(2-
H)-one
##STR00034##
[0532] A cooled (0.degree. C.) solution of methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)acetate (Cpd T, 800 mg, 0.487
mmol) in anhydrous N,N-dimethylformamide (70 mL) was treated with
sodium hydride (60 wt % dispersion in mineral oil, 125 mg, 3.12
mmol), then stirred at 10.degree. C. for 15 minutes. The mixture
was cooled again to 0.degree. C. and 3-iodotetrahydrofuran (463 mg,
2.34 mmol) was added. After stirring at room temperature for 12
hours, glacial acetic acid (2 drops) and water (10 mL) were added,
and the mixture extracted with ethyl acetate (3.times.20 mL). The
combined organic extracts were washed with brine (10 mL), dried
over sodium sulfate, concentrated, and purified by column
chromatography (silica gel, Petroleum ether/EtOAc=5:1.about.1:1) to
obtain
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichl-
oro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-3-yl)acet-
ic acid (1a, 500 mg, 56.3%) as a white solid; and methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-3-yl)acetate
(1b, 150 mg, 16.5%) as a yellow solid.
[0533] A solution of
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-3-yl)acetic
acid (1a, 650 mg, 1.14 mmol), potassium carbonate (315 mg, 2.28
mmol), and iodomethane (324 mg, 2.28 mmol) in N,N-dimethylformamide
(8 mL) was stirred at room temperature for 12 hours. The mixture
was diluted with water (20 mL) and ethyl acetate (50 mL). The
organic layer was separated and the aqueous layer was extracted
with ethyl acetate (2.times.15 mL). The organic layers were
combined, washed with brine (3.times.10 mL), dried over sodium
sulfate, filtered and concentrated in vacuo to give the crude
product, which was purified by column chromatography (silica gel,
Petroleum ether/EtOAc=1:1) to obtain methyl
2-(2-((2-(benzyloxy)-4,6-dimethyl
pyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-y-
l)-2-(tetrahydrofuran-3-yl)acetate (1b 600 mg, 90.1%) as a white
solid.
[0534] Lithium borohydride (28 mg, 1.29 mmol) was added in one
portion to a room temperature solution of methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-3-yl)acetate
(1b, 250 mg, 0.428 mmol) in anhydrous tetrahydrofuran (25 mL). The
resulting mixture was heated at 60.degree. C. for 2 hours. The
mixture was quenched with water (5 mL) and then extracted with
ethyl acetate (3.times.15 mL). The organic layers were combined,
washed with brine (5 mL), dried over sodium sulfate, filtered and
concentrated in vacuo to give the crude product, which was purified
by prep. TLC (Petroleum ether/EtOAc=1:1) to obtain
2-((2-(benzyloxy)-4,6-dimethyl
pyridin-3-yl)methyl)-5,8-dichloro-7-(2-hydroxy-1-(tetrahydrofuran-3-yl)et-
hyl)-3,4-dihydroisoquinolin-1(2H)-one (mixture of 4 stereoisomers,
160 mg, 67.2%) as a white solid. Combined batches (500 mg total) of
this stereoisomer mixture was resolved by preparative chiral SFC
(Chiralpak AD, 250.times.30 mm I.D., 5 .mu.m, mobile phase 35% EtOH
NH.sub.3 H.sub.2O, flow rate 50 mL/min) to obtain separated isomers
1c (peak one, 80 mg, 15.9%), 2c (peak two, 90 mg, 17.9%), 3c (peak
three, 110 mg, 21.9%) and 4c (peak four, 100 mg, 19.9%) as white
solids. Absolute stereochemistry of each isomer was not determined
at this stage.
[0535] A solution of
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(2-hydr-
oxy-1-(tetrahydrofuran-3-yl)ethyl)-3,4-dihydroisoquinolin-1(2H)-one
stereoisomer 1c (80 mg, 0.144 mmol) in dichloromethane (3 mL) and
trifluoroacetic acid (3 mL) was stirred at 35.degree. C. for 5
hours, and then evaporated to dryness. The residue was taken up in
methanol (10 mL), cooled to 10.degree. C., and potassium carbonate
(99.5 mg, 0.720 mmol) added. After stirring for 30 minutes at
10.degree. C., the reaction mixture was filtered, and the filter
pad washed with dichloromethane/methanol (10:1, 10 mL). The
filtrate was concentrated in vacuo and the residue, purified by
prep. TLC (CH2Cl2/MeOH=10:1, Rf=0.4 in CH2Cl2/MeOH=10:1) to obtain
Example 1 (38 mg, 57%) as a white solid.
[0536] By the same procedure, stereoisomer 2c (90 mg, 0.162 mmol)
afforded Example 2 (44 mg, 59%); stereoisomer 3c (110 mg, 0.198
mmol) afforded Example 3 (61 mg, 66%); and stereoisomer 4c (100 mg,
0.18 mmol) afforded Example 4 (35 mg, 41%); all as white
solids.
[0537] A small-molecule X-Ray crystal structure of Example 4 shows
it to have absolute (S,S) stereochemistry. Absolute (R,R)
stereochemistry was attributed to Example 1 because its .sup.1HNMR
spectrum is identical to that of Example 4. The .sup.1HNMR spectra
of Example 2 and Example 3 are identical to each other, and clearly
different from that of Example 4, suggesting they are the (R,S) and
(S,R) stereoisomers, though the absolute configuration of each was
not determined.
Example 1
[0538]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-{(1R)-2-hydroxy-1-[(3R)-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquino-
lin-1(2H)-one (absolute stereochemistry assigned based on crystal
structure of enantiomeric compound). .sup.1H NMR (400 MHz, CD3OD)
.delta. 7.63 (s, 1H), 6.12 (s, 1H), 4.77 (s, 2H), 3.94-3.90 (m,
1H), 3.81-3.80 (m, 3H), 3.59-3.57 (m, 2H), 3.51-3.49 (m, 2H),
3.17-3.10 (m, 1H), 2.98-2.95 (m, 2H), 2.71 (br s, 1H), 2.30 (s,
3H), 2.29-2.25 (m, 1H), 2.25 (s, 3H), 1.83-1.78 (m, 1H). MS: 465
[M+H].sup.+. Chiral analysis: 95.66% ee/de; retention time: 6.867
min; column: Chiralpak AD-3 150.times.4.6 mm I.D., 3 .mu.m; mobile
phase: ethanol (0.05% DEA) in CO2 from 5% to 40%; flow rate: 2.5
mL/min; wavelength 220 nm.
Example 2
[0539]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-{(1R*)-2-hydroxy-1-[(3S*)-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoqui-
nolin-1(2H)-one (relative stereochemistry known, absolute
stereochemistry undetermined). .sup.1H NMR (400 MHz, CD3OD) .delta.
7.62 (s, 1H), 6.11 (s, 1H), 4.76 (s, 2H), 4.13-4.11 (m, 1H),
3.78-3.75 (m, 1H), 3.69-3.68 (m, 2H), 3.61-3.59 (m, 3H), 3.51-3.50
(m, 2H), 2.98-2.95 (m, 2H), 2.65 (br s, 1H), 2.30 (s, 3H), 2.25 (s,
3H), 1.77-1.75 (m, 1H), 1.42-1.37 (m, 1H).). MS: 465 [M+H].sup.+.
Chiral analysis: 98.70% ee/de; retention time: 7.309 min; column:
Chiralpak AD-3 150.times.4.6 mm I.D., 3 .mu.m; mobile phase:
ethanol (0.05% DEA) in CO2 from 5% to 40%; flow rate: 2.5 mL/min;
wavelength 220 nm.
Example 3
[0540]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-{(1S*)-2-hydroxy-1-[(3R*)-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoqui-
nolin-1(2H)-one (relative stereochemistry known, absolute
stereochemistry undetermined). .sup.1H NMR (400 MHz, CD3OD) .delta.
7.62 (s, 1H), 6.11 (s, 1H), 4.76 (s, 2H), 4.12-4.11 (m, 1H),
3.80-3.78 (m, 1H), 3.69-3.67 (m, 3H), 3.67-3.62 (m, 2H), 3.61-3.50
(m, 2H), 2.98-2.95 (m, 2H), 2.65 (br s, 1H), 2.29 (s, 3H), 2.25 (s,
3H), 1.77-1.74 (m, 1H), 1.42-1.37 (m, 1H). MS: 465 [M+H].sup.+.
Chiral analysis: 96.48% ee/de; retention time: 8.021 min; column:
Chiralpak AD-3 150.times.4.6 mm I.D., 3 .mu.m; mobile phase:
ethanol (0.05% DEA) in CO2 from 5% to 40%; flow rate: 2.5 mL/min;
wavelength 220 nm.
Example 4
[0541]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-{(1S)-2-hydroxy-1-[(3S)-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquino-
lin-1(2H)-one (absolute stereochemistry determined by X-Ray crystal
structure). .sup.1H NMR (400 MHz, CD3OD) .delta. 7.64 (s, 1H), 6.13
(s, 1H), 4.78 (s, 2H), 3.95-3.90 (m, 1H), 3.83-3.81 (m, 3H),
3.60-3.55 (m, 2H), 3.55-3.52 (m, 2H), 3.32-3.19, (m, 1H), 2.99-2.96
(m, 2H), 2.75 (br s, 1H), 2.32 (s, 3H), 2.31-2.29 (m, 1H), 2.27 (s,
3H), 1.84-1.79 (m, 1H). MS: 465 [M+H].sup.+. Chiral analysis:
99.18% ee/de; retention time: 8.429 min; column: Chiralpak AD-3
150.times.4.6 mm I.D., 3 .mu.m; mobile phase: ethanol (0.05% DEA)
in CO2 from 5% to 40%; flow rate: 2.5 mL/min; wavelength 220
nm.
Method B
Example 5
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{1-[-
(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydroisoquinolin-1(2H)-
-one
Example 6
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1.-
xi.)-1-[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydroisoquinol-
in-1(2H)-one isomer A
Example 7
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1.-
xi.)-1-[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydroisoquinol-
in-1(2H)-one isomer B
##STR00035## ##STR00036##
[0543] To an ice bath-cooled solution of methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)-2-diazoacetate (Cpd U, 800 mg,
1.48 mmol) in anhydrous dichloromethane (10 mL) was added HBr (800
uL, 4.42 mmol, 33% wt in HOAc), causing gas evolution. The solution
was allowed to warm to room temperature and stirred overnight. The
reaction mixture was carefully quenched with saturated aqueous
sodium hydrogen carbonate, and extracted with ethyl acetate
(2.times.50 mL). The combined organic phases were dried over sodium
sulfate, concentrated to dryness, and purified by a silica gel
column with a gradient elution of 0.fwdarw.10% MeOH/EA to afford
racemic methyl
2-bromo-2-(5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)me-
thyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)acetate (5a, 655 mg,
88%) as a solid. MS: 501.00/502.05. .sup.1H NMR (400 MHz, CDCl3)
.delta. 7.91 (s, 1H), 6.18 (s, 1H), 6.03 (s, 1H), 4.76 (s, 2H),
3.81 (s, 3H), 3.68 (t, J=6.24 Hz, 2H), 2.98 (t, J=6.24 Hz, 2H),
2.45 (s, 3H), 2.37 (s, 3H).
[0544] A mixture of methyl
2-bromo-2-(5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)me-
thyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)acetate (5a, 162 mg,
0.323 mmol), (3R)-3-fluoropyrrolidine hydrochloride (144 mg, 1.15
mmol), N,N-diisopropylethylamine (0.35 mL, 2.01 mmol), and
anhydrous N,N-dimethylformamide (4 mL) was stirred at room
temperature overnight. The reaction mixture was partitioned between
ethyl acetate (20 mL) and water (20 mL). The organic phase was
separated, washed sequentially with water (20 mL) and brine (20
mL), dried over sodium sulfate, and concentrated to dryness to give
crude methyl
2-(5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1--
oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-((R)-3-fluoropyrrolidin-1-yl)ace-
tate, as a mixture of diastereomers (5b, 162 mg, 98% yield), which
was used in the next step without further purification. LCMS:
T=510.15/511.10/512.20.
[0545] The crude mixture of methyl
2-(5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1--
oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-((R)-3-fluoropyrrolidin-1-yl)ace-
tate diastereomers (5b, 152 mg, 0.298 mmol) in anhydrous
tetrahydrofuran (4.0 mL) was treated with lithium borohydride (2.0
M solution in THF, 0.45 mL, 0.90 mmol) followed by a few drops of
methanol. The process of addition was repeated 4 times, then the
reaction was quenched with 2 M NH.sub.4Cl (20 mL) and extracted
with ethyl acetate (2.times.20 mL). The combined organic phases
were dried over sodium sulfate, concentrated to dryness, and
purified by preparative HPLC to afford
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{1--
[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydroisoquinolin-1(2H-
)-one as a mixture of diastereomers at the benzylic carbon (Example
5, 32.2 mg, 22% yield over two steps). .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 11.66 (br. s., 1H), 7.80 (d, J=3.67 Hz, 1H), 6.00
(s, 1H), 5.15-5.43 (m, 1H), 4.85 (br. s., 1H), 4.69 (s, 2H),
4.03-4.13 (m, 1H), 3.66-3.84 (m, 2H), 3.50-3.63 (m, 2H), 3.02-3.10
(m, 1H), 2.94-3.02 (m, 2H), 2.71-2.90 (m, 2H), 2.42-2.55 (m, 1H),
2.28 (s, 3H), 2.24 (s, 3H), 2.09-2.23 (m, 1H), 1.87-2.08 (m, 1H).
MS: 482 [M+H].sup.+.
[0546] The mixture of diastereomers of
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{1--
[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydroisoquinolin-1(2H-
)-one (Example 5, 20.0 mg, 0.0415 mmol) was separated by chiral
preparative SFC on a Chiralcel OJ-3 4.6.times.100 mm 3 u column,
eluting with 10% MeOH/DEA @ 120 bar, 4 mL/min, affording, after
lyophilization, Example 6 (Peak 1, retention time 1.18 min, 5.65
mg, 28%) and Example 7 (Peak 2, retention time 1.42 min, 6.23 mg,
31%). The absolute configuration of the benzylic carbon in each
isomer was not determined.
Example 6
[0547]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-{(1.xi.)-1-[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydrois-
oquinolin-1(2H)-one--isomer A. MS: 482 [M+H].sup.+. Chiral
analysis: .about.88% de, [.alpha.]D=+62.1.degree. (c 0.01 MeOH)
Example 7
[0548]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-{(1.xi.)-1-[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydrois-
oquinolin-1(2H)-one--isomer B. MS: 482 [M+H].sup.+. Chiral
analysis: .about.98% de; [.alpha.]D=-58.9.degree. (c 0.01 MeOH)
Method C
Example 8
(+)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7--
{fluoro[1-(hydroxyacetyl)piperidin-4-yl]methyl}-3,4-dihydroisoquinolin-(2H-
)-one
Example 9
(-)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7--
{fluoro[1-(hydroxyacetyl)piperidin-4-yl]methyl}-3,4-dihydroisoquinolin-(2H-
)-one
##STR00037##
[0550] To a solution
2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3-
,4-dihydroisoquinolin-1(2H)-one (Cpd S, 311.0 mg, 0.598 mmol) in
tetrahydrofuran (5.0 mL) and 1,4-dioxane (0.5 mL) at -40.degree. C.
(in an acetonitrile/dry ice bath) was added iPrMgCl--LiCl (1.3 M in
THF, 0.850 mL, 1.10 mmol) and the reaction was stirred for 1 hour.
N-Boc-4-formylpiperidine (0.242 g, 1.14 mmol) was then added, and
the flask was warmed to 0.degree. C. in an ice bath. After 1 hour
at 0.degree. C., the solution was quenched with sat. aq. NH.sub.4Cl
and extracted with MTBE. The MTBE layer was concentrated, and the
resulting oil purified on silica gel (Isco RediSepRf, 12 g, 10-70%
gradient of ethyl acetate in heptane) to give racemic tert-butyl
4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-
-1,2,3,4-tetrahydroisoquinolin-7-yl)(hydroxy)methyl)piperidine-1-carboxyla-
te (8a, 0.229 g, 59%) as a white solid. .sup.1H NMR (400 MHz,
CD3OD) .delta. 7.66 (s, 1H), 7.36-7.44 (m, 2H), 7.18-7.31 (m, 3H),
6.71 (s, 1H), 5.42 (s, 2H), 5.04 (d, J=5.14 Hz, 1H), 4.83 (d,
J=1.96 Hz, 2H), 4.08 (d, J=12.72 Hz, 2H), 3.23 (t, J=6.24 Hz, 2H),
2.73 (t, J=6.11 Hz, 2H), 2.53-2.71 (m, 2H), 2.39 (s, 3H), 2.35 (s,
3H), 1.76-1.90 (m, 1H), 1.32-1.62 (m, 13H); MS 654, 656
[M+H].sup.+.
[0551] To a solution of racemic tert-butyl
4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-
-1,2,3,4-tetrahydroisoquinolin-7-yl)(hydroxy)methyl)
piperidine-1-carboxylate (8a, 88 mg, 0.13 mmol) in dichloromethane
(4 ML) cooled in a dry ice/acetone bath was added Deoxo-Fluor.TM.
(50% in THF, 0.165 mL, 0.39 mmol). After stirring at -78.degree. C.
for 5 minutes, the cooling bath was removed and the mixture stirred
for 5 minutes. The reaction was quenched with the addition of sat.
aq. NaHCO.sub.3, the layers were separated, and the dichloromethane
layer was concentrated. The resulting oil was purified on silica
gel (Biotage SNAP, HP-Sil, 10 g, 0-40% gradient of ethyl acetate in
heptane) to give racemic tert-butyl
4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-
-1,2,3,4-tetrahydroisoquinolin-7-yl)fluoromethyl)piperidine-1-carboxylate
(8b, 0.075 g, 85%) as a white, sticky solid. MS: 656, 658
[M+H].sup.+.
[0552] A solution of racemic tert-butyl
4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-
-1,2,3,4-tetrahydroisoquinolin-7-yl)fluoromethyl)piperidine-1-carboxylate
(8b, (0.075 g, 0.11 mmol) in trifluoroacetic acid (5.0 mL) was
heated to 50.degree. C. for 1 hour. The reaction mixture was
diluted with heptane and concentrated under vacuum. The residue was
dissolved in ethanol and concentrated again. The remaining white
solid was dried under vacuum to give crude, racemic
5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-7-(fl-
uoro(piperidin-4-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (8c
0.085 g) as the TFA salt. MS 466, 468 [M+H].sup.+. This material
was dissolved in dichloromethane (5 mL) and cooled to 0.degree. C.
Triethylamine (0.060 mL, 0.43 mmol) and then 2-acetoxyacetyl
chloride (0.017 mL, 0.16 mmol) were added, the mixture stirred for
30 minutes, and then a few drops of methanol were added to quench
the excess reagent. The solution was concentrated under vacuum, and
the residue was dissolved in methanol (5 mL) and treated with
potassium carbonate (0.100 g, 0.724 mmol). After 4 hours at room
temperature, the reaction was filtered, concentrated, and purified
by preparative chiral SFC (OJ-H, 21.times.250 mm column, 32 mL
MeOH: 8 mL CO.sub.2, 100 bar, 40 mL/min) to give separated
enantiomers Example 8 (peak 1, 9.6 mg, 15%) and Example 9 (peak 2,
8.1 mg, 13%) as white solids. The absolute stereochemistry of each
isomer was not determined, but optical rotation measurements were
obtained.
Example 8
[0553]
(+)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)met-
hyl]-7-{fluoro[1-(hydroxyacetyl)piperidin-4-yl]methyl}-3,4-dihydroisoquino-
lin-1(2H)-one. MS 524, 526 [M+H].sup.+. Optical rotation:
[.alpha.].sub.D=+9.9.degree. (c, 0.1, DMSO). Chiral analysis:
>99% ee; Retention time 8.13 min on Lux Cellulose-2
4.6.times.100 mm 3 u column, 60% MeOH @ 120 bar, 4 mL/min.
Example 9
[0554]
(-)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)met-
hyl]-7-{fluoro[1-(hydroxyacetyl)piperidin-4-yl]methyl}-3,4-dihydroisoquino-
lin-1(2H)-one. .sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 11.55
(s, 1H), 7.58 (d, J=4.95 Hz, 1H), 5.88 (s, 1H), 5.72-5.84 (m, 1H),
4.55 (q, J=13.75 Hz, 2H), 4.46 (br. s., 1H), 4.30-4.42 (m, 1H),
4.05-4.13 (m, 1H), 3.97-4.04 (m, 1H), 3.69 (t, J=13.39 Hz, 1H),
3.45 (t, J=5.78 Hz, 2H), 2.80-2.98 (m, 3H), 2.20 (d, J=18.71 Hz,
1H), 2.11 (s, 3H), 1.61 (br. s., 1H), 1.46 (br. s., 1H), 1.34-1.42
(m, 1H), 1.23 (s, 5H); MS 524, 526 [M+H].sup.+. Optical rotation:
[.alpha.].sub.D=-6.5.degree. (c, 0.1, DMSO). Chiral analysis:
.about.95% ee; Retention time 10.29 min on Lux Cellulose-2
4.6.times.100 mm 3 u column, 60% MeOH @ 120 bar, 4 mL/min.
Method D
Example 10
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(2-h-
ydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one--isomer
A
Example 11
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(2-h-
ydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one--isomer
B
##STR00038##
[0556] To a stirred solution of dry methanol (12 mg, 0.36 mmol) and
dirhodium tetraacetate (1.2 mg, 0.0028 mmol) in dichloromethane (5
mL) was added a solution of methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)-2-diazoacetate (Cpd U, 150 mg,
0.278 mmol) in dichloromethane (5 mL) dropwise over a period of 60
minutes at room temperature. After the addition, the reaction was
heated to reflux for 18 hours. The mixture was concentrated and
purified by flash chromatography (eluting with petroleum
ether/EtOAc=10:1, Rf.about.0.3) to afford racemic methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)-2-methoxyacetate (11a, 100 mg,
66%) as brown oil.
[0557] To a stirred solution of racemic methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)-2-methoxyacetate (11a, 100 mg,
0.184 mmol) in anhydrous tetrahydrofuran (10 mL) was added solid
lithium borohydride (12 mg, 0.55 mmol) in one portion at room
temperature. The resulting mixture was heated at 60.degree. C. for
1 hour. The mixture was quenched with water (15 mL) and extracted
with ethyl acetate (3.times.15 mL). The combined organic extracts
were washed with saturated aq. NaCl (15 mL), dried over sodium
sulfate, filtered and concentrated to afford the crude product (91
mg). Purification was accomplished using flash chromatography
(eluting with petroleum ether/EtOAc=1:1) to afford racemic
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(2-hydr-
oxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one (11b, 60 mg,
63%) as a white solid.
[0558] A solution of racemic
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(2-hydr-
oxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one (11b 60 mg,
0.12 mmol) in dichloromethane (2 mL) and trifluoroacetic acid (2
mL) was stirred at room temperature for 18 hours. Analysis by LC-MS
showed complete deprotection of the benzyl group, but some
trifluoroacetate ester at the pyridone oxygen, so the mixture was
concentrated and then methanol (10 mL) and potassium carbonate
(80.9 mg, 0.585 mmol) were added. The resulting mixture was stirred
at room temperature for 30 minutes, at which time analysis by LC-MS
showed complete deprotection of the TFA-ester. The mixture was
filtered, the collected solid was washed with DCM/MeOH (10:1, 10
mL) and the filtrate was concentrated. Purification was
accomplished using flash chromatography (eluting with
DCM/MeOH=10:1, Rf.about.0.6) to obtain a racemic
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(2--
hydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one (37 mg,
75%) as a white solid.
[0559] Combined batches of racemic
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(2--
hydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one (total 100
mg, 0.235 mmol) were separated by chiral SFC on a Chiralpak IC 250
mm.times.30 mm, 10 .mu.m column, eluting with 50% EtOH/NH.sub.4OH
at a flow rate of 70 mL/min. After lyophilization, Example 10 (peak
1, 33 mg, 33%) and Example 11 (peak 2, 35 mg, 35%) were obtained as
off-white solids. Absolute stereochemistry for each isomer was not
determined.
Example 10
[0560]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-(2-hydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one--isomer
A. .sup.1H NMR (400 MHz, CDCl3) .delta. 12.23 (br. s., 1H), 7.53
(s, 1H), 5.95 (s, 1H), 4.92-4.89 (m, 1H), 4.78 (s, 2H), 3.80-3.71
(m, 1H), 3.68-3.62 (m, 2H), 3.53-3.51 (m, 1H), 3.33 (s, 3H), 2.94
(t, J=6.0 Hz, 2H), 2.35 (s, 3H), 2.29 (s, 3H). MS: 425 [M+H].sup.+.
Chiral analysis: 100% ee, retention time 7.717 min, column:
Chiralpak IC-3 150.times.4.6 mm I.D., 3 .mu.m; mobile phase: 40%
ethanol (0.05% DEA) in CO2; flow rate: 2.35 mL/min.
Example 11
[0561]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-(2-hydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one--isomer
B. .sup.1H NMR (400 MHz, CDCl3) .delta. 12.15 (br. s., 1H), 7.53
(s, 1H), 5.95 (s, 1H), 4.92-4.89 (m, 1H), 4.78 (s, 2H), 3.80-3.71
(m, 1H), 3.68-3.63 (m, 2H), 3.55-3.51 (m, 1H), 3.34 (s, 3H), 2.94
(t, J=5.6 Hz, 2H), 2.35 (s, 3H), 2.29 (s, 3H). MS: 425 [M+H].sup.+.
Chiral analysis: 100% ee, retention time 11.063 min, column:
Chiralpak IC-3 150.times.4.6 mm I.D., 3 .mu.m; mobile phase: 40%
ethanol (0.05% DEA) in CO2; flow rate: 2.35 mL/min.
Method E
Example 12
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1R-
)-2-hydroxy-1-[(2S)-tetrahydrofuran-2-yl]ethyl}-3,4-dihydroisoquinolin-1(2-
H)-one
Example 13
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1R-
)-2-hydroxy-1-[(2R)-tetrahydrofuran-2-yl]ethyl}-3,4-dihydroisoquinolin-1(2-
H)-one
##STR00039##
[0563] To a solution methyl
2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)-2-diazoacetate (Cpd U, 1.78 g,
3.29 mmol) in anhydrous tetrahydrofuran (20 mL) was added a
solution of copper (II) triflate (120 mg, 0.332 mmol) and
(S)-(-)-2,2'-isopropylidene-bis(4-phenyl-2-oxazoline) (130 mg,
0.389 mmol) in anhydrous tetrahydrofuran (4 mL). The resulting
solution was heated to reflux overnight in an 80.degree. C. oil
bath. After cooling to room temperature, the reaction mixture was
concentrated to dryness and purified by a silica gel column with a
gradient elution of 0.fwdarw.40% EA/HEP to afford a mixture of
diastereomers with (S) geometry at the benzylic carbon
(stereochemical assignment by analogy to Jimenez-Oses, G. et al.,
J. Org. Chem. 2013, 78, 5851-5857), methyl
(2S)-2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-
-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-2-yl)acetate
(12a, 333 mg, 17%). MS:583.10/584.20 [M+H].sup.+. .sup.1H NMR (400
MHz, CDCl3) .delta. 7.69 (d, J=2.20 Hz, 1H), 7.45 (d, J=7.58 Hz,
2H), 7.30-7.39 (m, 3H), 6.64 (s, 1H), 5.45 (s, 2H), 4.80-4.92 (m,
2H), 4.41-4.59 (m, 1H), 3.79-4.02 (m, 1H), 3.72-3.80 (m, 1H), 3.70
(d, J=5.01 Hz, 3H), 3.23-3.31 (m, 2H), 2.69-2.76 (m, 2H), 2.44 (s,
3H), 2.34 (d, J=3.79 Hz, 3H), 1.93-2.18 (m, 1H), 1.80-1.94 (m, 2H),
1.55-1.80 (m, 2H).
[0564] [Under the same conditions, use of the enantiomeric
(R)-(-)-2,2'-isopropylidene-bis(4-phenyl-2-oxazoline) ligand
produces a mixture of diastereomers with (R) geometry at the
benzylic position, methyl
(2R)-2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dic-
hloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-2-yl)ac-
etate.]
[0565] Lithium borohydride (2.0 M solution in THF, 1.0 mL, 2.0
mmol) was added to a solution of methyl
(2S)-2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-
-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-2-yl)acetate
(12a 333 mg, 0.571 mmol) in anhydrous tetrahydrofuran (10 mL),
followed by a few drops of methanol. Gas was evolved. The mixture
was stirred at room temperature for 1 hour, then quenched with 10
mL 2 M NH4Cl, diluted with water, and extracted with ether
(3.times.50 mL). The combined organic phases were dried over sodium
sulfate, concentrated to dryness, to give crude
2-((2-(benzyloxy)-4,6-dimethyl
pyridin-3-yl)methyl)-5,8-dichloro-7-((1R)-2-hydroxy-1-(tetrahydrofuran-2--
yl)ethyl)-3,4-dihydroisoquinolin-1(2H)-one (12b, 280 mg, 88%), as a
mixture of diastereomers with (R) geometry at the benzylic carbon.
This mixture was used in the next step without further
purification. MS: 555.20/557.20.
[0566] A solution of the crude mixture of diastereomers,
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-((1R)-2-
-hydroxy-1-(tetrahydrofuran-2-yl)ethyl)-3,4-dihydroisoquinolin-1(2H)-one
(12b, 280 mg, 0.504 mmol) in trifluoroacetic acid (8 mL) was
stirred at 50.degree. C. for 1 hour. After removing excess
trifluoroacetic acid, the residue was dissolved in methanol (10 mL)
and treated with 4 M NaOH for 30 minutes at 50.degree. C. The
reaction mixture was partitioned between ethyl acetate (50 mL) and
water (50 mL). The organic phase was separated. The aqueous phase
was acidified to pH .about.2-3, and extracted with ethyl acetate
(2.times.50 mL). The combined organic phases were dried over sodium
sulfate, concentrated to dryness, and purified by chiral SFC
(Chiralpak AD-3 4.6.times.100 mm 3 u column; eluting with 5-60%
MeOH in 3 minutes, 120 bar, 4 mL/min) yielding the separated
diastereomeric products, Example 12 (peak 1, 32 mg, 14%) and
Example 13 (peak 2, 77 mg, 33%). Stereochemistry of the isolated
products were assigned by analogy to Jimenez-Oses, G. et al., J.
Org. Chem. 2013, 78, 5851-5857.
Example 12
[0567]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-{(1R)-2-hydroxy-1-[(2S)-tetrahydrofuran-2-yl]ethyl}-3,4-dihydroisoquino-
lin-1(2H)-one. .sup.1H NMR (400 MHz, CD3OD) .delta. 7.58 (s, 1H),
6.11 (s, 1H), 4.76 (s, 2H), 4.10-4.21 (m, 1H), 3.86-3.99 (m, 3H),
3.75-3.84 (m, 1H), 3.64-3.72 (m, 1H), 3.46-3.55 (m, 2H), 2.91-3.01
(m, 2H), 2.29 (s, 3H), 2.25 (s, 3H), 1.73-1.97 (m, 3H), 1.44-1.58
(m, 1H). MS: 465 {M+H].sup.+. Chiral analysis: 91% ee/de; retention
time 2.91 min on Chiralpak AD-3 4.6.times.100 mm 3.mu. column;
eluting with 5-60% MeOH in 3 minutes, 120 bar, 4 mL/min.
Example 13
[0568]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-{(1R)-2-hydroxy-1-[(2R)-tetrahydrofuran-2-yl]ethyl}-3,4-dihydroisoquino-
lin-1(2H)-one. .sup.1H NMR (400 MHz, CD3OD) .delta. 7.66 (s, 1H),
6.10 (s, 1H), 4.77 (s, 2H), 4.27 (dt, J=7.86, 6.16 Hz, 1H),
3.72-3.91 (m, 3H), 3.67 (t, J=6.79 Hz, 2H), 3.49 (t, J=6.24 Hz,
2H), 2.95 (t, J=6.17 Hz, 2H), 2.28 (s, 3H), 2.24 (s, 3H), 2.01-2.13
(m, 1H), 1.68-1.92 (m, 2H), 1.49-1.62 (m, 1H). MS: 465 {M+H].sup.+.
Chiral analysis: 93% ee/de; retention time 3.21 minutes on
Chiralpak AD-3 4.6.times.100 mm 3.mu. column; eluting with 5-60%
MeOH in 3 minutes, 120 bar, 4 mL/min.
Method F
Example 14
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(pro-
pan-2-yl)-3,4-dihydroisoquinolin-1(2H)-one
##STR00040##
[0570] A solution of
2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3-
,4-dihydroisoquinolin-1(2H)-one (Cpd S, 500 mg, 0.961 mmol),
triethylamine (0.30 mL, 2.2 mmol), and
[1.1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II)-dichlorometha-
ne complex (24 mg, 0.028 mmol) in 2-propanol (15 mL) under nitrogen
in a capped microwave tube was heated at 100.degree. C. for 1 hour
in a microwave reactor. After removal of the solvent, the product
was extracted into ether (3.times.10 mL) and the combined organic
extracts were washed with water (2.times.), dried over magnesium
sulfate, concentrated, and purified by flash chromatography (silica
gel, 0-60% EtOAc in heptane) to afford
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(prop-1-
-en-2-yl)-3,4-dihydroisoquinolin-1(2H)-one (14a, 240 mg, 52%) as a
white foam. .sup.1H NMR (400 MHz, CDCl3) .delta. 7.37 (d, J=6.8 Hz,
2H), 7.30-7.20 (m, 3H), 7.18 (d, J=2.6 Hz, 1H), 6.55 (s, 1H), 5.35
(s, 2H), 5.16 (d, J=1.5 Hz, 1H), 4.85 (d, J=1.5 Hz, 1H), 4.80 (s,
2H), 3.20 (t, J=6.2 Hz, 2H), 2.65 (t, J=6.2 Hz, 2H), 2.34 (s, 3H),
2.26 (s, 3H), 2.01 (s, 3H). MS: 481 [M+H].sup.+.
[0571] A solution of
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(prop-1-
-en-2-yl)-3,4-dihydroisoquinolin-1(2H)-one (14a, 75 mg, 0.16 mmol)
and platinum(IV) oxide (71 mg, 0.31 mmol) in ethyl acetate (5 mL)
was stirred under a hydrogen balloon for 2 hours. The catalyst was
filtered off and the solvent removed in vacuo. The crude product
was purified via supercritical fluid chromatography (SFC/ZymorSpher
HAP 150.times.21.2 mm column with 8% MeOH @ 100 bar, 58 mL/min) to
afford
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(pr-
opan-2-yl)-3,4-dihydroisoquinolin-1(2H)-one (Example 14 (7 mg,
10%), as a white solid. .sup.1H NMR (400 MHz, CDCl3) .delta. 7.35
(s, 1H), 5.90 (s, 1H), 4.78 (s, 2H), 3.63 (t, J=6.30 Hz, 2H),
3.5-3.6 (m, 1H), 2.90 (t, J=6.11 Hz, 2H), 2.35 (s, 3H), 2.24 (s,
3H), 1.24 (d, J=6.85 Hz, 6H). MS: 393 [M+H].
Method G
Example 15
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(1-m-
ethoxypropyl)-3,4-dihydroisoquinolin-1(2H)-one--isomer A
Example 16
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(1-m-
ethoxypropyl)-3,4-dihydroisoquinolin-1(2H)-one--isomer B
##STR00041## ##STR00042##
[0573] A mixture of
2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3-
,4-dihydroisoquinolin-1(2H)-one (Cpd S, 200 mg, 0.384 mmol),
(Z)-3-Hexenyl-3-boronic acid catechol ester (155 mg, 0.769 mmol)
and cesium fluoride (182 mg, 1.20 mmol) in dioxane (2 mL) and water
(0.4 mL) was degassed with nitrogen for 5 minutes.
Tetrakis(triphenylphosphine)palladium(0) (44.4 mg, 0.0384 mmol) was
added, the mixture degassed with nitrogen again, and then heated to
100.degree. C. for 4 hours. The mixture was diluted with water (15
mL) and extracted with EtOAc (3.times.15 mL). The combined organic
layers were washed with brine (20 mL), dried over sodium sulfate,
concentrated, and purified by column chromatography (silica gel,
Petroleum ether/EtOAc=10:1, Rf.about.0.5) to give
(E)-2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(he-
x-3-en-3-yl)-3,4-dihydroisoquinolin-1(2H)-one (15a, 150 mg, 74.5%)
as colorless oil. .sup.1H NMR (400 MHz, CDCl3) .delta. 7.45 (d,
J=7.7 Hz, 2H), 7.37-7.34 (m, 3H), 7.19 (s, 1H), 6.62 (s, 1H), 5.43
(s, 2H), 5.28 (t, J=6.8 Hz, 1H), 4.88 (s, 2H), 3.28 (t, J=6.4 Hz,
2H), 2.73 (t, J=6.4 Hz, 2H), 2.45-2.43 (m, 2H), 2.42 (s, 3H), 2.33
(s, 3H), 2.19 (quint, J=7.6 Hz, 2H), 1.04 (t, J=7.6 Hz, 3H),
0.89-0.86 (m, 4H).
[0574] A flow of ozone was bubbled through a stirred solution of
(E)-2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(he-
x-3-en-3-yl)-3,4-dihydroisoquinolin-1(2H)-one (15a, 864 mg, 1.65
mmol) in methanol (46 mL) at -78.degree. C. until a light purple
color was obtained (.about.10 minutes). Nitrogen was bubbled into
the solution until the purple color had disappeared, then
dimethylsulfide (0.4 mL) was added, and the solution stirred at
10.degree. C. for 3 hours. The reaction mixture was concentrated in
vacuum and the residue purified by column chromatography (silica
gel, petroleum ether/EtOAc=6:1, Rf.about.0.5) to give
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-propion-
yl-3,4-dihydroisoquinolin-1(2H)-one (15b, 346 mg, 42.1%) as yellow
oil.
[0575] Sodium borohydride (52.6 mg, 1.39 mmol) was added to a
cooled (0.degree. C.) solution of
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-propion-
yl-3,4-dihydroisoquinolin-1(2H)-one (15b, 346 mg, 0.696 mmol) in
methanol (20 mL), and stirring continued for 1 hour at 10.degree.
C. The reaction was quenched with Sat.NH4Cl (30 mL), extracted with
ethyl acetate (3.times.25 mL). The combined organic layers were
washed with brine (25 mL), dried over sodium sulfate, and
concentrated in vacuum. The residue was purified by column
chromatography (silica gel, Petroleum ether/EtOAc=2:1,
Rf.about.0.6), affording racemic 2-((2-(benzyloxy)-4,6-dimethyl
pyridin-3-yl)methyl)-5,8-dichloro-7-(1-hydroxypropyl)-3,4-dihydroisoquino-
lin-1(2H)-one (15c, 300 mg, 86.4%) as colorless oil.
[0576] Potassium tert-butoxide (111 mg, 0.985 mmol) and iodomethane
(140 mg, 0.985 mmol) were added to a cooled (10.degree. C.)
solution of
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-hydr-
oxypropyl)-3,4-dihydroisoquinolin-1(2H)-one (15c, 246 mg, 0.493
mmol) in N,N-dimethylformamide (25 mL), and stirred at 10.degree.
C. overnight. The reaction was quenched with water (30 mL),
extracted with EtOAc (3.times.35 mL). The combined organic layers
were washed with brine (35 mL), dried over sodium sulfate,
concentrated, and purified by column chromatography (silica gel,
Petroleum ether/EtOAc=5:1), yielding racemic
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-meth-
oxypropyl)-3,4-dihydroisoquinolin-1(2H)-one (15d, 148 mg, 58.5%) as
yellow oil.
[0577] Trifluoroacetic acid (9 mL) was added dropwise to a stirred
solution of
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-meth-
oxypropyl)-3,4-dihydroisoquinolin-1(2H)-one (15d, 218 mg, 0.423
mmol) in dichloromethane (9 mL) at 10.degree. C. The resulting
mixture was stirred at 25.degree. C. overnight. The mixture was
concentrated in vacuum and the residue purified by column
chromatography (silica gel, CH2Cl2/MeOH=10:1, Rf.about.0.55) to
give racemic
5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-7-(1--
methoxypropyl)-3,4-dihydroisoquinolin-1(2H)-one (15e, 150 mg,
83.8%) as pink oil. This racemic mixture was separated by
preparatory SFC [column: (AD (250 mm*30 mm, 5 um)), mobile phase:
25% MeOH NH.sub.3H.sub.2O, Flow rate: 50 mL/min, wavelength: 220
nm, workup: lyophilization] to give Isomer A (Example 15, 45.65 mg,
31.4%) as a white solid and Isomer B (Example 16, 31.8 mg, 21.2%)
as an off-white solid. The absolute stereochemistry of each isomer
was not determined.
Example 15
[0578]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-(1-methoxypropyl)-3,4-dihydroisoquinolin-1(2H)-one--isomer A.
.sup.1H NMR (400 MHz, CDCl3) .delta. 11.88 (br s, 1H), 7.53 (s,
1H), 5.94 (s, 1H), 4.78 (s, 2H), 4.69-4.66 (m, 1H), 3.65 (t, J=4.8
Hz, 2H), 3.24 (s, 3H), 2.93 (t, J=6.2 Hz, 2H), 2.36 (s, 3H), 2.29
(s, 3H), 1.76-1.72 (m, 1H), 1.65-1.60 (m, 1H), 0.96 (t, J=7.0 Hz,
3H). MS: 423 [M+H].sup.+. Chiral analysis: 100% ee; column:
Chiralpak AD-H 250.times.4.6 mm I.D., 5 um; retention time: 8.04
min; mobile phase: methanol (0.05% DEA) in CO2 from 5% to 40%; flow
rate: 2.5 mL/min; wavelength: 220 nm.
Example 16
[0579]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-(1-methoxypropyl)-3,4-dihydroisoquinolin-1(2H)-one--isomer B.
.sup.1H NMR (400 MHz, CDCl3) .delta. 10.88 (br s, 1H), 7.53 (s,
1H), 5.93 (s, 1H), 4.77 (s, 2H), 4.69-4.66 (m, 1H), 3.67-3.63 (m,
2H), 3.24 (s, 3H), 2.93 (t, J=5.8 Hz, 2H), 2.36 (s, 3H), 2.28 (s,
3H), 1.78-1.73 (m, 1H), 1.65-1.58 (m, 1H), 0.96 (t, J=7.2 Hz, 3H).
MS: 423 [M+H].sup.+. Chiral analysis: 99.6% ee; column: Chiralpak
AD-H 250.times.4.6 mm I.D., 5 um; retention time: 8.34 min; mobile
phase: methanol (0.05% DEA) in CO2 from 5% to 40%; flow rate: 2.5
mL/min; wavelength: 220 nm.
Method H
Example 75
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{1-[-
1-(hydroxyacetyl)azetidin-3-ylidene]ethyl}-3,4-dihydroisoquinolin-1(2H)-on-
e
##STR00043##
[0581] A solution of triphenylphosphine (15.83 g, 60.35 mmol) in
anhydrous dichloromethane (16 mL) was cooled to 0.degree. C. and
degassed by sparging with nitrogen for 5 minutes. Carbon
tetrabromide (9.98 g, 30.1 mmol) was added, and the solution
stirred at 0.degree. C. for 5 minutes before a solution of
3-oxo-azetidine-1-carboxylic acid tert-butyl ester (2.52 g, 14.7
mmol) in anhydrous dichloromethane (7 mL) was added dropwise via
syringe, over 1 minutes. After stirring at 0.degree. C. for 20
minutes, the reaction was allowed to stir at room temperature for
22.5 hours. Heptane (100 mL) was added and the resulting
precipitate removed by filtration. The filtrate was concentrated to
give 7.82 g off-white solid. This solid was stirred in 100 mL
heptane with sonication, and then residual solids removed by
suction filtration. The filtrate was concentrated to give
tert-butyl 3-(dibromomethylene)azetidine-1-carboxylate (75a, 4.59
g, 95% yield) as a white solid. .sup.1H NMR (400 MHz, CDCl3)
.delta. 4.32 (s, 4H), 1.46 (s, 9H). MS: 226, 228, 230
[M-Boc+H].sup.+.
[0582] To a solution of tert-butyl
3-(dibromomethylene)azetidine-1-carboxylate (75a, 542 mg, 1.66
mmol) in THF (16.6 mL) at -78.degree. C. was added n-butyllithium
(1.6M solution in hexanes, 1.86 mL, 2.98 mmol). After 30 minutes,
the reaction was treated with iodomethane (325 uL, 5.22 mmol) and
stirring continued at -78.degree. C. for one hour. The reaction was
quenched with sat. aq. NH.sub.4Cl and extracted with MTBE. The
organic layer was concentrated and purified on silica gel (Eluting
with 0-25% ethyl acetate in heptane) to give tert-butyl
3-(1-bromoethylidene)azetidine-1-carboxylate (75b 0.230 g, 53%
yield) as a clear oil. .sup.1H NMR (400 MHz, CDCl3) .delta.
4.38-4.43 (m, 2H), 4.31-4.37 (m, 2H), 2.14 (quint, J=1.74 Hz, 3H),
1.46 (s, 9H).
[0583] Isopropylmagnesium chloride lithium chloride complex (1.3M
solution in THF, 2.00 mL, 2.60 mmol, 2.00 mL) was added to a cooled
(-40.degree. C., acetonitrile/dry ice bath) solution of
2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3-
,4-dihydroisoquinolin-1(2H)-one (Cpd S, 657 mg, 1.26 mmol) in THF
(12.6 mL), and the mixture stirred for one hour. Tri-n-butyltin
chloride (600 uL, 72.2 mmol) was added, and the flask was warmed to
0.degree. C. in an ice bath for 30 minutes. The reaction was
quenched with sat. aq. NH.sub.4Cl and extracted with MTBE. The MTBE
layer was washed with brine, concentrated, and the resulting crude
oil purified on silica gel (eluting with 0-30% ethyl acetate in
heptane) to give
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(tribut-
ylstannyl)-3,4-dihydroisoquinolin-1(2H)-one (75c, 0.699 g, 76%) as
a clear, thick oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.42 (d, J=7.09 Hz, 2H), 7.37 (s, 1H), 7.22-7.34 (m, 3H), 6.74 (s,
1H), 5.37 (s, 2H), 4.69 (s, 2H), 3.22 (t, J=5.99 Hz, 2H), 2.72 (t,
J=5.87 Hz, 2H), 2.35 (s, 3H), 2.30 (s, 3H), 1.51 (quint, J=7.76 Hz,
6H), 1.08-1.35 (m, 12H), 0.85 (t, J=7.34 Hz, 9H). MS: 731
[M+H].sup.+=731.
[0584] A mixture of
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(tribut-
ylstannyl)-3,4-dihydroisoquinolin-1(2H)-one (75c, 251 mg, 0.344
mmol) and tert-butyl 3-(1-bromoethylidene)azetidine-1-carboxylate
(75b, 103 mg, 0.393 mmol) 1,4-dioxane (4.00 mL) was treated with
tetrakis(triphenylphosphino)palladium(0) (60.8 mg, 0.0526 mmol),
and copper (I) iodide (10.0 mg, 0.0525 mmol). Nitrogen was bubbled
through the mixture for 10 minutes, then the vial was sealed and
irradiated in a microwave reactor at 120.degree. C. for 2 hours.
Dioxane was removed under vacuum, and the resulting oil was
purified on silica gel (eluting with 0-40% ethyl acetate in
heptane) to give tert-butyl
3-(1-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-o-
xo-1,2,3,4-tetrahydroisoquinolin-7-yl)ethyl
idene)azetidine-1-carboxylate (75d, 49 mg, 23% yield) as a clear
gum. .sup.1H NMR (400 MHz, CDCl3) .delta. 7.43-7.49 (m, 2H),
7.30-7.39 (m, 3H), 7.21 (s, 1H), 6.65 (s, 1H), 5.47 (s, 2H), 4.87
(s, 2H), 4.57 (br. s., 2H), 4.24 (br. s., 2H), 3.29 (t, J=6.24 Hz,
2H), 2.73 (t, J=6.24 Hz, 2H), 2.45 (s, 3H), 2.36 (s, 3H), 1.87 (s,
3H), 1.45 (s, 9H). MS: 622, 624 [M+H].sup.+.
[0585] A solution of tert-butyl
3-(1-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-o-
xo-1,2,3,4-tetrahydroisoquinolin-7-yl)ethylidene)azetidine-1-carboxylate
(75d, 49 mg, 0.079 mmol) in trifluoroacetic acid (5 mL, 70 mmol)
was stirred at room temperature for 6 hours. The solution was
concentrated to dryness. The residue was dissolved in methanol and
purified by SCX column (Varian Bond elute SCX, 2 g, 100% MeOH to
3.5M NH.sub.3 in MeOH) to give crude
7-(1-(azetidin-3-ylidene)ethyl)-5,8-dichloro-2-((4,6-dimethyl-2-oxo-
-1,2-dihydropyridin-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one
(Example 74, 38 mg, 100% yield) as a clear gum, which was used
without further purification in the next step. MS: 432, 434
[M+H].sup.+.
[0586] To a cooled (0.degree. C.) solution of crude
7-(1-(azetidin-3-ylidene)ethyl)-5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-d-
ihydropyridin-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one
(Example 74, 24 mg, 0.056 mmol) in dichloromethane (3.0 mL) was
added triethylamine (10 uL, 0.072 mmol) and then acetoxy acetyl
chloride (6.5 uL, 0.060 mmol). The reaction was stirred for 30
minutes and then quenched with methanol. The mixture was
concentrated, re-dissolved in methanol (5 mL), treated with cesium
carbonate (45 mg, 0.14 mmol), and stirred at room temperature
overnight. The resulting solution was concentrated to dryness. The
residue was dissolved in DMF, filtered, and purified by preparative
HPLC to give
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{1--
[1-(hydroxyacetyl)azetidin-3-ylidene]ethyl}-3,4-dihydroisoquinolin-1(2H)-o-
ne (Example 75, 9.38 mg, 34% yield) as a white powder. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 11.54 (br. s., 1H), 7.51 (s, 1H),
5.89 (s, 1H), 4.81-4.99 (m, 2H), 4.51-4.60 (m, 3H), 4.48 (br. s.,
1H), 4.19 (br. s., 1H), 3.83-4.01 (m, 2H), 3.46 (t, J=6.11 Hz, 2H),
2.88 (t, J=5.87 Hz, 2H), 2.17 (s, 3H), 2.12 (s, 3H), 1.85 (br. s.,
3H). MS; 490, 492 [M+H].sup.+.
Method I
Example 34
(.+-.)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-{[1-(hydroxyacetyl)piperidin-4-yl](methoxy)methyl}-3,4-dihydroisoquinol-
in-(2H)-one
Example 35
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{[1--
(hydroxyacetyl)piperidin-4-yl](methoxy)methyl}-3,4-dihydroisoquinolin-1(2H-
)-one--Isomer B
Example 36
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{[1--
(hydroxyacetyl)piperidin-4-yl](methoxy)methyl}-3,4-dihydroisoquinolin-1(2H-
)-one--Isomer A
##STR00044## ##STR00045##
[0588] To a solution of
2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3-
,4-dihydroisoquinolin-1(2H)-one (Cpd S, 311.0 mg, 0.598 mmol) in
tetrahydrofuran (5.0 mL) and 1,4-dioxane (0.5 mL) at -40.degree. C.
(in an acetonitrile/dry ice bath) was added isopropylmagnesium
chloride lithium chloride complex (1.3 M in THF, 0.850 mL, 1.10
mmol) and the reaction was stirred for 1 hour.
N-Boc-4-formylpiperidine (0.242 g, 1.14 mmol) was then added, and
the flask was warmed to 0.degree. C. in an ice bath. After 1 hour
at 0.degree. C., the solution was quenched with sat. aq. NH.sub.4Cl
and extracted with MTBE. The MTBE layer was concentrated, and the
resulting oil purified on silica gel (Isco RediSepRf, 12 g, 10-70%
gradient of ethyl acetate in heptane) to give racemic tert-butyl
4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-
-1,2,3,4-tetrahydroisoquinolin-7-yl)(hydroxy)methyl)piperidine-1-carboxyla-
te (8a, 0.229 g, 59%) as a white solid. .sup.1H NMR (400 MHz,
CD3OD) .delta. 7.66 (s, 1H), 7.36-7.44 (m, 2H), 7.18-7.31 (m, 3H),
6.71 (s, 1H), 5.42 (s, 2H), 5.04 (d, J=5.14 Hz, 1H), 4.83 (d,
J=1.96 Hz, 2H), 4.08 (d, J=12.72 Hz, 2H), 3.23 (t, J=6.24 Hz, 2H),
2.73 (t, J=6.11 Hz, 2H), 2.53-2.71 (m, 2H), 2.39 (s, 3H), 2.35 (s,
3H), 1.76-1.90 (m, 1H), 1.32-1.62 (m, 13H); MS 654, 656
[M+H].sup.+.
[0589] A cooled (0.degree. C.) solution of racemic tert-butyl
4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-
-1,2,3,4-tetrahydroisoquinolin-7-yl)(hydroxy)methyl)piperidine-1-carboxyla-
te (8a, 91.0 mg, 0.139 mmol) in THF (3.0 mL) was treated with
iodomethane (34 mg, 0.24 mmol) and potassium tert-butoxide (0.155
mL of a 1.0M solution in THF, 0.155 mmol). Stirring was continued
at 0.degree. C. for 30 minutes, then the mixture partitioned
between brine and MTBE. The organic phase was concentrated to
dryness, to give crude racemic tert-butyl
4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-
-1,2,3,4-tetrahydroisoquinolin-7-yl)(methoxy)methyl)piperidine-1-carboxyla-
te (34b, 97 mg, 100%) as a gum. MS: 612, 614 [M+H-tBu].sup.+.
[0590] Trifluoroacetic acid (0.10 mL, 1.35 mmol) was added to a
room temperature solution of crude racemic tert-butyl
4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-
-1,2,3,4-tetrahydroisoquinolin-7-yl)(methoxy)methyl)piperidine-1-carboxyla-
te (34b, 97 mg, 0.139 mmol) in dichloromethane (5.0 mL). The
mixture was stirred at room temperature for 2 hours, at 35.degree.
C. for 4 hours, at room temperature overnight, then at 40.degree.
C. for 6 hours. The solution was diluted with heptane and
concentrated to dryness, leaving crude racemic
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(methox-
y(piperidin-4-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (34c, 124
mg, 100%) as a gum. MS: 568, 570 [M+H].sup.+.
[0591] To a cooled (0.degree. C.) solution of crude racemic
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(methox-
y(piperidin-4-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (34c, 124
mg, 0.139 mmol) in dichloromethane (3.0 mL) was added triethylamine
(75 uL, 0.54 mmol) and 2-acetoxyacetyl chloride (16 uL, 0.15 mmol).
The mixture was stirred at 0.degree. C. for 30 minutes.
Trifluoroacetic acid (2.0 mL) was then added and the mixture
stirred at room temperature for 1 hour, then at 40.degree. C. for 7
hours. The solution was concentrated to dryness and further dried
under high vacuum for 2 days, giving crude racemic
2-(4-((5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-y-
l)methyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)(methoxy)methyl)piperid-
in-1-yl)-2-oxoethyl acetate (34d, 204 mg, 100%) as a golden oil.
MS: 578, 580 [M+H].sup.+.
[0592] The crude racemic
2-(4-((5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-
)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)(methoxy)methyl)piperidin-1-yl)-
-2-oxoethyl acetate (34d, 204 mg, 0.139 mmol) was dissolved in a 7N
solution of ammonia in methanol (4 mL, 28 mmol NH3), stirred at
room temperature for 1 hour, then stirred at 40.degree. C. for 4
hours. The solution was concentrated to dryness and the residue
purified by preparative HPLC to give
(.+-.)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-
]-7-{[1-(hydroxyacetyl)piperidin-4-yl](methoxy)methyl}-3,4-dihydroisoquino-
lin-1(2H)-one (Example 34, 19.87 mg, 27% yield from 8a) as a white
solid. .sup.1H NMR (400 MHz, CD3OD) .delta. 7.44 (s, 1H), 6.01 (s,
1H), 4.66 (s, 2H), 4.56 (d, J=5.62 Hz, 1H), 4.35-4.45 (m, 1H),
4.02-4.17 (m, 2H), 3.57-3.68 (m, 1H), 3.43 (t, J=6.24 Hz, 2H), 3.10
(s, 3H), 2.86-2.92 (m, 2H), 2.75-2.85 (m, 1H), 2.40-2.54 (m, 1H),
2.20 (s, 3H), 2.15 (s, 3H), 1.78-1.87 (m, 1H), 1.60-1.69 (m, 1H),
1.18-1.47 (m, 3H). MS: 536, 538 [M+H].sup.+.
[0593] The racemate (Example 34) was further purified by chiral
preparative SFC, affording, after lyophilization, Example 35
(Isomer B, retention time 13.019 min, 9.09 mg, 12% yield from 8a)
and Example 36 (Isomer A, retention time 10.712 min, 8.36 mg, 11%
yield from 8a) as white solids. The absolute configuration of the
benzylic carbon in each isomer was not determined.
Example 35
[0594]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-{[1-(hydroxyacetyl)piperidin-4-yl](methoxy)methyl}-3,4-dihydroisoquinol-
in-1(2H)-one--Isomer B MS: 536, 538 [M+H].sup.+. Chiral analysis:
.about.97% ee, retention time 13.019 min on a Lux Cellulose-4
4.6.times.100 mm 3 u column, eluting with 50% MeOH, 120 bar, 4
mL/min.
Example 36
[0595]
5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-{[1-(hydroxyacetyl)piperidin-4-yl](methoxy)methyl}-3,4-dihydroisoquinol-
in-1(2H)-one--Isomer A MS: 536, 538 [M+H].sup.+. Chiral analysis:
>99% ee, retention time 10.712 min on a Lux Cellulose-4
4.6.times.100 mm 3 u column, eluting with 50% MeOH, 120 bar, 4
mL/min.
Example 81
5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]--
7-[(R)-methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-1(2H)-one
Example 82
5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]--
7-[(S)-methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-1(2H)-one
##STR00046##
[0597] To a stirred, room temperature solution of
oxetan-3-ylmethanol (2.20 g, 24.97 mmol) in dichloromethane (110
mL) was added solid pyridinium dichromate (5.87 g, 15.6 mmol) in
five portions. The resulting black mixture was stirred at room
temperature for 16 hours. The deep brown suspension was then
filtered through a silica gel pad and the filter cake rinsed with
dichloromethane (8.times.120 mL). The combined dichloromethane
filtrates were partially concentrated under reduced pressure at
room temperature (27.about.30.degree. C.) to afford
oxetane-3-carbaldehyde (81a, 3 g, .about.26% yield) as a colorless
solution, 18.7 wt % solution in dichloromethane by NMR. The
solution was dried over magnesium sulfate, filtered and used
immediately in the next step. 1H NMR (400 MHz, CDCl3) .delta. 9.95
(d, J=2.4 Hz, 1H), 4.87 (m, 4H), 3.81 (m, 1H).
[0598] A solution of
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-7-bromo-5,8-dich-
loro-3,4-dihydroisoquinolin-1(2H)-one (Cpd SS, 500 mg, 0.932 mmol)
in anhydrous THF (7 mL) was cooled to -65.degree. C., then
isopropylmagnesium chloride lithium chloride complex (1.3 M
solution in THF, 2.15 mL, 2.80 mmol) was added dropwise over 3
minutes. The resulting brown solution was stirred at -65.degree. C.
for 10 minutes, then warmed to -10.degree. C. for 30 minutes. To
this was oxetane-3-carbaldehyde (81a, .about.2.2 g, .about.4.8
mmol, .about.18.7 wt % solution in dichloromethane) dropwise over 2
minutes, causing the color to change to light yellow. Stirring was
continued at -5.degree. C. for 30 minutes. The reaction was
quenched with glacial acetic acid (0.5 mL) and diluted with ethyl
acetate (10 mL), then washed with sat. aq. NaHCO3/sat. aq. NaCl
(1/1 v/v, 3.times.15 mL). The organic layer was dried over sodium
sulfate, filtered, concentrated, and purified by silica gel
chromatography (eluting with 1/1 petroleum ether/ethyl acetate) to
give
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-5,8-dichloro-7-(-
hydroxy(oxetan-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (81b,
300 mg, 59% yield, racemate) as a white solid. MS: 543
[M+H].sup.+.
[0599] Iodomethane (133 mg, 0.938 mmol) was added dropwise to a
cooled (-5.degree. C.) suspension of
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-5,8-dichloro-7-(-
hydroxy(oxetan-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (81b,
300 mg, 0.552 mmol) in anhydrous THF (5 mL). Potassium
tert-butoxide (1.0M solution in THF, 0.938 mL, 0.938 mmol) was
added, and the mixture stirred at 0.degree. C. for 1 hour. The
reaction mixture was partitioned between sat. aq. NaCl (15 mL) and
MTBE (3.times.15 mL). The combined organic extracts were washed
with sat. aq. NaCl (30 mL), dried over sodium sulfate,
concentrated, and purified by silica gel chromatography (eluting
with 1/1 petroleum ether/ethyl acetate) to give racemic
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-5,8-dichloro-7-(-
methoxy(oxetan-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (81c,
280 mg, 91% yield) as a yellow gum. MS: 557 [M+H].sup.+.
[0600] A room-temperature mixture of
2-((2-(benzyloxy)-4-methoxy-6-methyl
pyridin-3-yl)methyl)-5,8-dichloro-7-(methoxy(oxetan-3-yl)methyl)-3,4-dihy-
droisoquinolin-1(2H)-one (81c, 100 mg, 0.179 mmol) and PtO2 (21 mg,
0.092 mmol) in ethyl acetate (4 mL) was stirred under a hydrogen
balloon for 3 days. The solution was filtered through a Celite pad.
The flask and filter pad were rinsed with ethyl acetate (2.times.10
mL). The combined filtrates were concentrated and purified by
preparative thin layer chromatography (silica gel, eluting with
10/1 dichloromethane/methanol) to give racemic
5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-[methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-1(2H)-one
(81d, 45 mg, 54% yield) as a white solid.
[0601] Multiple batches of racemic
5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-[methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-1(2H)-one
(81d, 140 mg total) were combined for chiral separation by
preparative SFC [Column: (R,R)Whelk O1 250 mm*30 mm, 5 um; mobile
phase: base-ETOH; wavelength: 220 nm; workup: lyophilization] to
give Example 81 (50.34 mg, 36% yield) as a gray solid, and, after
further purification by preparative TLC (silica gel, eluting with
10/1 dichloromethane/methanol), Example 82 (22.83 g, 16% yield) as
a brown solid. A small-molecule X-Ray crystal structure of Example
82 shows it to have absolute (S) stereochemistry, so absolute (R)
stereochemistry was attributed to its enantiomer, Example 81.
Example 81
[0602]
5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)m-
ethyl]-7-[(R)-methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-1(2H)-one-
. .sup.1H NMR (400 MHz, CDCl3) .delta. 12.34 (brs, 1H), 7.49 (s,
1H), 5.93 (s, 1H), 5.05 (d, J=6.0 Hz, 1H), 4.78-4.61 (m, 6H), 3.88
(s, 3H), 3.50-3.48 (m, 2H), 3.38-3.37 (m, 1H), 3.31 (s, 3H), 2.94
(t, J=6.2 Hz, 2H), 2.35 (s, 3H). MS: 489 [M+Na].sup.+. Chiral
analysis: 100% ee; retention time 9.85 min; column (R,R)Whelk O1,
250.times.4.6 mm I.D., 5 um; mobile phase 50% ethanol (0.05% DEA)
in CO2; wavelength 220 nm.
Example 82
[0603]
5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)m-
ethyl]-7-[(S)-methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-1(2H)-one-
. .sup.1H NMR (400 MHz, CDCl3) .delta. 12.38 (brs, 1H), 7.49 (s,
1H), 5.92 (s, 1H), 5.05 (d, J=6.0 Hz, 1H), 4.78-4.64 (m, 6H), 3.87
(s, 3H), 3.50-3.47 (m, 2H), 3.38-3.37 (m, 1H), 3.31 (s, 3H), 2.93
(t, J=6.2 Hz, 2H), 2.35 (s, 3H). MS: 467 [M+H].sup.+. Chiral
analysis: 98% ee; retention time 8.65 min; column: (R,R)Whelk O1,
250.times.4.6 mm I.D., 5 um; mobile phase 50% ethanol (0.05% DEA)
in CO2; wavelength 220 nm.
Example 83
8-chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(-
R)-methoxy[(3R)-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquinol-
in-1(2H)-one
Example 84
8-chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(-
R*)-methoxy[(3S*)-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquin-
olin-1 (2H)-one
Example 85
8-chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(-
S*)-methoxy[(3R*)-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquin-
olin-(2H)-one
Example 86
8-chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(-
S)-methoxy[(3S)-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquinol-
in-1(2H)-one
##STR00047##
[0605] Aqueous tetrahydrofuran-3-carboxaldehyde solution
(.about.4.0 mL of 50 wt % in water, 4.0 g) was extracted with
dichloromethane (2.times.2.5 mL). The combined organic layers were
cooled (15.degree. C.) and phosphorus pentoxide slowly added. The
resulting dark solution was filtered to remove solids and the
yellow filtrate (.about.16 wt % tetrahydrofuran-3-carboxaldehyde in
dichloromethane by NMR) was promptly used as described below.
[0606] Isopropylmagnesium chloride lithium chloride complex (1.3 M
solution in THF, 3.73 mL, 4.85 mmol) was added dropwise to a cooled
(-70.degree. C.) solution of
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-iodo--
5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd UU, 910 mg, 1.617
mmol) in anhydrous THF (10 mL) over 5 minutes. The resulting brown
mixture was stirred at -70.degree. C. for 30 minutes, then the
tetrahydrofuran-3-carboxaldehyde solution prepared above (3.98 g of
.about.16 wt % in dichloromethane, .about.636 mg
tetrahydrofuran-3-carboxaldehyde, .about.16.35 mmol) was added
dropwise over 5 minutes. Stirring was continued at -70.degree. C.
for 30 minutes. The reaction was quenched with glacial acetic acid
(0.5 mL) and diluted with ethyl acetate (80 mL), then washed with
sat. aq. NaHCO3/sat. aq. NaCl (1/1 v/v, 3.times.35 mL). The organic
layer was dried over sodium sulfate, filtered, concentrated, and
purified by silica gel chromatography (eluting with 1/1 petroleum
ether/ethyl acetate) to give
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-(hydr-
oxy(tetrahydrofuran-3-yl)methyl)-5-methyl-3,4-dihydroisoquinolin-1(2H)-one
(83a, 450 mg, 52% yield, mixture of 4 diastereomers) as a white
solid. .sup.1H NMR (400 MHz, CDCl3) .delta. 7.37-7.46 (m, 3H),
7.20-7.31 (m, 3H), 6.39 (s, 1H), 5.44 (s, 2H), 5.22-5.31 (m, 1H),
4.87 (s, 2H), 3.88-4.02 (m, 1H), 3.83 (s, 3H), 3.65-3.82 (m, 3H),
3.16 (t, J=5.99 Hz, 2H), 2.71-2.89 (m, 1H), 2.55 (t, J=6.17 Hz,
2H), 2.44 (s, 3H), 2.20 (s, 3H), 1.66-1.95 (m, 3H).
[0607] Iodomethane (225 mg, 1.58 mmol) was added dropwise to a
cooled (-5.degree. C.) suspension of
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-(hydr-
oxy(tetrahydrofuran-3-yl)methyl)-5-methyl-3,4-dihydroisoquinolin-1(2H)-one
(83a, 500 mg, 0.931 mmol in THF (15 mL), followed by potassium
tert-butoxide (1.0M solution in THF, 1.58 mL, 1.58 mmol). The
mixture was stirred at 0.degree. C. for 1 hour. Glacial acetic acid
(0.5 mL) and ethyl acetate (100 mL) were added, and the solution
washed with sat. aq. NaHCO3 (3.times.20 mL) and brine (30 mL). The
organic layer was dried over sodium sulfate, filtered,
concentrated, and purified by silica gel chromatography (eluting
with 1/1 petroleum ether/ethyl acetate), to give
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-(meth-
oxy(tetrahydrofuran-3-yl)methyl)-5-methyl-3,4-dihydroisoquinolin-1(2H)-one
as a mixture of 4 diastereomers (400 mg, 70% yield).
[0608] The stereoisomers were separated by preparative chiral SFC
(column: AD, 250*30 mm, 5 um, mobile phase: 30% IPA+NH3H2O 60
mL/min, wavelength: 220 nm, workup: lyophilization) to give 140 mg
of peak 12 (mixture of peak 1 and 2) and 120 mg of peak 34 (mixture
of peak 3 and 4) as white solids.
[0609] The peak 12 mixture (140 mg) was re-separated by preparative
SFC (column: AD, 250*30 mm, 5 um, mobile phase: 25% MeOH+NH3H2O 60
mL/min, wavelength: 220 nm, workup: lyophilization) to give
enantio-enriched peak 1 (60 mg, 81% chiral purity, further purified
as described below) and pure peak 2 (1610b, 60 mg, 12% yield, 96%
chiral purity, used without further purification).
[0610] The peak 34 mixture (120 mg) was re-separated by preparative
SFC (Column: AD, 250*30 mm, 5 um, mobile phase: 25% MeOH+NH3H2O 60
mL/min, wavelength: 220 nm, workup: lyophilization) to give pure
peak 3 (1613b, 50 mg, 9.8% yield, 95% chiral purity, used without
further purification) and enantio-enriched peak 4 (50 mg, 88%
chiral purity, further purified as described below).
[0611] The enantio-enriched peak 1 material (60 mg, 81% chiral
purity) was further purified by preparative SFC (column: AD, 250*30
mm, 5 um, mobile phase: 25% MeOH+NH3H2O 70 mL/min, wavelength: 220
nm, workup: lyophilization) to give pure peak 1 (83b, 45 mg, 8.7%
yield, 98% chiral purity).
[0612] The enantio-enriched peak 4 material (50 mg, 88% chiral
purity) was further purified by preparative SFC (column: AD, 250*30
mm, 5 um, mobile: 25% MeOH+NH3H2O 70 mL/min, wavelength: 220 nm,
workup: lyophilization) to give pure peak 4 (1613b, 45 mg, 8.7%
yield, 99% chiral purity). Absolute or relative stereochemistry of
each isomer was not determined at this stage.
[0613] A stirred solution of
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-(meth-
oxy(tetrahydrofuran-3-yl)methyl)-5-methyl-3,4-dihydroisoquinolin-1(2H)-one
peak 1 (83b, 45 mg, 0.082 mmol) in dichloromethane (2 mL) was
treated with TFA (2 mL) at room temperature. The mixture was heated
to 30.degree. C. for 16 hours, then the solution was diluted with
dichloromethane (20 mL) and concentrated to dryness. The residue
was purified by silica gel chromatography (eluting with 10/1
dichloromethane/methanol) to give
8-chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{-
(R)-methoxy[(3R)-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquino-
lin-1(2H)-one (Example 83, 18.55 mg, 50% yield, 100% ee) as a white
solid. A small molecule X-Ray crystal structure of Example 83
confirms it has absolute (R,R) configuration. .sup.1H NMR (400 MHz,
CDCl3) .delta. 12.36 (brs, 1H), 7.29 (s, 1H), 5.91 (s, 1H),
4.85-4.75 (m, 3H), 3.89-3.83 (m, 6H), 3.71-3.69 (m, 1H), 3.47-3.44
(m, 2H), 3.16 (s, 3H), 2.76-2.73 (m, 2H), 2.58-2.54 (m, 1H), 2.34
(s, 3H), 2.26 (s, 3H), 1.73-1.70 (m, 2H). MS: 461 [M+H].sup.+.
Chiral analysis: 100% ee; retention time 34.91 min; column:
Chiralpak IC 250.times.4.6 mm I.D., 5 um; mobile phase: 50% ethanol
(0.05% DEA) in CO2; flow rate: 2.0 mL/min; wavelength: 220 nm.
[0614] By the same procedure, the peak 2 stereoisomer (1610b, 60.0
mg, 0.109 mmol) afforded
8-chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{-
(R*)-methoxy[(3S*)-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoqui-
nolin-1(2H)-one (Example 84, 30 mg, 60% yield, 97% ee) as a white
solid. The absolute stereochemistry of this isomer was not
determined, but the .sup.1HNMR spectrum shows clear differences
from that of Example 83, suggesting that Example 84 is either the
R,S or S,R isomer. .sup.1H NMR (400 MHz, CDCl3) .delta. 12.33 (br.
s, 1H), 7.30 (s, 1H), 5.91 (s, 1H), 4.84-4.77 (m, 3H), 3.89-3.86
(m, 4H), 3.73-3.68 (m, 2H), 3.60-3.58 (m, 1H), 3.46-3.44 (m, 2H),
3.19 (s, 3H), 2.75-2.73 (m, 2H), 2.64-2.62 (m, 1H), 2.34 (s, 3H),
2.24 (s, 3H), 1.98-1.95 (m, 2H). MS: 461 [M+H].sup.+. Chiral
analysis: 97% ee, retention time 39.01 min; column: Chiralpak IC
250.times.4.6 mm I.D., 5 um; mobile phase: 50% ethanol (0.05% DEA)
in CO2; flow rate: 2.0 mL/min; wavelength: 220 nm.
[0615] By the same procedure, the peak 3 stereoisomer (85b, 50.0
mg, 0.091 mmol) afforded
8-chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{-
(S*)-methoxy[(3R*)-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoqui-
nolin-1(2H)-one (Example 85, 16.06 mg, 38% yield, 100% ee) as a
white solid. The absolute stereochemistry of this isomer was not
determined, but the .sup.1HNMR spectrum shows clear differences
from that of Example 83, and is identical to that of Example 84,
suggesting that Example 85 is either the S,R or R,S isomer. .sup.1H
NMR (400 MHz, CDCl3) .delta. 12.30 (br s, 1H), 7.30 (s, 1H), 5.91
(s, 1H), 4.84-4.77 (m, 3H), 3.89-3.86 (m, 4H), 3.74-3.66 (m, 2H),
3.59-3.58 (m, 1H), 3.46-3.44 (m, 2H), 3.19 (s, 3H), 2.75-2.73 (m,
2H), 2.65-2.63 (m, 1H), 2.34 (s, 3H), 2.24 (s, 3H), 1.98-1.95 (m,
2H). MS: 461 [M+H].sup.+. Chiral analysis: 100% ee, retention time
29.05 min; column: Chiralpak IC 250.times.4.6 mm I.D., 5 um; mobile
phase; 50% ethanol (0.05% DEA) in CO2; flow rate: 2.0 mL/min;
wavelength: 220 nm.
[0616] By the same procedure, the peak 4 stereoisomer (86b, 45.0
mg, 0.082 mmol) afforded
8-chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{-
(S)-methoxy[(3S)-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquino-
lin-1(2H)-one (Example 86, 14.79 mg, 39% yield, 100% ee) as a white
solid. Though it has a different retention time on the chiral
column, the .sup.1HNMR spectrum of this compound is identical to
that of Example 83, which was shown to have R,R configuration by
X-Ray crystal structure. This suggests that Example 86 is the S,S
stereoisomer. .sup.1H NMR (400 MHz, CDCl3) .delta. 12.29 (br. s,
1H), 7.30 (s, 1H), 5.91 (s, 1H), 4.84-4.75 (m, 3H), 3.89-3.83 (m,
6H), 3.71-3.69 (m, 1H), 3.47-3.44 (m, 2H), 3.16 (s, 3H), 2.76-2.73
(m, 2H), 2.58-2.56 (m, 1H), 2.34 (s, 3H), 2.25 (s, 3H), 1.73-1.70
(m, 2H). MS: 461 [M+H].sup.+. Chiral analysis: 100% ee, retention
time 32.28 min; column: Chiralpak IC 250.times.4.6 mm I.D., 5 um;
mobile phase: 50% ethanol (0.05% DEA) in CO2; flow rate: 2.0
mL/min; wavelength: 220 nm.
Method J
Example 87
5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]--
7-[1-(1-methylazetidin-3-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one--Isome-
r A
Example 88
5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]--
7-[1-(1-methylazetidin-3-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one--Isome-
r B
##STR00048## ##STR00049## ##STR00050##
[0618] A solution of 1-boc-azetidine-3-carboxylic acid (5.00 g,
24.8 mmol, and CDI (4.23 g, 26.1 mmol) in dichloromethane (100 mL)
was stirred at room temperature for 1 hour, then
N,O-dimethylhydroxylamine hydrochloride (4.0 g, 29.8 mmol) was
added and stirring continued at room temperature for 16 hours. The
resulting suspension was washed with water (3.times.30 mL), sat.
aq. NaHCO3 (3.times.30 mL), and brine (3.times.30 mL). The organic
layer was dried over sodium sulfate, filtered, and concentrated to
give tert-butyl 3-(methoxy(methyl)carbamoyl)azetidine-1-carboxylate
(87a, 5.1 g, 84% yield) as a colorless oil. .sup.1H NMR (400 MHz,
CDCl3) .delta. 4.14 (br s, 2H), 4.05 (t, J=8.6 Hz, 2H), 3.66 (s,
3H), 3.65 (m, 1H), 3.20 (s, 3H), 1.43 (s, 9H).
[0619] Methylmagnesium bromide (3M solution in THF, 10.4 mL, 31.3
mmol) was added dropwise to a cooled (0.degree. C.) solution of
tert-butyl 3-(methoxy(methyl)carbamoyl)azetidine-1-carboxylate
(87a, 5.1 g, 20.88 mmol) in anhydrous THF (100 mL). Stirring was
continued at 0.degree. C. for one hour, then at room temperature
for 16 hours. The mixture was cooled to 0.degree. C. and quenched
with sat. aq. NaHCO3 (35 mL), then extracted with ethyl acetate
(3.times.40 mL). The combined organic extracts were washed with
brine (3.times.40 mL), dried over sodium sulfate, filtered,
concentrated, and purified by silica gel chromatography (eluting
with petroleum ether/ethyl acetate from 10:1 to 3:1) to give
tert-butyl 3-acetylazetidine-1-carboxylate (87b, (3.20 g, 77%
yield) as light yellow oil. .sup.1H NMR (400 MHz, CDCl3) .delta.
4.05 (d, J=7.6 Hz, 4H), 3.41 (quint, J=7.6 Hz, 1H), 2.18 (s, 3H),
1.43 (s, 9H).
[0620] A solution of
2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-7-bromo-5,8-dich-
loro-3,4-dihydroisoquinolin-1(2H)-one (Cpd SS, 1.20 g, 2.238 mmol)
in anhydrous THF (15 mL) was cooled to -60.degree. C., then
isopropylmagnesium chloride lithium chloride complex (1.3 M
solution in THF, 5.16 mL, 6.71 mmol) was added dropwise via syringe
over 3 minutes. Stirring was continued at -60.degree. C. for 10
minutes, then at 0.degree. C. for 20 minutes. To this was added
tert-butyl 3-(methoxy(methyl)carbamoyl)azetidine-1-carboxylate
(87a, 892 mg, 4.48 mmol) dropwise, then the mixture stirred at
0.degree. C. for 1 hour. The mixture was quenched with glacial
acetic acid (1 mL) and diluted with ethyl acetate (100 mL). The
organic phase was washed with NaHCO3/brine (v/v=1/1, 3.times.50 mL)
and brine (50 mL), dried over sodium sulfate, and filtered. The
filtrate was concentrated to give the crude product (2.0 g, yellow
oil), which was purified by silica gel chromatography (eluting with
petroleum ether/ethyl acetate=1:1) to give racemic tert-butyl
3-(1-(2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-5,8-dichlo-
ro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-1-hydroxyethyl)azetidine-1-ca-
rboxylate (87c, 450 mg, 31% yield) as a white solid. MS: 656
[M+H].sup.+.
[0621] Trifluoromethanesulfonic acid (0.54 mL, 6.15 mmol) was added
dropwise to a cooled (0.degree. C.) solution of racemic tert-butyl
3-(1-(2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-5,8-dichlo-
ro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-1-hydroxyethyl)azetidine-1-ca-
rboxylate (87c, 450 mg, 0.685 mmol) in anhydrous dichloromethane
(10 mL). The mixture was stirred at 5-10.degree. C. for 1 hour,
then re-cooled to 0.degree. C. and more trifluoromethanesulfonic
acid (0.54 mL, 6.15 mmol) was added. After stirring at 2-5.degree.
C. for 12 hours, sat. aq. sodium bicarbonate was added to bring the
solution to pH .about.8. The mixture was concentrated to remove
dichloromethane, and the aqueous residue diluted with THF (20 mL).
Solid sodium bicarbonate (288 mg, 3.43 mmol) and di-tert-butyl
dicarbonate (448 mg, 2.06 mmol) were added and the mixture stirred
at 2-5.degree. C. for 16 hours, then let stand at 15.degree. C. for
18 hours. The solution was extracted with ethyl acetate (2.times.50
mL). The combined organic layers were washed with brine (2.times.20
mL), dried over sodium sulfate, and filtered. The filtrate was
concentrated to give the crude product (1 g, yellow solid), which
was purified first by silica gel chromatography (eluting with 10%
methanol in dichloromethane) and then re-purified by preparative
SFC (column: AD 250 mm*30 mm, 5 um; mobile phase: 35% Base-ETOH;
wavelength: 220 nm; workup: concentration) to give tert-butyl
3-(1-(5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)m-
ethyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)vinyl)azetidine-1-carboxyl-
ate (87d, 256 mg, 68.1%) as a yellow solid. MS: 548
[M+H].sup.+.
[0622] A suspension of tert-butyl
3-(1-(5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)m-
ethyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)vinyl)azetidine-1-carboxyl-
ate (87d, 236 mg, 0.43 mmol) and platinum oxide (80 mg, 0.35 mmol)
in ethyl acetate (15 mL) and methanol (5 mL) was stirred at room
temperature under a hydrogen balloon for 3 hours. After filtration
to remove solids, the filtrate was concentrated and purified by
preparative TLC (silica gel, dichloromethane/methanol=15:1 to give
racemic tert-butyl
3-(1-(5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)m-
ethyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)ethyl)azetidine-1-carboxyl-
ate (87e, 180 mg, 76% yield) as a white solid. MS: 549
[M+H].sup.+.
[0623] A solution of give racemic tert-butyl
3-(1-(5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)m-
ethyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)ethyl)azetidine-1-carboxyl-
ate (87e, 180 mg, 0.327 mmol) in dichloromethane (10 mL) was
stirred with HCl (4.0 M solution in methanol, 5 mL, 20 mmol) at
14.degree. C. for 30 minutes. The solution was concentrated to
dryness, and the residue dissolved in methanol (5 mL). Concentrated
NH4OH was added to bring the pH to .about.8, and the mixture was
again concentrated to dryness, leaving crude, racemic
7-(1-(azetidin-3-yl)ethyl)-5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2--
dihydropyridin-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (87f,
180 mg, 100%) as a white solid, which was used without further
purification.
[0624] Glacial acetic acid (0.1 mL) was added to a 15.degree. C.
solution of crude, racemic
7-(1-(azetidin-3-yl)ethyl)-5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2--
dihydropyridin-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (87f,
180 mg, 0.327 mmol) and formaldehyde (37 wt % in water, 79.5 mg,
0.980 mmol) in methanol (5 mL), and the mixture stirred at
15.degree. C. for 45 minutes. Sodium cyanoborohydride (41 mg, 0.653
mmol) was added and stirring continued at room temperature for 12
hours. The mixture was quenched with saturated NH4Cl solution (2
mL) and stirred at 15.degree. C. for 30 minutes, then concentrated
to remove the solvent. The residue was dissolved in
dichloromethane/methanol (v/v=10:1, 50 mL) and filtered. The
filtrate was concentrated and purified by preparative HPLC [column:
Phenomenex Gemini C18 250*50 10 u; mobile phase: from 4% to 34%
acetonitrile(with 0.225% formic acid) in water; wavelength: 220 nm;
workup: lyophilization] to give the formate salt of
(.+-.)-5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)-
methyl)-7-(1-(1-methylazetidin-3-yl)ethyl)-3,4-dihydroisoquinolin-1(2H)-on-
e (110 mg, 66%) as a white solid. The enantiomers of this racemic
salt were separated by preparative SFC [column: AD(250 mm*30 mm, 5
um); mobile phase: 30% base-ETOH; wavelength: 220 nm; workup:
lyophilization], and each enantiomer separately re-purified by
preparative HPLC [column: Phenomenex Gemini C18 250*50 10 u; mobile
phase: from 28% MeCN (0.05% ammonia) in water to 48% MeCN (0.05%
ammonia) in water; wavelength: 220 nm; workup: lyophilization],
affording isomer A (Example 87, 15.67 mg, 16% yield) and isomer B
(Example 88, 13.35 mg, 13% yield) as white solids. The absolute
stereochemistry was not determined for either isomer.
Example 87
[0625]
5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)m-
ethyl]-7-[1-(1-methylazetidin-3-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one-
--Isomer A. .sup.1H NMR (400 MHz, CD3OD) .delta. 7.44 (s, 1H), 6.27
(s, 1H), 4.73 (s, 2H), 3.91 (m, 3H), 3.77-3.68 (m, 2H), 3.41-3.37
(m, 3H), 3.16 (t, J=7.4 Hz, 1H), 2.95-2.92 (m, 2H), 2.90-2.83 (m,
2H), 2.39 (s, 3H), 2.34 (s, 3H), 1.16 (d, J=6.4 Hz, 3H). MS: 464
[M+H].sup.+. Chiral analysis: 99% ee; retention time 5.511 min on
Chiralpak AD-3 150.times.4.6 mm I.D., 3 um column [mobile phase: A:
CO2 B:ethanol (0.05% DEA); gradient: from 5% to 40% of B in 5.0 min
and hold 40% for 2.5 min, then 5% of B for 2.5 min; wavelength: 220
nm].
Example 88
[0626]
5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)m-
ethyl]-7-[1-(1-methylazetidin-3-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one-
--Isomer B. .sup.1H NMR (400 MHz, CD3OD) .delta. 7.43 (s, 1H), 6.26
(s, 1H), 4.73 (s, 2H), 3.91 (m, 3H), 3.77-3.68 (m, 2H), 3.41-3.37
(m, 3H), 3.12 (brs, 1H), 2.95-2.92 (m, 2H), 2.90-2.83 (m, 2H), 2.36
(s, 3H), 2.33 (s, 3H), 1.16 (d, J=6.4 Hz, 3H). MS: 464 [M+H].sup.+.
Chiral analysis: 100% ee; retention time 5.997 min on Chiralpak
AD-3 150.times.4.6 mm I.D., 3 um column [mobile phase: A: CO2
B:ethanol (0.05% DEA); gradient: from 5% to 40% of B in 5.0 min and
hold 40% for 2.5 min, then 5% of B for 2.5 min; wavelength: 220
nm].
Method K
Example 89
(.+-.)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-
-7-[1-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one
Example 90
(+)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7--
[1-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one
Example 91
(-)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7--
[1-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one
##STR00051## ##STR00052##
[0628] To a cooled (-40.degree. C.) solution of
2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3-
,4-dihydroisoquinolin-1(2H)-one (Cpd S, 5.00 g, 9.61 mmol) in
anhydrous THF (50 mL) and 1,4-dioxane (5 mL) was added
isopropylmagnesium chloride-lithium chloride complex (1.3 M
solution in THF, 22.2 mL, 28.8 mmol) via syringe. After stirring at
-40.degree. C. for 30 minutes, zinc chloride (1.0 M solution in
ether, 11.5 mL, 11.5 mmol) was added. Stirring was continued at
-40.degree. C. for 30 minutes, then
tetrakis(triphenylphosphine)palladium(0) (1.11 g, 0.961 mmol) and
acetyl chloride (1.51 g, 19.2 mmol) were added. The mixture was
stirred and allowed to warm to room temperature over 18 hours. The
reaction was quenched with sat. aq. NH4Cl (5 mL), diluted with
ethyl acetate (30 mL), and washed with sat. aq. NH4Cl (40 mL) and
brine (40 mL). The organic layer was dried over sodium sulfate,
filtered, concentrated, and purified by silica gel chromatography
(eluting with 0-30% ethyl acetate in petroleum ether) to give
7-acetyl-2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro--
3,4-dihydroisoquinolin-1(2H)-one (89a, 4.00 g, 78% yield) as a
yellow solid. .sup.1H NMR (400 MHz, CDCl3) .delta. 7.45 (d, J=6.8
Hz, 2H), 7.39 (s, 1H), 7.38-7.27 (m, 3H), 6.63 (s, 1H), 5.43 (s,
2H), 4.86 (s, 2H), 3.29 (t, J=6.3 Hz, 2H), 2.75 (t, J=6.3 Hz, 2H),
2.63 (s, 3H), 2.42 (s, 3H), 2.34 (s, 3H). MS: 483 [M+H].sup.+.
[0629] Sodium borohydride (47.0 mg, 1.24 mmol) was added to a room
temperature solution of
7-acetyl-2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro--
3,4-dihydroisoquinolin-1(2H)-one (89a, 200 mg, 0.414 mmol) in
methanol (5 mL). After stirring at room temperature for 30 minutes,
the reaction mixture was concentrated and purified by silica gel
chromatography (eluting with 0-30% ethyl acetate in petroleum
ether) to give
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-hydr-
oxyethyl)-3,4-dihydroisoquinolin-1(2H)-one (89b, 190 mg, 95% yield)
as a white solid.
[0630] A cooled (0.degree. C.) solution of
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-hydr-
oxyethyl)-3,4-dihydroisoquinolin-1(2H)-one (89b, 190 mg, 0.391
mmol) and triethylamine (119 mg, 1.17 mmol) in dichloromethane (5
mL) was treated with methanesulfonyl chloride (67.3 mg, 0.587
mmol), then stirred at 0.degree. C. for one hour. The reaction
mixture was diluted with dichloromethane (30 mL); washed
sequentially with sat. aq. NH4Cl, sat. aq. NaHCO3, and sat. aq.
NaCl; dried over sodium sulfate, and concentrated to dryness,
leaving crude racemic
1-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)ethyl methanesulfonate (89c, 225
mg, 100% yield) as a white solid, which was used immediately
without further purification.
[0631] A suspension of
1-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo--
1,2,3,4-tetrahydroisoquinolin-7-yl)ethyl methanesulfonate (89c, 100
mg, 0.177 mmol), morpholine (46.4 mg, 0.532 mmol), and potassium
carbonate (73.6 mg, 0.532 mmol) in acetonitrile (5 mL) was stirred
at reflux (85.degree. C.) for 2 hours. After cooling to room
temperature, the suspension was filtered to remove solids. The
filtrate was concentrated and purified by silica gel chromatography
(eluting with 0-30% ethyl acetate in petroleum ether) to give
racemic
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-morp-
holinoethyl)-3,4-dihydroisoquinolin-1(2H)-one (89d, 90 mg, 91%
yield, 80% pure by LCMS) as a gum. MS: 576 [M+Na].sup.+.
[0632] A solution of racemic
2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-morp-
holinoethyl)-3,4-dihydroisoquinolin-1(2H)-one (89d, 90 mg, 0.16
mmol) in dichloromethane (5 mL) and trifluoroacetic acid (2 mL) was
stirred at room temperature for 19 hours. The solution was
evaporated to dryness, and then the residue was dissolved in
toluene (10 mL) and basified to pH 8-9 by adding a few drops of
conc. NH4OH. The solution was concentrated and purified by
preparative HPLC to give
(.+-.)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-
]-7-[1-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one
(Example 89, 29.49 mg, 39% yield) as a white solid. .sup.1H NMR
(400 MHz, CDCl3) .delta. 7.74 (s, 1H), 5.94 (s, 1H), 4.87-4.70 (m,
2H), 4.00 (q, J=6.3 Hz, 1H), 3.76-3.59 (m, 6H), 2.99-2.81 (m, 2H),
2.53 (br. s., 2H), 2.36 (m, 5H), 2.29 (s, 3H), 1.24 (d, J=6.5 Hz,
3H). MS: 464 [M+H].sup.+.
[0633] The racemic material (Example 89) was further purified by
chiral SFC separation conditions to provide the compounds of
Example 90 and Example 91.
[0634] Additional compounds of the invention were prepared by
modifications of the methods exemplified herein. Selected compounds
prepared and corresponding characterization data are presented in
Table 1 below.
TABLE-US-00001 TABLE 1 Ex. Meth- .sup.1H NMR (ppm); Stereochemistry
No. Structure/IUPAC name od LCMs [M + H].sup.+ Note 1 ##STR00053##
A .sup.1H NMR (400 MHz, CD3OD) .delta. 7.63 (s, 1H), 6.12 (s, 1H),
4.77 (s, 2H), 3.94- 3.90 (m, 1H), 3.81- 3.80 (m, 3H), 3.59- 3.57
(m, 2H), 3.51- 3.49 (m, 2H), 3.17- 3.10 (m, 1H), 2.98- 2.95 (m,
2H), 2.71 (br s, 1H), 2.30 (s, 3H), 2.29-2.25 (m, 1H), 2.25 (s,
3H), 1.83- 1.78 (m, 1H); LCMs [M + H].sup.+ 465 R,R isomer;
stereo-chemistry determined from x-ray crystal structure of
enantiomeric compound (Ex. 4); Chiral purity: 95.66%; retention
time: 6.867 min; column: Chiralpak AD-3 150 .times. 4.6 mm I.D., 3
um; mobile phase: 5-40% ethanol (0.05% DEA) in CO2; flow rate: 2.5
mL/min 2 ##STR00054## A .sup.1H NMR (400 MHz, CD3OD) .delta. 7.62
(s, 1H), 6.11 (s, 1H), 4.76 (s, 2H), 4.13- 4.11 (m, 1H), 3.78- 3.75
(m, 1H), 3.69- 3.68 (m, 2H), 3.61- 3.59 (m, 3H), 3.51- 3.50 (m,
2H), 2.98- 2.95 (m, 2H), 2.65 (br s, 1H), 2.30 (s, 3H), 2.25 (s,
3H), 1.77- 1.75 (m, 1H), 1.42- 1.37 (m, 1H); LCMs [M + H].sup.+ 465
Single enantiomer, either R,S or S,R but absolute stereochemistry
unknown; Enantiomer of Ex. 3; Chiral purity: 98.70%; retention
time: 7.309 min; column: Chiralpak AD-3 150 .times. 4.6 mm I.D., 3
um; mobile phase: 5-40% ethanol (0.05% DEA) in CO2; flow rate: 2.5
mL/min 3 ##STR00055## A .sup.1H NMR (400 MHz, CD3OD) .delta. 7.62
(s, 1H), 6.11 (s, 1H), 4.76 (s, 2H), 4.12- 4.11 (m, 1H), 3.80- 3.78
(m, 1H), 3.69- 3.67 (m, 3H), 3.67- 3.62 (m, 2H), 3.61- 3.50 (m,
2H), 2.98- 2.95 (m, 2H), 2.65 (br s, 1H), 2.29 (s, 3H), 2.25 (s,
3H), 1.77- 1.74 (m, 1H), 1.42- 1.37 (m, 1H); LCMs [M + H].sup.+ 465
Single enantiomer, either S,R or R,S isomer but absolute
stereochemistry unknown; Enantiomer of Ex. 2 Chiral purity: 96.48%;
retention time: 8.021 min; column: Chiralpak AD-3 150 .times. 4.6
mm I.D., 3 um; mobile phase: 5-40% ethanol (0.05% DEA) in CO2; flow
rate: 2.5 mL/min 4 ##STR00056## A .sup.1H NMR (400 MHz, CD3OD)
.delta. 7.64 (s, 1H), 6.13 (s, 1H), 4.78 (m, 1H), 3.95- 3.90 (m,
1H), 3.83- 3.81 (m, 3H), 3.60- 3.55 (m, 2H), 3.55- 3.52 (m, 2H),
3.32- 3.19, (m, 1H), 2.99- 2.96 (m, 2H), 2.75 (br s, 1H), 2.32 (s,
3H), 2.31-2.29 (m, 1H), 2.27 (s, 3H), 1.84- 1.79 (m, 1H); LCMs [M +
H].sup.+ 465 Known to be S,S by X-ray crystal structure; Enantiomer
of Ex. 1; Chiral purity: 99.18%; retention time: 8.429 min; column:
Chiralpak AD-3 150 .times. 4.6 mm I.D., 3 um; mobile phase: 5-40%
ethanol (0.05% DEA) in CO2; flow rate: 2.5 mL/min 5 ##STR00057## B
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.66 (br. s., 1H), 7.80 (d,
J = 3.67 Hz, 1H), 6.00 (s, 1H), 5.15-5.43 (m, 1H), 4.85 (br. s.,
1H), 4.69 (s, 2H), 4.03-4.13 (m, 1H), 3.66-3.84 (m, 2H), 3.50-3.63
(m, 2H), 3.02-3.10 (m, 1H), 2.94-3.02 (m, 2H), 2.71-2.90 (m, 2H),
2.42-2.55 (m, 1H), 2.28 (s, 3H), 2.24 (s, 3H), 2.09-2.23 (m, 1H),
1.87-2.08 (m, 1H); LCMs [M + H].sup.+ 482 Mixture of diastereomers
containing (R)-3- fluoropyrrolidine Mixture separated to Ex. 6 and
Ex. 7 6 ##STR00058## B HNMR was not taken due to limited quantity.
See Ex. 5; LCMs [M + H].sup.+ 482 One component of the Ex. 5
mixture. Single diasteromer, containing (R)-3- fluoropyrrolidine,
other chiral center undetermined; >99% de (-), [.alpha.]D =
-51.3.degree.(c 0.01 MeOH) 1st peak; RT 1.18 min Chiralcel OJ-3 4.6
.times. 100 mm 3 u column; 10% MeOH/DEA @ 120 bar, 4 mL/min 7
##STR00059## B HNMR was not taken due to limited quantity. See Ex
5; LCMs [M + H].sup.+ 482 One component of the Ex. 5 mixture;
single diastereomer containing (R)-3- fluoropyrrolidine, other
chiral center undetermined; ~88% de (+) [.alpha.]D =
+62.1.degree.(c 0.01 MeOH) 2nd peak; RT 1.42 min Chiralcel OJ-3 4.6
.times. 100 mm 3 u column; 10% MeOH/DEA @ 120 bar, 4 mL/min; 8
##STR00060## C .sup.1H NMR (600 MHz, DMSO-d6) .delta. ppm 11.55 (s,
1 H), 7.58 (d, J = 4.95 Hz, 1 H), 5.88 (s, 1 H), 5.72- 5.84 (m, 1
H), 4.55 (q, J = 13.75 Hz, 2 H), 4.46 (br. s., 1 H), 4.30-4.42 (m,
1 H), 4.05-4.13 (m, 1 H), 3.97-4.04 (m, 1 H), 3.69 (t, J = 13.39
Hz, 1 H), 3.45 (t, J = 5.78 Hz, 2 H), 2.80-2.98 (m, 3 H), 2.20 (d,
J = 18.71 Hz, 1 H), 2.11 (s, 3 H), 1.61 (br. s., 1 H), 1.46 (br.
s., 1 H), 1.34-1.42 (m, 1 H), 1.23 (s, 5 H); LCMs [M + H].sup.+ 524
>99% ee (+), [.alpha.]D = +9.9.degree. (c 0.1 DMSO) First peak
off column: Lux Cellulose-2 4.6 .times. 100 mm 3 u column 60% MeOH
@ 120 bar, 4 mL/min Peak 1 @ 8.13 min (prep: OJ-H, 21 .times. 250
mm column, 32 mL MeOH: 8 mL CO2, 100 bar, 40 mL/min) 9 ##STR00061##
C .sup.1H NMR (600 MHz, DMSO-d6) .delta. ppm 11.55 (s, 1 H), 7.58
(d, J = 4.95 Hz, 1 H), 5.88 (s, 1 H), 5.72-5.84 (m, 1 H), 4.55 (q,
J = 13.75 Hz, 2 H), 4.46 (br. s., 1 H), 4.30-4.42 (m, 1 H),
4.05-4.13 (m, 1 H), 3.97-4.04 (m, 1 H), 3.69 (t, J = 13.39 Hz, 1
H), 3.45 (t, J = 5.78 Hz, 2 H), 2.80-2.98 (m, 3 H), 2.20 (d, J =
18.71 Hz, 1 H), 2.11 (s, 3 H), 1.61 (br. s., 1 H), 1.46 (br. s., 1
H), 1.34-1.42 (m, 1 H), 1.23 (s, 5 H); LCMs [M + H].sup.+ 524 ~95%
ee (-), [.alpha.]D = -6.5.degree. (c 0.1 DMSO) Second peak off
column: Lux Cellulose-2 4.6 .times. 100 mm 3 u column 60% MeOH @
120 bar, 4 mL/min Peak 2 @ 10.29 min (prep: OJ-H, 21 .times. 250 mm
column, 32 mL MeOH: 8 mL CO2, 100 bar, 40 mL/min) 10 ##STR00062## D
.sup.1H NMR (400 MHz, CDCl3) .delta. 12.23 (br. s., 1H), 7.53 (s,
1H), 5.95 (s, 1H), 4.92- 4.89 (m, 1H), 4.78 (s, 2H), 3.80-3.71 (m,
1H), 3.68-3.62 (m, 2H), 3.53-3.51 (m, 1H), 3.33 (s, 3H), 2.94 (t, J
= 6.0 Hz, 2H), 2.35 (s, 3H), 2.29 (s, 3H); LCMs [M + H].sup.+ 425
Single isomer, absolute stereochemistry unknown: Enantiomer of Ex.
11; 100% ee; retention time 7.717 min; column: Chiralpak IC-3 150
.times. 4.6 mm I.D., 3 um; mobile phase: 40% ethanol (0.05% DEA) in
CO2; flow rate: 2.35 mL/min 11 ##STR00063## D .sup.1H NMR (400 MHz,
CDCl3) .delta. 12.15 (br. s., 1H), 7.53 (s, 1H), 5.95 (s, 1H),
4.92- 4.89 (m, 1H), 4.78 (s, 2H), 3.80-3.71 (m, 1H), 3.68-3.63 (m,
2H), 3.55-3.51 (m, 1H), 3.34 (s, 3H), 2.94 (t, J = 5.6 Hz, 2H),
2.35 (s, 3H), 2.29 (s, 3H); LCMs [M + H].sup.+ 425 Single isomer,
absolute stereochemistry unknown; Enantiomer of Ex. 10; 100% ee;:
retention time 11.063 min; column: Chiralpak IC-3 150 .times. 4.6
mm I.D., 3 um; mobile phase: 40% ethanol (0.05% DEA) in CO2; flow
rate: 2.35 mL/min 12 ##STR00064## E .sup.1H NMR (400 MHz, CD3OD)
.delta. 7.58 (s, 1H), 6.11 (s, 1H), 4.76 (s, 2H), 4.10-4.21 (m,
1H), 3.86-3.99 (m, 3H), 3.75-3.84 (m, 1H), 3.64-3.72 (m, 1H),
3.46-3.55 (m, 2H), 2.91-3.01 (m, 2H), 2.29 (s, 3H), 2.25 (s, 3H),
1.73-1.97 (m, 3H), 1.44-1.58 (m, 1H); LCMs [M + H].sup.+ 465 Single
enantiomer: (R) at benzylic carbon and (S) at THF 91% ee 1st peak;
RT 2.91 min Chiralpak AD-3 4.6 .times. 100 mm 3 u column; 5-60%
MeOH in 3 minutes, 120 bar, 4 mL/min 13 ##STR00065## E .sup.1H NMR
(400 MHz, CD3OD) .delta. 7.66 (s, 1H), 6.10 (s, 1H), 4.77 (s, 2H),
4.27 (dt, J = 7.86, 6.16 Hz, 1H), 3.72-3.91 (m, 3H), 3.67 (t, J =
6.79 Hz, 2H), 3.49 (t, J = 6.24 Hz, 2H), 2.95 (t, J = 6.17 Hz, 2H),
2.28 (s, 3H), 2.24 (s, 3H), 2.01-2.13 (m, 1H), 1.68-1.92 (m, 2H),
1.49-1.62 (m, 1H); LCMs [M + H].sup.+ 465 Single enantiomer (R,R);
93% ee 2nd Peak; RT 3.21 min Chiralpak AD-3 4.6 .times. 100 mm 3 u
column; 5-60% MeOH in 3 minutes, 120 bar, 4 mL/min 14 ##STR00066##
F .sup.1H NMR (400 MHz, CDCl3) .delta. 7.35 (s, 1H), 5.90 (s, 1H),
4.78 (s, 2H), 3.63 (t, J = 6.30 Hz, 2H), 3.5- 3.6 (m, 1H), 2.90 (t,
J = 6.11 Hz, 2H), 2.35 (s, 3H), 2.24 (s, 3H), 1.24 (d, J = 6.85 Hz,
6H); LCMs [M + H].sup.+ 393 N/A 15 ##STR00067## G .sup.1H NMR (400
MHz, CDCl3) .delta. 11.88 (br. s., 1H), 7.53 (s, 1H), 5.94 (s, 1H),
4.78 (s, 2H), 4.69-4.66 (m, 1H), 3.65 (t, J = 4.8 Hz, 2H), 3.24 (s,
3H), 2.93 (t, J = 6.2 Hz, 2H), 2.36 (s, 3H), 2.29 (s, 3H), 1.76-
1.72 (m, 1H), 1.65- 1.60 (m, 1H), 0.96 (t, J = 7.0 Hz, 3H); LCMs [M
+ H].sup.+ 423 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 16; 100% Chiral Purity; 1st peak, RT 8.04 min;
column: Chiralpak AD-H 250 .times. 4.6 mm I.D., 5 um; mobile phase:
5-40% methanol (0.05% DEA) in CO2; flow rate: 2.5 mL/min 16
##STR00068## G .sup.1H NMR (400 MHz, CDCl3) .delta. 10.88 (br s,
1H), 7.53 (s, 1H), 5.93 (s, 1H), 4.77 (s, 2H), 4.69-4.66 (m, 1H),
3.67-3.63 (m, 2H), 3.24 (s, 3H), 2.93 (t, J = 5.8 Hz, 2H), 2.36 (s,
3H), 2.28 (s, 3H), 1.78- 1.73 (m, 1H), 1.65- 1.58 (m, 1H), 0.96 (t,
J = 7.2 Hz, 3H); LCMs [M + H].sup.+ 423 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 15; 99.6194% Chiral
Purity; 2nd peak, RT 8.34 min; column: Chiralpak AD-H 250 .times.
4.6 mm I.D., 5 um; mobile phase: 5-40% methanol (0.05% DEA) in CO2;
flow rate: 2.5 mL/min 17 ##STR00069## A .sup.1H NMR (400 MHz,
CDCl3) .delta. 11.54 (s, 1H), 7.37 (s, 1H), 5.93 (s, 1H), 4.78 (s,
2H), 3.79-3.71 (m, 3H), 3.64-3.61 (m, 2H), 2.91 (t, J = 6 Hz, 2H),
2.34 (s, 3H), 2.28 (s, 3H), 1.43 (s, 1H), 1.27 (t, J = 6.4 Hz, 3H);
LCMs [M + H].sup.+ 409 Racemic mixture 18 ##STR00070## A .sup.1H
NMR (400 MHz, CDCl3) .delta. 11.54 (s, 1H), 7.37 (s, 1H), 5.93 (s,
1H), 4.78 (s, 2H), 3.79-3.71 (m, 3H), 3.64-3.61 (m, 2H), 2.91 (t, J
= 6 Hz, 2H), 2.34 (s, 3H), 2.28 (s, 3H), 1.43 (s, 1H), 1.27 (t, J =
6.4 Hz, 3H); LCMs [M + H].sup.+ 409 Single isomer, known; (S)
stereochemistry determined from x- ray crystal structure; 1st peak
under the following SFC conditions: Chiralpak AS-H 4.6 .times. 100
mm 5 u column; 20% MeOH @ 120 bar CO2, 4 mL/min 19 ##STR00071## A
.sup.1H NMR (400 MHz, CDCl3) .delta. 11.54 (s, 1H), 7.37 (s, 1H),
5.93 (s, 1H), 4.78 (s, 2H), 3.79-3.71 (m, 3H), 3.64-3.61 (m, 2H),
2.91 (t, J = 6 Hz, 2H), 2.34 (s, 3H), 2.28 (s, 3H), 1.43 (s, 1H),
1.27 (t, J = 6.4 Hz, 3H); LCMs [M + H].sup.+ 409 Single isomer,
known; (R) stereochemistry determined from x-ray crystal structure
of enantiomeric compound (Ex. 18); 2nd peak under the following SFC
conditions: Chiralpak AS-H 4.6 .times. 100 mm 5 u column; 20% MeOH
@ 120 bar CO2, 4 mL/min 20 ##STR00072## A .sup.1H NMR (400 MHz,
CD3OD) .delta. 7.49 (s, 1H), 6.11 (s, 1H), 4.76 (s, 2H), 3.94 (s,
2H), 3.71-3.66 (m, 2H), 3.52-3.50 (m, 3H), 2.95 (t, J = 6.4 Hz,
2H), 2.29 (s, 3H), 2.25 (s, 3H), 1.90- 1.82 (m, 1H), 1.65- 1.55 (m,
1H), 0.85 (t, J = 7.6 Hz, 3H); LCMs [M + H].sup.+ 423 Racemic
mixture 21 ##STR00073## A .sup.1H NMR (400 MHz, CD3OD) .delta. 7.63
(s, 1H), 6.10 (s, 1H), 4.75 (s, 2H), 3.97 (sxt, J = 6.94 Hz, 1H),
3.79 (t, J = 5.38 Hz, 2H), 3.52 (t, J = 6.17 Hz, 2H), 3.33-3.40 (m,
1H), 3.00-3.19 (m, 3H), 2.92-3.00 (m, 2H), 2.72 (s, 3H), 2.30 (s,
3H), 2.25 (s, 3H), 1.33 (d, J = 6.85 Hz, 3H); LCMs [M + H].sup.+
466 Racemic mixture 22 ##STR00074## A .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 12.50 (br. s., 1H), 11.54 (br. s., 1H), 7.66 (s,
1H), 5.88 (s, 1H), 4.57 (s, 2H), 3.77 (s, 2H), 3.45 (t, J = 5.50
Hz, 2H), 2.88 (t, J = 5.07 Hz, 2H), 2.16 (s, 3H), 2.12 (s, 3H);
LCMs [M + H].sup.+ 409 N/A 23 ##STR00075## F .sup.1H NMR (400 MHz,
CD3OD) .delta. 7.48 (s, 1H), 6.12 (s, 1H), 4.77 (s, 2H), 4.50 (d, J
= 13.57 Hz, 1H), 4.14-4.29 (m, 2H), 3.73 (d, J = 13.45 Hz, 1H),
3.48-3.56 (m, 2H), 2.92-3.03 (m, 3H), 2.79 (d, J = 7.09 Hz, 2H),
2.66 (t, J = 12.53 Hz, 1H), 2.31 (s, 3H), 2.26 (s, 3H), 1.93-2.06
(m, 1H), 1.70 (d, J = 13.57 Hz, 2H), 1.21-1.34 (m, 2H); LCMs [M +
H].sup.+ 506 N/A 24 ##STR00076## J .sup.1H NMR (400 MHz, CD3OD)
.delta. 7.40 (s, 1H), 6.01 (s, 1H), 4.66 (s, 2H), 4.27-4.49 (m,
1H), 4.00-4.18 (m, 2H), 3.49- 3.73 (m, 1H), 3.41 (t, J = 6.24 Hz,
2H), 3.23- 3.30 (m, 1H), 2.75- 2.95 (m, 3H), 2.41-2.63 (m, 1H),
2.19 (s, 3H), 2.15 (s, 3H), 1.80-1.89 (m, 1H), 1.67-1.79 (m, 1H),
1.28-1.36 (m, 1H), 0.97-1.22 (m, 5H); LCMs [M + H].sup.+ 520
Racemic mixture 25 ##STR00077## G .sup.1H NMR (400 MHz, CDCl3)
.delta. 13.22 (br. s., 1H), 7.41 (s, 1H), 6.01 (s, 1H), 5.13 (dd, J
= 7.76, 3.36 Hz, 1H), 4.85 (d, J = 13.94 Hz, 1H), 4.53 (d, J =
13.94 Hz, 1H), 3.70 (dt, J = 12.50, 4.94 Hz, 1H), 3.41-3.53 (m,
1H), 2.79 (t, J = 5.99 Hz, 2H), 2.42 (s, 3H), 2.31 (s, 3H),
1.67-1.82 (m, 1H), 1.41-1.57 (m, 1H), 0.96 (t, J = 7.34 Hz, 3H);
LCMs [M + H].sup.+ 409 Racemic mixture 26 ##STR00078## A .sup.1H
NMR (400 MHz, CD3OD) .delta. 7.52 (s, 1H), 6.13 (s, 1H), 4.78 (s,
2H), 3.91- 3.84 (m, 1H), 3.53- 3.48 (m, 3H), 2.97 (t, J = 6.2 Hz,
2H), 2.31 (s, 3H), 2.27 (s, 3H), 1.29 (d, J = 7.2 Hz, 3H), 1.14 (d,
J = 6.0 Hz, 3H); LCMs [M + H].sup.+ 423 Racemic mixture of (2R,3S)
and (2S,3R) isomers separated enantiomers are Ex. 47 and Ex. 48 27
##STR00079## G .sup.1H NMR (400 MHz, CDCl3) .delta. 13.22 (br. s.,
1H), 7.41 (s, 1H), 6.01 (s, 1H), 5.13 (dd, J = 7.76, 3.36 Hz, 1H),
4.85 (d, J = 13.94 Hz, 1H), 4.53 (d, J = 13.94 Hz, 1H), 3.70 (dt, J
= 12.50, 4.94 Hz, 1H), 3.41- 3.53 (m, 1H), 2.79 (t, J = 5.99 Hz,
2H), 2.42 (s, 3H), 2.31 (s, 3H), 1.67-1.82 (m, 1H), 1.41-1.57 (m,
1H), 0.96 (t, J = 7.34 Hz, 3H); LCMs [M + H].sup.+ 409 Single
isomer, absolute stereochemistry unknown; Enantiomer of Ex. 28;
[.alpha.]D = +48.8.degree. (c 0.1 MeOH) >99% ee (+); RT 1.407
min; column: Chiralpak AS-3 4.6 .times. 100 mm 3 u; 20% MeOH/DEA @
120 bar CO2, 4 mL/min; 28 ##STR00080## G .sup.1H NMR (400 MHz,
CDCl3) .delta. 13.22 (br. s., 1H), 7.41 (s, 1H), 6.01 (s, 1H), 5.13
(dd, J = 7.76, 3.36 Hz, 1H), 4.85 (d, J = 13.94 Hz, 1H), 4.53 (d, J
= 13.94 Hz, 1H), 3.70 (dt, J = 12.50, 4.94 Hz, 1H), 3.41- 3.53 (m,
1H), 2.79 (t, J = 5.99 Hz, 2H), 2.42 (s, 3H), 2.31 (s, 3H),
1.67-1.82 (m, 1H), 1.41-1.57 (m, 1H), 0.96 (t, J = 7.34 Hz, 3H);
LCMs [M + H].sup.+ 409 Single isomer, absolute stereochemistry
unknown; Enantiomer of Ex. 27; [.alpha.]D = -47.5.degree. (c 0.1
MeOH) ~99% ee (-); RT 1.893 min; column: Chiralpak AS-3 4.6 .times.
100 mm 3 u; 20% MeOH/DEA @ 120 bar CO2, 4 mL/min; 29 ##STR00081## A
.sup.1H NMR (400 MHz, CD3OD) .delta. 7.49 (s, 1H), 6.11 (s, 1H),
4.76 (s, 2H), 3.71- 3.66 (m, 2H), 3.52- 3.50 (m, 3H), 2.95 (t, J =
6.4 Hz, 2H), 2.29 (s, 3H), 2.25 (s, 3H), 1.90-1.82 (m, 1H),
1.65-1.55 (m, 1H), 0.85 (t, J = 7.6 Hz, 3H); LCMs [M + H].sup.+ 423
Single isomer, absolute stereochemistry unknown; Enantiomer of Ex.
30; 1st peak under the following SFC conditions: Chiralpak AD-3 4.6
.times. 100 mm 3 u column; 30% MeOH @ 120 bar CO2, 4 mL/min 30
##STR00082## A .sup.1H NMR (400 MHz, CD3OD) .delta. 7.49 (s, 1H),
6.11 (s, 1H), 4.76 (s, 2H), 3.71-3.66 (m, 2H), 3.52-3.50 (m, 3H),
2.95 (t, J = 6.4 Hz, 2H), 2.29 (s, 3H), 2.25 (s, 3H), 1.90-1.82 (m,
1H), 1.65-1.55 (m, 1H), 0.85 (t, J = 7.6 Hz, 3H); LCMs [M +
H].sup.+ 423 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 29; 2nd peak under the following SFC conditions:
Chiralpak AD-3 4.6 .times. 100 mm 3 u column; 30% MeOH @ 120 bar
CO2, 4 mL/min 31 ##STR00083## A .sup.1H NMR (400 MHz, CD3OD)
.delta. 11.65 (bs, 1H),
7.43 (s, 1H), 5.95 (s, 1H), 4.77 (s, 2H), 4.04-3.87 (m, 4H),
3.68-3.40 (m, 3H), 3.31-3.30 (m, 2H), 2.92 (t, J = 6.4 Hz, 2H),
2.37 (s, 3H), 2.29 (s, 3H), 2.05- 1.82 (m, 2H), 1.32- 1.28 (m, 4H);
LCMs [M + H].sup.+ 479 Racemic mixture 32 ##STR00084## G .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 11.55 (br. s., 1H), 7.77 (s, 1H),
5.89 (s, 1H), 4.61-4.68 (m, 1H), 4.58 (s, 2H), 3.42- 3.50 (m, 2H),
2.87- 2.95 (m, 2H), 2.16 (s, 3H), 2.12 (s, 3H), 1.56-1.63 (m, 3H);
LCMs [M + H].sup.+ 404 Racemic mixture 33 ##STR00085## A .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 7.53 (s, 1H), 5.92 (s, 1H), 4.57 (s,
2H), 3.48- 3.47 (m, 1H), 3.41 (t, J = 6.2 Hz, 2H), 3.34 (t, J = 6.6
Hz, 2H), 2.85 (t, J = 6.0 Hz, 2H), 2.16 (s, 3H), 2.13 (s, 3H),
1.78- 1.68 (m, 2H), 1.15 (d, J = 6.8 Hz, 3H); LCMs [M + H].sup.+
423 Racemic mixture 34 ##STR00086## I .sup.1H NMR (400 MHz, CD3OD)
.delta. 7.44 (s, 1H), 6.01 (s, 1H), 4.66 (s, 2H), 4.56 (d, J = 5.62
Hz, 1H), 4.35- 4.45 (m, 1H), 4.02- 4.17 (m, 2H), 3.57- 3.68 (m,
1H), 3.43 (t, J = 6.24 Hz, 2H), 3.10 (s, 3H), 2.86-2.92 (m, 2H),
2.75-2.85 (m, 1H), 2.40-2.54 (m, 1H), 2.20 (s, 3H), 2.15 (s, 3H),
1.78- 1.87 (m, 1H), 1.60- 1.69 (m, 1H), 1.18- 1.47 (m, 3H); LCMs [M
+ H].sup.+ 536 Racemic mixture 35 ##STR00087## I NMR of Racemate,
Ex. 34: .sup.1H NMR (400 MHz, CD3OD) .delta. 7.44 (s, 1H), 6.01 (s,
1H), 4.66 (s, 2H), 4.56 (d, J = 5.62 Hz, 1H), 4.35-4.45 (m, 1H),
4.02-4.17 (m, 2H), 3.57-3.68 (m, 1H), 3.43 (t, J = 6.24 Hz, 2H),
3.10 (s, 3H), 2.86-2.92 (m, 2H), 2.75-2.85 (m, 1H), 2.40-2.54 (m,
1H), 2.20 (s, 3H), 2.15 (s, 3H), 1.78-1.87 (m, 1H), 1.60-1.69 (m,
1H), 1.18-1.47 (m, 3H); LCMs [M + H].sup.+ 536 Single isomer,
absolute stereochemistry unknown; Enantiomer of Ex. 36; ~97% ee;
retention time 13.019 min; Lux Cellulose-4 4.6 .times. 100 mm 3 u
column; 50% MeOH @ 120 bar CO2, 4 mL/min 36 ##STR00088## I NMR of
Racemate, Ex. 34: .sup.1H NMR (400 MHz, CD3OD) .delta. 7.44 (s,
1H), 6.01 (s, 1H), 4.66 (s, 2H), 4.56 (d, J = 5.62 Hz, 1H),
4.35-4.45 (m, 1H), 4.02-4.17 (m, 2H), 3.57-3.68 (m, 1H), 3.43 (t, J
= 6.24 Hz, 2H), 3.10 (s, 3H), 2.86-2.92 (m, 2H), 2.75-2.85 (m, 1H),
2.40-2.54 (m, 1H), 2.20 (s, 3H), 2.15 (s, 3H), 1.78-1.87 (m, 1H),
1.60-1.69 (m, 1H), 1.18-1.47 (m, 3H); LCMs [M + H].sup.+ 536 Single
isomer, absolute stereochemistry unknown; Enantiomer of Ex. 35;
>99% ee; retention time 10.712 min; Lux Cellulose-4 4.6 .times.
100 mm 3 u column; 50% MeOH @ 120 bar CO2, 4 mL/min 37 ##STR00089##
A .sup.1H NMR (400 MHz, CD3OD) .delta. 11.65 (bs, 1H), 7.43 (s,
1H), 5.95 (s, 1H), 4.77 (s, 2H), 4.04-3.87 (m, 4H), 4H), 3.68-3.40
(m, 3H), 3.31-3.30 (m, 2H), 2.92 (t, J = 6.4 Hz, 2H), 2.37 (s, 3H),
2.29 (s, 3H), 2.05- 1.82 (m, 2H), 1.32- 1.28 (m, 4H); LCMs [M +
H].sup.+ 479 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 38; 1st peak under the following SFC conditions:
Chiralpak AD-3 4.6 .times. 100 mm 3 u column; 30% MeOH @ 120 bar
CO2, 4 mL/min 38 ##STR00090## A .sup.1H NMR (400 MHz, CD3OD)
.delta. 11.65 (bs, 1H), 7.43 (s, 1H), 5.95 (s, 1H), 4.77 (s, 2H),
4.04-3.87 (m, 4H), 3.68- 3.40 (m, 3H), 3.31-3.30 (m, 2H), 2.92 (t,
J = 6.4 Hz, 2H), 2.37 (s, 3H), 2.29 (s, 3H), 2.05-1.82 (m, 2H),
1.32-1.28 (m, 4H); LCMs [M + H].sup.+ 479 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 37; 2nd peak under the
following SFC conditions: Chiralpak AD-3 4.6 .times. 100 mm 3 u
column; 30% MeOH @ 120 bar CO2, 4 mL/min 39 ##STR00091## A .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 7.53 (s, 1H), 5.92 (s, 1H), 4.57 (s,
2H), 3.48- 3.47 (m, 1H), 3.41 (t, J = 6.2 Hz, 2H), 3.34 (t, J = 6.6
Hz, 2H), 2.85 (t, J = 6.0 Hz, 2H), 2.16 (s, 3H), 2.13 (s, 3H),
1.78- 1.68 (m, 2H), 1.15 (d, J = 6.8 Hz, 3H); LCMs [M + H].sup.+
423 Single enantiomer, absolute stereochemistry unknown; 1st peak
under the following SFC conditions: Lux Cellulose-4 4.6 .times. 100
mm 3 u column; 50% MeOH @ 120 bar, 4 mL/min 40 ##STR00092## A
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 7.53 (s, 1H), 5.92 (s, 1H),
4.57 (s, 2H), 3.48-3.47 (m, 1H), 3.41 (t, J = 6.2 Hz, 2H), 3.34 (t,
J = 6.6 Hz, 2H), 2.85 (t, J = 6.0 Hz, 2H), 2.16 (s, 3H), 2.13 (s,
3H), 1.78- 1.68 (m, 2H), 1.15 (d, J = 6.8 Hz, 3H); LCMs [M +
H].sup.+ 423 Single enantiomer, absolute stereochemistry unknown;
2nd peak under the following SFC conditions: Lux Cellulose-4 4.6
.times. 100 mm 3 u column; 50% MeOH @ 120 bar, 4 mL/min 41
##STR00093## G .sup.1H NMR (400 MHz, CDCl3) .delta. 12.02 (br. s.,
1H), 7.78 (s, 1H), 6.28 (s, 1H), 4.74 (s, 2H), 4.58 (dq, J = 3.06,
6.48 Hz, 1H), 4.24 (qd, J = 6.21, 7.79 Hz, 1H), 3.68 (t, J = 6.11
Hz, 2H), 3.54 (dd, J = 2.93, 8.07 Hz, 1H), 2.97 (t, J = 6.11 Hz,
2H), 2.52 (s, 3H), 2.40 (s, 3H), 1.18 (d, J = 6.36 Hz, 3H), 1.05
(d, J = 6.36 Hz, 3H); LCMs [M + H].sup.+ 423 Racemic mixture of
2,4-anti diols, stereochemistry at 3-position unknown. 42
##STR00094## G .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.54 (br.
s., 1H), 7.74 (s, 1H), 5.88 (s, 1H), 4.57 (s, 2H), 4.54 (br. s.,
2H), 4.07 (quin, J = 5.81 Hz, 2H), 3.44 (t, J = 6.11 Hz, 2H), 3.24
(t, J = 5.87 Hz, 1H), 2.86 (t, J = 5.99 Hz, 2H), 2.17 (s, 3H), 2.12
(s, 3H), 1.02 (d, J = 6.11 Hz, 6H); LCMs [M + H].sup.+ 453 Single
achiral/meso 2,4- syn diol, stereochemistry at 3-position unknown
43 ##STR00095## A .sup.1H NMR (700 MHz, DMSO-d6) .delta. 8.84 (d, J
= 1.51 Hz, 1H), 7.60 (s, 1H), 6.65 (d, J = 1.51 Hz, 1H), 5.88 (s,
1H), 5.14 (br. s, 1H), 4.88 (t, J = 6.99 Hz, 1H), 4.57 (s, 2H),
3.93-4.00 (m, 2H), 3.43 (t, J = 6.24 Hz, 2H), 2.86 (t, J = 6.24 Hz,
2H), 2.15 (s, 3H), 2.12 (s, 3H); LCMs [M + H].sup.+ 462 Racemic
Mixture 44 ##STR00096## G .sup.1H NMR (400 MHz, DMSO-d6) .delta.
11.53 (br. s., 1H), 7.52 (s, 1H), 5.88 (s, 1H), 4.78 (q, J = 6.24
Hz, 1H), 4.57 (s, 2H), 3.45 (t, J = 6.24 Hz, 2H), 3.18 (s, 3H),
2.89 (t, J = 6.11 Hz, 2H), 2.16 (s, 3H), 2.12 (s, 3H), 1.32 (d, J =
6.36 Hz, 3H); LCMs [M + H].sup.+ 409 Racemic mixture 45
##STR00097## E .sup.1H NMR (400 MHz, CD3OD) .delta. 7.58 (s, 1H),
6.10 (s, 1H), 4.76 (s, 2H), 4.09- 4.22 (m, 1H), 3.86- 4.00 (m, 3H),
3.75- 3.84 (m, 1H), 3.62- 3.72 (m, 1H), 3.45- 3.55 (m, 2H), 2.95
(t, J = 6.17 Hz, 2H), 2.29 (s, 3H), 2.25 (s, 3H), 1.72-1.98 (m,
3H), 1.44-1.58 (m, 1H); LCMs [M + H].sup.+ 465 Single enantiomer, S
at benzyl, R at THF 91% ee 1st peak; RT 2.91 min Chiralpak AD-3 4.6
.times. 100 mm 3 u column, 5-60% MeOH in 3 minutes, 120 bar, 4
mL/min 46 ##STR00098## E .sup.1H NMR (400 MHz, CD3OD) .delta. 7.66
(s, 1H), 6.10 (s, 1H), 4.76 (s, 2H), 4.23- 4.32 (m, 1H), 3.73- 3.91
(m, 3H), 3.67 (t, J = 6.79 Hz, 2H), 3.50 (t, J = 6.24 Hz, 2H), 2.95
(t, J = 6.17 Hz, 2H), 2.29 (s, 3H), 2.25 (s, 3H), 2.01- 2.11 (m,
1H), 1.80- 1.92 (m, 1H), 1.69- 1.80 (m, 1H), 1.49- 1.62 (m, 1H);
LCMs [M + H].sup.+ 465 Single enantiomer, S,S 92% ee 2nd peak; RT
3.21 min Chiralpak AD-3 4.6 .times. 100 mm 3 u column, 5-60% MeOH
in 3 minutes, 120 bar, 4 mL/min 47 ##STR00099## A .sup.1H NMR (400
MHz, CD3OD) .delta. 7.52 (s, 1H), 6.13 (s, 1H), 4.78 (s, 2H), 3.89-
3.86 (m, 1H), 3.54- 3.48 (m, 3H), 2.97 (t, J = 6.2 Hz, 2H), 2.31
(s, 3H), 2.27 (s, 3H), 1.29 (d, J = 7.2 Hz, 3H), 1.14 (d, J = 6.4
Hz, 3H); LCMs [M + H].sup.+ 423 Single enantiomer; relative
stereochemistry known; absolute stereochemistry unknown; 99.08% ee;
1st peak, RT 9.67 min Column: Chiralpak AD-H 250 .times. 4.6 mm
I.D., 5 um; mobile phase: 5-40% isopropanol (0.05% DEA) in CO2;
flow rate: 2.5 mL/min 48 ##STR00100## A .sup.1H NMR (400 MHz,
CD3OD) .delta. 7.52 (s, 1H), 6.13 (s, 1H), 4.78 (s, 2H), 3.91- 3.84
(m, 1H), 3.53-3.48 (m, 3H), 2.97 (t, J = 6.0 Hz, 2H), 2.31 (s, 3H),
2.27 (s, 3H), 1.29 (d, J = 7.2 Hz, 3H), 1.14 (d, J = 6.0 Hz, 3H);
LCMs [M + H].sup.+ 423 Single enantiomer; relative stereochemistry
known; absolute stereochemistry unknown; 96.73% ee; 2nd peak, RT
10.21 min Column: Chiralpak AD-H 250 .times. 4.6 mm I.D., 5 um;
mobile phase: 5-40% isopropanol (0.05% DEA) in CO2; flow rate: 2.5
mL/min 49 ##STR00101## D .sup.1H NMR (400 MHz, CDCl3) .delta. 7.51
(s, 1H), 5.93 (s, 1H), 5.32 (s, 1H), 4.76 (s, 2H), 3.75-3.60 (m,
2H), 3.39 (s, 3H), 3.00-2.90 (m, 2H), 2.36 (s, 3H), 2.27 (s, 3H),
2.19 (s, 3H), 3H), 2.19 (s, 3H), 1.26 (s, 1H); LCMs [M + H].sup.+
437 Racemic mixture 50 ##STR00102## I .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 11.54 (br. s., 1H), 7.45 (s, 1H), 5.88 (s, 1H),
4.56 (s, 3H), 3.74- 3.89 (m, 2H), 3.46 (t, J = 6.11 Hz, 2H), 3.14-
3.24 (m, 2H), 3.13 (s, 3H), 2.89 (t, J = 6.11 Hz, 2H), 2.17 (s,
3H), 2.12 (s, 3H), 1.76- 1.90 (m, 1H), 1.32- 1.56 (m, 3H), 1.17-
1.27 (m, 1H); LCMs [M + H].sup.+ 479 [.alpha.]D = -56.4.degree. (c
0.1, MeOH) ~96% ee (-); retention time 3.17 min; Lux Cellulose-4
4.6 .times. 100 mm 3 u column; mobile phase: 50% MeOH @ 120 bar, 4
mL/min 51 ##STR00103## I .sup.1H NMR (400 MHz, DMSO-d6) .delta.
11.54 (br. s., 1H), 7.45 (s, 1H), 5.89 (s, 1H), 4.56 (s, 3H), 3.75-
3.89 (m, 2H), 3.46 (t, J = 6.11 Hz, 2H), 3.14- 3.23 (m, 2H), 3.13
(s, 3H), 2.89 (t, J = 6.24 Hz, 2H), 2.17 (s, 3H), 2.12 (s, 3H),
1.76- 1.89 (m, 1H), 1.32- 1.57 (m, 3H), 1.16- 1.26 (m, 1H); LCMs [M
+ H].sup.+ 479 [.alpha.]D = +80.9.degree. (c 0.1, MeOH) ~99% ee
(+); retention time 4.15 min; Lux Cellulose-4 4.6 .times. 100 mm 3
u column; mobile phase 50% MeOH @ 120 bar CO2, 4 mL/min 52
##STR00104## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.54 (br.
s., 1H), 7.62 (s, 1H), 5.88 (s, 1H), 4.65 (s, 1H), 4.62 (s, 1H),
4.56 (s, 2H), 3.61-3.69 (m, 1H), 3.43-3.59 (m, 5H), 3.12 (s, 3H),
2.86- 2.92 (m, 2H), 2.18 (s, 3H), 2.12 (s, 3H), 1.57-1.75 (m, 3H),
0.92 (d, J = 12.72 Hz, 1H); LCMs [M + H].sup.+ 495 [.alpha.]D =
-51.3.degree. (c 0.1, MeOH) >99% ee (-); retention time 2.51
min; Lux Cellulose-4 4.6 .times. 100 mm 3 u column; mobile phase:
50% MeOH @ 120 bar CO2; 4 mL/min 53 ##STR00105## I .sup.1H NMR (400
MHz, DMSO-d6) .delta. 11.54 (br. s., 1H), 7.62 (s, 1H), 5.88 (s,
1H), 4.65 (s, 1H), 4.62 (s, 1H), 4.56 (s, 2H), 3.62-3.68 (m, 1H),
3.43-3.59 (m, 5H), 3.12 (s, 3H), 2.85- 2.93 (m, 2H), 2.18 (s, 3H),
2.12 (s, 3H), 1.57-1.74 (m, 3H), 0.92 (d, J = 13.45 Hz, 1H); LCMs
[M + H].sup.+ 495 [.alpha.]D = +73.8.degree. (c 0.1, MeOH) >99%
ee (+); retention time 3.85 min; Lux Cellulose-4 4.6 .times. 100 mm
3 u column; mobile phase: 50% MeOH @ 120 bar CO2, 4 mL/min 54
##STR00106## A .sup.1H NMR (400 MHz, CDCl3) .delta. 11.80- 13.19
(m, 1H), 7.49 (s, 1H), 6.01 (s, 1H), 4.78 (s, 2H), 4.09 (t, J =
7.76 Hz, 1H), 3.82- 3.90 (m, 1H), 3.73- 3.81 (m, 2H), 3.64- 3.73
(m, 2H), 3.57- 3.64 (m, 1H), 2.87- 3.03 (m, 2H), 2.73- 2.87 (m,
1H), 2.59- 2.73 (m, 2H), 2.38 (s, 3H), 2.32 (s, 3H), 1.84-1.97 (m,
1H), 1.34-1.47 (m, 1H); LCMs [M + H].sup.+ 474 single enantiomer
from chiral reagents; absolute stereochemistry S,S 55 ##STR00107##
A .sup.1H NMR (400 MHz, CD3OD) .delta. 7.59 (s, 1H), 7.21-6.85 (m,
1H), 6.21 (s, 1H), 4.71 (s, 2H), 3.98- 3.96 (m, 1H), 3.85- 3.81 (m,
3H), 3.52- 3.49 (m, 5H), 2.98 (t, J = 6.2 Hz, 2H), 2.31 (s, 3H),
1.92-1.89 (m, 2H), 1.46-1.43 (m, 1H), 1.29-1.23 (m, 3H); LCMs [M +
H].sup.+ 531 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 56: 98.46% ee; retention time: 3.767 min; column:
Chiralpak AS-H 150*4.6 mm I.D., 5 um; mobile phase: 5-40% ethanol
(0.05% DEA) in CO2; flow rate: 3 mL/min 56 ##STR00108## A .sup.1H
NMR (400 MHz, CD3OD) .delta. 7.59 (s, 1H), 7.21-6.85 (m, 1H), 6.21
(s, 1H), 4.71 (s, 2H), 3.96- 3.94 (m, 1H), 3.85- 3.81 (m, 3H),
3.52- 3.43 (m, 5H), 2.98 (t, J = 6.2 Hz, 2H), 2.31 (s, 3H),
1.92-1.89 (m, 2H), 1.46-1.42 (m, 1H), 1.29-1.23 (m, 3H); LCMs [M +
H].sup.+ 531 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 55: 99.02% ee; retention time: 3.585 min; column:
Chiralpak AS-H 150*4.6 mm I.D., 5 um; mobile phase: 5-40% ethanol
(0.05% DEA) in CO2; flow rate: 3 mL/min 57 ##STR00109## B .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 11.50 (br. s., 1H), 7.68 (s, 1H),
5.89 (s, 1H), 4.65 (br. s., 1H), 4.58 (s, 2H), 3.90 (t, J = 4.52
Hz, 1H), 3.56- 3.76 (m, 2H), 3.40- 3.49 (m, 2H), 2.87 (t, J = 6.11
Hz, 2H), 2.33- 2.44 (m, 2H), 2.17 (s, 3H), 2.13 (s, 3H), 1.67 (br.
s., 4H), two Hs obscured by DMSO peak; LCMs [M + H].sup.+ 464
Racemic mixture 58 ##STR00110## B .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 11.51 (br. s., 1H), 7.68 (d, J = 3.55 Hz, 1H), 5.89 (s,
1H), 5.03-5.31 (m, 1H), 4.75 (br. s., 1H), 4.58 (s, 2H), 3.92-4.01
(m, 1H), 3.54-3.73 (m, 2H), 3.38-3.52 (m, 2H), 2.91-2.99 (m, 1H),
2.84-2.91 (m, 2H), 2.59-2.78 (m, 2H), 2.29-2.43 (m, 1H), 2.17 (s,
3H), 2.13 (s, 3H), 1.99-2.12 (m, 1H), 1.76-1.97 (m, 1H); LCMs [M +
H].sup.+ 482 Mixture of diastereomers containing (S)-3-
fluoropyrrolidine Mixture separated to give Ex. 62 and Ex. 63 59
##STR00111## B .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.51 (br.
s., 1H), 7.66 (s, 1H), 5.88 (s, 1H), 4.83 (br. s., 1H), 4.57 (s,
2H), 4.03 (t, J = 4.52 Hz, 1H), 3.54- 3.70 (m, 2H), 3.38-3.50 (m,
2H), 3.04 (dt, J = 14.37, 11.10 Hz, 1H), 2.79-2.94 (m, 3H),
2.62-2.78 (m, 2H), 2.18-2.30 (m, 2H), 2.16 (s, 3H), 2.12 (s, 3H);
LCMs [M + H].sup.+ 500 Racemic mixture 60 ##STR00112## B .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 11.54 (br. s., 1H), 7.72 (s, 1H),
5.89 (s, 1H), 4.71 (br. s., 1H), 4.58 (s, 2H), 3.95 (t, J = 4.65
Hz, 1H), 3.68- 3.78 (m, 1H), 3.51- 3.67 (m, 5H), 3.45 (t, J = 6.54
Hz, 2H), 2.87 (t, J = 6.11 Hz, 2H), 2.54 (br. s., 2H), 2.27- 2.39
(m, 2H), 2.17 (s, 3H), 2.13 (s, 3H); LCMs [M + H].sup.+ 480 Racemic
mixture 61 ##STR00113## I .sup.1H NMR (400 MHz, CD3OD) .delta.
11.40 (br. s., 1H), 7.46 (s, 1H), 5.95 (s, 1H), 4.82- 4.73 (m, 2H),
4.64- 4.57 (m, 2H), 3.68 (t, J = 5.4 Hz, 3H), 3.47 (s, 2H), 3.20
(s, 3H), 3.09-3.02 (m, 1H), 2.95 (t, J = 6 Hz, 2H), 2.52-2.48 (m,
1H), 2.37 (s, 3H), 2.29 (s, 3H), 1.89-1.87 (m, 1H), 1.75-1.68 (m,
2H), 1.61-1.51 (m, 2H); LCMs [M + H].sup.+ 545 Single isomer,
absolute stereochemistry unknown; Enantiomer of Ex. 71; 96.42% ee;
retention time: 9.135 min; column: Chiralpak AS-H 250 .times. 4.6
mm I.D., 5 um; mobile phase: 5-40% methanol (0.05% DEA) in CO2;
flow rate: 2.35 mL/min 62 ##STR00114## B HNMR was not taken due to
limited quantity. See Ex. 58; LCMs [M + H].sup.+ 482 One component
of the Ex. 58 mixture. Single diasteromer containing (S)-3-
fluoropyrrolidine, other chiral center undetermined; [.alpha.]D =
-58.9.degree. (c 0.01 MeOH) ~98% de (-); 1st peak; RT 1.233 min;
Chiralcel OJ- 3 4.6 .times. 100 mm 3 u column; 10% MeOH/DEA @ 120
bar, 4 mL/min 63 ##STR00115## B HNMR was not taken due to limited
quantity. See Ex. 58; LCMs [M + H].sup.+ 482 One component of the
Ex. 58 mixture. Single diasteromer containing (S)-3-
fluoropyrrolidine, other chiral center
undetermined; ~90% de (+); 2nd peak; RT 1.489 min; Chiralcel OJ- 3
4.6 .times. 100 mm 3 u column; 10% MeOH/DEA @ 120 bar, 4 mL/min 64
##STR00116## A .sup.1H NMR (400 MHz, CD3OD) .delta. 7.50 (d, J =
7.83 Hz, 1H), 7.20 (d, J = 7.83 Hz, 1H), 6.13 (s, 1H), 4.79 (s,
2H), 3.95 (dt, J = 3.67, 8.31 Hz, 1H), 3.76-3.87 (m, 3H), 3.58 (t,
J = 7.95 Hz, 2H), 3.43-3.52 (m, 2H), 3.18 (t, J = 8.44 Hz, 1H),
2.87 (dd, J = 5.14, 7.09 Hz, 2H), 2.75 (dd, J = 7.83, 15.89 Hz,
1H), 2.33-2.25 (m, 1H), 2.30 (s, 3H), 2.26 (s, 3H), 1.83 (qd, J =
8.60, 12.10 Hz, 1H); LCMs [M + H].sup.+ 431 Racemic Mixture of R,R
and S,S isomers (Assigned by analogy to Ex. 4, which had a crystal
structure showing S,S stereochemistry) 65 ##STR00117## A .sup.1H
NMR (400 MHz, CD3OD) .delta. 7.48 (d, J = 8.07 Hz, 1H), 7.19 (d, J
= 7.83 Hz, 1H), 6.11 (s, 1H), 4.78 (s, 2H), 4.13 (t, J = 7.95 Hz,
1H), 3.78 (dt, J = 4.16, 8.31 Hz, 1H), 3.55-3.73 (m, 5H), 3.47 (t,
J = 5.26 Hz, 2H), 2.86 (dd, J = 4.40, 6.85 Hz, 2H), 2.60-2.74 (m,
1H), 2.29 (s, 3H), 2.25 (s, 3H), 1.68-1.80 (m, 1H), 1.40 (qd, J =
8.56, 12.23 Hz, 1H); LCMs [M + H].sup.+ 431 Racemic Mixture of R,S
and S,R isomers 66 ##STR00118## B .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 11.51 (br. s., 1H), 7.66 (s, 1H), 5.88 (s, 1H), 4.83 (br.
s., 1H), 4.57 (s, 2H), 4.03 (t, J = 4.52 Hz, 1H), 3.54-3.70 (m,
2H), 3.38-3.50 (m, 2H), 3.04 (dt, J = 14.37, 11.10 Hz, 1H), 2.79-
2.94 (m, 3H), 2.62- 2.78 (m, 2H), 2.18-2.30 (m, 2H), 2.16 (s, 3H),
2.12 (s, 3H); LCMs [M + H].sup.+ 500 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 67; [.alpha.]D =
-65.35.degree. (c 0.01 MeOH) >99% ee (-); 1st peak; RT 2.120
min; Chiralpak IC- 3 4.6 .times. 100 mm 3 u column; 40% MeOH @ 120
bar, 4 mL/min 67 ##STR00119## B .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 11.51 (br. s., 1H), 7.66 (s, 1H), 5.88 (s, 1H), 4.83 (br.
s., 1H), 4.57 (s, 2H), 4.03 (t, J = 4.52 Hz, 1H), 3.54-3.70 (m,
2H), 3.38-3.50 (m, 2H), 3.04 (dt, J = 14.37, 11.10 Hz, 1H),
2.79-2.94 (m, 3H), 2.62-2.78 (m, 2H), 2.18-2.30 (m, 2H), 2.16 (s,
3H), 2.12 (s, 3H); LCMs [M + H].sup.+ 500 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 66; [.alpha.]D =
+92.77.degree. (c 0.01 MeOH) ~99% ee (+); 2nd peak; RT 2.866 min;
Chiralpak IC- 3 4.6 .times. 100 mm 3 u column; 40% MeOH @ 120 bar,
4 mL/min 68 ##STR00120## B .sup.1H NMR (400 MHz, DMSO-d6) .delta.
11.54 (br. s., 1H), 7.72 (s, 1H), 5.89 (s, 1H), 4.71 (br. s., 1H),
4.58 (s, 2H), 3.95 (t, J = 4.65 Hz, 1H), 3.68- 3.78 (m, 1H), 3.51-
3.67 (m, 5H), 3.45 (t, J = 6.54 Hz, 2H), 2.87 (t, J = 6.11 Hz, 2H),
2.54 (br. s., 2H), 2.27- 2.39 (m, 2H), 2.17 (s, 3H), 2.13 (s, 3H);
LCMs [M + H].sup.+ 480 Single isomer, absolute stereochemistry
unknown; Enantiomer of Ex. 69; [.alpha.]D = -52.63.degree. (c 0.01
MeOH) >99% ee (-); 1st peak; RT 3.039 min; Chiralpak AD- 3 4.6
.times. 100 mm 3 u column; 20% MeOH/DEA @ 120 bar, 4 mL/min 69
##STR00121## B .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.54 (br.
s., 1H), 7.72 (s, 1H), 5.89 (s, 1H), 4.71 (br. s., 1H), 4.58 (s,
2H), 3.95 (t, J = 4.65 Hz, 1H), 3.68- 3.78 (m, 1H), 3.51- 3.67 (m,
5H), 3.45 (t, J = 6.54 Hz, 2H), 2.87 (t, J = 6.11 Hz, 2H), 2.54
(br. s., 2H), 2.27- 2.39 (m, 2H), 2.17 (s, 3H), 2.13 (s, 3H); LCMs
[M + H].sup.+ 480 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 68; [.alpha.]D = +75.09.degree. (c 0.01 MeOH)
~97.6% ee (+); 2nd peak; RT 4.327 min; Chiralpak AD-3 4.6 .times.
100 mm 3 u column; 20% MeOH/DEA @ 120 bar, 4 mL/min; 70
##STR00122## I .sup.1H NMR (400 MHz, CD3OD) .delta. 7.54 (s, 1H),
6.11 (s, 1H), 4.76 (s, 2H), 4.66 (d, J = 5.62 Hz, 1H), 4.43- 4.56
(m, 1H), 4.11- 4.28 (m, 2H), 3.66- 3.80 (m, 1H), 3.53 (t, J = 6.11
Hz, 2H), 2.99 (t, J = 6.11 Hz, 2H), 2.84-2.96 (m, 1H), 2.50-2.64
(m, 1H), 2.30 (s, 3H), 2.25 (s, 3H), 1.80-2.01 (m, 1H), 1.74 (dd, J
= 2.20, 13.20 Hz, 1H), 1.28-1.58 (m, 3H); LCMs [M + H].sup.+ 539
Racemic mixture 71 ##STR00123## I .sup.1H NMR (400 MHz, CD3OD)
.delta. 11.61 (br. s., 1H), 7.46 (s, 1H), 5.95 (s, 1H), 4.82- 4.73
(m, 2H), 4.64- 4.57 (m, 2H), 3.68 (t, J = 5.4 Hz, 3H), 3.47 (s,
2H), 3.20 (s, 3H), 3.09-3.02 (m, 1H), 2.95 (t, J = 6 Hz, 2H),
2.52-2.49 (m, 1H), 2.37 (s, 3H), 2.29 (s, 3H), 1.89-1.87 (m, 1H),
1.75-1.68 (m, 2H), 1.62-1.51 (m, 2H); LCMs [M + H].sup.+ 545 Single
isomer, absolute stereochemistry unknown; Enantiomer of Ex. 61;
97.43% ee; retention time: 8.742 min; column: Chiralpak AS-H 250
.times. 4.6 mm I.D., 5 um; mobile phase: 5-40% methanol (0.05% DEA)
in CO2; flow rate: 2.35 mL/min 72 ##STR00124## I .sup.1H NMR (400
MHz, CDCl3) .delta. 11.52 (br s, 1H), 7.99 (s, 1H), 7.47 (s, 1H),
5.96 (s, 1H), 4.77 (s, 2H), 4.63 (d, J = 4.8 Hz, 1H), 4.42 (d, J =
11.6 Hz, 1H), 3.68 (t, J = 6 Hz, 2H), 3.61 (t, J = 13.2 Hz, 1H),
3.20 (s, 3H), 2.97-2.94 (m, 3H), 2.49-2.47 (m, 1H), 2.38 (s, 3H),
2.29 (s, 3H), 1.89- 1.87 (m, 1H), 1.36- 1.26 (m, 4H); LCMs [M +
H].sup.+ 506 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 73; 94.43% ee; retention time: 8.623 min; column:
Chiralpak AS-H 250 .times. 4.6 mm I.D., 5 um; mobile phase: 5-40%
methanol (0.05% DEA) in CO2; flow rate: 2.35 mL/min 73 ##STR00125##
I .sup.1H NMR (400 MHz, CDCl3) .delta. 11.29 (br s, 1H), 7.99 (s,
1H), 7.47 (s, 1H), 5.95 (s, 1H), 4.77 (s, 2H), 4.63 (d, J = 4.8 Hz,
1H), 4.42 (d, J = 12.0 Hz, 1H), 3.68 (t, J = 6 Hz, 2H), 3.61 (t, J
= 12.8 Hz, 1H), 3.20 (s, 3H), 2.97-2.94 (m, 3H), 2.49-2.47 (m, 1H),
2.37 (s, 3H), 2.29 (s, 3H), 1.89- 1.87 (m, 1H), 1.36- 1.26 (m, 4H);
LCMs [M + H].sup.+ 506 Single isomer absolute stereochemistry
unknown; Enantiomer of Ex. 72; 92.36% ee; retention time: 9.05 min;
column: Chiralpak AS-H 250 .times. 4.6 mm I.D., 5 um; mobile phase:
5-40% methanol (0.05% DEA) in CO2; flow rate: 2.35 mL/min 74
##STR00126## H N/A; LCMs [M + H].sup.+ 432 N/A 75 ##STR00127## H
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.54 (br. s., 1H), 7.51 (s,
1H), 5.89 (s, 1H), 4.81-4.99 (m, 2H), 4.51-4.60 (m, 3H), 4.48 (br.
s., 1H), 4.19 (br. s., 1H), 3.83-4.01 (m, 2H), 3.46 (t, J = 6.11
Hz, 2H), 2.88 (t, J = 5.87 Hz, 2H), 2.17 (s, 3H), 2.12 (s, 3H),
1.85 (br. s., 3H); LCMs [M + H].sup.+ 490 N/A 76 ##STR00128## H
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.46 (br. s., 1H), 7.44 (s,
0.56H), 7.43 (s, 0.44H), 5.81 (s, 1H), 4.72 (br. s., 0.88H), 4.49
(s, 2H), 4.40 (br. s., 1.12H), 4.36 (br. s., 1.12H), 4.05 (br. s.,
0.88H), 3.39 (t, J = 6.24 Hz, 2H), 2.81 (t, J = 6.11 Hz, 2H), 2.10
(s, 3H), 2.05 (s, 3H), 1.78 (br. s., 3H), 1.76 (s, 1.32H), 1.68 (s,
1.68H) Rotamers (~4:5 ratio); LCMs [M + H].sup.+ 474 N/A 77
##STR00129## H .sup.1H NMR (400 MHz, CDCl3) .delta. 11.72 (br. s.,
1H), 6.98 (s, 1H), 5.93 (s, 1H), 4.79 (s, 2H), 4.65 (br. s., 2H),
4.29 (br. s., 2H), 3.62 (t, J = 6.15 Hz, 2H), 2.89 (s, 3H), 2.73
(t, J = 6.15 Hz, 2H), 2.36 (s, 3H), 2.23-2.30 (m, 3H), 2.21 (s,
3H), 1.87 (s, 3H); LCMs [M + H].sup.+ 490 N/A 78 ##STR00130## H
.sup.1H NMR (400 MHz, CD3OD) .delta. 7.17 (s, 1H), 6.28 (s, 1H),
4.74 (s, 2H), 4.68 (br. s., 2H), 4.24-4.33 (m, 2H), 3.91 (s, 3H),
3.38 (t, J = 6.27 Hz, 2H), 2.99 (s, 3H), 2.80 (t, J = 6.27 Hz, 2H),
2.34 (s, 3H), 2.27 (s, 3H), 1.91 (t, J = 1.51 Hz, 3H); [M +
Na].sup.+ 528 N/A 79 ##STR00131## H .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 11.58 (brs, 1H), 7.84 (d, J = 5.6 Hz, 1H), 5.89 (s, 1H),
5.15-5.00 (m, 1H), 5.00-4.95 (m, 1H), 4.80-4.75 (m, 1H), 4.65-4.60
(m, 1H), 4.57 (s, 2H), 4.50-4.45 (m, 1H), 4.00-3.95 (m, 2H), 3.47
(t, J = 6.0 Hz, 2H), 2.93 (t, J = 4.8 Hz, 2H), 2.16 (s, 3H), 2.12
(s, 3H); [M + H].sup.+ 294 N/A 80 ##STR00132## H .sup.1H NMR (400
MHz, CD3OD) .delta. 7.36 (s, 1H), 6.18 (s, 1H), 4.78-4.83 (m, 2H),
3.60 (t, J = 6.24 Hz, 2H), 2.98-3.12 (m, 4H), 2.91 (t, J = 5.14 Hz,
2H), 2.58-2.69 (m, 1H), 2.47-2.58 (m, 1H), 2.37 (s, 3H), 2.32 (s,
3H), 2.05 (d, J = 7.58 Hz, 2H), 1.97- 2.01 (m, 3H); [M + H].sup.+
460 N/A 81 ##STR00133## I .sup.1H NMR (400 MHz, CDCl3) .delta.
12.34 (brs, 1H), 7.49 (s, 1H), 5.93 (s, 1H), 5.05 (d, J = 6.0 Hz,
1H), 4.61- 4.78 (m, 6H), 3.88 (s, 3H), 3.48-3.50 (m, 2H), 3.37-3.38
(m, 1H), 3.31 (s, 3H), 2.94 (t, J = 6.2 Hz, 2H), 2.35 (s, 3H); [M +
Na].sup.+ 489 R isomer; stereochemistry determined from X-ray
crystal structure of enantiomeric compound Ex. 82; 100% ee;
retention time 9.85 min; Column: (R,R) Whelk O1, 250 .times. 4.6 mm
I.D., 5 um; Mobile phase: 50% ethanol (0.05% DEA) in CO2;
wavelength 220 nm 82 ##STR00134## I .sup.1H NMR (400 MHz, CDCl3)
.delta. 12.38 (brs, 1H), 7.49 (s, 1H), 5.92 (s, 1H), 5.05 (d, J =
6.0 Hz, 1H), 4.64- 4.78 (m, 6H), 3.87 (s, 3H), 3.47-3.50 (m, 2H),
3.37-3.38 (m, 1H), 3.31 (s, 3H), 2.93 (t, J = 6.2 Hz, 2H), 2.35 (s,
3H); [M + H].sup.+ 467 Known to be S isomer by X-ray crystal
structure; Enantiomer of Ex. 81; 98% ee; retention time 8.65 min;
column: (R,R)Whelk O1, 250 .times. 4.6 mm I.D., 5 um; mobile phase:
50% ethanol (0.05% DEA) in CO2; wavelength 220 nm 83 ##STR00135## I
.sup.1H NMR (400 MHz, CDCl3) .delta. 12.36 (brs, 1H), 7.29 (s, 1H),
5.91 (s, 1H), 4.75- 4.85 (m, 3H), 3.83- 3.89 (m, 6H), 3.69- 3.71
(m, 1H), 3.44- 3.47 (m, 2H), 3.16 (s, 3H), 2.73-2.76 (m, 2H),
2.54-2.58 (m, 1H), 2.34 (s, 3H), 2.26 (s, 3H), 1.70- 1.73 (m, 2H);
[M + H].sup.+ 461 Known to be R,R isomer by X-ray crystal
structure; Enantiomer of Ex. 86; Diastereomer of Ex. 84 and Ex. 85;
100% ee; retention time 34.91 min; column: Chiralpak IC 250 .times.
4.6 mm I.D., 5 um; mobile phase: 50% ethanol (0.05% DEA) in CO2;
flow rate: 2.0 mL/min 84 ##STR00136## I .sup.1H NMR (400 MHz,
CDCl3) .delta. 12.33 (brs, 1H), 7.30 (s, 1H), 5.91 (s, 1H), 4.77-
4.84 (m, 3H), 3.86- 3.89 (m, 4H), 3.68- 3.73 (m, 2H), 3.58- 3.60
(m, 1H), 3.44- 3.46 (m, 2H), 3.19 (s, 3H), 2.73-2.75 (m, 2H),
2.62-2.64 (m, 1H), 2.34 (s, 3H), 2.24 (s, 3H), 1.95- 1.98 (m, 2H);
[M + H].sup.+ 461 Single enantiomer, either R,S or S,R, but
absolute stereochemistry unknown; Enantiomer of Ex. 85;
Diastereomer of Ex. 83 and Ex. 86; 97% ee; retention time 39.01
min; column: Chiralpak IC 250 .times. 4.6 mm I.D., 5 um; mobile
phase: 50% ethanol (0.05% DEA) in CO2; flow rate: 2.0 mL/min 85
##STR00137## I .sup.1H NMR (400 MHz, CDCl3) .delta. 12.30 (brs,
1H), 7.30 (s, 1H), 5.91 (s, 1H), 4.77- 4.84 (m, 3H), 3.86- 3.89 (m,
4H), 3.66- 3.74 (m, 2H), 3.58- 3.59 (m, 1H), 3.44- 3.46 (m, 2H),
3.19 (s, 3H), 2.73-2.75 (m, 2H), 2.63-2.65 (m, 1H), 2.34 (s, 3H),
2.24 (s, 3H), 1.95- 1.98 (m, 2H); [M + H].sup.+ 461 Single
enantiomer, either R,S or S,R, but absolute stereochemistry
unknown; Enantiomer of Ex. 84; Diastereomer of Ex. 83 and Ex. 86;
100% ee; retention time 29.05 min; column: Chiralpak IC 250 .times.
4.6 mm I.D., 5 um; mobile phase: 50% ethanol (0.05% DEA) in CO2;
flow rate: 2.0 mL/min 86 ##STR00138## I .sup.1H NMR (400 MHz,
CDCl3) .delta. 12.29 (brs, 1H), 7.30 (s, 1H), 5.91 (s, 1H), 4.75-
4.84 (m, 3H), 3.83- 3.89 (m, 6H), 3.69- 3.71 (m, 1H), 3.44- 3.47
(m, 2H), 3.16 (s, 3H), 2.73-2.76 (m, 2H), 2.56-2.58 (m, 1H), 2.34
(s, 3H), 2.25 (s, 3H), 1.70- 1.73 (m, 2H); [M + H].sup.+ 461 S,S
isomer; stereochemisty determined by X- ray crystal structure of
enantiomeric compound Ex. 83; Diastereomer of Ex. 84 and Ex. 85;
100% ee; retention time 32.28 min; column: Chiralpak IC 250 .times.
4.6 mm I.D., 5 um; mobile phase: 50% ethanol (0.05% DEA) in CO2;
flow rate: 2.0 mL/min 87 ##STR00139## J .sup.1H NMR (400 MHz,
CD3OD) .delta. 7.44 (s, 1H), 6.27 (s, 1H), 4.73 (s, 2H), 3.91 (s,
3H), 3.67-3.78 (m, 2H), 3.36-3.42 (m, 3H), 3.16 (t, J = 7.40 Hz,
1H), 2.90-2.96 (m, 2H), 2.82-2.90 (m, 2H), 2.39 (s, 3H), 2.33 (s,
3H), 1.16 (d, J = 6.78 Hz, 3H); [M + H].sup.+ 464 Single isomer,
absolute stereochemistry unknown; Enantiomer of Ex. 88; 99% ee;
retention time 5.511 mi; Column: Chiralpak AD-3 150 .times. 4.6 mm
I.D., 3 um; mobile phase; ethanol (0.05% DEA) in CO2 88
##STR00140## J .sup.1H NMR (400 MHz, CD3OD) .delta. 7.43 (s, 1H),
6.26 (s, 1H), 4.73 (s, 2H), 3.91 (s, 3H), 3.64-3.75 (m, 2H),
3.34-3.42 (m, 3H), 3.12 (br. s., 1H), 2.93 (t, J = 6.02 Hz, 2H),
2.83 (br. s., 2H), 2.35-2.40 (m, 3H), 2.33 (s, 3H), 1.16 (d, J =
6.78 Hz, 3H); [M + H].sup.+ 464 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 87; 100% ee; retention
time 55.997 min; Column: Chiralpak AD-3 150 .times. 4.6 mm I.D., 3
um; mobile phase; ethanol (0.05% DEA) in CO2 89 ##STR00141## K
.sup.1H NMR (400 MHz, CDCl3) .delta. 7.74 (s, 1H), 5.94 (s, 1H),
4.87-4.70 (m, 2H), 4.00 (q, J = 6.3 Hz, 1H), 3.76-3.59 (m, 6H),
2.99-2.81 (m, 2H), 2.53 (br. s., 2H), 2.36 (m, 5H), 2.29 (s, 3H),
1.24 (d, J = 6.5 Hz, 3H); [M + H].sup.+ 464 racemic mixture 90
##STR00142## K [M + H].sup.+ 464 (+) isomer of Ex. 89 91
##STR00143## K [M + H].sup.+ 464 (-) isomer of Ex. 89 92
##STR00144## A .sup.1H NMR (400 MHz, CDCl3) .delta. 12.31 (br. s.,
1H), 7.42 (s, 1H), 6.72 (t, J = 72 Hz, 1H), 6.09 (s, 1H), 4.70 (s,
2H), 3.82- 3.96 (m, 4H), 3.61- 3.65 (m, 4H), 3.20 (t, J = 8.4 Hz,
2H), 2.97 (t, J = 6.2 Hz, 2H), 2.70-2.72 (m, 1H), 2.35 (s, 3H),
2.21- 2.23 (m, 1H), 1.75- 1.81 (m, 1H); [M + H].sup.+ 517 Single
isomer, absolute stereochemistry unknown; Enantiomer of Ex. 93;
Diastereomer of Ex. 94 and Ex. 95; 100% ee; retention time 5.37
min; column: Chiralcel OJ-H 250 .times. 4.6 mm I.D., 5 um; mobile
phase: methanol (0.05% DEA) CO2 from 5% to 40%; flow rate; 2.5
mL/min 93 ##STR00145## A .sup.1H NMR (400 MHz, CDCl3) .delta. 12.25
(br. s., 1H), 7.42 (s, 1H), 6.72 (t, J = 72 Hz, 1H), 6.09 (s, 1H),
4.70 (s, 2H), 3.82- 3.89 (m, 4H), 3.61- 3.66 (m, 4H), 3.20 (t, J =
8.4 Hz, 2H), 2.98 (t, J = 6.2 Hz, 2H), 2.70-2.72 (m, 1H), 2.35 (s,
3H), 2.21- 2.23 (m, 1H), 1.76- 1.81 (m, 1H); [M + H].sup.+ 517
Single isomer, absolute stereochemistry unknown; Enantiomer of Ex.
92; Diastereomer of Ex. 94 and Ex. 95; 100% ee; retention time 5.54
min; column: Chiralcel OJ-H 250 .times. 4.6 mm I.D., 5 um; mobile
phase: methanol (0.05% DEA) in CO2 from 5% to 40%; flow rate; 2.5
mL/min 94 ##STR00146## A .sup.1H NMR (400 MHz, CDCl3) .delta. 12.12
(br. s., 1H), 7.45 (s, 1H), 6.73 (t, J = 72 Hz, 1H), 6.10 (s, 1H),
4.71 (t, J = 13.3 Hz, 2H), 4.07-4.10 (m, 1H), 3.82-3.70 (m, 4H),
3.60-3.65 (m, 4H), 2.99 (t, J = 6.2 Hz, 2H), 2.63-2.65 (m, 1H),
2.35 (s, 3H), 1.78-1.8 (m, 1H), 1.40-1.45 (m, 1H); [M + H].sup.+
517 Single isomer, absolute stereochemistry unknown; Enantiomer of
Ex. 95; Diastereomer of Ex. 92 and Ex. 93; 100% ee; retention time
5.80 min; column: Chiralcel OJ-H 250 .times. 4.6 mm I.D., 5 um;
mobile phase: methanol (0.05% DEA) in CO2 from 5% to 40%; flow
rate; 2.5 mL/min 95 ##STR00147## A .sup.1H NMR (400 MHz, CDCl3)
.delta. 12.36 (br. s., 1H), 7.45 (s, 1H), 6.72 (t, J = 73 Hz, 1H),
6.09 (s, 1H), 4.71 (q, J = 13.9 Hz, 2H), 4.06-4.08 (m, 1H),
3.70-3.83 (m, 4H), 3.59-3.63 (m, 4H), 2.98 (t, J = 5.4 Hz, 2H),
2.63-2.65 (m, 1H), 2.35 (s, 3H), 1.78-1.8 (m,
1H), 1.39-1.45 (m, 1H); [M + H].sup.+ 517 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 94; Diastereomer of Ex.
92 and Ex. 93; 100% ee; retention time 5.98 min; column: Chiralcel
OJ-H 250 .times. 4.6 mm I.D., 5 um; mobile phase: methanol (0.05%
DEA) in CO2 from 5% to 40%; flow rate; 2.5 mL/min 96 ##STR00148## A
.sup.1H NMR (400 MHz, CD3OD) .delta. 7.48 (d, J = 7.34 Hz, 1H),
7.18 (d, J = 7.83 Hz, 1H), 6.10 (s, 1H), 4.77 (s, 2H), 3.89-3.98
(m, 1H), 3.74-3.86 (m, 3H), 3.57 (t, J = 7.70 Hz, 2H), 3.42-3.51
(m, 2H), 3.16 (t, J = 8.31 Hz, 1H), 2.82-2.89 (m, 2H), 2.64-2.80
(m, 1H), 2.32-2.23 (m, 1H), 2.28 (s, 3H), 2.24 (s, 3H), 1.74-1.89
(m, 1H); [M + H].sup.+ 431 [.alpha.].sub.D = +51.7.degree. (c 0.2
MeOH); (+) isomer of Ex. 64 racemate; either R,R or S,S isomer;
absolute stereochemistry undetermined; Enantiomer of Ex. 97 97
##STR00149## A .sup.1H NMR (400 MHz, CD3OD) .delta. 7.48 (d, J =
7.58 Hz, 1H), 7.18 (d, J = 7.83 Hz, 1H), 6.10 (s, 1H), 4.77 (s,
2H), 3.93 (dt, J = 3.79, 8.13 Hz, 1H), 3.74- 3.86 (m, 3H), 3.57 (t,
J = 7.83 Hz, 2H), 3.43- 3.51 (m, 2H), 3.16 (t, J = 8.44 Hz, 1H),
2.85 (t, J = 5.75 Hz, 2H), 2.65-2.80 (m, 1H), 2.32-2.23 (m, 1H),
2.28 (s, 3H), 2.24 (s, 3H), 1.75-1.89 (m, 1H); [M + H].sup.+ 431
[.alpha.].sub.D = -27.4.degree. (c 0.1 MeOH); (-) isomer of Ex. 64
racemate; either R,R or S,S isomer; absolute stereochemistry
undeteremined; Enantiomer of Ex. 96 98 ##STR00150## A .sup.1H NMR
(400 MHz, CD3OD) .delta. 7.48 (d, J = 7.83 Hz, 1H), 7.19 (d, J =
7.82 Hz, 1H), 6.10 (s, 1H), 4.78 (s, 2H), 4.13 (t, J = 7.83 Hz,
1H), 3.78 (dt, J = 4.16, 8.31 Hz, 1H), 3.55-3.73 (m, 5H), 3.43-3.50
(m, 2H), 2.82-2.89 (m, 2H), 2.61-2.74 (m, 1H), 2.29 (s, 3H), 2.24
(s, 3H), 1.69-1.79 (m, 1H), 1.34-1.45 (m, 1H); [M + H].sup.+ 431
[.alpha.].sub.D = +20.5.degree. (c 0.1 MeOH); (+) isomer of Ex. 65
racemate; either R,S or S,R isomer; absolute stereochemistry
undetermined; Enantiomer of Ex. 99 99 ##STR00151## A .sup.1H NMR
(400 MHz, CD3OD) .delta. 7.48 (d, J = 7.83 Hz, 1H), 7.19 (d, J =
8.07 Hz, 1H), 6.10 (s, 1H), 4.78 (s, 2H), 4.13 (t, J = 7.83 Hz,
1H), 3.78 (dt, J = 4.16, 8.31 Hz, 1H), 3.56-3.74 (m, 5H), 3.42-3.52
(m, 2H), 2.82-2.89 (m, 2H), 2.59-2.75 (m, 1H), 2.29 (s, 3H), 2.24
(s, 3H), 1.68-1.79 (m, 1H), 1.40 (qd, J = 8.57, 12.20 Hz, 1H); [M +
H].sup.+ 431 [.alpha.].sub.D = -33.1.degree. (c 0.1 MeOH); (-)
isomer of Ex. 65 racemate; either R,S or S,R isomer; absolute
stereochemistry undetermined; Enantiomer of Ex. 98 100 ##STR00152##
A .sup.1H NMR (700 MHz, DMSO-d6) .delta. 7.27 (br. s., 1H), 5.89
(s, 1H), 4.50-4.61 (m, 2H), 3.84 (d, J = 8.8 Hz, 1H), 3.21-3.74 (m,
7H), 3.15 (td, J = 10.76, 3.59 Hz, 1H), 2.67 (t, J = 5.81 Hz, 2H),
2.19 (s, 3H), 2.15 (s, 3H), 2.12 (s, 3H), 1.82-1.91 (m, 1H), 1.78
(d, J = 12.64 Hz, 1H), 1.20-1.31 (m, 1H), 1.05-1.15 (m, 2H); [M +
H].sup.+ 459 [.alpha.].sub.D = +10.8.degree. (c 0.1 MeOH); 99% ee;
absolute stereochemistry undetermined; Enantiomer of Ex. 101 101
##STR00153## A .sup.1H NMR (700 MHz, DMSO-d6) .delta. 7.27 (br. s.,
1H), 5.89 (s, 1H), 4.50-4.63 (m, 2H), 3.84 (d, J = 11.0 Hz, 1H),
3.20-3.75 (m, 7H), 3.15 (td, J = 10.89, 3.67 Hz, 1H), 2.67 (t, J =
5.89 Hz, 2H), 2.19 (s, 3H), 2.15 (s, 3H), 2.12 (s, 3H), 1.81-1.91
(m, 1H), 1.78 (d, J = 12.98 Hz, 1H), 1.20-1.30 (m, 1H), 1.06-1.15
(m, 2H); [M + H].sup.+ 459 [.alpha.].sub.D = -9.7.degree. (c 0.1
MeOH); >99% ee; absolute stereochemistry undetermined;
Enantiomer of Ex. 100 102 ##STR00154## D .sup.1H NMR (400 MHz,
CDCl3) .delta. 11.52 (br. s., 1H), 7.54 (s, 1H), 5.96 (s, 1H), 4.88
(br. s., 1H), 4.77 (s, 2H), 4.08-4.11 (m, 1H), 3.65-3.70 (m, 2H),
3.31 (s, 3H), 2.99- 2.91 (m, 2H), 2.37 (s, 3H), 2.29 (s, 3H), 1.05
(d, J = 6.4 Hz, 3H); [M + H].sup.+ 439 Mixture of 4 possible
diastereomers 103 ##STR00155## D .sup.1H NMR (400 MHz, CD3OD)
.delta. 7.52 (s, 1H), 6.00 (s, 1H), 4.97 (dd, J = 7.15, 2.87 Hz,
1H), 4.66 (s, 2H), 4.00-4.05 (m, 1H), 3.83-3.91 (m, 1H), 3.72 (td,
J = 8.34, 4.10 Hz, 1H), 3.56- 3.66 (m, 3H), 3.37- 3.45 (m, 3H),
2.88 (t, J = 6.24 Hz, 2H), 2.19 (s, 3H), 2.15 (s, 3H), 2.05-2.13
(m, 1H), 1.85-1.97 (m, 1H); [M + H].sup.+ 481 Single isomer, (R) at
THF center, other chiral center undetermined; Diastereomer of Ex.
104; 99% de; retention time 1.088 min on Chiralcel OJ-3 4.6 .times.
100 mm 3 u column; 10% MeOH @ 120 bar, 4 mL/min 104 ##STR00156## D
.sup.1H NMR (400 MHz, CD3OD) .delta. 7.57 (s, 1H), 6.00 (s, 1H),
4.91 (dd, J = 7.03, 2.87 Hz, 1H), 4.65 (s, 2H), 4.04-4.11 (m, 1H),
3.90 (d, J = 9.66 Hz, 1H), 3.78 (q, J = 8.07 Hz, 1H), 3.62- 3.69
(m, 2H), 3.59 (dd, J = 11.86, 2.93 Hz, 1H), 3.37-3.45 (m, 3H), 2.88
(t, J = 6.24 Hz, 2H), 2.19 (s, 3H), 2.15 (s, 3H), 1.74-1.96 (m,
2H); [M + H].sup.+ 481 Single isomer, (R) at THF center, other
chiral center undetermined; Diastereomer of Ex. 103; 94% de;
retention time 1.558 min on Chiralcel OJ-3 4.6 .times. 100 mm 3 u
column; 10% MeOH @ 120 bar, 4 mL/min 105 ##STR00157## E .sup.1H NMR
(400 MHz, CD3OD) .delta. 7.67 (s, 1H), 6.27 (s, 1H), 4.74 (s, 2H),
4.28-4.29 (m, 1H), 3.90-3.79 (m, 6H), 3.69 (t, J = 6.8 Hz, 2H),
3.40 (t, J = 6.0 Hz, 2H), 2.94 (t, J = 6.0 Hz, 2H), 2.34 (s, 3H),
2.10-2.07 (m, 1H), 1.90-1.86 (m, 1H), 1.77-1.75 (m, 1H), 1.60-1.56
(m, 1H); [M + H].sup.+ 481 1R,2R isomer; Diastereomer of Ex. 106;
retention time: 3.480 min; column: Chiralpak AD-3 150 .times. 4.6
mm I.D., 3 um; mobile phase 40% ethanol (0.05% DEA) in CO2 106
##STR00158## E .sup.1H NMR (400 MHz, CD3OD) .delta. 7.60 (s, 1H),
6.27 (s, 1H), 4.74 (s, 2H), 4.20- 4.18 (m, 1H), 3.97- 3.92 (m, 6H),
3.91- 3.80 (m, 1H), 3.70 (s, 1H), 3.40 (t, J = 5.6 Hz, 2H), 2.95
(t, J = 6.0 Hz, 2H), 2.34 (s, 3H), 1.96-1.82 (m, 3H), 1.55-1.50 (m,
1H); [M + H].sup.+ 481 1R,2R isomer; Diastereomer of Ex. 105;
retention time: 2.507 min; column: Chiralpak AD-3 150 .times. 4.6
mm I.D., 3 um; mobile phase 40% ethanol (0.05% DEA) in CO2 107
##STR00159## G .sup.1H NMR (400 MHz, CD3OD) .delta. 7.56 (s, 1H),
6.27 (s, 1H), 4.73 (s, 2H), 4.70- 4.72 (m, 1H), 3.91 (s, 3H),
3.40-3.43 (m, 2H), 3.26 (s, 3H), 2.95-2.99 (m, 2H), 2.34 (s, 3H),
1.74- 1.79 (m, 1H), 1.62- 1.67 (m, 1H), 0.99 (t, J = 7.2 Hz, 3H);
[M + H].sup.+ 439 Single isomer absolute stereochemistry unknown;
Enantiomer of Ex. 108; 100% ee; retention time 11.41 min; column:
Pheno Lux Cellulose-2, 150 .times. 4.6 mm I.D., 5 um; mobile phase:
50% MeOH (0.05% DEA) in CO2; Flow rate: 2.0 mL/min 108 ##STR00160##
G .sup.1H NMR (400 MHz, CD3OD) .delta. 7.56 (s, 1H), 6.27 (s, 1H),
4.74 (s, 2H), 4.70- 4.72 (m, 1H), 3.91 (s, 3H), 3.40-3.43 (m, 2H),
3.26 (s, 3H), 2.95-2.99 (m, 2H), 2.34 (s, 3H), 1.73- 1.77 (m, 1H),
1.62- 1.67 (m, 1H), 0.99 (t, J = 7.4 Hz, 3H); [M + H].sup.+ 439
Single isomer, absolute stereochemistry unknown; Enantiomer of Ex.
107; 99% ee; retention time 15.01 min; column: Pheno Lux
Cellulose-2, 150 .times. 4.6 mm I.D., 5 um; mobile phase: 50% MeOH
(0.05% DEA) in CO2; Flow rate: 2.0 mL/min 109 ##STR00161## I
.sup.1H NMR (400 MHz, CDCl3) .delta. 12.03 (br. s., 1H), 7.46 (s,
1H), 5.95 (s, 1H), 4.74-4.82 (m, 2H), 4.64-4.65 (m, 2H), 3.77-3.79
(m, 1H), 3.68 (t, J = 5.8 Hz, 2H), 3.19 (s, 3H), 2.90-2.96 (m, 3H),
2.41-2.47 (m, 1H), 2.37 (s, 3H), 2.29 (s, 3H), 2.07 (s, 3H),
1.83-1.86 (m, 1H), 1.52-1.57 (m, 2H), 1.34-1.41 (m, 2H); [M +
H].sup.+ 520 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 110; 100% ee; retention time: 10.42 min; column:
Chiralpak AD-H 250 .times. 4.6 mm I.D., 5 um; mobile phase: 5-40%
methanol (0.05% DEA) in CO2; flow rate: 2.5 mL/min 110 ##STR00162##
I .sup.1H NMR (400 MHz, CDCl3) .delta. 12.01 (br. s., 1H), 7.47 (s,
1H), 5.95 (s, 1H), 4.74-4.82 (m, 2H), 4.62-4.65 (m, 2H), 3.76-3.79
(m, 1H), 3.69 (t, J = 6 Hz, 2H), 3.19 (s, 3H), 2.90- 2.96 (m, 3H),
2.41- 2.47 (m, 1H), 2.37 (s, 3H), 2.29 (s, 3H), 2.07 (s, 3H), 1.83-
1.86 (m, 1H), 1.52- 1.58 (m, 2H), 1.34- 1.40 (m, 2H); [M + H].sup.+
520 Single isomer, absolute stereochemistry unknown; Enantiomer of
Ex. 109; 93% ee; retention time: 11.08 min; column: Chiralpak AD-H
250 .times. 4.6 mm I.D., 5 um; mobile phase: 5-40% methanol (0.05%
DEA) in CO2; flow rate: 2.5 mL/min 111 ##STR00163## I .sup.1H NMR
(400 MHz, CDCl3) .delta. 12.06 (s, 1H), 7.44 (d, J = 3.2 Hz, 1H),
5.93 (s, 1H), 4.78 (s, 2H), 4.62- 4.64 (m, 2H), 4.10- 4.14 (m, 2H),
3.88 (s, 3H), 3.72 (s, 1H), 3.47-3.53 (m, 3H), 3.20 (s, 3H), 2.84-
2.96 (m, 3H), 2.46- 2.60 (m, 1H), 2.35 (s, 3H), 1.70-1.72 (m, 1H),
1.58-1.59 (m, 2H), 1.33-1.46 (m, 2H); [M + H].sup.+ 574 Single
isomer, absolute stereochemistry unknown; Enantiomer of Ex. 112;
100% ee; retention time: 9.21 min; column: Chiralpak AD-3 150
.times. 4.6 mm I.D., 3 um; mobile phase: 30% ethanol (0.1%
ethanolamine) in CO2 112 ##STR00164## I .sup.1H NMR (400 MHz,
CDCl3) .delta. 11.98 (s, 1H), 7.44 (d, J = 3.6 Hz, 1H), 5.93 (s,
1H), 4.77-4.81 (m, 2H), 4.59-4.64 (m, 2H), 4.10-4.14 (m, 2H), 3.88
(s, 3H), 3.72 (s, 1H), 3.47-3.53 (m, 3H), 3.20 (s, 3H), 2.83-2.96
(m, 3H), 2.46-2.60 (m, 1H), 2.35 (s, 3H), 1.89- 1.90 (m, 1H), 1.58-
1.59 (m, 2H), 1.33- 1.48 (m, 2H); [M + Na].sup.+ 574 Single isomer,
absolute stereochemistry unknown; Enantiomer of Ex. 111; 100% ee;
retention time: 11.45 min; column: Chiralpak AD-3 150 .times. 4.6
mm I.D., 3 um; mobile phase: 30% ethanol (0.1% ethanolamine) in CO2
113 ##STR00165## I .sup.1H NMR (400 MHz, CD3OD) .delta. 7.63 (s,
1H), 6.13 (s, 1H), 4.78 (s, 2H), 4.09- 4.17 (m, 2H), 3.67 (brs,
1H), 3.52-3.60 (m, 3H), 3.50-3.50 (m, 1H), 3.22-3.25 (m, 3H),
3.00-3.03 (m, 2H), 2.54-2.74 (m, 1H), 2.32 (s, 3H), 2.27 (s, 3H),
1.73- 2.07 (m, 3H); [M + H].sup.+ 522 mixture of diastereomers 114
##STR00166## I .sup.1H NMR (400 MHz, CD3OD) .delta. 7.54 (s, 1H),
6.11 (s, 1H), 4.75-4.81 (m, 2H), 4.60-4.68 (m, 1H), 3.53 (t, J =
6.2 Hz, 2H), 3.31-3.37 (m, 2H), 3.20 (s, 3H), 2.98-2.99 (m, 2H),
2.59-2.66 (m, 2H), 2.30 (s, 3H), 2.25 (s, 3H), 2.06-2.15 (m, 1H),
1.71-1.92 (m, 4H); [M + H].sup.+ 511 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 115; 100% ee; retention
time: 6.190 min; column: Chiralcel OJ-H 250 .times. 4.6 mm I.D., 5
um; mobile phase: 5-40% methanol (0.05% DEA) in CO2 115
##STR00167## I .sup.1H NMR (400 MHz, CD3OD) .delta. 7.54 (s, 1H),
6.11 (s, 1H), 4.75-4.76 (m, 2H), 4.66-4.68 (m, 1H), 3.52 (t, J =
6.4 Hz, 2H), 3.37-3.51 (m, 2H), 3.20 (s, 3H), 2.99 (t, J = 6.0 Hz,
2H), 2.60-2.68 (m, 2H), 2.30 (s, 3H), 2.25 (s, 3H), 2.06- 2.09 (m,
1H), 1.71- 1.96 (m, 4H); [M + H].sup.+ 511 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 114; 98% ee; retention
time: 6.995 min; column: Chiralcel OJ-H 250 .times. 4.6 mm I.D., 5
um; mobile phase: 5-40% methanol (0.05% DEA) in CO2 116
##STR00168## I .sup.1H NMR (400 MHz, CD3OD) .delta. 7.56 (s, 1H),
6.11 (s, 1H), 4.83 (s, 2H), 4.70- 4.75 (m, 1H), 3.52 (t, J = 6.2
Hz, 2H), 3.23 (s, 3H), 2.99-3.20 (m, 6H), 2.30 (s, 3H), 2.24 (s,
3H), 1.97- 1.99 (m, 2H), 1.91- 1.95 (m, 3H); [M + H].sup.+ 527
Single isomer, absolute stereochemistry unknown; Enantiomer of Ex.
117; 100% ee; retention time: 1.170 min; column: Chiralcel OJ-3
50*4.6 mm I.D., 3 um; mobile phase: 5-40% methanol (0.05% DEA) in
CO2 117 ##STR00169## I .sup.1H NMR (400 MHz, CD3OD) .delta. 7.56
(s, 1H), 6.12 (s, 1H), 4.81 (s, 2H), 4.71- 4.79 (m, 1H), 3.53 (t, J
= 6.0 Hz, 2H), 3.23 (s, 3H), 2.99-3.20 (m, 6H), 2.30 (s, 3H), 2.25
(s, 3H), 2.10- 2.16 (m, 2H), 1.92- 2.00 (m, 3H); [M + H].sup.+ 527
Single isomer, absolute stereochemistry unknown; Enantiomer of Ex.
116; 92% ee; retention time: 1.364 min; column: Chiralcel OJ-3 50*
4.6 mm I.D., 3 um; mobile phase: 5-40% methanol (0.05% DEA) in CO2
118 ##STR00170## I .sup.1H NMR (400 MHz, CDCl3) .delta. 12.40 (s,
1H), 7.99 (s, 1H), 7.45 (d, J = 2.0 Hz, 1H), 5.94 (s, 1H),
4.78-4.82 (m, 2H), 4.63 (d, J = 4.8 Hz, 1H), 4.40-4.44 (m, 1H),
3.88 (s, 3H), 3.59- 3.62 (m, 1H), 3.49-3.52 (m, 2H), 3.20 (s, 3H),
2.91-2.96 (m, 3H), 2.46- 2.50 (m, 1H), 2.36 (s, 3H), 1.88-1.89 (m,
1H), 1.26-1.57 (m, 4H); [M + H].sup.+ 522 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 119; 100% ee; retention
time: 7.866 min; column: Chiralpak AS-H 250 .times. 4.6 mm I.D., 5
um; mobile phase: 20% methanol (0.1% ethanolamine) in CO2 119
##STR00171## I .sup.1H NMR (400 MHz, CDCl3) .delta. 12.29 (s, 1H),
8.00 (s, 1H), 7.45 (d, J = 2.4 Hz, 1H), 5.93 (s, 1H), 4.74-4.78 (m,
2H), 4.64 (d, J = 5.2 Hz, 1H), 4.41-4.44 (m, 1H), 3.88 (s, 3H),
3.59-3.63 (m, 1H), 3.51-3.52 (m, 2H), 3.20 (s, 3H), 2.93- 3.01 (m,
3H), 2.47- 2.53 (m, 1H), 2.35 (s, 3H), 1.89-1.90 (m, 1H), 1.75-1.77
(m, 1H), 1.26-1.57 (m, 3H); [M + H].sup.+ 522 Single isomer,
absolute stereochemistry unknown; Enantiomer of Ex. 118: 94% ee;
retention time: 9.458 min; column: Chiralpak AS-H 250 .times. 4.6
mm I.D., 5 um; mobile phase: 20% methanol (0.1% ethanolamine) in
CO2 120 ##STR00172## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.58
(brs, 1H), 7.49 (s, 1H), 5.90 (s, 1H), 4.85 (d, J = 18.8 Hz, 1H),
4.57 (s, 2H), 3.51-3.46 (m, 2H), 3.16 (s, 3H), 2.91 (t, J = 6 Hz,
2H), 2.58- 2.64 (m, 2H), 2.19 (s, 3H), 2.14 (s, 3H), 2.13 (s, 3H),
1.93- 2.03 (m, 3H), 1.68- 1.87 (m, 2H), 1.36 (t, J = 10.8 Hz, 1H);
[M + H].sup.+ 510 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 121; 90% ee; retention time: 7.5333 min; column:
AD-H 250 .times. 4.6 mm I.D., 5 um; mobile phase: 5-40% isopropanol
(0.05% DEA) in CO2 121 ##STR00173## I .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 11.59 (s, 1H), 10.77 (brs, 1H), 7.51 (s, 1H), 5.90
(s, 1H), 4.93 (d, J = 18.4 Hz, 1H), 4.57 (s, 2H), 3.50-3.52 (m,
3H), 3.19 (s, 3H), 2.92-3.05 (m, 4H), 2.72 (s, 3H), 2.13- 2.34 (m,
9H), 1.65 (brs, 1H); [M + Na].sup.+ 532 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 120; 91% ee; retention
time: 7.810 min; column: AD-H 250 .times. 4.6 mm I.D., 5 um; mobile
phase: 5-40% isopropanol (0.05% DEA) in CO2 122 ##STR00174## I
.sup.1H NMR (400 MHz, CDCl3) .delta. 12.40 (brs, 1H), 7.45 (d, J =
2 Hz, 1H), 5.94 (s, 1H), 4.77-4.79 (m, 2H), 4.63 (d, J = 5.2 Hz,
2H), 3.88 (s, 3H), 3.80-3.83 (m, 1H), 3.51 (s, 2H), 3.20 (s, 3H),
2.93-2.96 (m, 3H), 2.36-2.38 (m, 4H), 2.07 (s, 3H), 1.71-1.72 (m,
1H), 1.42-1.52 (m, 4H); [M + H].sup.+ 536 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 123; 100% ee; retention
time: 9.93 min; column: Chiralpak AD-H 250 .times. 4.6 mm I.D., 5
um; mobile phase: 5-40% methanol (0.05% DEA) in CO2 123
##STR00175## I .sup.1H NMR (400 MHz, CDCl3) .delta. 12.26 (brs,
1H), 7.45 (d, J = 2.4 Hz, 1H), 5.95 (s, 1H), 4.73-4.82 (m, 2H),
4.63 (d, J = 4.8 Hz, 2H), 3.89 (s, 3H), 3.79-3.82 (m, 1H), 3.51 (s,
2H), 3.20 (s, 3H), 2.91-2.96 (m, 3H), 2.36-2.44 (m, 4H), 2.07 (s,
3H), 1.38-1.71 (m, 5H); [M + H].sup.+ 536 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 122; 97% ee; retention
time: 10.60 min; column: Chiralpak AD-H 250 .times. 4.6 mm I.D., 5
um; mobile phase: 5-40% methanol (0.05% DEA) in CO2 124
##STR00176## I .sup.1H NMR (400 MHz, CDCl3) .delta. 12.35 (s,
1H),
7.53 (s, 1H), 5.93 (s, 1H), 4.78- 4.84 (m, 3H), 3.72- 3.88 (m, 6H),
3.51- 3.70 (m, 1H), 3.48- 3.50 (m, 2H), 3.18 (s, 3H), 2.95-2.96 (m,
2H), 2.55-2.60 (m, 1H), 2.35 (s, 3H), 1.70-1.76 (m, 2H); [M +
H].sup.+ 481 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 126; Diastereomer of Ex. 125 and Ex. 127; 92% ee;
retention time: 12.59 min; column: Chiralpak AD-H 250 .times. 4.6
mm I.D., 5 um; mobile phase: 40/60 hexane (0.1% DEA)/isopropanol
(0.1% ethanolamine) 125 ##STR00177## I .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 11.46 (s, 1H), 7.57 (s, 1H), 6.10 (s, 1H), 4.72
(d, J = 8.8 Hz, 1H), 4.51 (s, 2H), 3.78 (s, 3H), 3.74-3.69 (m, 3H),
3.59-3.57 (m, 1H), 3.30-3.29 (m, 2H), 3.09 (s, 3H), 2.89- 2.87 (m,
2H), 2.51- 2.50 (m, 1H), 2.19 (s, 3H), 1.51-1.47 (m, 2H); [M +
H].sup.+ 481 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 127; Diastereomer of Ex. 124 and Ex. 126; 98% ee;
retention time: 13.43 min; column: Chiralpak AD-H 250 .times. 4.6
mm I.D., 5 um; mobile phase: 70/30 hexane (0.1% DEA)/ isopropanol
(0.1% ethanolamine) 126 ##STR00178## I .sup.1H NMR (400 MHz, CDCl3)
.delta. 12.62 (s, 1H), 7.52 (s, 1H), 5.93 (s, 1H), 4.77- 4.83 (m,
3H), 3.87- 3.91 (m, 4H), 3.72- 3.76 (m, 2H), 3.62- 3.63 (m, 1H),
3.48- 3.51 (m, 2H), 3.22 (s, 3H), 2.94-2.96 (m, 2H), 2.63-2.65 (m,
1H), 2.35 (s, 3H), 1.95-1.96 (m, 1H), 1.80-1.82 (m, 1H); [M +
H].sup.+ 481 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 124; Diastereomer of Ex. 125 and Ex. 127; 100%
ee; retention time: 13.65 min; column: Chiralpak AD-H 250 .times.
4.6 mm I.D., 5 um; mobile phase: 40/60 hexane (0.1% DEA)/
isopropanol (0.1% ethanolamine) 127 ##STR00179## I .sup.1H NMR (400
MHz, DMSO-d6) .delta. 11.44 (s, 1H), 7.53 (s, 1H), 6.09 (s, 1H),
4.70 (d, J = 6.4 Hz, 1H), 4.50 (s, 2H), 3.70-3.77 (m, 4H),
3.58-3.60 (m, 2H), 3.37-3.41 (m, 2H), 3.13 (s, 3H), 2.86-2.88 (m,
2H), 2.57-2.59 (m, 1H), 2.18 (s, 3H), 1.70- 1.85 (m, 2H); [M +
H].sup.+ 481 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 125; Diastereomer of Ex. 124 and Ex. 126; 99% ee;
retention time: 14.76 min; column: Chiralpak AD-H 250 .times. 4.6
mm I.D., 5 um; mobile phase: 70/30 hexane (0.1% DEA)/ isopropanol
(0.1% ethanolamine) 128 ##STR00180## I .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 11.57 (s, 1H), 7.56 (s, 1H), 5.89 (s, 1H), 4.85-
4.92 (m, 2H), 4.57 (s, 2H), 4.09-4.19 (m, 1H), 3.97-4.04 (m, 1H),
3.81-3.90 (m, 3H), 3.71-3.78 (m, 1H), 3.46 (t, J = 6.40 Hz, 2H),
3.22 (s, 3H), 2.95-3.05 (m, 1H), 2.90 (t, J = 5.9 Hz, 2H), 2.17 (s,
3H), 2.13 (s, 3H); [M + Na].sup.+ 530 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 129; 100% ee; retention
time: 8.161 min; column: Chiralcel OD-H 150 .times. 4.6 mm I.D., 5
um; mobile phase: 5-40% methanol (0.05% DEA) in CO2 129
##STR00181## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.46 (brs,
1H), 7.56 (s, 1H), 5.90 (s, 1H), 5.07-4.74 (m, 2H), 4.58 (s, 2H),
4.18- 4.10 (m, 1H), 4.02- 3.98 (m, 1H), 3.93- 3.81 (m, 3H), 3.77-
3.71 (m, 1H), 3.46 (t, J = 6 Hz, 2H), 3.22 (s, 3H), 3.03-2.96 (m,
1H), 2.90 (t, J = 6 Hz, 2H), 2.17 (s, 3H), 2.13 (s, 3H); [M +
Na].sup.+ 530 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 128; 96% ee; retention time: 8.511 min; column:
Chiralcel OD-H 150 .times. 4.6 mm I.D., 5 um; mobile phase: 5-40%
methanol (0.05% DEA) in CO2 130 ##STR00182## I .sup.1H NMR (400
MHz, CDCl3) .delta. 11.75 (brs, 1H), 7.24 (s, 1H), 5.94 (s, 1H),
4.78- 4.83 (m, 2H), 4.69 (d, J = 5.20 Hz, 1H), 3.63 (t, J = 6.00
Hz, 2H), 3.18 (s, 3H), 3.02- 3.10 (m, 2H), 2.79- 2.94 (m, 2H), 2.75
(t, J = 6.00 Hz, 2H), 2.36 (s, 3H), 2.28 (s, 3H), 2.26 (s, 3H),
2.03- 2.22 (m, 3H), 1.86- 2.02 (m, 2H); [M + H].sup.+ 507 Single
isomer, absolute stereochemistry unknown; Enantiomer of Ex. 131;
99% ee; retention time: 3.455 min; column: Chiralpak AD-H 250
.times. 4.6 mm I.D., 5 um; mobile phase; 40% methanol (0.05% DEA)
in CO2 131 ##STR00183## I .sup.1H NMR (400 MHz, CDCl3) .delta.
11.75 (brs, 1H), 7.24 (s, 1H), 5.94 (s, 1H), 4.78- 4.83 (m, 2H),
4.69 (d, J = 5.20 Hz, 1H), 3.63 (t, J = 6.00 Hz, 2H), 3.18 (s, 3H),
3.02- 3.10 (m, 2H), 2.79- 2.94 (m, 2H), 2.75 (t, J = 6.00 Hz, 2H),
2.36 (s, 3H), 2.28 (s, 3H), 2.26 (s, 3H), 2.03- 2.22 (m, 3H), 1.86-
2.02 (m, 2H); [M + H].sup.+ 507 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 130; 99% ee; retention
time: 6.221 min; column: Chiralpak AD-H 250 .times. 4.6 mm I.D., 5
um; mobile phase; 40% methanol (0.05% DEA) in CO2 132 ##STR00184##
I .sup.1H NMR (400 MHz, CDCl3) .delta. 12.11 (brs, 1H), 7.47 (d, J
= 8 Hz, 1H), 7.11 (d, J = 7.6 Hz, 1H), 5.95 (s, 1H), 4.81-4.83 (m,
3H), 3.83-3.89 (m, 1H), 3.70-3.73 (m, 2H), 3.61-3.64 (m, 3H), 3.19
(s, 3H), 2.82- 2.85 (m, 2H), 2.64- 2.66 (m, 1H), 2.37 (s, 3H), 2.29
(s, 3H), 1.95-1.97 (m, 1H), 1.81-1.84 (m, 1H); [M + H].sup.+ 431
Single isomer, absolute stereochemistry unknown; Enantiomer of Ex.
133; Diastereomer of Ex. 135 and Ex. 134; 99% ee; retention time:
6.84 min; column: Chiralpak AD-H 250 .times. 4.6 mm I.D., 5 um;
mobile phase: 5-40% methanol (0.05% DEA) in CO2 133 ##STR00185## I
.sup.1H NMR (400 MHz, CDCl3) .delta. 12.18 (brs, 1H), 7.47 (d, J =
7.6 Hz, 1H), 7.11 (d, J = 8 Hz, 1H), 5.96 (s, 1H), 4.81-4.83 (m,
3H), 3.87-3.89 (m, 1H), 3.70-3.73 (m, 2H), 3.61-3.64 (m, 3H), 3.19
(s, 3H), 2.82- 2.85 (m, 2H), 2.64- 2.65 (m, 1H), 2.37 (s, 3H), 2.29
(s, 3H), 1.95-1.97 (m, 1H), 1.80-1.6 (m, 1H); [M + H].sup.+ 431
Single isomer, absolute stereochemistry unknown; Enantiomer of Ex.
132; Diastereomer of Ex. 135 and Ex. 134; 100% ee; retention time:
7.13 min; column: Chiralpak AD-H 250 .times. 4.6 mm I.D., 5 um;
mobile phase: 5-40% methanol (0.05% DEA) in CO2 134 ##STR00186## I
.sup.1H NMR (400 MHz, CDCl3) .delta. 11.84 (brs, 1H), 7.47 (d, J =
8 Hz, 1H), 7.12 (d, J = 8 Hz, 1H), 5.96 (s, 1H), 4.80-4.88 (m, 3H),
3.71-3.88 (m, 3H), 3.63-3.65 (m, 3H), 3.16 (s, 3H), 2.83- 2.86 (m,
2H), 2.51- 2.60 (m, 1H), 2.38 (s, 3H), 2.29 (s, 3H), 1.69-1.74 (m,
2H); [M + H].sup.+ 431 Single isomer, absolute stereochemistry
unknown; enantiomer of Ex. 135; diastereomer of Ex. 132 and Ex.
133; 100% ee; retention time: 7.21 min; column: Chiralpak AD-H 250
.times. 4.6 mm I.D., 5 um; mobile phase: 5-40% methanol (0.05% DEA)
in CO2; wavelength: 220 nm 135 ##STR00187## I .sup.1H NMR (400 MHz,
CDCl3) .delta. 11.71 (brs, 1H), 7.47 (d, J = 7.6 Hz, 1H), 7.12 (d,
J = 7.6 Hz, 1H), 5.95 (s, 1H), 4.80-4.88 (m, 3H), 3.83-3.88 (m,
3H), 3.64-3.72 (m, 3H), 3.16 (s, 3H), 2.83-2.86 (m, 2H), 2.55-2.60
(m, 1H), 2.38 (s, 3H), 2.28 (s, 3H), 1.69-1.74 (m, 2H); [M +
H].sup.+ 431 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 134; Diastereomer of Ex. 132 and Ex. 133; 99% ee;
retention time: 7.32 min; column: Chiralpak AD-H 250 .times. 4.6 mm
I.D., 5 um; mobile phase: 5-40% methanol (0.05% DEA) in CO2 136
##STR00188## I .sup.1H NMR (400 MHz, CD3OD) .delta. 7.58 (s, 1H),
6.16 (s, 1H), 4.81 (s, 2H), 4.72 (d, J = 4.65 Hz, 1H), 3.58 (t, J =
6.24 Hz, 2H), 3.26 (s, 3H), 2.99- 3.10 (m, 4H), 2.40 (s, 3H), 2.35
(s, 3H), 2.30 (s, 3H), 2.17 (d, J = 11.86 Hz, 2H), 1.68-1.85 (m,
3H), 1.54 (d, J = 7.70 Hz, 2H); [M + H].sup.+ 492 [.alpha.]D22 =
+70.1.degree. (c = 0.2, MeOH); Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 137; ~99% ee; retention
time 10.03 min; column: Lux Cellulose-4 4.6 .times. 100 mm 3 u;
mobile phase: 50% MeOH/DEA in CO2, 4 mL/min 137 ##STR00189## I
.sup.1H NMR (400 MHz, CD3OD) .delta. 7.58 (s, 1H), 6.16 (s, 1H),
4.81 (s, 2H), 4.72 (d, J = 4.65 Hz, 1H), 3.58 (t, J = 6.24 Hz, 2H),
3.26 (s, 3H), 2.99- 3.10 (m, 4H), 2.40 (s, 3H), 2.35 (s, 3H), 2.30
(s, 3H), 2.17 (d, J = 11.86 Hz, 2H), 1.68-1.85 (m, 3H), 1.54 (d, J
= 7.70 Hz, 2H); [M + H].sup.+ 492 [.alpha.]D22 = -59.5.degree. (c =
0.2, MeOH); Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 136; >99% ee; retention time 7.25 min; column:
Lux Cellulose-4 4.6 .times. 100 mm 3 u; mobile phase: 50% MeOH/DEA
in CO2, 4 mL/min 138 ##STR00190## I .sup.1H NMR (400 MHz, CD3OD)
.delta. 7.77 (d, J = 4.00 Hz, 1H), 6.11 (s, 1H), 4.97-4.98 (m, 2H),
4.75 (s, 2H), 4.22-4.27 (m, 2H), 3.98-4.09 (m, 5H), 3.84-3.86 (m,
1H), 3.52 (t, J = 6.40 Hz, 2H), 3.08-3.12 (m, 1H), 2.97-3.00 (m,
2H), 2.30 (s, 3H), 2.25 (s, 3H); [M + H].sup.+ 554 Single isomer,
absolute stereochemistry unknown; Enantiomer of Ex. 139; 96% ee;
retention time: 6.104 min; column: Chiralcel OD-3 150 .times. 4.6
mm I.D., 3 um; mobile phase: 5-40% methanol (0.05% DEA) in CO2;
flow rate: 2.5 mL/min 139 ##STR00191## I .sup.1H NMR (400 MHz,
CD3OD) .delta. 7.77 (d, J = 4.00 Hz, 1H), 6.11 (s, 1H), 4.92-4.98
(m, 2H), 4.57 (s, 2H), 4.22-4.27 (m, 2H), 3.98-4.08 (m, 5H),
3.84-3.86 (m, 1H), 3.52 (t, J = 6.40 Hz, 2H), 3.08-3.12 (m, 1H),
2.98-3.00 (m, 2H), 2.30 (s, 3H), 2.25 (s, 3H); [M + H].sup.+ 554
Single isomer, absolute stereochemistry unknown; Enantiomer of Ex.
138; 98% ee; retention time: 6.403 min; column: Chiralcel OD-3 150
.times. 4.6 mm I.D., 3 um; mobile phase: 5-40% methanol (0.05% DEA)
in CO2; flow rate: 2.5 mL/min 140 ##STR00192## I .sup.1H NMR (400
MHz, DMSO-d6) .delta. 11.55 (brs, 1H), 7.47 (s, 1H), 5.89 (s, 1H),
4.66 (d, J = 7.2 Hz, 1H), 4.57 (s, 2H), 4.38 (t, J = 5.6 Hz, 1H),
3.40-3.50 (m, 4H), 3.09 (s, 3H), 2.85-2.95 (m, 2H), 2.17 (s, 3H),
2.12 (s, 3H), 1.80-1.85 (m, 1H), 0.78 (d, J = 6.8 Hz, 3H); [M +
Na].sup.+ 475 Single (2R) isomer, other stereocenter unknown;
Enantiomer of Ex. 142; Diastereomer of Ex. 143 and Ex. 141; 95% ee;
retention time 6.36 min; column: Chiralpak AD-H 250 .times. 4.6 mm
I.D., 5 um; mobile phase: 5-40% methanol (0.05% DEA) in CO2 141
##STR00193## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.58 (brs,
1H), 7.42 (s, 1H), 5.89 (s, 1H), 4.83 (d, J = 2.8 Hz, 1H), 4.65 (t,
J = 5.2 Hz, 1H), 4.57 (s, 2H), 3.40-3.48 (m, 3H), 3.27-3.28 (m,
1H), 3.15 (s, 3H), 2.89 (t, J = 6.0 Hz, 2H), 2.16 (s, 3H), 2.12 (s,
3H), 1.85-1.89 (m, 1H), 0.66 (d, J = 6.8 Hz, 3H); [M + Na].sup.+
475 Single (2R) isomer, other stereocenter unknown; Enantiomer of
Ex. 143; Diasteromer of Ex. 140 and Ex. 142; 99% ee; retention time
7.69 min; column: Chiralpak AD-H 250 .times. 4.6 mm I.D., 5 um;
mobile phase: 5-40% isopropanol (0.05% DEA) in CO2 142 ##STR00194##
I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.56 (brs, 1H), 7.47 (s,
1H), 5.89 (s, 1H), 4.66 (d, J = 7.2 Hz, 1H), 4.56 (s, 2H),
3.40-3.50 (m, 5H), 3.09 (s, 3H), 2.89 (t, J = 5.6 Hz, 2H), 2.17 (s,
3H), 2.12 (s, 3H), 1.80-1.85 (m, 1H), 0.78 (d, J = 6.4 Hz, 3H); [M
+ Na].sup.+ 475 Single (2S) isomer, other stereocenter unknown;
Enantiomer of Ex. 140; Diastereomer of Ex. 143 and Ex. 141; 100%
ee; retention time: 6.64 min; column: Chiralpak AD-H 250 .times.
4.6 mm I.D., 5 um; mobile phase: 5-40% methanol (0.05% DEA) in CO2
143 ##STR00195## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.57
(brs, 1H), 7.42 (s, 1H), 5.89 (s, 1H), 4.83 (d, J = 2.8 Hz, 1H),
4.57 (s, 2H), 3.40-3.50 (m, 5H), 3.16 (s, 3H), 2.89 (t, J = 6.4 Hz,
2H), 2.16 (s, 3H), 2.12 (s, 3H), 1.86-1.89 (m, 1H), 0.66 (d, J =
6.8 Hz, 3H); [M + H].sup.+ 453 Single (2S) isomer, other
stereocenter unknown; Enantiomer of Ex. 141; Diastereomer of Ex.
142 and Ex. 140; 98% ee; retention time: 7.43 min; column:
Chiralpak AD-H 250 .times. 4.6 mm I.D., 5 um; mobile phase: 5-40%
isopropanol (0.05% DEA) in CO2 144 ##STR00196## I .sup.1H NMR (400
MHz, CDCl3) .delta. 12.06 (brs, 1H), 7.26 (s, 1H), 5.92 (s, 1H),
5.11 (d, J = 6.8 Hz, 1H), 4.75- 4.79 (m, 3H), 4.62- 4.68 (m, 3H),
3.87 (s, 3H), 3.37-3.47 (m, 3H), 3.28 (s, 3H), 2.74 (d, J = 3.2 Hz,
2H), 2.34 (s, 3H), 2.24 (s, 3H); [M + H].sup.+ 447 Single isomer,
absolute stereochemistry unknown; Enantiomer of Ex. 145; 100% ee;
retention time: 5.041 min; column: Chiralcel OD-3 150 .times. 4.6
mm I.D., 3 um; mobile phase: 5-40% ethanol (0.05% DEA) in CO2 145
##STR00197## I .sup.1H NMR (400 MHz, CDCl3) .delta. 7.26 (s, 1H),
5.92 (s, 1H), 5.11-5.12 (m, 1H), 4.76-4.78 (m, 3H), 4.62-4.68 (m,
3H), 3.87 (s, 3H), 3.37- 3.47 (m, 3H), 3.28 (s, 3H), 2.74-2.75 (m,
2H), 2.33 (s, 3H), 2.24 (s, 3H); [M + H].sup.+ 447 Single isomer,
absolute stereochemistry unknown; Enantiomer of Ex. 144; 99% ee;
retention time: 5.168 min; column: Chiralcel OD-3 150 .times. 4.6
mm I.D., 3 um; mobile phase: 5-40% ethanol (0.05% DEA) in CO2 146
##STR00198## I .sup.1H NMR (400 MHz, CDCl3) .delta. 11.45 (brs,
1H), 7.55 (s, 1H), 5.88 (s, 1H), 4.70- 4.73 (m, 2H), 4.58 (d, J =
4.40 Hz, 1H), 3.57- 3.60 (m, 2H), 3.30- 3.33 (m, 2H), 3.14 (s, 3H),
2.86-2.89 (m, 2H), 2.63 (s, 3H), 2.48-2.53 (m, 2H), 2.29 (s, 3H),
2.22 (s, 3H), 1.98-2.02 (m, 1H), 1.83-1.88 (m, 1H), 1.74-1.79 (m,
2H), 1.58-1.63 (m, 1H); [M + H].sup.+ 538 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 147; 100% ee; retention
time; 4.177 min; column: Chiralcel OD-3 100 .times. 4.6 mm I.D., 3
um; mobile phase: 5-40% ethanol (0.05% DEA) in CO2 147 ##STR00199##
I .sup.1H NMR (400 MHz, CDCl3) .delta. 11.82 (brs, 1H), 7.57 (s,
1H), 5.87 (s, 1H), 4.69 (s, 2H), 4.58 (d, J = 4.40 Hz, 1H), 3.57-
3.60- (m, 2H), 3.12 (s, 3H), 3.01-3.07 (m, 2H), 2.85-2.88 (m, 2H),
2.38 (s, 3H), 2.29 (s, 3H), 2.22 (s, 3H), 2.08-2.13 (m, 2H),
1.82-1.88 (m, 1H), 1.69-1.75 (m, 2H), 1.58-1.63 (m, 1H), 1.40-1.46
(m, 1H); [M + H].sup.+ 538 Single isomer, absolute stereochemistry
unknown; Enantiomer of Ex. 146; 100% ee; retention time; 4.202 min;
column: Chiralcel OD-3 100 .times. 4.6 mm I.D., 3 um; mobile phase:
5-40% ethanol (0.05% DEA) in CO2 148 ##STR00200## I .sup.1H NMR
(400 MHz, CDCl3) .delta. 11.25 (br. s., 1H), 7.43 (s, 1H), 5.95 (s,
1H), 4.71- 4.85 (m, 3H), 3.60- 3.76 (m, 2H), 3.27 (s, 3H),
3.18-3.25 (m, 1H), 2.88-3.16 (m, 5H), 2.79 (m, 1H), 2.37 (s, 3H),
2.29 (s, 3H), 2.16-2.27 (m, 2H); [M + Na].sup.+ 535 Single isomer,
absolute stereochemistry unknown; Enantiomer of Ex. 151;
Diastereomer of Ex. 149 and Ex. 150; 97% ee; retention time: 1.865
min; column: Chiralcel OJ-3 100 .times. 4.6 mm I.D., 3 um; mobile
phase: 15% methanol (0.05% DEA) in CO2 149 ##STR00201## I .sup.1H
NMR (400 MHz, CDCl3) .delta. 10.79 (br. s., 1H), 7.48 (s, 1H), 5.93
(s, 1H), 4.70- 4.84 (m, 3H), 3.61- 3.74 (m, 2H), 3.26 (s, 3H),
3.18-3.25 (m, 1H), 3.14-3.16 (m, 2H), 2.96 (t, J = 6.27 Hz, 2H),
2.74-2.93 (m, 2H), 2.36 (s, 3H), 2.27 (s, 3H), 1.94- 2.19 (m, 2H);
[M + Na].sup.+ 535 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 150; Diastereomer of Ex. 148 and Ex. 151; 100%
ee; retention time: 1.949 min; column: Chiralcel OJ-3 100 .times.
4.6 mm I.D., 3 um; mobile phase: 15% methanol (0.05% DEA) in
CO2
150 ##STR00202## I .sup.1H NMR (400 MHz, CDCl3) .delta. 11.61 (br.
s., 1H), 7.49 (s, 1H), 5.95 (s, 1H), 4.68- 4.86 (m, 3H), 3.58- 3.77
(m, 2H), 3.27 (s, 3H), 3.19-3.25 (m, 1H), 3.14-3.17 (m, 2H), 2.96
(t, J = 6.15 Hz, 2H), 2.75-2.94 (m, 2H), 2.37 (s, 3H), 2.29 (s,
3H), 1.96- 2.21 (m, 2H); [M + Na].sup.+ 535 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 149; Diastereomer of Ex.
148 and Ex. 151; 97% ee; retention time: 2.253 min; column:
Chiralcel OJ-3 100 .times. 4.6 mm I.D., 3 um; mobile phase: 15%
methanol (0.05% DEA) in CO2 151 ##STR00203## I .sup.1H NMR (400
MHz, CDCl3) .delta. 11.72 (br. s., 1H), 7.43 (s, 1H), 5.96 (s, 1H),
4.71- 4.86 (m, 3H), 3.58- 3.78 (m, 2H), 3.22- 3.31 (m, 4H), 2.87-
3.19 (m, 5H), 2.73- 2.85 (m, 1H), 2.36 (s, 3H), 2.29 (s, 3H),
2.16-2.27 (m, 2H); [M + Na].sup.+ 535 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 148; Diastereomer of Ex.
149 and Ex. 150; 100% ee; retention time: 2.596 min; column:
Chiralcel OJ-3 100 .times. 4.6 mm I.D., 3 um; mobile phase: 15%
methanol (0.05% DEA) in CO2 152 ##STR00204## I .sup.1H NMR (400
MHz, CDCl3) .delta. 7.47 (s, 1H), 5.96 (s, 1H), 4.93 (d, J = 4.8,
1H), 4.77 (s, 2H), 4.22- 4.28 (m, 2H), 4.05- 4.08 (m, 1H), 3.67-
3.79 (m, 3H), 3.32 (s, 3H), 2.92-2.98 (m, 3H), 2.37 (s, 3H), 2.29
(s, 3H); [M + H].sup.+ 499 Single isomer, absolute stereochemistry
unknown; Enantiomer of Ex. 153; 100% ee; retention time 5.106 min;
column: Chiralpak AD-3 100 .times. 4.6 mm I.D., 3 um; mobile phase:
5-40% ethanol (0.05% DEA) in CO2 153 ##STR00205## I .sup.1H NMR
(400 MHz, CDCl3) .delta. 7.47 (s, 1H), 5.96 (s, 1H), 4.93 (d, J =
4.8, 1H), 4.77 (s, 2H), 4.22- 4.28 (m, 2H), 4.05- 4.08 (m, 1H),
3.67- 3.78 (m, 3H), 3.32 (s, 3H), 2.92-2.98 (m, 3H), 2.37 (s, 3H),
2.29 (s, 3H); [M + H].sup.+ 499 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 152; 100% ee; retention
time 4.69 min; column: Chiralpak AD-3 100 .times. 4.6 mm I.D., 3
um; mobile phase: 5-40% ethanol (0.05% DEA) in CO2 154 ##STR00206##
I .sup.1H NMR (400 MHz, CD3OD) .delta. 7.35 (s, 1H), 6.13 (s, 1H),
4.79 (s, 2H), 4.56- 4.70 (m, 6H), 3.42- 3.53 (m, 3H), 3.19 (s, 3H),
2.72-2.87 (m, 5H), 2.31 (s, 6H), 2.25-2.28 (m, 3H), 1.64-1.83 (m,
5H); [M + H].sup.+ 514 Single isomer, absolute stereochemistry
unknown; Enantiomer of Ex. 155; 99% ee; retention time: 4.001 min;
column: Chiralpak AD-3 100 .times. 4.6 mm I.D., 3 um; mobile phase:
5-40% methanol (0.05% DEA) in CO2 155 ##STR00207## I .sup.1H NMR
(400 MHz, CD3OD) .delta. 7.35 (s, 1H), 6.13 (s, 1H), 4.79 (s, 2H),
4.54- 4.69 (m, 6H), 3.41- 3.51 (m, 4H), 3.19 (s, 3H), 2.73-2.86 (m,
4H), 2.30 (s, 6H), 2.26 (s, 3H), 1.64- 1.86 (m, 5H); [M + H].sup.+
514 Single isomer, absolute stereochemistry unknown; Enantiomer of
Ex. 154; 96% ee; retention time: 4.312 min; column: Chiralpak AD-3
100 .times. 4.6 mm I.D., 3 um; mobile phase: 5-40% methanol (0.05%
DEA) in CO2 156 ##STR00208## I .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 11.57 (brs, 1H), 7.44 (s, 1H), 5.89 (s, 1H), 4.50-4.65 (m,
3H), 4.45-4.50 (m, 2H), 4.35-4.40 (m, 2H), 3.46 (t, J = 6.0 Hz,
2H), 3.30-3.35 (m, 3H), 3.12 (s, 3H), 2.89 (t, J = 6.0 Hz, 2H),
2.60-2.70 (m, 2H), 2.17 (s, 3H), 2.12 (s, 3H), 1.50- 1.65 (m, 4H),
1.35- 1.45 (m, 1H); [M + H].sup.+ 534 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 157; 100% ee; retention
time: 2.263 min; column: Chiralpak AY 100 .times. 4.6 mm I.D., 3
um; mobile phase: 5-40% methanol (0.05% DEA) in CO2 157
##STR00209## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.58 (brs,
1H), 7.44 (s, 1H), 5.89 (s, 1H), 4.50-4.65 (m, 3H), 4.45-4.50 (m,
2H), 4.35-4.40 (m, 2H), 3.46 (t, J = 6.0 Hz, 2H), 3.30-3.35 (m,
3H), 3.12 (s, 3H), 2.89 (t, J = 5.6 Hz, 2H), 2.60-2.70 (m, 2H),
2.17 (s, 3H), 2.12 (s, 3H), 1.50- 1.65 (m, 4H), 1.35- 1.45 (m, 1H);
[M + H].sup.+ 534 Single isomer absolute stereochemistry unknown;
Enantiomer of Ex. 156; 100% ee; retention time: 3.339 min; column:
Chiralpak AY 100 .times. 4.6 mm I.D., 3 um; mobile phase: 5-40%
methanol (0.05% DEA) in CO2 158 ##STR00210## I .sup.1H NMR (400
MHz, CDCl3) .delta. 11.26 (brs, 1H), 7.49 (s, 1H), 5.94 (s, 1H),
5.03 (d, J = 4.4 Hz, 1H), 4.77 (s, 2H), 4.21-4.26 (m, 2H),
4.03-4.04 (m, 1H), 3.68-3.76 (m, 3H), 3.40-3.48 (m, 2H), 2.94-2.97
(m, 3H), 2.37 (s, 3H), 2.28 (s, 3H), 1.25 (t, J = 7 Hz, 3H); [M +
H].sup.+ 513 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 159; 100% ee; retention time: 2.535 min; column:
Chiralcel OJ-3 100 .times. 4.6 mm I.D., 3 um; mobile phase: 5-40%
ethanol (0.05% DEA) in CO2 159 ##STR00211## I .sup.1H NMR (400 MHz,
CDCl3) .delta. 11.35 (brs, 1H), 7.49 (s, 1H), 5.95 (s, 1H), 5.02
(d, J = 4.8 Hz, 1H), 4.77- 4.8 (m, 2H), 4.21- 4.26 (m, 2H), 4.03-
4.04 (m, 1H), 3.68- 3.76 (m, 3H), 3.40- 3.48 (m, 2H), 2.94- 2.97
(m, 3H), 2.37 (s, 3H), 2.28 (s, 3H), 1.25 (t, J = 6.8 Hz, 3H); [M +
H].sup.+ 513 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 158; 100% ee; retention time: 22.782 min; column:
Chiralcel OJ-3 100 .times. 4.6 mm I.D., 3 um; mobile phase: 5-40%
ethanol (0.05% DEA) in CO2 160 ##STR00212## I .sup.1H NMR (400 MHz,
CD3OD) .delta. 7.61 (s, 1H), 6.10 (s, 1H), 4.76 (s, 2H), 3.83- 3.91
(m, 1H), 3.67- 3.76 (m, 2H), 3.58 (t, J = 7.83 Hz, 1H), 3.53 (t, J
= 6.24 Hz, 2H), 3.22 (s, 3H), 2.99 (t, J = 6.24 Hz, 2H), 2.64 (m,
1H), 2.30 (s, 3H), 2.25 (s, 3H), 1.93- 2.05 (m, 1H), 1.78- 1.89 (m,
1H) One proton obscured by solvent; [M + H].sup.+ 465
[.alpha.].sub.D = -48.0.degree. (c 0.1 MeOH), >99% ee; Absolute
and relative stereochemistry undetermined; Enantiomer of Ex. 161;
Diastereomer of Ex. 162 and 163 161 ##STR00213## I .sup.1H NMR (400
MHz, CD3OD) .delta. 7.61 (s, 1H), 6.10 (s, 1H), 4.76 (s, 2H), 3.83-
3.92 (m, 1H), 3.67- 3.76 (m, 2H), 3.58 (t, J = 7.82 Hz, 1H), 3.53
(t, J = 6.24 Hz, 2H), 3.22 (s, 3H), 2.99 (t, J = 6.11 Hz, 2H), 2.64
(m, 1H), 2.30 (s, 3H), 2.25 (s, 3H), 1.93- 2.04 (m, 1H), 1.78- 1.92
(m, 1H) One proton obscured by solvent; [M + H].sup.+ 465
[.alpha.].sub.D = +74.7.degree. (c 0.1 MeOH), 90% ee, Absolute and
relative stereochemistry undetermined; Enantiomer of Ex. 160;
Diastereomer of Ex. 162 and Ex. 163 162 ##STR00214## I .sup.1H NMR
(400 MHz, CD3OD) .delta. 7.61 (s, 1H), 6.11 (s, 1H), 4.76 (s, 2H),
3.76- 3.90 (m, 3H), 3.70 (q, J = 7.50 Hz, 1H), 3.53 (t, J = 6.24
Hz, 2H), 3.18 (s, 3H), 2.97- 3.03 (m, 2H), 2.54- 2.66 (m, 1H), 2.30
(s, 3H), 2.25 (s, 3H), 1.62-1.81 (m, 2H) One proton obscured by
solvent; [M + H].sup.+ 465 [.alpha.].sub.D = -68.8.degree. (c 0.1
MeOH); >99% ee; Absolue and relative stereochemistry
undetermined; Enantiomer of Ex. 163; Diastereomer Ex. 160 and Ex.
161 163 ##STR00215## I .sup.1H NMR (400 MHz, CD3OD) .delta. 7.61
(s, 1H), 6.11 (s, 1H), 4.76 (s, 2H), 3.76- 3.91 (m, 3H), 3.70 (q, J
= 7.58 Hz, 1H), 3.53 (t, J = 6.24 Hz, 2H), 3.18 (s, 3H), 2.96- 3.03
(m, 2H), 2.60 (sxt, J = 7.38 Hz, 1H), 2.30 (s, 3H), 2.25 (s, 3H),
1.63-1.81 (m, 2H) One proton obscured by solvent; [M + H].sup.+ 465
[.alpha.].sub.D = +59.4.degree. (c 0.1 MeOH); >99% ee; Absolute
and relative stereochemistry undetermined; Enantiomer of Ex. 162;
Diastereomer of Ex. 160 and Ex. 161 164 ##STR00216## I .sup.1H NMR
(400 MHz, CD3OD) .delta. 7.54 (s, 1H), 6.11 (s, 1H), 4.76 (s, 2H),
4.66 (d, J = 5.62 Hz, 1H), 4.44- 4.56 (m, 1H), 4.12- 4.27 (m, 2H),
3.66- 3.78 (m, 1H), 3.53 (t, J = 6.24 Hz, 2H), 2.99 (t, J = 6.11
Hz, 2H), 2.85-2.96 (m, 1H), 2.51-2.66 (m, 1H), 2.30 (s, 3H), 2.25
(s, 3H), 1.86-2.00 (m, 1H), 1.69-1.79 (m, 1H), 1.31-1.54 (m, 3H);
[M + H].sup.+ 539 [.alpha.].sub.D = -36.9.degree. (c 0.1 MeOH);
Absolute stereochemistry undetermined; Enantiomer of Ex. 165; (-)
isomer of Ex. 70 racemate 165 ##STR00217## I .sup.1H NMR (400 MHz,
CD3OD) .delta. 7.54 (s, 1H), 6.11 (s, 1H), 4.76 (s, 2H), 4.66 (d, J
= 5.62 Hz, 1H), 4.43- 4.56 (m, 1H), 4.12- 4.27 (m, 2H), 3.67- 3.78
(m, 1H), 3.53 (t, J = 6.11 Hz, 2H), 2.99 (t, J = 6.11 Hz, 2H),
2.84-2.96 (m, 1H), 2.49-2.67 (m, 1H), 2.30 (s, 3H), 2.25 (s, 3H),
1.86-2.01 (m, 1H), 1.69-1.79 (m, 1H), 1.32-1.56 (m, 3H); [M +
H].sup.+ 539 [.alpha.].sub.D = +139.1.degree. (c 0.1 MeOH);
Absolute stereochemistry undetermined; Enantiomer of Ex. 164; (+)
isomer of Ex. 70 racemate 166 ##STR00218## I .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 7.42 (s, 1H), 5.91 (s, 1H), 4.55 (s, 2H), 4.51 (d,
J = 5.50 Hz, 1H), 3.43- 3.45 (m, 2H), 3.10 (s, 3H), 2.87 (t, J =
5.59 Hz, 2H), 2.16 (s, 3H), 2.12 (s, 3H), 1.45-1.74 (m, 2H),
1.20-1.40 (m, 3H) Four protons obscured by solvent; [M + H].sup.+
478 [.alpha.].sub.D = +73.0.degree. (c 0.1 MeOH); 97% ee; Absolute
stereochemistry undetermined; Enantiomer of Ex. 167 167
##STR00219## I .sup.1H NMR (700 MHz, DMSO-d6) .delta. 7.43 (s, 1H),
5.89 (s, 1H), 4.54-4.58 (m, 2H), 4.52 (d, J = 5.72 Hz, 1H), 3.46
(t, J = 5.94 Hz, 2H), 3.11 (s, 3H), 2.83-3.00 (m, 3H), 2.17 (s,
3H), 2.12 (s, 3H), 1.62-1.72 (m, 1H), 1.51-1.62 (m, 1H), 1.29-1.42
(m, 1H), 1.19-1.28 (m, 2H). Two protons obscured by solvent; [M +
H].sup.+ 478 [.alpha.].sub.D = -59.7.degree. (c 0.1 MeOH); 99% ee;
Absolute stereochemistry undetermined; Enantiomer of Ex. 166 168
##STR00220## I .sup.1H NMR (700 MHz, DMSO-d6) .delta. 11.55 (br.
s., 1H), 7.26 (s, 1H), 5.88 (s, 1H), 4.52-4.62 (m, 3H), 4.44 (br.
s., 1H), 4.28-4.39 (m, 1H), 4.04-4.11 (m, 1H), 3.96-4.04 (m, 1H),
3.57-3.71 (m, 1H), 3.40 (br. s., 2H), 3.08 (s, 3H), 2.82 (t, J =
11.88 Hz, 1H), 2.72 (t, J = 5.94 Hz, 2H), 2.43- 2.48 (m, 1H), 2.24
(s, 3H), 2.16 (s, 3H), 2.12 (s, 3H), 1.79-1.88 (m, 1H), 1.65-1.74
(m, 1H), 1.15-1.37 (m, 3H); [M + H].sup.+ 516 [.alpha.].sub.D =
+60.2.degree. (c 0.1 MeOH); 99% ee; Absolute stereochemistry
undetermined; Enantiomer of Ex. 169 169 ##STR00221## I .sup.1H NMR
(700 MHz, DMSO-d6) .delta. 11.55 (br. s., 1H), 7.26 (s, 1H), 5.88
(s, 1H), 4.51-4.60 (m, 3H), 4.44 (br. s., 1H), 4.29-4.39 (m, 1H),
4.04-4.10 (m, 1H), 3.95-4.04 (m, 1H), 3.58-3.70 (m, 1H), 3.40 (br.
s., 2H), 3.08 (s, 3H), 2.82 (t, J = 11.99 Hz, 1H), 2.72 (t, J =
5.94 Hz, 2H), 2.43- 2.47 (m, 1H), 2.24 (s, 3H), 2.16 (s, 3H), 2.12
(s, 3H), 1.79-1.87 (m, 1H), 1.66-1.72 (m, 1H), 1.16-1.36 (m, 3H);
[M + H].sup.+ 516 [.alpha.].sub.D = -31.2.degree. (c 0.1 MeOH):
~80% ee; Absolute stereochemistry undetermined; Enantiomer of Ex.
168 170 ##STR00222## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.54
(br. s., 1H), 7.55 (s, 1H), 5.88 (s, 1H), 5.00 (dd, J = 4.89, 5.87
Hz, 1H), 4.91 (d, J = 4.89 Hz, 1H), 4.57 (s, 2H), 4.02-4.24 (m,
3H), 3.78-3.89 (m, 1H), 3.66-3.75 (m, 1H), 3.46 (t, J = 5.99 Hz,
2H), 3.21 (s, 3H), 2.94-3.04 (m, 1H), 2.90 (t, J = 6.11 Hz, 2H),
2.17 (s, 3H), 2.12 (s, 3H), 1.15 (d, J = 6.85 Hz, 3H); [M +
H].sup.+ 522 [.alpha.].sub.D = +123.5.degree. (c = 0.1, MeOH);
>99% de; Single diastereomer containing (R)-2- hydroxy-
propanamide; other chiral center undetermined; Enantiomer of Ex.
173 Diastereomer of Ex. 171 and Ex. 172 171 ##STR00223## I .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 11.54 (br. s., 1H), 7.55 (s, 1H),
5.89 (s, 1H), 4.97 (dd, J = 6.05, 15.34 Hz, 1H), 4.91 (t, J = 6.42
Hz, 1H), 4.57 (s, 2H), 4.17- 4.29 (m, 1H), 4.00-4.14 (m, 2H),
3.70-3.86 (m, 2H), 3.46 (t, J = 5.62 Hz, 2H), 3.21 (d, J = 1.96 Hz,
3H), 2.93-3.03 (m, 1H), 2.90 (t, J = 5.99 Hz, 2H), 2.17 (s, 3H),
2.12 (s, 3H), 1.14 (dd, J = 4.34, 6.66 Hz, 3H); [M + H].sup.+ 522
[.alpha.].sub.D = -110.6.degree. (c = 0.1, MeOH); >99% de;
Single diastereomer containing (R)-2- hydroxy- propanamide; other
chiral center undetermined; Enantiomer of Ex. 172 Diastereomer of
Ex. 170 and Ex. 173 172 ##STR00224## I .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 11.53 (br. s., 1H), 7.55 (s, 1H), 5.88 (s, 1H),
4.96 (dd, J = 6.11, 15.41 Hz, 1H), 4.90 (t, J = 6.36 Hz, 1H), 4.56
(s, 2H), 4.17-4.27 (m, 1H), 4.00-4.12 (m, 2H), 3.68- 3.87 (m, 2H),
3.46 (t, J = 5.87 Hz, 2H), 3.21 (d, J = 1.96 Hz, 3H), 2.93- 3.02
(m, 1H), 2.89 (t, J = 6.11 Hz, 2H), 2.16 (s, 3H), 2.12 (s, 3H),
1.14 (dd, J = 4.16, 6.60 Hz, 3H); [M + H].sup.+ 522 [.alpha.].sub.D
= +69.5.degree. (c = 0.1, MeOH); >99% de; Single diastereomer
containing (S)-2- hydroxy- propanamide; other chiral center
undetermined; Enantiomer of Ex. 171 Diastereomer of Ex. 173 and Ex.
170 173 ##STR00225## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.54
(br. s., 1H), 7.54 (s, 1H), 5.89 (s, 1H), 5.00 (t, J = 5.26 Hz,
1H), 4.91 (d, J = 4.89 Hz, 1H), 4.57 (s, 2H), 4.01-4.26 (m, 3H),
3.77-3.90 (m, 1H), 3.66-3.74 (m, 1H), 3.46 (t, J = 5.87 Hz, 2H),
3.21 (s, 3H), 2.93- 3.03 (m, 1H), 2.90 (t, J = 6.11 Hz, 2H), 2.17
(s, 3H), 2.12 (s, 3H), 1.15 (d, J = 6.36 Hz, 3H); [M + H].sup.+ 522
[.alpha.].sub.D = -105.1.degree. (c = 0.1, MeOH); ~88% de; Single
diastereomer containing (S)-2- hydroxy- propanamide; other chiral
center undetermined; Enantiomer of Ex. 170 Diasteromer of Ex. 172
and Ex. 171 174 ##STR00226## I .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 11.48 (br. s., 1H), 7.47 (s, 1H), 5.88 (s, 1H), 4.89 (d, J
= 7.09 Hz, 1H), 4.56 (s, 2H), 3.46 (t, J = 6.11 Hz, 2H), 3.10-3.19
(m, 4H), 2.97-3.09 (m, 2H), 2.79-2.94 (m, 3H), 2.62-2.71 (m, 1H),
2.17 (s, 3H), 2.15 (s, 3H), 2.12 (s, 3H); [M + H].sup.+ 464
[.alpha.].sub.D = +88.5.degree. (c = 0.1, MeOH); >99% ee;
Absolute stereochemistry undetermined; Enantiomer of Ex. 175 175
##STR00227## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.48 (br.
s., 1H), 7.47 (s, 1H), 5.88 (s, 1H), 4.89 (d, J = 7.09 Hz, 1H),
4.56 (s, 2H), 3.46 (t, J = 6.11 Hz, 2H), 3.10-3.19 (m, 4H),
2.97-3.09 (m, 2H), 2.79-2.94 (m, 3H), 2.62-2.71 (m, 1H), 2.17 (s,
3H), 2.15 (s, 3H), 2.12 (s, 3H); [M + H].sup.+ 464 [.alpha.].sub.D
= -70.2.degree. (c = 0.1, MeOH); >99% ee; Absolute
stereochemistry undetermined; Enantiomer of Ex. 174 176
##STR00228## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.54 (br.
s., 1H), 7.61 (s, 1H), 5.88 (s, 1H), 4.87 (d, J = 5.14 Hz, 1H),
4.57 (s, 2H), 3.97 (t, J = 7.46 Hz, 1H), 3.75-3.87 (m, 2H), 3.67
(t, J = 8.31 Hz, 1H), 3.45 (t, J = 6.24 Hz, 2H), 3.23 (s, 3H), 2.99
(s, 3H), 2.93-3.03 (m, 1H), 2.89 (t, J = 5.99 Hz, 2H), 2.17 (s,
3H), 2.12 (s, 3H); [M + H].sup.+ 528 [.alpha.].sub.D =
+104.4.degree. (c = 0.2, MeOH); >99% ee; Absolute
stereochemistry undetermined; Enantiomer of Ex. 177 177
##STR00229## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.54 (br.
s., 1H), 7.61 (s, 1H), 5.88 (s, 1H), 4.87 (d, J = 5.14 Hz, 1H),
4.57 (s, 2H), 3.97 (t, J = 7.34 Hz, 1H), 3.76-3.86 (m, 2H), 3.67
(t, J = 8.31 Hz, 1H), 3.45 (t, J = 6.11 Hz, 2H), 3.23 (s, 3H), 2.99
(s, 3H), 2.94-3.02 (m, 1H), 2.89 (t, J = 6.11 Hz, 2H), 2.17 (s,
3H), 2.12 (s, 3H); [M + H].sup.+ 528 [.alpha.].sub.D =
-111.7.degree. (c = 0.1, MeOH); ~97% ee; Absolute stereochemistry
undetermined; Enantiomer of Ex. 176 178 ##STR00230## I .sup.1H NMR
(400 MHz, CD3OD) .delta. 7.37 (s, 1H), 6.10 (s, 1H), 4.99 (d, J =
5.50 Hz, 1H), 4.77 (s, 2H), 3.79-3.95 (m, 2H), 3.55-3.72 (m, 2H),
3.46 (t, J = 5.99 Hz, 2H), 3.29 (s, 3H), 3.16 (dd, J = 7.76, 14.12
Hz, 1H), 2.81 (t, J = 5.93 Hz, 2H), 2.29 (s, 6H), 2.25 (s, 3H); [M
+ H].sup.+ 430 racemic mixture 179 ##STR00231## I .sup.1H NMR (400
MHz, DMSO-d6) .delta. 11.54 (br. s.,
1H), 7.55 (s, 1H), 5.89 (s, 1H), 4.74 (d, J = 5.87 Hz, 1H), 4.56
(s, 2H), 3.47 (t, J = 6.11 Hz, 2H), 3.16 (s, 3H), 3.09 (dd, J =
6.36, 9.54 Hz, 1H), 2.90 (t, J = 5.87 Hz, 2H), 2.70- 2.78 (m, 1H),
2.67 (s, 3H), 2.26-2.35 (m, 1H), 2.17 (s, 3H), 2.07-2.16 (m, 4H)
One proton under water peak; [M + H].sup.+ 492 [.alpha.].sub.D =
+51.2.degree. (c = 0.1, MeOH); >99% de; Absolute and relative
stereochemistry undetermined; Enantiomer of Ex. 180 180
##STR00232## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.54 (s,
1H), 7.55 (s, 1H), 5.89 (s, 1H), 4.74 (d, J = 6.11 Hz, 1H), 4.56
(s, 2H), 3.47 (t, J = 6.24 Hz, 2H), 3.16 (s, 3H), 3.09 (dd, J =
6.24, 9.66 Hz, 1H), 2.90 (t, J = 6.11 Hz, 2H), 2.69-2.77 (m, 1H),
2.67 (s, 3H), 2.26-2.34 (m, 1H), 2.18 (s, 3H), 2.08-2.16 (m, 4H)
One proton under water peak; [M + H].sup.+ 492 [.alpha.].sub.D =
-69.5.degree. (c = 0.1, MeOH); ~99% de; Absolute and relative
stereochemistry undetermined; Enantiomer of Ex. 179 181
##STR00233## I .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.52 (br.
s., 1H), 7.29 (s, 1H), 5.88 (s, 1H), 4.91 (d, J = 6.85 Hz, 1H),
4.56 (s, 2H), 4.52 (t, J = 6.48 Hz, 2H), 4.28-4.38 (m, 2H),
3.60-3.74 (m, 1H), 3.40 (t, J = 6.11 Hz, 2H), 3.16-3.25 (m, 2H),
3.06- 3.15 (m, 4H), 2.97-3.06 (m, 1H), 2.68-2.79 (m, 3H), 2.22 (s,
3H), 2.17 (s, 3H), 2.12 (s, 3H); [M + H].sup.+ 486 [.alpha.].sub.D
= +110.8.degree. (c = 0.1, MeOH); ~99% ee; Absolute stereochemistry
undetermined; Enantiomer of Ex. 182 182 ##STR00234## I .sup.1H NMR
(400 MHz, DMSO-d6) .delta. 11.53 (br. s., 1H), 7.29 (s, 1H), 5.88
(s, 1H), 4.91 (d, J = 7.09 Hz, 1H), 4.56 (s, 2H), 4.52 (t, J = 6.48
Hz, 2H), 4.33 (td, J = 5.93, 8.19 Hz, 2H), 3.66 (quin, J = 5.93 Hz,
1H), 3.40 (t, J = 5.99 Hz, 2H), 3.15-3.23 (m, 2H), 3.13 (s, 3H),
3.09 (t, J = 7.46 Hz, 1H), 3.00 (t, J = 6.97 Hz, 1H), 2.68-2.78 (m,
3H), 2.22 (s, 3H), 2.17 (s, 3H), 2.12 (s, 3H); [M + H].sup.+ 486
[.alpha.].sub.D = -85.3.degree. (c = 0.1, MeOH); >99% ee;
Absolute stereochemistry undetermined; Enantiomer of Ex. 181 183
##STR00235## J .sup.1H NMR (400 MHz, CDCl3) .delta. 11.37 (brs,
1H), 7.17 (s, 1H), 5.94 (s, 1H), 4.77 (s, 2H), 4.19- 4.27 (m, 1H),
3.92- 4.00 (m, 4H), 3.57- 3.74 (m, 4H), 3.16- 3.22 (m, 1H), 3.01-
2.88 (m 3H), 2.36 (s, 3H), 2.27 (s, 3H), 1.19 (d, J = 6.01 Hz, 3H);
[M + H].sup.+ 492 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 184; 96% ee; retention time: 2.459 min; column:
Chiralcel OJ-3 100 .times. 4.6 mm I.D., 3 um; mobile phase: 5-40%
methanol (0.05% DEA) in CO2 184 ##STR00236## J .sup.1H NMR (400
MHz, CDCl3) .delta. 11.70 (brs, 1H), 7.17 (s, 1H), 5.94 (s, 1H),
4.77 (s, 2H), 4.22- 4.38 (m, 1H), 3.89- 4.00 (m, 4H), 3.55- 3.75
(m, 4H), 3.16- 3.23 (m, 1H), 2.99- 2.86 (m, 3H), 2.36 (s, 3H), 2.28
(s, 3H), 1.19 (d, J = 6.01 Hz, 3H); [M + H].sup.+ 492 Single
isomer, absolute stereochemistry unknown; Enantiomer of Ex. 183;
94% ee; retention time: 2.661 min; column: Chiralcel OJ-3 100
.times. 4.6 mm I.D., 3 um; mobile phase: 5-40% methanol (0.05% DEA)
in CO2 185 ##STR00237## J .sup.1H NMR (400 MHz, DMSO-d6) .delta.
11.59 (br. s., 1H), 7.55 (s, 1H), 5.89 (br. s., 1H), 4.57 (br. s.,
2H), 3.97 (br. s., 2H), 3.77 (d, J = 12.30 Hz, 2H), 3.62 (br. s.,
2H), 3.44-3.47 (m, 2H), 2.98 (s, 3H), 2.85 (br. s., 2H), 2.15 (d, J
= 14.05 Hz, 6H), 1.13 (d, J = 6.02 Hz, 3H); [M + Na].sup.+ 534
Single isomer, absolute stereochemistry unknown; Enantiomer of Ex.
186; 100% ee; retention time 5.299 min; column: Chiralpak AD-3 100
.times. 4.6 mm I.D., 3 um; mobile phase: 5-40% ethanol (0.05% DEA)
in CO2 186 ##STR00238## J .sup.1H NMR (400 MHz, DMSO-d6) .delta.
11.57 (br. s., 1H), 7.55 (s, 1H), 5.89 (s, 1H), 4.57 (s, 2H), 3.97
(t, J = 8.03 Hz, 2H), 3.74- 3.83 (m, 2H), 3.62 (d, J = 9.29 Hz,
2H), 3.44 (br. s., 2H), 2.99 (s, 3H), 2.87 (d, J = 6.02 Hz, 2H),
2.15 (d, J = 16.56 Hz, 6H), 1.13 (d, J = 6.78 Hz, 3H); [M +
Na].sup.+ 534 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 185; 99% ee; retention time 5.807 min; column:
Chiralpak AD-3 100 .times. 4.6 mm I.D., 3 um; mobile phase: 5-40%
ethanol (0.05% DEA) in CO2 187 ##STR00239## J .sup.1H NMR (400 MHz,
CD3OD) .delta. 7.50 (br. s., 1H), 6.14 (s, 1H), 4.77 (s, 2H), 4.13
(d, J = 7.53 Hz, 2H), 3.77- 4.02 (m, 3H), 3.68 (d, J = 10.29 Hz,
1H), 3.53 (t, J = 6.02 Hz, 2H), 2.88-3.01 (m, 5H), 2.32 (s, 3H),
2.27 (s, 3H), 1.22 (d, J = 6.78 Hz, 3H); [M + H].sup.+ 448 Single
isomer, absolute stereochemistry unknown; Enantiomer of Ex. 188;
100% ee; retention time 5.295 min; column: Chiralpak AD-3 150
.times. 4.6 mm I.D., 3 um; mobile phase: 5-40% methanol (0.05% DEA)
in CO2 188 ##STR00240## J .sup.1H NMR (400 MHz, CD3OD) .delta. 7.48
(s, 1H), 6.14 (s, 1H), 4.77 (s, 2H), 4.09 (t, J = 8.41 Hz, 1H),
3.73- 3.89 (m, 3H), 3.53 (t, J = 6.27 Hz, 2H), 3.44 (t, J = 8.53
Hz, 1H), 3.07-3.19 (m, 1H), 2.97 (q, J = 5.77 Hz, 2H), 2.74 (s,
3H), 2.32 (s, 3H), 2.27 (s, 3H), 1.20 (d, J = 6.78 Hz, 3H); [M +
H].sup.+ 448 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 187; 100% ee; retention time 5.420 min; column:
Chiralpak AD-3 150 .times. 4.6 mm I.D., 3 um; mobile phase: 5-40%
methanol (0.05% DEA) in CO2 189 ##STR00241## J .sup.1H NMR (400
MHz, CDCl3) .delta. 7.12 (s, 1H), 5.92 (s, 1H), 5.33-5.37 (m, 1H),
4.89-4.93 (m, 1H), 4.36-4.39 (m, 1H), 4.13-4.17 (m, 1H), 3.82-3.91
(m, 3H), 3.65-3.70 (m, 3H), 3.19-3.23 (m, 1H), 2.88-2.91 (m, 2H),
2.40 (s, 3H), 2.30 (s, 3H), 1.15 (d, J = 6.80 Hz, 3H); [M +
Na].sup.+ 456 Single isomer, absolute stereochemisty unknown;
Enantiomer of Ex. 190; 100% ee; retention time 6.673 min; column:
Chiralcel OD-3 150 .times. 4.6 mm I.D., 3 um; mobile phase: 5-40%
ethanol (0.05% DEA) in CO2 190 ##STR00242## J .sup.1H NMR (400 MHz,
CDCl3) .delta. 7.15 (s, 1H), 5.95 (s, 1H), 5.33-5.37 (m, 1H),
4.82-4.92 (m, 1H), 4.38-4.42 (m, 1H), 4.13-4.17 (m, 1H), 3.82- 3.91
(m, 3H), 3.65-3.70 (m, 3H), 3.19-3.23 (m, 1H), 2.88-2.91 (m, 2H),
2.40 (s, 3H), 2.30 (s, 3H), 1.15 (d, J = 6.80 Hz, 3H); [M +
H].sup.+ 434 Single isomer, absolute stereochemistry unknown;
Enantiomer Ex. 189; 97% ee; retention time 5.996 min; column:
Chiralcel OD-3 150 .times. 4.6 mm I.D., 3 um; mobile phase: 5-40%
ethanol (0.05% DEA) in CO2 191 ##STR00243## J .sup.1H NMR (400 MHz,
CD3OD) .delta. 7.23 (s, 1H), 6.27 (s, 1H), 4.69-4.78 (m, 2H), 4.10
(t, J = 8.03 Hz, 1H), 3.91 (s, 3H), 3.74-3.85 (m, 3H), 3.46 (dd, J
= 6.78, 8.03 Hz, 1H), 3.34- 3.38 (m, 2H), 3.02 (dd, J = 6.78, 17.32
Hz, 1H), 2.93 (s, 3H), 2.77 (t, J = 6.27 Hz, 2H), 2.33 (s, 3H),
2.28 (s, 3H), 1.20 (d, J = 6.78 Hz, 3H); [M + H].sup.+ 508 Single
isomer, absolute stereochemistry unknown; Enantiomer of Ex. 192;
96% ee; retention time 4.765 min; column: Chiralpak AD-3 100
.times. 4.6 mm I.D., 3 um; mobile phase: 5-40% ethanol (0.05% DEA)
in CO2 192 ##STR00244## J .sup.1H NMR (400 MHz, CD3OD) .delta. 7.23
(s, 1H), 6.27 (s, 1H), 4.74 (d, J = 1.25 Hz, 2H), 4.10 (t, J = 8.16
Hz, 1H), 3.90 (s, 3H), 3.74-3.86 (m, 3H), 3.46 (dd, J = 6.65, 7.91
Hz, 1H), 3.34-3.38 (m, 2H), 3.05 (m, 1H), 2.93 (s, 3H), 2.74-2.81
(m, 2H), 2.33 (s, 3H), 2.28 (s, 3H), 1.20 (d, J = 6.78 Hz, 3H); [M
+ Na].sup.+ 530 Single isomer, absolute stereochemistry unknown;
Enantiomer of Ex. 191; 100% ee; retention time 5.207 min; column:
Chiralpak AD-3 100 .times. 4.6 mm I.D., 3 um; mobile phase: 5-40%
ethanol (0.05% DEA) in CO2 193 ##STR00245## J .sup.1H NMR (400 MHz,
CDCl3) .delta. 11.33 (br. s., 1H), 6.96 (s, 1H), 5.93 (s, 1H),
4.72- 4.87 (m, 2H), 4.10 (t, J = 8.03 Hz, 1H), 3.69- 3.84 (m, 3H),
3.60 (t, J = 6.02 Hz, 2H), 3.50 (t, J = 7.28 Hz, 1H), 2.85-2.95 (m,
1H), 2.84 (s, 3H), 2.71 (t, J = 6.02 Hz, 2H), 2.36 (s, 3H), 2.27
(s, 3H), 2.21 (s, 3H), 1.17 (d, J = 6.78 Hz, 3H); [M + H].sup.+ 492
Single isomer, absolute stereochemisty unknown; Enantiomer of Ex.
194; 100% ee; retention time 5.024 min; column: Chiralpak AD-3 100
.times. 4.6 mm I.D. 3 um; mobile phase: 5-40% ethanol (0.05% DEA)
in CO2 194 ##STR00246## J .sup.1H NMR (400 MHz, CDCl3) .delta.
11.82 (br. s., 1H), 6.96 (s, 1H), 5.93 (s, 1H), 4.72-4.87 (m, 2H),
4.09 (t, J = 8.03 Hz, 1H), 3.67- 3.84 (m, 3H), 3.60 (t, J = 6.02
Hz, 2H), 3.50 (t, J = 7.28 Hz, 1H), 2.89 (d, J = 9.29 Hz, 1H), 2.84
(s, 3H), 2.71 (t, J = 6.15 Hz, 2H), 2.36 (s, 3H), 2.28 (s, 3H),
2.21 (s, 3H), 1.17 (d, J = 6.78 Hz, 3H); [M + H].sup.+ 492 Single
isomer, absolute stereochemistry unknown; Enantiomer of Ex. 193;
95% ee; retention time 5.219 min; column: Chiralpak AD-3 100
.times. 4.6 mm I.D., 3 um; mobile phase: 5-40% ethanol (0.05% DEA)
in CO2 195 ##STR00247## J .sup.1H NMR (400 MHz, CDCl3) .delta.
11.67 (brs, 1H), 6.91 (s, 1H), 5.94 (s, 1H), 4.86 (t, J = 6.8 Hz,
1H), 4.80 (2H, s), 4.55-4.65 (m, 2H), 4.30 (t, J = 6.0 Hz, 1H),
3.90- 4.00 (m, 1H), 3.60 (t, J = 6.0 Hz, 2H), 3.25- 3.40 (m, 1H),
2.70 (t, J = 6.0 Hz, 2H), 2.69 (s, 3H), 2.37 (s, 3H), 2.28 (s, 3H),
1.13 (d, J = 6.8 Hz, 3H); [M + H].sup.+ 415 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 196; 99% ee; retention
time: 2.854 min; column: OJ-3 100 .times. 4.6 mm I.D., 3 um; mobile
phase: 5-40% methanol (0.05% DEA) in CO2 196 ##STR00248## J .sup.1H
NMR (400 MHz, CDCl3) .delta. 11.72 (brs, 1H), 6.91 (s, 1H), 5.94
(s, 1H), 4.86 (t, J = 6.8 Hz, 1H), 4.80 (s, 2H), 4.55-4.65 (m, 2H),
4.31 (t, J = 6.4 Hz, 1H), 3.90-4.00 (m, 1H), 3.60 (t, J = 6.0 Hz,
2H), 3.25- 3.40 (m, 1H), 2.70 (t, J = 6.0 Hz, 2H), 2.69 (s, 3H),
2.37 (s, 3H), 2.28 (s, 3H), 1.13 (d, J = 6.8 Hz, 3H); [M + H].sup.+
415 Single isomer, absolute stereochemistry unknown; Enantiomer of
Ex. 195; 99% ee; retention time: 3.078 min; column: OJ-3 100
.times. 4.6 mm I.D., 3 um; mobile phase: 5-40% methanol (0.05% DEA)
in CO2 197 ##STR00249## J .sup.1H NMR (400 MHz, CDCl3) .delta.
12.75 (brs, 1H), 6.89 (s, 1H), 5.91 (s, 1H), 4.80-4.88 (m, 3H),
4.57-4.61 (m, 2H), 4.28 (t, J = 6.4 Hz, 1H), 3.86-3.93 (m, 4H),
3.31- 3.42 (m, 3H), 2.70 (t, J = 6.0 Hz, 2H), 2.34 (s, 3H), 2.20
(s, 3H), 1.13 (d, J = 6.8 Hz, 3H); [M + H].sup.+ 431 Single isomer,
absolute stereochemistry unknown; Enantiomer of Ex. 198; 97% ee;
retention time: 1.521 min; column: Chiralpak AD-3 150 .times. 4.6
mm I.D., 3 um; mobile phase: 40% isopropanol (0.05% DEA) in CO2 198
##STR00250## J .sup.1H NMR (400 MHz, CDCl3) .delta. 12.39 (brs,
1H), 6.90 (s, 1H), 5.92 (s, 1H), 4.81-4.89 (m, 3H), 4.57-4.61 (m,
2H), 4.30 (t, J = 6.0 Hz, 1H), 3.87-3.94 (m, 4H), 3.31- 3.43 (m,
3H), 2.71 (t, J = 5.2 Hz, 2H), 2.34 (s, 3H), 2.22 (s, 3H), 1.13 (d,
J = 6.8 Hz, 3H); [M + H].sup.+ 431 Single isomer, absolute
stereochemistry unknown; Enantiomer of Ex. 197; 94% ee; retention
time: 2.081 min; column: Chiralpak AD-3 150 .times. 4.6 mm I.D., 3
um; mobile phase: 40% isopropanol (0.05% DEA) in CO2 199
##STR00251## J .sup.1H NMR (400 MHz, CD3OD) .delta. 7.67 (s, 1H),
6.10 (s, 1H), 4.76 (s, 2H), 3.77- 3.90 (m, 2H), 3.45- 3.55 (m, 2H),
2.96 (t, J = 6.24 Hz, 2H), 2.79- 2.86 (m, 1H), 2.28 (s, 3H), 2.25
(s, 3H), 1.11-1.22 (m, 1H), 0.64-0.74 (m, 1H), 0.35-0.50 (m, 2H),
0.02-0.11 (m, 1H); [M + H].sup.+ 436 racemic mixture 200
##STR00252## J .sup.1H NMR (400 MHz, CDCl3) .delta. 12.19 (s, 1H),
7.23 (d, J = 3.2 Hz, 1H), 5.91 (s, 1H), 4.82-4.73 (m, 2H),
4.70-4.50 (m, 1H), 4.15-4.11 (m, 2H), 3.87 (s, 3H), 3.70- 3.66 (m,
1H), 3.54- 3.40 (m, 4H), 2.92- 2.86 (m, 3H), 2.64- 2.58 (m, 1H),
2.34 (s, 3H), 1.84-1.62 (m, 2H), 1.50-1.40 (m, 1H), 1.26-1.19 (m,
5H); [M + H].sup.+ 536 Single isomer, absolute stereochemistry
unknown; Enantiomer of Ex. 201; 90% ee; retention time 26.03 min;
column: Chiralpak AD-H 250 .times. 4.6 mm I.D., 5 u; mobile phase:
50/50 hexane (0.1% DEA)/ethanol (0.1% ethanolamine) 201
##STR00253## J .sup.1H NMR (400 MHz, CDCl3) .delta. 12.03 (s, 1H),
7.23 (d, J = 3.2 Hz, 1H), 5.92 (s, 1H), 4.82-4.73 (m, 2H),
4.70-4.50 (m, 1H), 4.14-4.11 (m, 2H), 3.87 (s, 3H), 3.70- 3.66 (m,
1H), 3.47- 3.40 (m, 4H), 2.92- 2.86 (m, 3H), 2.64- 2.61 (m, 1H),
2.34 (s, 3H), 1.84-1.62 (m, 2H), 1.50-1.40 (m, 1H), 1.26-1.19 (m,
5H); [M + H].sup.+ 536 Single isomer, absolute stereochemistry
unknown; Enantiomer Ex. 200; 97% ee; retention time: 51.00 min;
column: Chiralpak AD-H 250 .times. 4.6 mm I.D., 5 um; mobile phase:
50/50 hexane (0.1% DEA)/ethanol (0.1% ethanolamine); 202
##STR00254## K .sup.1H NMR (400 MHz, CDCl3) .delta. 7.72 (s, 1H),
5.93 (s, 1H), 4.84-4.71 (m, 2H), 4.01 (q, J = 6.3 Hz, 1H), 3.63 (t,
J = 6.1 Hz, 2H), 2.90 (d, J = 3.5 Hz, 2H), 2.42 (br. s., 6H), 2.35
(s, 3H), 2.28 (s, 6H), 1.79 (br. s., 2H), 1.24 (d, J = 6.5 Hz, 3H);
[M + H].sup.+ 477 racemic mixture 203 ##STR00255## K .sup.1H NMR
(400 MHz, CDCl3) .delta. 12.34 (br. s., 1H), 7.43 (s, 1H), 5.95 (s,
1H), 4.61-4.88 (m, 4H), 4.48 (d, J = 10.04 Hz, 1H), 4.25-4.34 (m,
1H), 4.21 (t, J = 6.90 Hz, 1H), 3.82-3.95 (m, 3H), 3.49 (d, J =
1.76 Hz, 3H), 2.89 (t, J = 6.02 Hz, 2H), 2.35 (s, 3H), 2.13 (s,
6H); [M + H].sup.+ 480 Single isomer, absolute stereochemistry
unknown; Enantiomer of Ex. 204; 88% ee; retention time 3.994 min;
column: Chiralpak AD-3 100 .times. 4.6 mm I.D., 3 um; mobile phase:
5-40% isopropanol (0.05% DEA) in CO2 204 ##STR00256## K .sup.1H NMR
(400 MHz, CDCl3) .delta. 12.32 (brs, 1H), 7.46 (brs, 1H), 5.96 (s,
1H), 4.57- 4.88 (m, 4H), 4.50 (brs, 1H), 4.28 (t, J = 6.90 Hz, 1H),
4.19 (d, J = 6.27 Hz, 1H), 3.88 (s, 3H), 3.52 (br. s., 3H), 2.88
(brs, 2H), 2.37 (brs, 4H), 2.15 (brs, 5H); [M + H].sup.+ 480 Single
isomer, absolute stereochemistry unknown; Enantiomer of Ex. 203;
97% ee; retention time 4.325 min; column: Chiralpak AD-3 100
.times. 4.6 mm I.D., 3 um; mobile phase: 5-40% isopropanol (0.05%
DEA) in CO2 205 ##STR00257## K .sup.1H NMR (400 MHz, CDCl3) .delta.
7.72 (s, 1H), 5.94 (s, 1H), 4.85-4.69 (m, 2H), 4.02 (q, J = 6.4 Hz,
2H), 3.62 (t, J = 6.4 Hz, 2H), 2.90 (br. s., 6H), 2.55 (br. s.,
2H), 2.42-2.31 (m, 5H), 2.29 (s, 3H), 1.24 (d, J = 6.5 Hz, 3H); [M
+ H].sup.+ 463 racemic mixture
Biological Assays and Data
Purification of WT and Mutant EZH2 Y641N
[0635] WT and mutant EZH2 were purified using the same procedure.
The genes for EZH2, EED, SUZ12, and RBBP4 proteins were cloned into
pBacPAK9 vectors (Clontech). RBBP4 was FLAG tagged on the
N-terminal end. The baculovirus expressions of these proteins were
used to co-infect SF9 insect cells. Insect cell pellets were lysed
in a buffer containing 25 mM Tris pH8.0, 300 mM NaCl, 0.5 mM TCEP,
complete EDTA-free protease inhibitor (Roche), 0.1% NP-40. The
supernatant from the lysate was incubated with FLAG.RTM. M2
antibody resin (Sigma). The resin was washed on the chromatography
column and eluted with 0.2 mg/ml FLAG peptide. The elute was
incubated with omnicleave nucleases (Epicentre Technologies) at
4.degree. C. overnight, then concentrated and loaded onto a
Superdex 200 (GE Healthcare) column. The Superdex 200 column was
eluted with 25 mM Tris pH8.0, 150 mM NaCl, 0.5 mM TCEP. Fractions
containing the PRC2 complex were pooled.
Nucleosome Assay Protocol:
[0636] (The same protocol was used for the WT and mutant EZH2
Y6412N assays)
[0637] A. Compound preparation [0638] 1. Prepare 10 mM stock
solutions in 100% DMSO from solid material [0639] 2. Serial dilute
10 mM compound stocks either 2 or 3-fold in 100% DMSO to generate
compounds for 11 point dose response
[0640] B. Reagent preparation [0641] 1. Prepare 1.times. assay
buffer containing 100 mM Tris pH 8.5, 4 mM DTT and 0.01% Tween-20
[0642] 2. Dilute purified HeLa oligonucleosomes and recombinant
histone H1 (New England Biolabs) in assay buffer to 1.67.times..
[0643] 3. Dilute PRC2 4 protein complex (EZH2, EED, SUZ12, RbAp48)
to 3.5.times. in assay buffer [0644] 4. Prepare 10.times..sup.3H
SAM solution in assay buffer using 0.94 .mu.Ci/well of radioactive
SAM (Perkin Elmer) and sufficient non-labeled SAM (Sigma) for 1.5 M
final concentration. [0645] 5. Dilute TCA to 20% in DI water
[0646] C. Enzyme reaction [0647] 1. Final reaction conditions are
PRC2 4-protein complex at 4 nM when using WT EZH2 or 6 nM when
using Y641N mutant EZH2, 1.5 .mu.M SAM, 25 .mu.g/mL
oligonucleosomes, 50 nM rH1 in a 50 .mu.l reaction volume. [0648]
2. Add 1 .mu.l of diluted compound to the assay plate (96-well
V-bottom polypropylene plates) or 1 .mu.l of DMSO for control
wells. [0649] 3. Add 30 .mu.l of nucleosomes to the assay plate
[0650] 4. Add 14 .mu.l of either WT or Y641N mutant PRC2 4 protein
complex to the assay plate [0651] 5. Add 5 .mu.l of .sup.3H SAM to
start the reaction. [0652] 6. Stop the reaction after 60 minutes
with the addition of 100 .mu.l of 20% TCA [0653] 7. Transfer 150
.mu.l of quenched reaction into a prepared filterplate (Millipore
#MSIPN4B10) [0654] 8. Apply vacuum to the filterplate to filter the
reaction mix through the membrane. [0655] 9. Wash the filterplate
with 5.times.200 .mu.l of PBS, blot dry and dry in an oven for 30
minutes [0656] 10. Add 50 .mu.l of microscint-20 scintillation
fluid (Perkin Elmer) to each well, wait 30 minutes and count on a
liquid scintillation counter. [0657] 11. Some compounds were tested
under high SAM conditions. In this case, the assay is as described
above except that the reaction contains 15 uM SAM. SAM is added to
the assay as a 3.3.times. stock with a total of 14.5 uCi/well.
[0658] D. Data analysis [0659] 1. IC.sub.50 values were determined
by fitting the data to a 4-parameter IC.sub.50 equation using
proprietary curve fitting software. [0660] 2. For compounds tested
under high SAM conditions, K.sub.i.sup.app values were obtained by
fitting the dose response curve to a model for competitive
inhibition using proprietary curve fitting software.
[0661] Preparation of HeLa Oligonucleosomes:
[0662] Reagents [0663] Cell Pellet: 15 L HeLa S3 (Accelgen)+6 L
HeLa S3 (in house) [0664] Mnase (Worthington Biochemicals)
[0665] Equipment [0666] SW-28 Rotor [0667] Dounce Homogenizer/B
Pestle
[0668] Buffers [0669] Lysis: 20 mM Hepes pH 7.5, 0.25M Sucrose, 3
mM MgCl.sub.2, 0.5% Nonidet P-40, 0.5 mM TCEP, 1 Roche Protease
Tablet [0670] B: 20 mM Hepes pH7.5, 3 mM MgCl.sub.2, 0.5 mM EDTA,
0.5 mM TCEP, 1 Roche Protease Tablet [0671] MSB: 20 mM Hepes pH7.5,
0.4 M NaCl, 1 mM EDTA, 5% v/v Glycerol, 0.5 mM TCEP, 0.2 mM PMSF
[0672] LSB: 20 mM Hepes pH7.5, 0.1M NaCl, 1 mM EDTA, 0.5 mM TCEP,
0.2 mM PMSF [0673] NG: 20 mM Hepes pH7.5, 1 mM EDTA, 0.4 m NaCl,
0.2 mM PMSF, 0.5 mM TCEP [0674] Storage: 20 mM Hepes pH7.5, 1 mM
EDTA, 10% Glycerol, 0.2 mM PMSF, 0.5 mM TCEP
Protocol
[0675] A. Nuclei [0676] 1. Resuspend .about.10 L pellet in
2.times.40 mL lysis using dounce homogenizer [0677] 2. Spin
3000.times.g 15' [0678] 3. Repeat 2 more times [0679] 4. Resuspend
pellet in 2.times.40 mL B [0680] 5. Spin 3000.times.g 15'
[0681] B. Nuclei Resuspension [0682] 1. Resuspend pellet in
2.times.40 mL MSB. Spin 5000.times.g 20' [0683] 2. Resuspend pellet
in 2.times.15 mL HSB [0684] 3. Pool and Homogenize 40 Strokes to
shear DNA [0685] 4. Pellet 10000.times.g 20' [0686] 5. Dialyze O/N
4.degree. C. in LSB except for Batch A which was Dialyzed LSB at 50
nM NaCl for 3 hr
[0687] C. Mnase Digestion [0688] Test Mnase digestion (200 ul)
[0689] 1. Warm to 37.degree. C. for 5' [0690] 2. Add CaCl.sub.2 to
3 mM and add 10 U of Mnase [0691] 3. 37.degree. C. 30' taking 25
.mu.L sample every 5' [0692] 4. Process reaction with 1 .mu.L 0.5M
EDTA, 40 .mu.L H.sub.2O, 15 .mu.L 10% SDS, 10 .mu.L 5M NaCl, and
100 .mu.L phenol-chloroform vortexing after each addition [0693] 5.
Spin 5' 13 k [0694] 6. Run 5 .mu.L of Aqueous phase on 1% agarose
gel [0695] 7. Take time that yields .about.2 kb fragments [0696] 8.
Selected 15' for A & B and 20' for C & D for scale up
[0697] Added NaCl to 0.6M
[0698] D. Sucrose Gradient 1 [0699] 1. Poured 6.times.34 mL
gradient from 5 to 35% sucrose in NG using AKTA purifier in 38.5 mL
pollyallomer tubes [0700] 2. Lead .about.4.0 mL on top of MN1
digest [0701] 3. Spin 26 k 16 hr 4.degree. C. [0702] 4. Take 2 mL
fractions from top [0703] 5. Run on Page Gel [0704] 6. Dialyze
Fractions 7-14 0/N 4.degree. C. in 4 L LSB except Batch D which had
2.times.2 hr [0705] 7. Repeat 3.times.
[0706] E. Final [0707] 1. Pool all and concentrate in Amicon
(somewhat cloudy) [0708] 2. Added 10% Glycerol [0709] 3. Spun 5K
15' [0710] 4. 1.8 mg/mL at 80 mL for 144 mg Total
Biological Activity
[0711] Biological activity of selected examples in the EZH2
nucleosome assay are provided in Tables 3. Data are presented as WT
and Mutant Y641N EZH2 IC.sub.50 value (.mu.M) or K.sub.i.sup.app
(.mu.M) as indicated.
TABLE-US-00002 TABLE 2 WT EZH2 EZH2 Mutant WT EZH2 Nucleosome Y641N
Nucleosome assay Nucleosome assay (10X SAM) assay Ex. No. IC.sub.50
(.mu.M) Ki (.mu.M) IC.sub.50 (.mu.M) 1 0.001 0.003 2 0.002 0.004 3
0.066 0.302 4 0.316 1.954 5 0.033 0.189 6 0.019 0.136 7 0.171 1.617
8 0.652 5.016 9 0.005 0.035 10 0.326 1.980 11 0.038 0.174 12 0.004
0.016 13 0.003 0.018 14 0.008 0.040 15 0.003 0.019 16 0.004 0.040
17 0.035 0.125 18 0.126 0.115 0.995 19 0.013 0.010 0.132 20 0.011
0.053 21 0.015 0.109 22 7.817 95.165 23 0.009 0.067 24 0.004 0.008
25 0.084 0.693 26 0.027 0.057 27 0.037 0.300 28 0.126 1.894 29
0.017 0.107 30 0.004 0.020 31 0.004 0.006 32 0.093 1.133 33 0.012
0.070 35 0.002 0.000 0.002 36 0.105 0.136 0.443 37 0.001 0.002 38
0.378 0.740 39 0.040 0.242 40 0.006 0.024 41 0.012 0.082 42 0.012
0.102 43 0.039 0.348 44 0.022 0.144 45 0.103 0.345 46 0.121 0.423
47 0.138 0.847 48 0.008 0.050 49 0.215 0.996 50 0.077 0.129 51
0.000 0.002 52 0.298 1.038 53 0.001 0.017 54 0.004 0.012 55 0.162
0.843 56 0.000 0.004 57 0.406 1.352 58 0.057 0.513 59 0.025 0.155
60 0.031 0.180 61 0.031 0.127 62 0.035 0.262 63 0.528 4.104 64
0.007 0.043 65 0.006 0.047 66 0.009 0.081 67 1.599 19.543 68 0.007
0.074 69 0.417 4.308 70 0.000 0.006 71 0.000 0.003 72 0.041 0.128
73 0.000 0.002 74 0.005 0.051 75 0.002 0.020 76 0.005 0.043 77
0.022 0.132 78 0.004 0.040 79 0.017 0.133 80 0.001 0.007 81 0.0002
0.003 82 0.006 0.131 83 0.0004 0.004 84 0.0002 0.002 85 0.052 0.238
86 0.061 0.637 87 0.00001 0.002 88 0.028 0.122 89 0.008 0.088 90
0.459 5.634 91 0.001 0.023 92 0.001 0.009 93 0.159 1.332 94 0.323
4.870 95 0.001 0.012 96 0.204 2.99 97 0.003 0.027 98 1.22 8.72 99
0.003 0.017 100 0.309 3.73 101 0.0003 0.007 102 0.018 0.178 103
0.052 0.320 104 0.673 7.77 105 0.0008 0.013 106 0.0006 0.008 107
0.0019 0.018 108 0.0017 0.016 109 0.086 0.911 110 0.00004 0.002 111
0.00002 0.001 112 0.021 0.125 113 0.0008 0.005 114 0.016 0.152 115
0.0003 0.005 116 0.071 0.340 117 0.0004 0.005 118 0.0001 0.003 119
0.015 0.100 120 0.002 0.014 121 0.029 0.255 122 0.073 0.236 123
0.0002 0.005 124 0.058 0.892 125 0.0009 0.008 126 0.056 0.165 127
0.0002 0.0022 128 0.184 1.167 129 0.0008 0.008 130 0.220 1.99 131
0.0007 0.005 132 0.001 0.013 133 0.837 8.635 134 0.131 1.926 135
0.002 0.014 136 0.0001 0.003 137 0.133 1.166 138 0.046 0.249 139
0.0005 0.0085 140 0.103 1.17 141 0.002 0.016 142 0.008 0.109 143
0.276 1.77 144 0.0002 0.005 145 0.070 0.272 146 0.0001 0.0035 147
0.128 0.543 148 0.056 0.439 149 0.539 2.267 150 0.0001 0.003 151
0.0008 0.009 152 0.0003 0.004 153 0.416 3.10 154 0.0446 0.553 155
0.0013 0.027 156 0.375 1.711 157 0.0006 0.009 158 0.578 5.13 159
0.0016 0.017 160 0.021 0.086 161 0.0004 0.004 162 0.148 2.971 163
0.0004 0.004 164 0.200 1.971 165 0.0003 0.005 166 0.0002 0.002 167
0.062 0.850 168 0.0007 0.008 169 0.0151 0.070 170 0.0008 0.014 171
0.328 2.63 172 0.0008 0.012 173 0.016 0.206 174 0.0006 0.013 175
0.085 1.00 176 0.0003 0.005 177 0.049 0.282 178 0.003 0.020 179
0.001 0.009 180 0.390 2.16 181 0.001 0.041 182 2.23 9.15 183 0.056
0.517 184 0.0006 0.005 185 0.001 0.012 186 0.014 0.240 187 0.001
0.010 188 0.014 0.151 189 0.0007 0.006 190 0.016 0.100 191 0.0003
0.005 192 0.117 0.605 193 0.001 0.010 194 0.044 0.309 195 0.046
0.216 196 0.0002 0.003 197 0.033 0.062 198 0.00001 0.001 199 0.003
0.037 200 0.0001 0.002 201 0.007 0.037 202 0.016 0.207 203 0.055
0.204 204 0.002 0.062 205 0.0007 0.008
[0712] All publications and patent applications cited in the
specification are herein incorporated by reference in their
entirety. Although the foregoing invention has been described in
some detail by way of illustration and example, it will be readily
apparent to those of ordinary skill in the art in light of the
teachings of this invention that certain changes and modifications
may be made thereto without departing from the spirit or scope of
the appended claims.
* * * * *