U.S. patent number RE47,428 [Application Number 15/878,663] was granted by the patent office on 2019-06-11 for modulators of methyl modifying enzymes, compositions and uses thereof.
This patent grant is currently assigned to Constellation Pharmaceuticals, Inc.. The grantee listed for this patent is Constellation Pharmaceuticals, Inc.. Invention is credited to Brian K. Albrecht, James Edmund Audia, Les A. Dakin, Victor S. Gehling, Jean-Christophe Harmange, Christopher G. Nasveschuk, Rishi G. Vaswani.
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United States Patent |
RE47,428 |
Albrecht , et al. |
June 11, 2019 |
Modulators of methyl modifying enzymes, compositions and uses
thereof
Abstract
Agents for modulating methyl modifying enzymes, compositions and
uses thereof are provided herein.
Inventors: |
Albrecht; Brian K. (Cambridge,
MA), Audia; James Edmund (Cambridge, MA), Dakin; Les
A. (Natick, MA), Gehling; Victor S. (Somerville, MA),
Harmange; Jean-Christophe (Andover, MA), Nasveschuk;
Christopher G. (Stoneham, MA), Vaswani; Rishi G.
(Lexington, MA) |
Applicant: |
Name |
City |
State |
Country |
Type |
Constellation Pharmaceuticals, Inc. |
Cambridge |
MA |
US |
|
|
Assignee: |
Constellation Pharmaceuticals,
Inc. (Cambridge, MA)
|
Family
ID: |
48948171 |
Appl.
No.: |
15/878,663 |
Filed: |
January 24, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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14766632 |
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PCT/US2014/015706 |
Feb 11, 2014 |
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Reissue of: |
14839273 |
Aug 28, 2015 |
9469646 |
Oct 18, 2016 |
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Foreign Application Priority Data
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Feb 11, 2013 [WO] |
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PCT/US2013/025639 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P
13/12 (20180101); C07D 401/14 (20130101); A61P
43/00 (20180101); C07D 471/04 (20130101); C07D
401/12 (20130101); A61K 31/4545 (20130101); C07D
409/06 (20130101); A61P 13/08 (20180101); C07D
409/06 (20130101); A61P 25/00 (20180101); C07D
401/14 (20130101); A61P 35/00 (20180101); C07D
417/04 (20130101); A61P 15/00 (20180101); C07D
417/14 (20130101); A61P 17/00 (20180101); A61P
11/00 (20180101); A61P 1/04 (20180101); A61P
1/16 (20180101); C07D 405/14 (20130101); A61P
13/10 (20180101); C07D 487/04 (20130101); A61K
31/4545 (20130101); C07D 417/14 (20130101); C07D
471/04 (20130101); C07D 405/14 (20130101); C07D
417/04 (20130101); C07D 487/04 (20130101); C07D
401/12 (20130101) |
Current International
Class: |
C07D
401/12 (20060101); C07D 417/04 (20060101); C07D
417/14 (20060101); C07D 405/14 (20060101); C07D
471/04 (20060101); C07D 409/06 (20060101); A61K
31/4545 (20060101); C07D 401/14 (20060101); C07D
487/04 (20060101) |
References Cited
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WO |
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2015/200650 |
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Dec 2015 |
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WO |
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|
Primary Examiner: Campell; Bruce R
Attorney, Agent or Firm: McCarter & English, LLP Davis;
Steven G. DeGrazia; Michael J.
Parent Case Text
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a .Iadd.reissue application of U.S. Pat. No.
9,469,646, which is a .Iaddend.continuation of U.S. application
Ser. No. 14/766,632, filed Aug. 7, 2015, which is a 35 U.S.C.
.sctn.371 national stage filing of International Application No.
PCT/US2014/015706, filed Feb. 11, 2014, which claims priority to
International Application No. PCT/US2013/025639, filed Feb. 11,
2013.Iadd., the entire contents of each of which are incorporated
by reference herein.Iaddend..
Claims
The invention claimed is:
1. A compound of the formula: ##STR00090## or a pharmaceutically
acceptable salt thereof.
2. The compound of claim 1, wherein the compound is of the formula:
##STR00091## or a pharmaceutically acceptable salt thereof.
3. A pharmaceutical composition comprising a compound of the
formula: ##STR00092## or a pharmaceutically acceptable salt
thereof; and a pharmaceutically acceptable carrier.
4. The pharmaceutical composition of claim 3, wherein the compound
is of the formula: ##STR00093## or a pharmaceutically acceptable
salt thereof.
Description
BACKGROUND OF THE INVENTION
Eukaryotic chromatin is composed of macromolecular complexes called
nucleosomes. A nucleosome has 147 base pairs of DNA wrapped around
a protein octamer having two subunits of each of histone protein
H2A, H2B, H3, and H4. Histone proteins are subject to
post-translational modifications which in turn affect chromatin
structure and gene expression. One type of post-translational
modification found on histones is methylation of lysine and
arginine residues. Histone methylation plays a critical role in the
regulation of gene expression in eukaryotes. Methylation affects
chromatin structure and has been linked to both activation and
repression of transcription (Zhang and Reinberg, Genes Dev.
15:2343-2360, 2001). Enzymes that catalyze attachment and removal
of methyl groups from histones are implicated in gene silencing,
embryonic development, cell proliferation, and other processes.
SUMMARY OF THE INVENTION
The present disclosure encompasses the recognition that methyl
modifying enzymes are an attractive target for modulation, given
their role in the regulation of diverse biological processes. It
has now been found that compounds of this invention, and
pharmaceutically acceptable compositions thereof, are effective as
agents that stimulate activity of histone methyl modifying enzymes,
including histone methylases and histone demethylases. Such
compounds have the general formula II:
##STR00001## or a pharmaceutically acceptable salt thereof, wherein
each variable is as defined herein.
Compounds of the present invention, and pharmaceutically acceptable
compositions thereof, are useful for treating a variety of
diseases, disorders or conditions, associated with a methyl
modifying enzyme. Such diseases, disorders, or conditions include
those described herein.
Compounds provided by this invention are also useful for the study
of methyl modifying enzymes in biological and pathological
phenomena; the study of intracellular signal transduction pathways
mediated by methyl modifying enzymes and the comparative evaluation
of new methyl modifying enzyme modulators.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1. Exemplary compounds of Formula II.
DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS
1. General Description of Compounds of the Invention
In certain embodiments, the present invention provides a compound
of Formula II:
##STR00002## or a pharmaceutically acceptable salt thereof,
wherein:
A is CH or N;
R.sup.1a is selected from --C.sub.1-C.sub.2 alkyl and
--O--(C.sub.1-C.sub.2 alkyl), wherein R.sup.1a is optionally
substituted with one or more fluoro;
R.sup.4a is selected from --(C.sub.1-C.sub.4
alkylene)-O--(C.sub.1-C.sub.3 alkyl), 1-substituted-piperidin-4-yl,
C.sub.3-C.sub.6 cycloalkyl optionally substituted with one or more
fluoro, and tetrahydropyranyl; and
R.sup.13 is selected from hydrogen, halo, phenyl, pyridinyl, and
--O--(C.sub.1-C.sub.4 alkyl).
2. Compounds and Definitions
Definitions of specific functional groups and chemical terms are
described in more detail below. For purposes of this invention, the
chemical elements are identified in accordance with the Periodic
Table of the Elements, CAS version, Handbook of Chemistry and
Physics, 75.sup.th Ed., inside cover, and specific functional
groups are generally defined as described therein. Additionally,
general principles of organic chemistry, as well as specific
functional moieties and reactivity, are described in Organic
Chemistry, Thomas Sorrell, University Science Books, Sausalito,
1999; Smith and March March's Advanced Organic Chemistry, 5.sup.th
Edition, John Wiley & Sons, Inc., New York, 2001; Larock,
Comprehensive Organic Transformations, VCH Publishers, Inc., New
York, 1989; Carruthers, Some Modern Methods of Organic Synthesis,
3.sup.rd Edition, Cambridge University Press, Cambridge, 1987; the
entire contents of each of which are incorporated herein by
reference.
Unless otherwise stated, structures depicted herein are also meant
to include all isomeric (e.g., enantiomeric, diastereomeric, and
geometric (or conformational)) forms of the structure; for example,
the R and S configurations for each asymmetric center, Z and E
double bond isomers, and Z and E conformational isomers. Therefore,
single stereochemical isomers as well as enantiomeric,
diastereomeric, and geometric (or conformational) mixtures of the
present compounds are within the scope of the invention. Unless
otherwise stated, all tautomeric forms of the compounds of the
invention are within the scope of the invention. Additionally,
unless otherwise stated, structures depicted herein are also meant
to include compounds that differ only in the presence of one or
more isotopically enriched atoms. For example, compounds having the
present structures including the replacement of hydrogen by
deuterium or tritium, or the replacement of a carbon by a .sup.13C-
or .sup.14C-enriched carbon are within the scope of this invention.
Such compounds are useful, for example, as analytical tools, as
probes in biological assays, or as therapeutic agents in accordance
with the present invention.
Where a particular enantiomer is preferred, it may, in some
embodiments be provided substantially free of the corresponding
enantiomer, and may also be referred to as "optically enriched."
"Optically-enriched," as used herein, means that the compound is
made up of a significantly greater proportion of one enantiomer. In
certain embodiments the compound is made up of at least about 90%
by weight of a preferred enantiomer. In other embodiments the
compound is made up of at least about 95%, 98%, or 99% by weight of
a preferred enantiomer. Preferred enantiomers may be isolated from
racemic mixtures by any method known to those skilled in the art,
including chiral high pressure liquid chromatography (HPLC) and the
formation and crystallization of chiral salts or prepared by
asymmetric syntheses. See, for example, Jacques et al.,
Enantiomers, Racemates and Resolutions (Wiley Interscience, New
York, 1981); Wilen, et al., Tetrahedron 33:2725 (1977); Eliel, E.
L. Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962);
Wilen, S. H. Tables of Resolving Agents and Optical Resolutions p.
268 (E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind.
1972).
A wavy bond
at a chiral center in a chemical structure is used to denote
compounds of the invention that are optically pure, but whose
optical rotation has not been determined. A straight bond at a
chiral center indicates a racemic mixture although, as stated
above, the invention also includes all possible isomeric forms of
the racemate.
The terms "halo" and "halogen" as used herein refer to an atom
selected from fluorine (fluoro, --F), chlorine (chloro, --Cl),
bromine (bromo, --Br), and iodine (iodo, --I).
The term "alkylene" refers to a bivalent alkyl group. An "alkylene
chain" is a polymethylene group, i.e., --(CH.sub.2).sub.n--,
wherein n is a positive integer, preferably from 1 to 6, from 1 to
4, from 1 to 3, from 1 to 2, or from 2 to 3. A substituted alkylene
chain is a polymethylene group in which one or more methylene
hydrogen atoms are replaced with a substituent. Suitable
substituents include those described below for a substituted
aliphatic group.
The term "alkyl," as used herein, refers to a monovalent saturated,
straight- or branched-chain hydrocarbon radical derived from an
aliphatic moiety containing between one and six carbon atoms by
removal of a single hydrogen atom. In some embodiments, alkyl
contains 1-5 carbon atoms. In another embodiment, alkyl contains
1-4 carbon atoms. In still other embodiments, alkyl contains 1-3
carbon atoms. In yet another embodiment, alkyl contains 1-2
carbons. Examples of alkyl radicals include, but are not limited
to, methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl,
sec-butyl, sec-pentyl, iso-pentyl, tert-butyl, n-pentyl, neopentyl,
n-hexyl, sec-hexyl, and the like.
As described herein, compounds of the invention may contain
"optionally substituted" moieties. In general, the term
"substituted", whether preceded by the term "optionally" or not,
means that one or more hydrogens of the designated moiety are
replaced with a suitable substituent. Unless otherwise indicated,
an "optionally substituted" group may have a suitable substituent
at each substitutable position of the group, and when more than one
position in any given structure may be substituted with more than
one substituent selected from a specified group, the substituent
may be either the same or different at each position. Combinations
of substituents envisioned under this invention are preferably
those that result in the formation of stable or chemically feasible
compounds. The term "stable", as used herein, refers to compounds
that are not substantially altered when subjected to conditions to
allow for their production, detection, and, in certain embodiments,
their recovery, purification, and use for one or more of the
purposes disclosed herein.
Suitable monovalent substituents on a substitutable carbon atom of
an "optionally substituted" group are independently halogen;
--(CH.sub.2).sub.0-4R.sup..smallcircle.;
--(CH.sub.2).sub.0-4OR.sup..smallcircle.;
--O--(CH.sub.2).sub.0-4C(O)OR.sup..smallcircle.;
--(CH.sub.2).sub.0-4CH(OR.sup..smallcircle.).sub.2;
--(CH.sub.2).sub.0-4SR.sup..smallcircle.; --(CH.sub.2).sub.0-4Ph,
which may be substituted with R.sup..smallcircle.;
--(CH.sub.2).sub.0-4O(CH.sub.2).sub.0-1Ph which may be substituted
with R.sup..smallcircle.; --CH.dbd.CHPh, which may be substituted
with R.sup..smallcircle.; --NO.sub.2; --CN; --N.sub.3;
--(CH.sub.2).sub.0-4N(R.sup..smallcircle.).sub.2;
--(CH.sub.2).sub.0-4N(R.sup..smallcircle.)C(O)R.sup..smallcircle.;
--N(R.sup..smallcircle.)C(S)R.sup..smallcircle.;
--(CH.sub.2).sub.0-4N(R.sup..smallcircle.)C(O)NR.sup..smallcircle..sub.2;
--N(R.sup..smallcircle.)C(S)NR.sup..smallcircle..sub.2;
--(CH.sub.2).sub.0-4N(R.sup..smallcircle.)C(O)OR.sup..smallcircle.;
--N(R.sup..smallcircle.)N(R.sup..smallcircle.)C(O)R.sup..smallcircle.;
--N(R.sup..smallcircle.)N(R.sup..smallcircle.)C(O)NR.sup..smallcircle..su-
b.2; --N(R.sup..smallcircle.)
N(R.sup..smallcircle.)C(O)OR.sup..smallcircle.;
--(CH.sub.2).sub.0-4C(O)R.sup..smallcircle.;
--C(S)R.sup..smallcircle.;
--(CH.sub.2).sub.0-4C(O)OR.sup..smallcircle.;
--(CH.sub.2).sub.0-4C(O)SR.sup..smallcircle.;
--(CH.sub.2).sub.0-4C(O)OSiR.sup..smallcircle..sub.3;
--(CH.sub.2).sub.0-4OC(O)R.sup..smallcircle.;
--OC(O)(CH.sub.2).sub.0-4SR--; --SC(S)SR.sup..smallcircle.;
--(CH.sub.2).sub.0-4SC(O)R.sup..smallcircle.;
--(CH.sub.2).sub.0-4C(O)NR.sup..smallcircle..sub.2;
--C(S)NR.sup..smallcircle..sub.2; --C(S)SR.sup..smallcircle.;
--SC(S)SR.sup..smallcircle.;
--(CH.sub.2).sub.0-4OC(O)NR.sup..smallcircle..sub.2;
--C(O)N(OR.sup..smallcircle.) R.sup..smallcircle.;
--C(O)C(O)R.sup..smallcircle.;
--C(O)CH.sub.2C(O)R.sup..smallcircle.;
--C(NOR.sup..smallcircle.)R.sup..smallcircle.;
--(CH.sub.2).sub.0-4SSR.sup..smallcircle.;
--(CH.sub.2).sub.0-4S(O).sub.2R.sup..smallcircle.;
--(CH.sub.2).sub.0-4S(O).sub.2OR.sup..smallcircle.;
--(CH.sub.2).sub.0-4OS(O).sub.2R.sup..smallcircle.;
--S(O).sub.2NR.sup..smallcircle..sub.2;
--(CH.sub.2).sub.0-4S(O)R.sup..smallcircle.;
--N(R.sup..smallcircle.)S(O).sub.2NR.sup..smallcircle..sub.2;
--N(R.sup..smallcircle.)S(O).sub.2R.sup..smallcircle.;
--N(OR.sup..smallcircle.)R.sup..smallcircle.;
--C(NH)NR.sup..smallcircle..sub.2; --P(O).sub.2R.sup..smallcircle.;
--P(O)R.sup..smallcircle..sub.2; --OP(O)R.sup..smallcircle..sub.2;
--OP(O)(OR.sup..smallcircle.).sub.2; --SiR.sup..smallcircle..sub.3;
--(C.sub.1-4 straight or
branched)alkylene)O--N(R.sup..smallcircle.).sub.2; or --(C.sub.1-4
straight or branched alkylene)C(O)O--N(R.sup..smallcircle.).sub.2,
wherein each R.sup..smallcircle. may be substituted as defined
below and is independently hydrogen, C.sub.1-6 aliphatic,
--CH.sub.2Ph, --O(CH.sub.2).sub.0-1Ph, or a 5-6-membered saturated,
partially unsaturated, or aryl ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, or sulfur, or,
notwithstanding the definition above, two independent occurrences
of R.sup..smallcircle., taken together with their intervening
atom(s), form a 3-12-membered saturated, partially unsaturated, or
aryl mono- or bicyclic ring having 0-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur, which may be substituted
as defined below.
Suitable substituents on a substitutable nitrogen of an "optionally
substituted" group include --R.sup..dagger.,
--NR.sup..dagger..sub.2, --C(O)R.sup..dagger.,
--C(O)OR.sup..dagger., --C(O)C(O)R.sup..dagger.,
--C(O)CH.sub.2C(O)R.sup..dagger., --S(O).sub.2R.sup..dagger.,
--S(O).sub.2NR.sup..dagger..sub.2, --C(S)NR.sup..dagger..sub.2,
--C(NH)NR.sup..dagger..sub.2, or
--N(R.sup..dagger.)S(O).sub.2R.sup..dagger.; wherein each
R.sup..dagger. is independently hydrogen, C.sub.1-6 aliphatic which
may be substituted as defined below, unsubstituted --OPh, or an
unsubstituted 5-6-membered saturated, partially unsaturated, or
aryl ring having 0-4 heteroatoms independently selected from
nitrogen, oxygen, or sulfur, or, notwithstanding the definition
above, two independent occurrences of R.sup..dagger., taken
together with their intervening atom(s) form an unsubstituted
3-12-membered saturated, partially unsaturated, or aryl mono- or
bicyclic ring having 0-4 heteroatoms independently selected from
nitrogen, oxygen, or sulfur.
Suitable substituents on the aliphatic group of R.sup..dagger. are
independently halogen, --R.sup..cndot., -(halo R.sup..cndot.),
--OH, --OR.sup..cndot., --O(halo R.sup..cndot.), --CN, --C(O)OH,
--C(O)O R.sup..cndot., --NH.sub.2, --NHR.sup..cndot.,
--NR.sup..cndot..sub.2, or --NO.sub.2, wherein each R.sup..cndot.
is unsubstituted or where preceded by "halo" is substituted only
with one or more halogens, and is independently C.sub.1-4aliphatic,
--CH.sub.2Ph, --O(CH.sub.2).sub.0-1Ph, or a 5-6-membered saturated,
partially unsaturated, or aryl ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, or sulfur.
As used herein, the term "inhibitor" is defined as a compound that
binds to and/or inhibits a target S-adenosylmethionine (SAM)
utilizing enzyme with measurable affinity. In certain embodiments,
an inhibitor has an IC.sub.50 and/or binding constant of less about
50 .mu.M, less than about 1 .mu.M, less than about 500 nM, less
than about 100 nM, or less than about 10 nM.
The terms "measurable affinity" and "measurably inhibit," as used
herein, means a measurable change in activity of at least one SAM
utilizing enzyme between a sample comprising a provided compound,
or composition thereof, and at least one SAM dependent enzyme, and
an equivalent sample comprising at least one SAM dependent enzyme,
in the absence of said compound, or composition thereof.
3. Description of Exemplary Compounds
In some embodiments of Formula II, R.sup.1a is selected from
--OCH.sub.3, --CH.sub.3, --OCHF.sub.2, and --CH.sub.2CH.sub.3.
In some embodiments of Formula II, R.sup.4a is selected from
--CH.sub.2OCH.sub.3, --CH(CH.sub.3)OCH.sub.3,
4,4-difluorocyclohexyl, cyclopropyl, tetrayhyrdopyran-4-yl,
1-(t-butoxycarbonyl)-piperidin-4-yl,
1-(isobutoxycarbonyl)-piperidin-4-yl,
1-(isopropoxycarbonyl)-piperidin-4-yl,
1-(2-fluoroethyl)-piperidin-4-yl,
1-(2,2-difluoroethyl)-piperidin-4-yl,
1-(2,2,2-trifluoroethyl)-piperidin-4-yl,
1-(2-hydroxyisobutyl)-piperidin-4-yl,
1-(hydroxyisopropylcarbonyl)-piperidin-4-yl,
1-(ethoxycarbonylmethyl)-piperidin-4-yl,
1-(isopropylcarbonyl)-piperidin-4-yl, 1-methylpiperidin-4-yl,
1-(methylsulfonyl)-piperidin-4-yl,
1-(ethylsulfonyl)-piperidin-4-yl,
1-(isopropylsulfonyl)-piperidin-4-yl, 1-(phenyl)-piperidin-4-yl,
1-(oxetan-3-yl)piperidin-4-yl, 1-(pyridin-2-yl)-piperidin-4-yl, and
1-(pyrimidin-2-yl)-piperidin-4-yl.
In some embodiments of Formula II, R.sup.13 is selected from
hydrogen, chloro, fluoro, --OCH(CH.sub.3).sub.2, phenyl, and
pyridin-2-yl.
Exemplary compounds of Formula II are set forth in FIG. 1. In some
cases two (or more) of the compounds in FIG. 1 having one (or more)
wavy bonds will have the exact same structure. Because the wavy
bond represents a chiral center of undetermined optical rotation,
such compounds will be understood to be separate and distinct
optical isomers of one another. FIG. 1 is annotated to indicate
those sets of two or more compounds that have the same depicted
structure, but are of different stereochemistry.
4. Uses, Formulation and Administration
Pharmaceutically Acceptable Compositions
According to another embodiment, the invention provides a
composition comprising a compound of this invention or a
pharmaceutically acceptable derivative thereof and a
pharmaceutically acceptable carrier, adjuvant, or vehicle. The
amount of compound in compositions of this invention is such that
is effective to measurably modulate a histone methyl modifying
enzyme, or a mutant thereof, in a biological sample or in a
patient. In certain embodiments, the amount of compound in
compositions of this invention is such that is effective to
measurably modulate a histone methyl modifying enzyme, or a mutant
thereof, in a biological sample or in a patient.
In certain embodiments, a composition of this invention is
formulated for administration to a patient in need of such
composition. In some embodiments, a composition of this invention
is formulated for oral administration to a patient.
The term "patient," as used herein, means an animal, preferably a
mammal, and most preferably a human.
The term "pharmaceutically acceptable carrier, adjuvant, or
vehicle" refers to a non-toxic carrier, adjuvant, or vehicle that
does not destroy the pharmacological activity of the compound with
which it is formulated. Pharmaceutically acceptable carriers,
adjuvants or vehicles that may be used in the compositions of this
invention include, but are not limited to, ion exchangers, alumina,
aluminum stearate, lecithin, serum proteins, such as human serum
albumin, buffer substances such as phosphates, glycine, sorbic
acid, potassium sorbate, partial glyceride mixtures of saturated
vegetable fatty acids, water, salts or electrolytes, such as
protamine sulfate, disodium hydrogen phosphate, potassium hydrogen
phosphate, sodium chloride, zinc salts, colloidal silica, magnesium
trisilicate, polyvinyl pyrrolidone, cellulose-based substances,
polyethylene glycol, sodium carboxymethylcellulose, polyacrylates,
waxes, polyethylenepolyoxypropylene-block polymers, polyethylene
glycol and wool fat.
A "pharmaceutically acceptable derivative" means any non-toxic
salt, ester, salt of an ester or other derivative of a compound of
this invention that, upon administration to a recipient, is capable
of providing, either directly or indirectly, a compound of this
invention or an inhibitorily active metabolite or residue
thereof.
Compositions of the present invention may be administered orally,
parenterally, by inhalation spray, topically, rectally, nasally,
buccally, vaginally or via an implanted reservoir. The term
"parenteral" as used herein includes subcutaneous, intravenous,
intramuscular, intra-articular, intra-synovial, intrasternal,
intrathecal, intrahepatic, intralesional and intracranial injection
or infusion techniques. Preferably, the compositions are
administered orally, intraperitoneally or intravenously. Sterile
injectable forms of the compositions of this invention may be
aqueous or oleaginous suspension. These suspensions may be
formulated according to techniques known in the art using suitable
dispersing or wetting agents and suspending agents. The sterile
injectable preparation may also be a sterile injectable solution or
suspension in a non-toxic parenterally acceptable diluent or
solvent, for example as a solution in 1,3-butanediol. Among the
acceptable vehicles and solvents that may be employed are water,
Ringer's solution and isotonic sodium chloride solution. In
addition, sterile, fixed oils are conventionally employed as a
solvent or suspending medium.
For this purpose, any bland fixed oil may be employed including
synthetic mono- or di-glycerides. Fatty acids, such as oleic acid
and its glyceride derivatives are useful in the preparation of
injectables, as are natural pharmaceutically-acceptable oils, such
as olive oil or castor oil, especially in their polyoxyethylated
versions. These oil solutions or suspensions may also contain a
long-chain alcohol diluent or dispersant, such as carboxymethyl
cellulose or similar dispersing agents that are commonly used in
the formulation of pharmaceutically acceptable dosage forms
including emulsions and suspensions. Other commonly used
surfactants, such as Tweens, Spans and other emulsifying agents or
bioavailability enhancers which are commonly used in the
manufacture of pharmaceutically acceptable solid, liquid, or other
dosage forms may also be used for the purposes of formulation.
Pharmaceutically acceptable compositions of this invention may be
orally administered in any orally acceptable dosage form including,
but not limited to, capsules, tablets, aqueous suspensions or
solutions. In the case of tablets for oral use, carriers commonly
used include lactose and corn starch. Lubricating agents, such as
magnesium stearate, are also typically added. For oral
administration in a capsule form, useful diluents include lactose
and dried cornstarch. When aqueous suspensions are required for
oral use, the active ingredient is combined with emulsifying and
suspending agents. If desired, certain sweetening, flavoring or
coloring agents may also be added.
Alternatively, pharmaceutically acceptable compositions of this
invention may be administered in the form of suppositories for
rectal administration. These can be prepared by mixing the agent
with a suitable non-irritating excipient that is solid at room
temperature but liquid at rectal temperature and therefore will
melt in the rectum to release the drug. Such materials include
cocoa butter, beeswax and polyethylene glycols.
Pharmaceutically acceptable compositions of this invention may also
be administered topically, especially when the target of treatment
includes areas or organs readily accessible by topical application,
including diseases of the eye, the skin, or the lower intestinal
tract. Suitable topical formulations are readily prepared for each
of these areas or organs.
Topical application for the lower intestinal tract can be effected
in a rectal suppository formulation (see above) or in a suitable
enema formulation. Topically-transdermal patches may also be
used.
For topical applications, provided pharmaceutically acceptable
compositions may be formulated in a suitable ointment containing
the active component suspended or dissolved in one or more
carriers. Carriers for topical administration of compounds of this
invention include, but are not limited to, mineral oil, liquid
petrolatum, white petrolatum, propylene glycol, polyoxyethylene,
polyoxypropylene compound, emulsifying wax and water.
Alternatively, provided pharmaceutically acceptable compositions
can be formulated in a suitable lotion or cream containing the
active components suspended or dissolved in one or more
pharmaceutically acceptable carriers. Suitable carriers include,
but are not limited to, mineral oil, sorbitan monostearate,
polysorbate 60, cetyl esters wax, cetearyl alcohol,
2-octyldodecanol, benzyl alcohol and water.
For ophthalmic use, provided pharmaceutically acceptable
compositions may be formulated as micronized suspensions in
isotonic, pH adjusted sterile saline, or, preferably, as solutions
in isotonic, pH adjusted sterile saline, either with or without a
preservative such as benzylalkonium chloride. Alternatively, for
ophthalmic uses, the pharmaceutically acceptable compositions may
be formulated in an ointment such as petrolatum.
Pharmaceutically acceptable compositions of this invention may also
be administered by nasal aerosol or inhalation. Such compositions
are prepared according to techniques well-known in the art of
pharmaceutical formulation and may be prepared as solutions in
saline, employing benzyl alcohol or other suitable preservatives,
absorption promoters to enhance bioavailability, fluorocarbons,
and/or other conventional solubilizing or dispersing agents.
Most preferably, pharmaceutically acceptable compositions of this
invention are formulated for oral administration. Such formulations
may be administered with or without food. In some embodiments,
pharmaceutically acceptable compositions of this invention are
administered without food. In other embodiments, pharmaceutically
acceptable compositions of this invention are administered with
food.
The amount of compounds of the present invention that may be
combined with the carrier materials to produce a composition in a
single dosage form will vary depending upon the host treated and
the particular mode of administration. Preferably, provided
compositions should be formulated so that a dosage of between
0.01-100 mg/kg body weight/day of the inhibitor can be administered
to a patient receiving these compositions.
It should also be understood that a specific dosage and treatment
regimen for any particular patient will depend upon a variety of
factors, including the activity of the specific compound employed,
the age, body weight, general health, sex, diet, time of
administration, rate of excretion, drug combination, and the
judgment of the treating physician and the severity of the
particular disease being treated. The amount of a compound of the
present invention in the composition will also depend upon the
particular compound in the composition.
Uses of Compounds and Pharmaceutically Acceptable Compositions
Compounds and compositions described herein are generally useful
for the modulating of activity of one or more enzymes involved in
epigenetic regulation.
Epigenetics is the study of heritable changes in gene expression
caused by mechanisms other than changes in the underlying DNA
sequence. Molecular mechanisms that play a role in epigenetic
regulation include DNA methylation and chromatin/histone
modifications. Histone methylation, in particular, is critical in
many epigenetic phenomena.
Chromatin, the organized assemblage of nuclear DNA and histone
proteins, is the basis for a multitude of vital nuclear processes
including regulation of transcription, replication, DNA-damage
repair and progression through the cell cycle. A number of factors,
such as chromatin-modifying enzymes, have been identified that play
an important role in maintaining the dynamic equilibrium of
chromatin (Margueron, et al. (2005) Curr. Opin. Genet. Dev.
15:163-176).
Histones are the chief protein components of chromatin. They act as
spools around which DNA winds, and they play a role in gene
regulation. There are a total of six classes of histones (H1, H2A,
H2B, H3, H4, and H5) organized into two super classes: core
histones (H2A, H2B, H3, and H4) and linker histones (H1 and H5).
The basic unit of chromatin is the nucleosome, which consists of
about 147 base pairs of DNA wrapped around the histone octamer,
consisting of two copies each of the core histones H2A, H2B, H3,
and H4 (Luger, et al. (1997) Nature 389:251-260).
Histones, particularly residues of the amino termini of histones H3
and H4 and the amino and carboxyl termini of histones H2A, H2B and
H1, are susceptible to a variety of post-translational
modifications including acetylation, methylation, phosphorylation,
ribosylation, sumoylation, ubiquitination, citrullination,
deimination, and biotinylation. The core of histones H2A and H3 can
also be modified. Histone modifications are integral to diverse
biological processes such as gene regulation, DNA repair, and
chromosome condensation.
The present disclosure provides compounds and compositions for
modulating activity of histone methyl modifying enzymes. Histone
methyl modifying enzymes are key regulators of cellular and
developmental processes. Histone methyl modifying enzymes may be
characterized as either histone methyl transferases or histone
demethylases. Histone demethylase enzymes have modules that mediate
binding to methylated residues. For example, multiple demethylases
contain a Tudor domain (e.g., JMJD2C/GASC1) or a PHD domain (e.g.,
JARID1C/SMCX, PHF8).
The lysine specificities of many histone methyltransferases have
been characterized. For example SET7/9, SMYD3, and MLL1-5 are
specific for H3K4. SUV39H1, DIM-5, and G9a are specific for H3K9.
SET8 is specific for H4K20.
DOT1 is an example of a non-SET domain containing histone
methylase. DOT1 methylates H3 on lysine 79.
Just as histone methylases have been shown to regulate
transcriptional activity, chromatin structure, and gene silencing,
demethylases have also been discovered which impact gene
expression. LSD1 was the first histone lysine demethylase to be
characterized. This enzyme displays homology to FAD-dependent amine
oxidases and acts as a transcriptional corepressor of neuronal
genes (Shi et al., Cell 119:941-953, 2004). Additional demethylases
defining separate demethylase families have been discovered,
including JHDM1 (or KDM2), JHDM2 (or KDM3), JMJD2 (or KDM4), JARID
(or KDM5), JMJD3 (or KDM6), and JMJD6 families (Lan et al., Curr.
Opin. Cell Biol. 20(3):316-325, 2008).
Demethylases act on specific lysine residues within substrate
sequences and discriminate between the degree of methylation
present on a given residue. For example, LSD1 removes mono- or
dimethyl-groups from H3K4. Members of the JARID1A-D family remove
trimethyl groups from H3K4. UTX and JMJD3 demethylate H3K27,
counteracting effects of EZH2 methylase activity. Substrate
specificities of other demethylases have been characterized (see
Shi, Nat. Rev. 8:829-833, 2007).
One class of histone methylases is characterized by the presence of
a SET domain, named after proteins that share the domain,
Su(var)3-9, enhancer of zeste [E(Z)], and trithorax. A SET domain
includes about 130 amino acids. SET domain-containing methylase
families include SUV39H1, SET1, SET2, EZH2, RIZ1, SMYD3, SUV4-20H1,
SET7/9, and PR-SET7/SET8 families (reviewed in Dillon et al.,
Genome Biol. 6:227, 2005). Members of a family typically include
similar sequence motifs in the vicinity of and within the SET
domain. The human genome encodes over 50 SET domain-containing
histone protein methylases, any of which can be used in an assay
described herein.
EZH2 is an example of a human SET-domain containing methylase. EZH2
associates with EED (Embryonic Ectoderm Development) and SUZ12
(suppressor of zeste 12 homolog) to form a complex known as PRC2
(Polycomb Group Repressive Complex 2) having the ability to
trimethylate histone H3 at lysine 27 (Cao and Zhang, Mol. Cell
15:57-67, 2004). PRC2 complexes can also include RBAP46 and RBAP48
subunits.
The oncogenic activities of EZH2 have been shown by a number of
studies. In cell line experiments, over-expression of EZH2 induces
cell invasion, growth in soft agar, and motility while knockdown of
EZH2 inhibits cell proliferation and cell invasion (Kleer et al.,
2003, Proc. Nat. Acad. Sci. USA 100:11606-11611; Varambally et al.,
(2002), "The polycomb group protein EZH2 is involved in progression
of prostate cancer," Nature 419, 624-629). It has been shown that
EZH2 represses the expression of several tumor supressors,
including E-cadherin, DAB2IP and RUNX3 among others. In xenograft
models, EZH2 knockdown inhibits tumor growth and metastasis.
Recently, it has been shown that down modulation of EZH2 in murine
models blocks prostate cancer metastasis (Min et al., "An
oncogene-tumor suppressor cascade drives metastatic prostate cancer
by coordinately activating Ras and nuclear factor-kappaB," Nat Med.
2010 March; 16(3):286-94). EZH2 overexpression is associated with
aggressiveness of certain cancers such as breast cancer (Kleer et
al., Proc. Nat. Acad. Sci. USA 100:11606-11611, 2003). Recent
studies also suggest that prostate cancer specific oncogenic fusion
gene TMPRSS2-ERG induces repressive epigenetic programs via direct
activation of EZH2 (Yu et al., "An Integrated Network of Androgen
Receptor, Polycomb, and TMPRSS2-ERG Gene Fusions in Prostate Cancer
Progression," Cancer Cell. 2010 May 18; 17(5):443-454).
In some embodiments, compounds of the present invention modulate
the activity of one or more enzymes involved in epigenetic
regulation. In some embodiments, compounds of the present invention
modulate the activity of a histone methyl modifying enzyme, or a
mutant thereof. In some embodiments, compounds of the present
invention modulate EZH2 activity. In some embodiments, compounds of
the present invention down-regulate or suppress the activity of
EZH2. In some embodiments, compounds of the present invention are
antagonists of EZH2 activity.
In some embodiments, compounds and compositions of the present
invention are useful in treating diseases and/or disorders
associated with a histone methyl modifying enzyme. Accordingly, in
some embodiments, the present invention provides a method of
modulating a disease and/or disorder associated with a histone
methyl modifying enzyme. In some embodiments, the present invention
provides a method of treating a subject suffering from a disease
and/or disorder associated with a histone methyl modifying enzyme
comprising the step of administering a compound or composition of
Formula II.
In some embodiments, compounds and compositions of the present
invention are useful in treating diseases and/or disorders
associated with overexpression of EZH2. In some embodiments, the
present invention provides a method of treating a subject suffering
from a disease and/or disorder associated with overexpression of
EZH2 comprising the step of administering a compound or composition
of Formula II. In some embodiments, the above method additionally
comprises the preliminary step of determining if the subject is
overexpressing EZH2.
In some embodiments, compounds and compositions of the present
invention are useful in treating diseases and/or disorders
associated with cellular proliferation. In some embodiments,
compounds and compositions of the present invention are useful in
treating diseases and/or disorders associated with misregulation of
cell cycle or DNA repair. In some embodiments, compounds and
compositions of the present invention are useful in treating
cancer. Exemplary types of cancer include breast cancer, prostate
cancer, colon cancer, renal cell carcinoma, glioblastoma multiforme
cancer, bladder cancer, melanoma, bronchial cancer, lymphoma and
liver cancer.
The study of EZH2 deletions, missense and frameshift mutations
suggest that EZH2 functions as a tumor suppressor in blood
disorders such as myelodysplastic syndromes (MDS) and myeloid
malignancies (Ernst et al., Nat Genet. 2010 August; 42(8):722-6;
Nikoloski et al., Nat Genet. 2010 August; 42(8):665-7).
Accordingly, in some embodiments, compounds and compositions of the
present invention are useful in treating diseases and/or disorders
associated with the presence of a mutant form of EZH2. In some
embodiments, compounds and compositions of the present invention
are useful in treating diseases and/or disorders associated with
the presence of Y641N EZH2. In some embodiment, the disease or
disorder associated with the presence of a mutant form of EZH2 is a
human B cell lymphoma. In some embodiments, the disease and/or
disorder associated with the presence of Y641N EZH2 is follicular
lymphoma or diffuse large-B-cell lymphoma. In some embodiments,
compounds or compositions of the present invention are useful in
treating blood disorders, such as myelodysplastic syndromes,
leukemia, anemia and cytopenia. Sneeringer et al., "Coordinated
activities of wild-type plus mutant EZH2 drive tumor-associated
hypertrimethylation of lysine 27 on histone H3 (H3K27) in human
B-cell lymphomas," Proceedings of the National Academy of Sciences,
PNAS Early Edition published ahead of print on Nov. 15, 2010.
In some embodiments, the present invention provides a method of
reducing the activity of EZH2 in a subject comprising the step of
administering a compound or composition of Formula II. In some
embodiments, the present invention provides a method of reducing
the activity of wide-type EZH2 in a subject comprising the step of
administering a compound or composition of Formula II. In some
embodiments, the present invention provides a method of reducing
the activity of a mutant form of EZH2 in a subject comprising the
step of administering a compound or composition of Formula II. In
some embodiments, the present invention provides a method of
reducing the activity of a mutant form of EZH2 in a subject
comprising the step of administering a compound or composition of
Formula II, wherein the mutant form of EZH2 is Y641N EZH2. In some
embodiments, the present invention provides a method of treating a
subject suffering from a disease and/or disorder associated with
EZH2 comprising the step of administering a compound or composition
of Formula II. In some embodiments, the present invention provides
a method of treating a subject suffering from a disease and/or
disorder associated with wide-type EZH2 comprising the step of
administering a compound or composition of Formula II. In some
embodiments, the present invention provides a method of treating a
subject suffering from a disease and/or disorder associated with a
mutant form of EZH2 comprising the step of administering a compound
or composition of Formula II. In some embodiments, the present
invention provides a method of treating a subject suffering from a
disease and/or disorder associated with a mutant form of EZH2
comprising the step of administering a compound or composition of
Formula II, wherein the mutant form of EZH2 is Y641N EZH2. In some
embodiments, the above method additionally comprises the
preliminary step of determining if the subject is expressing a
mutant form of EZH2, such as Y641N EZH2 In some embodiments, the
present invention provides a method of reducing the activity of a
mutant form of EZH2, such as Y641N EZH2, in a subject in need
thereof comprising the step of administering a compound or
composition of Formula II. In some embodiments, the present
invention provides a method of treating a subject suffering from a
disease and/or disorder associated with a mutant form of EZH2
comprising the step of administering a compound or composition of
Formula II. In some embodiments, the above method additionally
comprises the preliminary step of determining if the subject is
expressing a mutant form of EZH2, such as Y641N EZH2. In some
embodiments, that determination is made by determining if the
subject has increased levels of histone H3 Lys-27-specific
trimethylation (H3K27me3), as compared to a subject known not to
express a mutant form of EZH2.
EQUIVALENTS
The representative examples that follow are intended to help
illustrate the invention, and are not intended to, nor should they
be construed to, limit the scope of the invention. Indeed, various
modifications of the invention and many further embodiments
thereof, in addition to those shown and described herein, will
become apparent to those skilled in the art from the full contents
of this document, including the examples that follow and the
references to the scientific and patent literature cited herein. It
should further be appreciated that the contents of those cited
references are incorporated herein by reference to help illustrate
the state of the art.
It will be appreciated that for compound preparations described
herein, when reverse phase HPLC is used to purify a compound, a
compound may exist as an acid addition salt. In some embodiments, a
compound may exist as a formic acid or mono-, di-, or
tri-trifluoroacetic acid salt.
It will further be appreciated that the present invention
contemplates individual compounds described herein. Where
individual compounds exemplified are isolated and/or characterized
as a salt, for example, as a trifluoroacetic acid salt, the present
invention contemplates a free base of the salt, as well as other
pharmaceutically acceptable salts of the free base.
The following examples contain important additional information,
exemplification and guidance that can be adapted to the practice of
this invention in its various embodiments and the equivalents
thereof.
Procedures for preparing the compounds exemplified below, as well
as additional compounds/intermediates in the synthetic schemes can
be found in International Application No. PCT/US2013/025639, the
contents of which are incorporated herein by reference.
EXAMPLES
As depicted in the Examples below, in certain exemplary
embodiments, compounds are prepared according to the following
general procedures. It will be appreciated that, although the
synthetic methods and Schemes depict the synthesis of certain
compounds of the present invention, the following methods and other
methods known to one of ordinary skill in the art can be applied to
all compounds and subclasses and species of each of these
compounds, as described herein.
Unless otherwise noted, all solvents, chemicals, and reagents were
obtained commercially and used without purification. The .sup.1H
NMR spectra were obtained in CDCl.sub.3, d.sub.6-DMSO, CD.sub.3OD,
or d.sub.6-acetone at 25.degree. C. at 300 MHz on an OXFORD
(Varian) with chemical shift (.delta., ppm) reported relative to
TMS as an internal standard. HPLC-MS chromatograms and spectra were
obtained with Shimadzu LCMS-2020 system. Chiral analysis and
purification were obtained with Yilite P270.
Example 1
Synthesis of Compounds 327 and 346 and Related Compounds and
Intermediates
The title compounds of this Example and other related compounds
were prepared according to the following general scheme. In
addition, where indicated, modifications of this scheme are
disclosed for the synthesis of still additional related compounds
of the invention and intermediates thereof.
##STR00003## ##STR00004##
Step 1: (S,E)-tert-butyl
4-(((tert-butylsulfinyl)imino)methyl)piperidine-1-carboxylate
##STR00005## To a round bottomed flask charged with a magnetic stir
bar was added (S)-2-methylpropane-2-sulfinamide (20.46 g, 169
mmol), tert-butyl 4-formylpiperidine-1-carboxylate (30 g, 141
mmol), DCM (300 mL), and Ti(OEt).sub.4 (59.0 ml, 281 mmol). The
solution was stirred at room temperature for 3 h before it was
quenched with brine (80 mL). The solution was stirred for 30
minutes before filtering. The filter cake was washed with DCM and
the filtrate was placed in a separatory funnel and washed with
water. The organics layer was dried over Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. The crude residue solidified
to the title compound (29 g, 92 mmol, 65.1% yield) m/z 217.
The intermediate shown in the following table was prepared
according to the general procedure outlined in Step 1 using the
appropriate starting materials and modifications.
TABLE-US-00001 Name Structure m/z (S,E)-2-methyl-N-
((tetrahydro-2H- pyran-4-yl)methylene) propane-2-sulfinamide
##STR00006##
Step 2: Tert-butyl 4-((S)-1-((R or
S)-1,1-dimethylethylsulfinamido)ethyl) piperidine-1-carboxylate
##STR00007## To a round bottomed flask charged with a magnetic stir
bar was added (S,E)-tert-butyl
4-((tert-butylsulfinylimino)methyl)piperidine-1-carboxylate (36.4
g, 115 mmol), DCM (400 mL), and the solution was cooled to
0.degree. C. in an ice bath with stirring. To this solution was
added MeMgBr (77 ml, 230 mmol) (3M in diethyl ether) and the
reaction stirred for 4 h while warming to room temperature. The
reaction was carefully quenched via the addition of saturated
aqueous NH.sub.4Cl. The solid were broken up by the addition of 1N
HCl. The layers were separated and the aqueous phase was extracted
with DCM. The combined organics phase was dried over
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo to afford the
title compound (29 g, >9:1 dr) which is used without further
purification in the next step.
The intermediate shown in the following table was prepared
according to the general procedure outlined in Step 2 using the
appropriate starting materials and modifications.
TABLE-US-00002 Name Structure m/z (S)-2-methyl-N-((R or S)-1-
(tetrahydro-2H-pyran-4- yl)ethyl)propane- 2-sulfinamide
##STR00008## 234
Step 3: (R or S)-tert-butyl
4-(1-aminoethyl)piperidine-1-carboxylate
##STR00009## To a 1 L round bottomed flask charged with a magnetic
stir bar was added crude tert-butyl
4-((S)-1-((S)-1,1-dimethylethylsulfinamido)ethyl)piperidine-1-carboxylate
(29 g) was taken up in MeOH (200 mL) before addition of a 4 N
solution of HCl in 1,4-dioxane (24.06 ml, 96 mmol). The resulting
solution was then stirred at room temperature for 1 h at rt. The
methanol was then removed in vacuo to afford viscous oil which was
treated with sat'd aqueous NaHCO.sub.3 (.about.500 mL) and
extracted with ethyl acetate (2.times.500 mL). This organic phase
was combined, dried with MgSO.sub.4, filtered, and solvent was then
removed in vacuo affording the title compound (22 g) which was used
without further purification.
The intermediate shown in the following table was prepared
according to the general procedure outlined in Step 3 using the
appropriate starting materials.
TABLE-US-00003 Name Structure m/z (R or S)-1-(tetrahydro-
2H-pyran-4-yl)ethanamine ##STR00010## 130
Step 4: Methyl 2-(2-bromophenyl)-3-oxobutanoate
##STR00011## A round bottomed flask was charged with a magnetic
stir bar and methyl 2-(2-bromophenyl)acetate (25 g, 109 mmol) and
THF (50 mL). This solution was cooled to -78.degree. C. before drop
wise addition of a 1M solution of LiHMDS in THF (218 ml, 218 mmol).
The reaction was stirred for 30 min at -78.degree. C. before
addition of 1-(1H-imidazol-1-yl)ethanone (14.42 g, 131 mmol)
dissolved in a mixture of THF:DMF (112 mL THF, 24 mL DMF). The
solution was stirred for 1 h before quenching with sat'd aqueous
NH.sub.4Cl (.about.250 mL) and diluting with EtOAc. The layers were
separated and the aqueous phase was extracted with EtOAc
(.about.2.times.250 mL). The combined organic extract was washed
with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated
in vacuo. The crude residue was purified via silica gel
chromatography using an eluent of ethyl acetate/hexanes (10:1) to
afford methyl 2-(2-bromophenyl)-3-oxobutanoate (32.5 g, 102 mmol,
93% yield).
The intermediates shown in the following table were prepared
according to the general procedure outlined in Step 4 using the
appropriate starting materials.
TABLE-US-00004 Name Structure m/z methyl 2-(2-bromo-4-
chlorophenyl)-3-oxobutanoate ##STR00012## 304 methyl 2-(2-bromo-4-
methoxyphenyl)-3- oxobutanoate ##STR00013## 302 methyl
2-(2-bromo-4- fluorophenyl)-3-oxobutanoate ##STR00014## 289
Step 5: (R or S, E and Z)-tert-butyl
4-(1-(3-(2-bromophenyl)-4-methoxy-4-oxobut-2-en-2-ylamino)ethyl)piperidin-
e-1-carboxylate
##STR00015## To a round bottomed flask was added (R or
S)-tert-butyl 4-(1-aminoethyl)piperidine-1-carboxylate (9.35 g,
40.9 mmol), EtOH (75 mL), and methyl
2-(2-bromophenyl)-3-oxobutanoate (7.40 g, 27.3 mmol) (from Step 4).
To this solution was added AcOH (1.563 ml, 27.3 mmol) and the
reaction was heated overnight at 85.degree. C. before cooling to
room temperature and concentrating. The crude residue was purified
via silica gel chromatography (330 g, 100% hexanes to 25% EA in
hexanes) to afford the title compound (6.45 g, 13.40 mmol, 49.1%
yield).
The intermediates shown in the following table were prepared
according to the general procedure outlined in Step 5 using the
appropriate starting materials.
TABLE-US-00005 Name Structure m/z (R or S,Z)-methyl 2-
(2-bromophenyl)-3- ((1-(tetrahydro-2H- pyran-4-yl)ethyl)
amino)but-2-enoate ##STR00016## 383 (R or S,Z)-methyl 2-
(2-bromo-4- chlorophenyl)-3- ((1-(tetrahydro- 2H-pyran-
4-yl)ethyl)amino) but-2-enoate ##STR00017## 417 (R or S,Z)-methyl
2- (2-bromo-4- chlorophenyl)- 3-((1-(tetrahydro-2H-
pyran-4-yl)ethyl) amino)but-2-enoate ##STR00018## 417 (R or
S,Z)-methyl 2- (2-bromo-4- fluorophenyl)- 3-((1-(tetrahydro-2H-
pyran-4-yl)ethyl) amino)but-2-enoate ##STR00019## 401
Step 6: (R or S)-methyl
1-(1-(1-(tert-butoxycarbonyl)piperidin-4-yl)ethyl)-2-methyl-1H-indole-3-c-
arboxylate
##STR00020## A 250 mL round bottom flask was charged with a
magnetic stir bar, (R or S,Z)-tert-butyl
4-(1-(3-(2-bromophenyl)-4-methoxy-4-oxobut-2-en-2-ylamino)ethyl)piperidin-
e-1-carboxylate (3.33 g, 6.92 mmol), RuPhos Pre-catalyst II
(Methanesulfonato(2-dicyclohexylphosphino-2',6'-di-i-propoxy-1,1'-bipheny-
l)(2-amino-1,1'-biphenyl-2-yl) palladium(II)) (0.463 g, 0.553
mmol), dicyclohexyl(2',6'-diisopropoxybiphenyl-2-yl)phosphine
(0.387 g, 0.830 mmol), anhydrous 1,4-dioxane (27.7 ml, 6.92 mmol),
and sodium methoxide (0.561 g, 10.38 mmol). The reaction mixture
was purged and back-filled with nitrogen and heated to 100.degree.
C. with stirring overnight before being allowed to cool to rt. The
reaction was diluted with ethyl acetate (.about.100 ml) and the
mixture was filtered through a bed of diatomaceous earth. The
filtrate was pre-absorbed onto silica gel (.about.30 g) and
purified via silica gel chromatography (120 g) using ethyl
acetate/hexanes (1:1) as eluent to afford the title compound (2.01
g, 4.77 mmol, 68.9% yield).
The intermediates shown in the following table were prepared
according to the general procedure outlined in Step 6 using the
appropriate starting materials.
TABLE-US-00006 Name Structure m/z (R or S)-methyl 2-
methyl-1-(1-(tetrahydro- 2H-pyran- 4-yl)ethyl)-1H-indole-3-
carboxylate ##STR00021## 302 (R or S)-methyl 6-
chloro-2-methyl-1-(1- (tetrahydro-2H-pyran-4-
yl)ethyl)-1H-indole-3- carboxylate ##STR00022## 337 (R or S)-methyl
6- methoxy-2-methyl-1-(1- (tetrahydro-2H-pyran-4-
yl)ethyl)-1H-indole-3- carboxylate ##STR00023## 332 (R or S)-methyl
6-fluoro- 2-methyl-1-(1- (tetrahydro-2H-pyran-4-
yl)ethyl)-1H-indole-3- carboxylate ##STR00024## 320
Step 7: (R or
S)-2-methyl-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-indole-3-carboxylic
acid
##STR00025## A 1 L round bottom flask was charged with a magnetic
stir bar, (R or S)-methyl
2-methyl-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-indole-3-carboxylate
(11.60 g, 38.5 mmol), ethanol (96 ml, 38.5 mmol), and 6 N aqueous
NaOH (64.1 ml, 385 mmol). The flask was fitted with a reflux
condenser and heated to reflux for 6 h before being allowed to cool
to rt. The volatiles were removed in vacuo and the resulting
mixture was poured into 10% HCl (.about.300 mL). A precipitate
formed which was collected via vacuum filtration using a Buchner
funnel. The filter cake was rinsed with an additional portion of
water (.about.200 mL), collected, and dried under vacuum to afford
the title compound (10.87 g, 35.9 mmol, 93% yield) as an off-white
solid.
The intermediates shown in the following table were prepared
according to the general procedure outlined in Step 7 using the
appropriate starting materials.
TABLE-US-00007 Name Structure m/z (R or S)-2- methyl-1-(1-
(tetrahydro- 2H-pyran-4- yl)ethyl)- 1H-indole-3- carboxylic acid
##STR00026## 287 (R or S)-6- chloro-2-methyl- 1-(1-(tetrahydro-
2H-pyran-4- yl)ethyl)-1H- indole-3- carboxylic acid ##STR00027##
321 (R or S)-6- methoxy-2- methyl-1-(1- (tetrahydro- 2H-pyran-
4-yl)ethyl)- 1H-indole-3- carboxylic acid ##STR00028## 317 (R or
S)-6- fluoro-2- methyl-1- (1-(tetrahydro- 2H-pyran-4- yl)ethyl)-1H-
indole-3- carboxylic acid ##STR00029## 306 (R or S)-2- methyl-6-
(pyridin- 3-yl)-1-(1- (tetrahydro-2H- pyran-4-yl) ethyl)-1H-indole-
3-carboxylic acid ##STR00030## 365
Step 8: (R or S)-tert-butyl
4-(1-(3-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methylcarbamoy-
l)-2-methyl-1H-indol-1-yl)ethyl)piperidine-1-carboxylate (Compound
327)
##STR00031## A 250 mL round bottom flask was charged with a
magnetic stir bar, (R or
S)-1-(1-(1-(tert-butoxycarbonyl)piperidin-4-yl)ethyl)-2-methyl-1H-indole--
3-carboxylic acid (1.950 g, 5.05 mmol),
3-(aminomethyl)-4-methoxy-6-methylpyridin-2(1H)-one hydrochloride
(2.065 g, 10.09 mmol), DMF (25.2 ml, 5.05 mmol), Hunig's base (3.52
ml, 20.18 mmol). The reaction mixture was cooled to 0.degree. C.
and COMU (2.16 g, 5.05 mmol) was added. The reaction was allowed to
stir overnight to room temperature. The reaction mixture was
diluted with water and extracted with EtOAc. The combined organic
extract was washed with brine, dried with MgSO.sub.4, filtered and
conc. in vacuo to afford the crude material which was purified via
silica gel chromatography (120 g) using MeOH/ethyl acetate (1:5) as
eluent to afford the title compound (1.86 g, 3.29 mmol, 65.3%
yield). LCMS 537 (M+1).sup.+1H NMR (400 MHz, DMSO-d.sub.6)
.delta.=11.83-11.71 (m, 1H), 7.80 (br. s., 1H), 7.73 (d, J=7.6 Hz,
1H), 7.62 (d, J=7.8 Hz, 1H), 7.06 (td, J=7.1, 14.4 Hz, 2H), 6.21
(s, 1H), 4.32 (br. s., 2H), 4.16 (br. s., 1H), 4.02 (br. s., 1H),
3.85 (s, 3H), 3.75 (br. s., 1H), 2.70 (br. s., 1H), 2.58 (s, 3H),
2.37 (br. s., 1H), 2.21 (s, 3H), 1.90 (d, J=12.9 Hz, 1H), 1.53 (d,
J=6.9 Hz, 3H), 1.35 (s, 10H), 1.21 (br. s., 1H), 0.89 (d, J=8.7 Hz,
1H), 0.67 (d, J=11.8 Hz, 1H).
The compounds shown in the following table were prepared according
to the general procedure outlined in Step 8 using the appropriate
starting materials. The structures of the compounds are shown in
FIG. 1.
TABLE-US-00008 Compound Number Name .sup.1H NMR m/z 435 (R or
S)-tert-butyl 4-(1-(3-(((4,6- 521 dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)carbamoyl)-2-methyl-
1H-indol-1-yl)ethyl)piperidine-1- carboxylate 436 (R or
S)-tert-butyl 4-(1-(3-(((4- 573 (difluoromethoxy)-6-methyl-2-
oxo-1,2-dihydropyridin-3- yl)methyl)carbamoyl)-2-methyl-
1H-indol-1-yl)ethyl)piperidine-1- carboxylate 437 (R or
S)-tert-butyl 4-(1-(3-(((4- 535 ethyl-6-methyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)carbamoyl)-2-methyl-
1H-indol-1-yl)ethyl)piperidine-1- carboxylate 298 (R or
S)-N-((4-methoxy-6- (400 MHz, DMSO-d6) .delta. 11.60 (s, 438
methyl-2-oxo-1,2-dihydropyridin- 1H), 7.73-7.62 (m, 3H), 7.60 (d,
3-yl)methyl)-2-methyl-1-(1- 2H) 7.07-7.05 (m, 2H), 6.15 (s, 1H)
(tetrahydro-2H-pyran-4-yl)ethyl)- 4.33 (s, 1H), 4.21-4.11 (m, 1H),
1H-indole-3-carboxamide 3.92 (br. d., 1H), 3.65 (d, 1H), 3.34- 3.32
(m, 1H), 3.02 (t, 1H), 2.61 (s, 3H), 2.48-2.44 (m, 1H), 2.20 (s,
3H), 1.84-1.81 (m, 1H), 1.54 (d, 3H), 1.40-1.38 (m, 12H), 1.25-1.22
(m, 1H), 1.08-1.04 (m, 1H), 0.86 (br. s., 1H), 0.58 (br. d., 1H)
300 (R or S)-6-fluoro-N-((4-methoxy- (400 MHz, DMSO-d6) .delta. =
11.57 456 6-methyl-2-oxo-1,2- (br. s., 1 H), 7.75-7.67 (m, 2 H),
dihydropyridin-3-yl)methyl)-2- 7.48 (d, J = 10.7 Hz, 1 H), 6.90 (t,
methyl-1-(1-(tetrahydro-2H- J = 8.5 Hz, 1 H), 6.13 (s, 1 H), 4.29
pyran-4-yl)ethyl)-1H-indole-3- (d, J = 4.5 Hz, 2 H), 4.12 (br. s.,
1 carboxamide H), 3.94-3.87 (m, 1 H), 3.83 (s, 3 H), 3.64 (dd, J =
3.6, 10.9 Hz, 1 H), 3.35 (br. s., 1 H), 3.05 (br. s., 1 H), 2.56
(s, 3 H), 2.45-2.37 (m, 1 H), 2.18 (s, 3 H), 1.81 (d, J = 12.7 Hz,
1 H), 1.50 (d, J = 6.9 Hz, 3 H), 1.40- 1.29 (m, 1 H), 1.11-0.99 (m,
1 H), 0.61 (br. s., 1 H) 314 (R or S)-6-chloro-N-((4-methoxy- (400
MHz, DMSO-d6) .delta. 11.57 (s, 1 472 6-methyl-2-oxo-1,2 H), 7.75
(s, 2 H), 7.66 (d, J = 8.9 dihydropyridin-3-yl)methyl)-2- Hz, 1 H),
7.08 (d, J = 8.5 Hz, 1 H), methyl-1-(1-(tetrahydro-2H- 6.14 (s, 1
H), 4.30 (d, J = 4.5 Hz, 2 pyran-4-yl)ethyl)-1H-indole-3- H),
4.21-4.05 (m, 2 H), 3.91 (d, carboxamide J = 11.4 Hz, 1 H), 3.85
(s, 3 H), 3.65 (d, J = 10.5 Hz, 1 H), 3.02 (t, J = 11.3 Hz, 1 H),
2.58 (s, 3 H), 2.46- 2.31 (m, 1 H), 2.19 (s, 3 H), 1.82 (d, J =
12.0 Hz, 1 H), 1.59-1.45 (m, 4 H), 1.44-1.29 (m, 1 H), 0.57 (d, J =
12.9 Hz, 1 H) 321 (R or S)-6-methoxy-N-((4- (400 MHz, DMSO-d6)
.delta. = 11.59 (s, 468 methoxy-6-methyl-2-oxo-1,2- 1 H), 7.67-7.59
(m, 2 H), 7.03 (s, dihydropyridin-3-yl)methyl)-2- 1 H), 6.75-6.68
(m, 1 H), 6.14 (s, methyl-1-(1-(tetrahydro-2H- 1 H), 4.30 (d, J
=5.1 Hz, 2 H), 4.10 pyran-4-yl)ethyl)-1H-indole-3- (dd, J = 7.5,
10.4 Hz, 1 H), 3.91 carboxamide (dd, J = 3.0, 11.3 Hz, 1 H), 3.83
(s, 3 H), 3.80-3.76 (m, 3 H), 3.68- 3.60 (m, 1 H), 3.38-3.32 (m, 1
H), 3.10-3.00 (m, 1 H), 2.56 (s, 3 H), 2.19 (s, 3 H), 1.83 (d, J =
12.7 Hz, 1 H), 1.55-1.43 (m, 4 H), 1.34 (br. s., 1 H), 1.10-0.96
(m, 1 H), 0.62 (d, J = 13.4 Hz, 1 H) 335 (R or S)-N-((4- 474
(difluoromethoxy)-6-methyl-2- oxo-1,2-dihydropyridin-3-
yl)methyl)-2-methyl-1-(1- (tetrahydro-2H-pyran-4-yl)ethyl)-
1H-indole-3-carboxamide 394 (R or S)-N-((4,6-dimethyl-2-oxo- 422
1,2-dihydropyridin-3-yl)methyl)- 2-methyl-1-(1-(tetrahydro-2H-
pyran-4-yl)ethyl)-1H-indole-3- carboxamide 442 (R or
S)-6-chloro-N-((4,6- 456 dimethyl-2-oxo-1,2-
dihydropyridin-3-yl)methyl)-2- methyl-1-(1-(tetrahydro-2H-
pyran-4-yl)ethyl)-1H-indole-3- carboxamide
Step 9: (R or
S)--N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-
-1-(1-(piperidin-4-yl)ethyl)-1H-indole-3-carboxamide hydrochloride
(Compound 326)
##STR00032## A 250 mL round bottom flask was charged with a
magnetic stir bar, (R or S)-tert-butyl
4-(1-(3-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methylcarbamoy-
l)-2-methyl-1H-indol-1-yl)ethyl)piperidine-1-carboxylate (Compound
327) (1.850 g, 3.45 mmol), MeOH (13.79 ml, 3.45 mmol), and HCl
(2.59 ml, 10.34 mmol) (4 N in dioxane). The reaction was allowed to
stir at rt for 6 h before being conc. in vacuo to afford the title
compound (1.65 g, 3.14 mmol, 91% yield). LCMS 437 (M+1).sup.+.
The compound shown in the following table was prepared according to
the general procedure outlined in Step 9 using the appropriate
starting materials. The structure of this compound is shown in FIG.
1.
TABLE-US-00009 Compound Number Name .sup.1H NMR m/z 376 (R or
S)-1-(1-(1- (400 MHz, DMSO-d.sub.6) .delta. 12.27-12.10 (m, 1 476
(azetidin-3-yl)piperidin- H), 11.96-11.72 (m, 1 H), 9.80 (br. s., 1
4-yl)ethyl)-N-((4,6- H), 9.19 (br. s., 2 H), 7.89-7.67 (m, 2 H),
dimethyl-2-oxo-1,2- 7.62 (d, J = 7.6 Hz, 1 H), 7.09 (quin, J = 6.6
dihydropyridin-3- Hz, 2 H), 5.99 (s, 1 H), 4.59-4.36 (m, 3 H),
yl)methyl)-2-methyl-1H- 4.24-3.95 (m, 2 H), 3.48 (d, J = 13.2 Hz, 1
indole-3-carboxamide H), 3.17 (d, J = 12.0 Hz, 1 H), 2.87 (br. s.,
1 hydrochloride H), 2.70 (br. s., 2 H), 2.58 (s, 3 H), 2.34- 2.25
(m, 3 H), 2.19-2.10 (m, 3 H), 1.75 (d, J = 12.3 Hz, 1 H), 1.57 (d,
J = 6.7 Hz, 3 H), 1.47 (d, J = 12.7 Hz, 2 H), 1.33-1.21 (m, 2 H),
0.85 (d, J = 13.6 Hz, 1 H)
Step 10: (R or S)-isopropyl
4-(1-(3-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methylcarbamoy-
l)-2-methyl-1H-indol-1-yl)ethyl)piperidine-1-carboxylate (Compound
346)
##STR00033## A 250 mL round bottom flask was charged with a
magnetic stir bar, (R or
S)--N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-
-1-(1-(piperidin-4-yl)ethyl)-1H-indole-3-carboxamide hydrochloride
(0.467 g, 0.987 mmol), DMF (2.468 ml, 0.987 mmol), THF (2.468 ml,
0.987 mmol), and N-ethyl-N-isopropylpropan-2-amine (0.638 g, 4.94
mmol). The reaction was cooled to 0.degree. C. and isopropyl
carbonochloridate (0.160 ml, 1.086 mmol) was added drop wise via
syringe. The reaction was allowed to stir for 2 h to rt and was
then treated with 5 N LiOH for 1 h to remove any acylated pyridone.
This material was extracted with ethyl acetate, washed with brine,
dried with MgSO.sub.4 and filtered and conc. in vacuo. The
resulting material was purified via silica gel chromatography (50
g) using ethyl acetate/MeOH (5:1) as eluent to afford pure title
compound as a pale yellow solid (0.300 g, 0.545 mmol, 55.2% yield).
LCMS 523 (M+1).sup.+; .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.59
(br. s., 1H), 7.74 (d, J=7.8 Hz, 1H), 7.69 (t, J=4.9 Hz, 1H), 7.62
(d, J=7.8 Hz, 1H), 7.13-7.01 (m, 2H), 6.15 (s, 1H), 4.78-4.67 (m,
1H), 4.32 (d, J=4.9 Hz, 2H), 4.23-4.12 (m, 1H), 4.12-4.02 (m, 1H),
3.84 (s, 3H), 3.82-3.74 (m, 1H), 2.79-2.66 (m, 1H), 2.58 (s, 3H),
2.46-2.34 (m, 2H), 2.20 (s, 3H), 1.96-1.88 (m, 1H), 1.58-1.46 (m,
4H), 1.15 (d, J=6.0 Hz, 6H), 0.95-0.89 (m, 1H), 0.74-0.65 (m,
1H).
The compounds shown in the following table were prepared according
to the general procedure outlined in Step 10 using the appropriate
starting materials. The structures of the compounds are shown in
FIG. 1.
TABLE-US-00010 Compound Number Name .sup.1H NMR m/z 336 (R or
S)-N-((4-methoxy-6-methyl-2- (400 MHz, DMSO-d.sub.6) .delta. =
11.59 515 oxo-1,2-dihydropyridin-3- (s, 1 H), 7.78-7.66 (m, 2 H),
yl)methyl)-2-methyl-1-(1-(1- 7.64-7.57 (m, 1 H), 7.06 (s, 2
(methylsulfonyl)piperidin-4- H), 6.14 (s, 1 H), 4.31 (d, J =
yl)ethyl)-1H-indole-3-carboxamide 4.9 Hz, 2 H), 4.25-4.15 (m, 1 H),
3.83 (s, 3 H), 3.63 (s, 1 H), 3.40-3.33 (m, 1 H), 2.79 (s, 3 H),
2.75-2.65 (m, 1 H), 2.60 (s, 3 H), 2.45-2.27 (m, 1 H), 2.19 (s, 3
H), 2.06-1.98 (m, 1 H), 1.55 (d, J = 6.9 Hz, 3 H), 1.45- 1.36 (m, 1
H), 1.28-1.18 (m, 1 H), 1.14-1.03 (m, 1 H), 0.83- 0.74 (m, 1 H) 337
(R or S)-1-(1-(1-(2-hydroxy-2- (400 MHz, DMSO-d.sub.6) .delta.
11.58 523 methylpropanoyl)piperidin-4- (br. s., 1 H), 7.77-7.67 (m,
2 yl)ethyl)-N-((4-methoxy-6-methyl-2- H), 7.66-7.60 (m, 1 H), 7.06
(s, oxo-1,2-dihydropyridin-3- 2 H), 6.14 (s, 1 H), 5.32-5.23
yl)methyl)-2-methyl-1H-indole-3- (m, 1 H), 4.31 (d, J = 4.5 Hz, 2
carboxamide H), 4.19-4.10 (m, 1 H), 3.83 (s, 3 H), 2.75-2.62 (m, 2
H), 2.58 (s, 3 H), 2.19 (s, 4 H), 2.00- 1.90 (m, 2 H), 1.54 (d, J =
6.7 Hz, 3 H), 1.32-1.18 (m, 8 H), 0.87-0.78 (m, 1 H), 0.77-0.67 (m,
1 H) 342 (R or S)-1-(1-(1-isobutyrylpiperidin- (400 MHz,
DMSO-d.sub.6) .delta. = 11.59 507
4-yl)ethyl)-N-((4-methoxy-6-methyl- (s, 1 H), 7.75 (d, J = 7.4 Hz,
1 2-oxo-1,2-dihydropyridin-3- H), 7.72-7.67 (m, 1 H), 7.64
yl)methyl)-2-methyl-1H-indole-3- (d, J = 8.0 Hz, 1 H), 7.14-7.01
carboxamide (m, 2 H), 6.15 (s, 1 H), 4.58- 4.46 (m, 1 H), 4.32 (d,
J = 4.9 Hz, 2 H), 4.09-3.99 (m, 1 H), 3.84 (s, 3 H), 3.81-3.72 (m,
1 H), 3.08-2.97 (m, 1 H), 2.92- 2.81 (m, 1 H), 2.78-2.65 (m, 3 H),
2.59 (br. s., 3 H), 2.20 (s, 3 H), 2.03-1.90 (m, 1 H), 1.59- 1.47
(m, 4 H), 1.02-0.86 (m, 6 H), 0.78-0.69 (m, 1 H) 344 (R or
S)-N-((4-(difluoromethoxy)-6- (400 MHz, DMSO-d.sub.6) .delta.
12.02- 551 methyl-2-oxo-1,2-dihydropyridin-3- 11.95 (m, 1 H), 7.74
(d, J = 8.0 yl)methyl)-2-methyl-1-(1-(1- Hz, 1 H), 7.66-7.57 (m, 2
H), (methylsulfonyl)piperidin-4- 7.11-7.00 (m, 2 H), 6.08 (s, 1
yl)ethyl)-1H-indole-3-carboxamide H), 4.32 (d, J = 4.5 Hz, 2 H),
4.18 (d, J = 7.1 Hz, 1 H), 3.64 (d, J = 12.3 Hz, 1 H), 3.36 (d, J =
12.0 Hz, 1 H), 2.79 (s, 3 H), 2.75-2.65 (m, 2 H), 2.58 (s, 3 H),
2.45-2.27 (m, 2 H), 2.20 (s, 3 H), 2.07-1.98 (m, 1 H), 1.55 (d, J =
6.9 Hz, 3 H), 1.40 (d, J = 8.2 Hz, 1 H), 1.10 (d, J = 8.9 Hz, 1 H),
0.79 (d, J = 12.5 Hz, 1 H) 345 (R or S)-1-(1-(1- (400 MHz,
DMSO-d.sub.6) .delta. 11.57 529
(ethylsulfonyl)piperidin-4-yl)ethyl)- (s, 1 H), 7.75 (d, J = 8.0
Hz, 1 N-((4-methoxy-6-methyl-2-oxo-1,2- H), 7.69 (t, J = 5.0 Hz, 1
H), dihydropyridin-3-yl)methyl)-2- 7.62 (d, J = 7.4 Hz, 1 H), 7.06
methyl-1H-indole-3-carboxamide (d, J = 7.1 Hz, 2 H), 6.15 (s, 1 H),
4.32 (d, J = 5.1 Hz, 2 H), 4.25-4.15 (m, 1 H), 3.84 (s, 3 H),
3.73-3.65 (m, 1 H), 3.45- 3.36 (m, 1 H), 3.02-2.93 (m, J = 7.8 Hz,
2 H), 2.87-2.77 (m, 1 H), 2.75-2.66 (m, 1 H), 2.60 (s, 3 H),
2.42-2.30 (m, 1 H), 2.20 (s, 3 H), 2.06-1.97 (m, 1 H), 1.58-1.48
(m, 4 H), 1.42-1.31 (m, 1 H), 1.17 (t, J = 7.5 Hz, 3 H), 1.13-1.00
(m, 1 H), 0.83- 0.73 (m, 1 H) 355 (R or S)-1-(1-(4- (400 MHz,
DMSO-d.sub.6) .delta. 11.59 543
(isopropylsulfonyl)cyclohexyl)ethyl)- (s, 1H), 7.76-7.69 (m, 2H),
7.62 N-((4-methoxy-6-methyl-2-oxo-1,2- (d, 1H), 7.10-7.03 (m, 2H),
6.15 dihydropyridin-3-yl)methyl)-2- (s, 1H), 4.32 (d, 2H),
4.29-4.26 methyl-1H-indole-3-carboxamide (m, 2H), 3.84 (s, 3H),
3.72 (br. d., 1H), 3.45 (br. d., 1H), 3.26 (tt, 1H), 2.91 (dt, 1H),
2.60 (s, 3H), 2.20 (s, 3H), 1.97 (br. d., 1H), 1.54 (d, 3H),
1.35-1.24 (m, 2H), 1.18 (d, 3H), 1.16 (d, 3H), 1.05-0.78 (m, 2H)
357 (R or S)-isobutyl 4-(1-(3-(((4- (400 MHz, DMSO-d.sub.6) .delta.
537 methoxy-6-methyl-2-oxo-1,2- 11.60 (br. s., 1H), 7.75-7.60 (m,
dihydropyridin-3- 3H), 7.10-7.03 (m, 2H), 6.15 (s,
yl)methyl)carbamoyl)-2-methyl-1H- 1H) 4.33 (d, 1H), 4.13-4.06 (m,
indol-1-yl)ethyl)piperidine-1- 1H), 3.84 (s, 3H), 3.74 (d, 1H),
carboxylate 2.80-2.60 (m, 3H), 2.58 (s, 1H), 2.50-2.42 (m, 2H),
1.96-1.90 (m, 1H), 1.54 (d, 3H), 1.25-1.22 (m, 1H), 0.98-0.72 (m,
6H) 368 (R or S)-N-((4,6-dimethyl-2-oxo-1,2- (400 MHz,
DMSO-d.sub.6) .delta. 11.59 513
dihydropyridin-3-yl)methyl)-1-(1-(1- (s, 1 H), 7.78-7.71 (m, 1 H),
(ethylsulfonyl)piperidin-4-yl)ethyl)- 7.66-7.57 (m, 2 H), 7.07 (s,
2 2-methyl-1H-indole-3-carboxamide H), 5.89 (s, 1 H), 4.32 (s, 2
H), 4.25-4.15 (m, 1 H), 3.65-3.59 (m, 1 H), 3.19-3.10 (m, 1 H),
2.98 (d, J = 7.4 Hz, 2 H), 2.87- 2.77 (m, 1 H), 2.72-2.65 (m, 1 H),
2.58 (s, 3 H), 2.27 (s, 3 H), 2.12 (s, 3 H), 1.55 (d, J = 6.9 Hz, 4
H), 1.42-1.33 (m, 2 H), 1.17 (t, J = 7.4 Hz, 3 H), 1.12- 1.00 (m, 1
H), 0.84-0.74 (m, 1 H) 382 (R or S)-N-((4,6-dimethyl-2-oxo-1,2-
(400 MHz, DMSO-d.sub.6) .delta. 11.60 499
dihydropyridin-3-yl)methyl)-2- (s, 1 H), 7.75 (d, J = 7.1 Hz, 1
methyl-1-(1-(1- H), 7.65-7.58 (m, 2 H), 7.12-
(methylsulfonyl)piperidin-4- 7.02 (m, 2 H), 5.89 (s, 1 H),
yl)ethyl)-1H-indole-3-carboxamide 4.38-4.25 (m, 2 H), 4.20 (dd, J =
7.0, 10.6 Hz, 1 H), 2.80 (s, 3 H), 2.76-2.67 (m, 2 H), 2.59 (s, 3
H), 2.46-2.31 (m, 2 H), 2.27 (s, 3 H), 2.12 (s, 3 H), 1.55 (d, J =
6.9 Hz, 3 H), 1.51 (br. s., 1 H), 1.47-1.34 (m, 1 H), 1.29- 1.21
(m, 1 H), 1.17-1.04 (m, 1 H), 0.80 (d, J = 12.9 Hz, 1 H)
Example 2
Synthesis of (R or
S)-1-(1-(1-isopropylpiperidin-4-yl)ethyl)-N-((4-methoxy-6-methyl-2-oxo-1,-
2-dihydropyridin-3-yl)methyl)-2-methyl-1H-indole-3-carboxamide
(Compound 358)
##STR00034## A 25 mL vial was charged with a magnetic stir bar, (R
or
S)--N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-
-1-(1-(piperidin-4-yl)ethyl)-1H-indole-3-carboxamide hydrochloride,
THF (2.114 ml, 0.211 mmol), propan-2-one (0.061 g, 1.057 mmol), and
sodium triacetoxyborohydride (0.224 g, 1.057 mmol). The reaction
was allowed to stir at rt for 12 h. The reaction was inverse
quenched onto sat'd aqueous NaHCO.sub.3, extracted with ethyl
acetate and conc. in vacuo. The resulting material was treated with
10 mL 7 N ammonia in MeOH and was conc in vacuo to yield material
which was purified via silica gel chromatography (10 g) using
DCM/MeOH/NH.sub.4OH (90:1:0.1) as eluent to afford 33 mg, (0.065
mmol, 31.0% yield) of the title compound as a white solid.). LCMS
479 (M+1).sup.+; .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.59 (s.,
1H), 7.64-7.82 (m, 2H), 7.59 (d, 1H), 6.95-7.17 (m, 2H), 6.15 (s,
1H), 4.32 (d, 2H), 4.04-4.24 (m, 1H), 3.84 (.quadrature., 3H),
2.77-2.93 (.quadrature., 2H), 2.68 (.quadrature., 1H), 2.60
(.quadrature., 3H), 2.20 (.quadrature., 3H), 2.08-2.15
(.quadrature., 1H), 1.92 (.quadrature., 1H), 1.83 (.quadrature.p.
.quadrature.., 1H), 1.54 (.quadrature., 3H), 1.27-1.43
(.quadrature., 2H), 0.91 (.quadrature., 6H), 0.71-0.67
(.quadrature., 2H).
The compounds shown in the following table were prepared according
to the general procedure outlined in this Example using the
appropriate starting materials. The structures of the compounds are
shown in FIG. 1.
TABLE-US-00011 Compound Number Name .sup.1H NMR m/z 341 (R or
S)-N-((4-methoxy- (400 MHz, DMSO-d.sub.6) .delta. 11.58 (s, 1 H),
493 6-methyl-2-oxo-1,2- 7.76-7.65 (m, 2 H), 7.59 (d, J = 7.8 Hz, 1
dihydropyridin-3- H), 7.10-6.99 (m, 2 H), 6.14 (s, 1 H), 4.49
yl)methyl)-2-methyl-1- (t, J = 6.4 Hz, 1 H), 4.43 (t, J = 6.5 Hz, 1
(1-(1-(oxetan-3- H), 4.37 (t, J = 6.1 Hz, 1 H), 4.34-4.28 (m,
yl)piperidin-4-yl)ethyl)- 3 H), 4.21-4.10 (m, 1 H), 3.83 (s, 3 H),
1H-indole-3-carboxamide 3.30-3.23 (m, 1 H), 2.75 (br. s., 1 H),
2.71- 2.64 (m, 1 H), 2.60 (s, 3 H), 2.19 (s, 4 H), 1.90 (br. s., 1
H), 1.75 (br. s., 1 H), 1.53 (d, J = 6.9 Hz, 3 H), 1.42 (br. s., 2
H), 1.11- 0.98 (m, 1 H), 0.72-0.63 (m, 1 H) 343 (R or
S)-N-((4-methoxy- (400 MHz, DMSO-d.sub.6) .delta. 11.58 (s, 1 H),
451 6-methyl-2-oxo-1,2- 7.76-7.65 (m, 2 H), 7.59 (d, J = 7.6 Hz, 1
dihydropyridin-3- H), 7.11-6.99 (m, 2 H), 6.14 (s, 1 H), 4.31
yOmethyl)-2-methyl-1- (d, J = 5.1 Hz, 2 H), 4.13 (br. s., 1 H),
3.83 (1-(1-methylpiperidin-4- (s, 3 H), 2.83 (d, J = 10.0 Hz, 1 H),
2.61- yl)ethyl)-1H-indole-3- 2.52 (m, 5 H), 2.19 (s, 3 H), 2.09 (s,
4 H), carboxamide 1.88 (d, J = 10.7 Hz, 2 H), 1.53 (d, J = 6.7 Hz,
3 H), 1.34 (br. s., 1 H), 1.02 (d, J = 8.2 Hz, 1 H), 0.66 (br. s.,
1 H) 359 (R or S)-N-((4-methoxy- (400 MHz, DMSO-d.sub.6) .delta.
11.59 (s, 1 H), 495 6-methyl-2-oxo-1,2- 7.77-7.66 (m, 2 H), 7.60
(d, J = 7.8 Hz, 1 dihydropyridin-3- H), 7.12-7.01 (m, 2 H), 6.15
(s, 1 H), 4.32 yl)methyl)-1-(1-(1-(2- (d, J = 4.9 Hz, 2 H), 4.13
(d, J = 7.1 Hz, 1 methoxyethyl)piperidin- H), 3.85 (s, 3 H), 3.36
(t, J = 5.9 Hz, 2 H), 4-yl)ethyl)-2-methyl-1H- 3.19 (s, 3 H), 2.94
(d, J = 10.5 Hz, 1 H), indole-3-carboxamide 2.71-2.56 (m, 5 H),
2.43-2.32 (m, 2 H), 2.24-2.12 (m, 4 H), 1.54 (d, J = 6.9 Hz, 4 H),
1.39-1.27 (m, 2 H), 1.02 (d, J = 8.7 Hz, 1 H), 0.65 (d, J = 12.7
Hz, 1 H) 360 (R or S)-1-(1-(1- (400 MHz, DMSO-d.sub.6) .delta.
11.79-11.45 (m, 1 465 ethylpiperidin-4- H), 7.78-7.65 (m, 2 H),
7.59 (d, J = 7.8 yl)ethyl)-N-((4- Hz, 1 H), 7.14-6.99 (m, 2 H),
6.15 (s, 1 H), methoxy-6-methyl-2- 4.32 (d, J = 4.9 Hz, 2 H),
4.20-4.08 (m, 1 oxo-1,2-dihydropyridin- H), 3.84 (s, 3 H),
2.98-2.89 (m, 1 H), 2.71- 3-yl)methyl)-2-methyl- 2.61 (m, 2 H),
2.59 (s, 3 H), 2.27-2.21 1H-indole-3- (m, 2 H), 2.20 (s, 3 H),
1.94-1.80 (m, 2 H), carboxamide 1.54 (s, 4 H), 1.38-1.28 (m, 1 H),
1.06- 0.98 (m, 1 H), 0.93 (t, J = 7.1 Hz, 3 H), 0.71-0.63 (m, 1 H)
363 (R or S)-ethyl 2-(4-(1-(3- (400 MHz, DMSO-d.sub.6) .delta.
11.59 (br. s., 1 H), 523 (((4-methoxy-6-methyl- 7.81-7.65 (m, 2 H),
7.60 (d, J = 7.4 Hz, 1 2-oxo-1,2- H), 7.16-6.98 (m, 2 H), 6.15 (s,
1 H), 4.32 dihydropyridin-3- (d, J = 4.9 Hz, 2 H), 4.23-4.11 (m, 1
H), yl)methyl)carbamoyl)-2- 4.04 (q, J = 7.0 Hz, 2 H), 3.84 (s, 3
H), 2.95- methyl-1H-indol-1- 2.86 (m, 1 H), 2.60 (s, 5 H), 2.20 (s,
4 yl)ethyl)piperidin-1- H), 1.94-1.79 (m, 2 H), 1.54 (d, J = 6.9
Hz, 4 yl)acetate H), 1.41-1.32 (m, 1 H), 1.15 (t, J = 7.1 Hz, 3 H),
1.04 (d, J = 6.0 Hz, 2 H), 0.71-0.61 (m, 1 H) 366 (R or
S)-N-((4-ethyl-6- (400 MHz, DMSO-d.sub.6) .delta. 11.63 (s, 1 H),
449 methyl-2-oxo-1,2- 7.74 (d, J = 7.6 Hz, 1 H), 7.65-7.56 (m, 2
dihydropyridin-3- H), 7.12-7.01 (m, 2 H), 5.94 (s, 1 H), 4.34
yl)methyl)-2-methyl-1- (t, J = 5.1 Hz, 2 H), 4.19-4.09 (m, 1 H),
(1-(1-methylpiperidin-4- 2.88 (br. s., 1 H), 2.71-2.56 (m, 6 H),
2.14 yl)ethyl)-1H-indole-3- (s, 7 H), 1.91 (d, J = 12.5 Hz, 1 H),
1.54 (d, carboxamide J = 6.9 Hz, 4 H), 1.41-1.31 (m, 2 H), 1.14 (t,
J = 7.6 Hz, 3 H), 1.05 (d, J = 9.1 Hz, 1 H), 0.68 (d, J = 12.7 Hz,
1 H) 367 (R or S)-N-((4,6- (400 MHz, DMSO-d.sub.6) .delta. 11.59
(s, 1 H), 435 dimethyl-2-oxo-1,2- 7.74 (d, J = 6.9 Hz, 1 H),
7.65-7.56 (m, 2 dihydropyridin-3- H), 7.12-7.01 (m, 2 H), 5.89 (s,
1 H), 4.38- yl)methyl)-2-methyl-1- 4.25 (m, 2 H), 4.20-4.09 (m, 1
H), 2.95 (1-(1-methylpiperidin-4- (br. s., 1 H), 2.68 (br. s., 2
H), 2.58 (s, 3 H), yl)ethyl)-1H-indole-3- 2.27 (s, 3 H), 2.21 (br.
s., 3 H), 2.12 (s, 3 carboxamide H), 1.94 (d, J = 13.8 Hz, 1 H),
1.54 (d, J = 6.9 Hz, 4 H), 1.44-1.31 (m, 2 H), 1.07 (d, J = 12.5
Hz, 1 H), 0.71 (d, J = 13.2 Hz, 1 H) 375 (R or S)-N-((4,6- (400
MHz, DMSO-d.sub.6) .delta. 11.59 (br. s., 1 H), 477
dimethyl-2-oxo-1,2- 7.73 (d, J = 7.6 Hz, 1 H), 7.65-7.55 (m, 2
dihydropyridin-3- H), 7.12-7.00 (m, 2 H), 5.89 (s, 1 H), 4.53-
yl)methyl)-2-methyl-1- 4.48 (m, 1 H), 4.47-4.42 (m, 1 H), 4.38
(1-(1-(oxetan-3- (s, 1 H), 4.31 (t, J = 5.2 Hz, 3 H), 4.21-
yl)piperidin-4-yl)ethyl)- 4.10 (m, 1 H), 3.31-3.24 (m, 2 H), 2.81-
1H-indole-3- 2.64 (m, 2 H), 2.59 (s, 3 H), 2.26 (s, 3 H),
carboxamide 2.23-2.16 (m, 1 H), 2.12 (s, 3 H), 1.98- 1.85 (m, 1 H),
1.81-1.70 (m, 1 H), 1.54 (d, J = 6.9 Hz, 3 H), 1.51-1.22 (m, 1 H),
1.12- 0.96 (m, 2 H), 0.73-0.64 (m, 1 H) 380 (R or S)-N-((4-ethyl-6-
(400 MHz, DMSO-d.sub.6) .delta. 11.63 (br. s., 1 H), 491
methyl-2-oxo-1,2- 7.74 (d, J = 7.36 Hz, 1 H), 7.60 (d, J = 8.47
dihydropyridin-3- Hz, 2 H), 7.06 (quin, J = 7.13 Hz, 3 H),
yl)methyl)-2-methyl-1- 5.94 (s, 1 H), 4.51 (t, J = 6.47 Hz, 1 H),
(1-(1-(oxetan-3- 4.46 (t, J = 6.35 Hz, 1 H), 4.40 (t, J = 6.13
yl)piperidin-4-yl)ethyl)- Hz, 1 H), 4.37-4.30 (m, 2 H), 4.28-4.11
1H-indole-3- (m, 1 H), 3.57 (s, 1 H), 3.34 (br. s., 2 H),
carboxamide 2.81 (d, J = 10.70 Hz, 1 H), 2.67 (d, J = 14.94 Hz, 1
H), 2.64-2.57 (m, 4 H), 2.21 (d, J =10.93 Hz, 1 H), 2.14 (s, 3 H),
1.93 (d, J =12.49 Hz, 1 H), 1.83 (t, J = 11.37 Hz, 1 H), 1.54 (d, J
= 6.91 Hz, 3 H), 1.37 (d, J = 10.48 Hz, 1 H), 1.25 (q, J = 6.91 Hz,
1 H), 1.14 (t, J = 7.58 Hz, 3 H), 1.06 (d, J = 9.81 Hz, 1 H), 0.70
(d, J = 12.49 Hz, 1 H) 381 (R or S)-N-((4,6- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 11.60 505 dimethyl-2-oxo-1,2- (br. s., 1 H),
7.74 (d, J = 7.13 Hz, 1 H), 7.66- dihydropyridin-3- 7.50 (m, 2 H),
7.15-6.99 (m, 2 H), 5.89 yl)methyl)-2-methyl-1- (s, 1 H), 4.40-4.24
(m, 2 H), 4.21-4.07 (1-(1-(tetrahydro-2H- (m, 1 H), 3.95-3.78 (m, 2
H), 3.57 (s, 1 H), pyran-4-yl)piperidin-4- 3.32-3.17 (m, 3 H), 2.68
(br. s., 1 H), 2.58 yl)ethyl)-1H-indole-3- (s, 3 H), 2.33 (br. s.,
2 H), 2.27 (s, 3 H), carboxamide 2.12 (s, 3 H), 2.00-1.88 (m, 2 H),
1.75 (d, J = 12.04 Hz, 1 H), 1.62 (br. s., 2 H), 1.54 (d, J = 6.91
Hz, 3 H), 1.46-1.30 (m, 2 H), 1.01 (br. s., 1 H), 0.72 (br. s., 1
H) 440 (R or S)-tert-butyl 3-(4- 576 (1-(3-(((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)carbamoyl)-
2-methyl-1H-indol-1- yl)ethyl)piperidin-1- yl)azetidine-1-
carboxylate 377 (R or S)-N-((4,6- (400 MHz, DMSO-d.sub.6)
.delta.11.63-11.56 (m, 1 490 dimethyl-2-oxo-1,2- H), 7.76-7.70 (m,
1 H ), 7.64-7.55 (m, 2 dihydropyridin-3- H), 7.05 (s, 2 H), 5.89
(s, 1 H), 4.56 (s, 4 yl)methyl)-2-methyl-1- H), 4.31 (s, 2 H),
4.19-4.09 (m, 1 H), 3.36 (1-(1-(1-methylazetidin- (d, J = 4.9 Hz, 1
H), 2.77-2.56 (m, 5 H), 3-yl)piperidin-4- 2.26 (s, 3 H), 2.18 (s, 3
H), 2.12 (s, 3 H), yl)ethyl)-1H-indole-3- 1.94-1.85 (m, 1 H),
1.78-1.67 (m, 1 H), carboxamide 1.53 (d, J = 6.9 Hz, 3 H),
1.50-1.45 (m, 1 H), 1.44-1.22 (m, 2 H), 1.07-0.93 (m, 1 H),
0.71-0.61 (m, 1 H)
Example 3
Synthesis of (R or
S)-1-(1-(1-(2-fluoroethyl)piperidin-4-yl)ethyl)-N-((4-methoxy-6-methyl-2--
oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-1H-indole-3-carboxamide
(Compound 356)
##STR00035## A 25 mL vial was charged with a magnetic stir bar, (R
or
S)--N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-
-1-(1-(piperidin-4-yl)ethyl)-1H-indole-3-carboxamide hydrochloride
(0.062 g, 0.131 mmol), K.sub.2CO.sub.3 (0.072 g, 0.524 mmol), MeCN
(0.655 ml, 0.131 mmol), DMF (0.262 ml, 0.131 mmol) and
1-bromo-2-fluoroethane (0.020 ml, 0.262 mmol). The reaction was
capped and heated to 82.degree. C. with stirring for 4 h. The
reaction was allowed to cool to rt, filtered, and the filtrate was
pre-absorbed onto silica gel (12 g). The material was purified via
SiO.sub.2 chromatography (25 g) using DCM/MeOH/Et.sub.3N
(85:15:0.5) as eluent to afford the title compound as an off white
solid (30 mg, 0.059 mmol, 45.1% yield). LCMS 483 (M+1).sup.+;
.sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.59 (s, 1H), 7.75-7.68 (m,
2H), 7.60 (d, 1H) 7.09-7.03 (m, 2H), 6.15 (s, 1H) 4.53-4.51 (m,
1H), 4.42-4.39 (m, 1H), 4.32 (d, 2H), 4.24-4.2 (m, 1H), 3.84 (s,
3H), 2.98 (br. d., 1H), 2.70-2.49 (m, 4H), 2.60 (s, 3H), 2.20 (s,
3H), 2.01 (dt, 1H), 1.92-1.90 (m, 1H), 1.75-1.71 (m, 1H), 1.54 (d,
3H), 1.38-1.36 (m, 1H), 1.02-0.98 (m, 1H), 0.7-0.66 (br. d.,
1H).
The compounds shown in the following table were prepared according
to the general procedure outlined in this Example using the
appropriate starting materials. The structures of the compounds are
shown in FIG. 1.
TABLE-US-00012 Compound Number Name .sup.1H NMR m/z 362 (R or
S)-1-(1-(1-(2,2- (400 MHz, DMSO-d.sub.6) .delta. = 11.60 (br. s., 1
501 difluoroethyl)piperidin- H), 7.77-7.66 (m, 2 H), 7.60 (d, J =
7.6 Hz, 4-yl)ethyl)-N-((4- 1 H), 7.14-7.00 (m, 2H ), 6.15 (s, 1 H),
methoxy-6-methyl-2- 6.06 (t, J = 55.7 Hz, 1 H), 4.32 (d, J = 4.9
oxo-1,2-dihydropyridin- Hz, 2 H), 4.15 (br. s., 1 H), 3.84 (s, 3
H), 3-yl)methyl)-2-methyl- 3.03-2.93 (m, 2 H), 2.73-2.62 (m, 3 H),
1H-indole-3- 2.60 (s, 3 H), 2.26-2.10 (m, 4 H), 1.93- carboxamide
1.79 (m, 1 H), 1.59-1.46 (m, 4 H), 1.41- 1.29 (m, 1 H), 1.11-0.97
(m, 1 H), 0.67 (br. s., 1 H) 378 (R or S)-N-((4-methoxy- (400 MHz,
DMSO-d.sub.6) .delta. = 11.60 (br. s., 1 H), 533 6-methyl-2-oxo-1,2
7.78-7.66 (m, 2 H), 7.60 (d, J = 8.2 Hz, 1 dihydropyridin-3- H),
7.13-7.00 (m, 2 H), 6.15 (s, 1 H), 4.32 yl)methyl)-2-methyl-1- (d,
J = 4.9 Hz, 2 H), 4.22-4.09 (m, 1 H), (1-(1-(3,3,3- 3.84 (s, 3 H),
3.03-2.91 (m, 1 H), 2.73- trifluoropropyl)piperidin- 2.64 (m, 1 H),
2.60 (s, 3 H), 2.48-2.31 (m, 4-yl)ethyl)-1H-indole-3- 5 H), 2.20
(s, 3 H), 2.01-1.85 (m, 2 H), carboxamide 1.58-1.46 (m, 4 H),
1.36-1.29 (m, 1 H), 1.08-0.98 (m, 1 H), 0.73-0.62 (m, 1 H) 365 (R
or S)-N-((4-methoxy- (500 MHz, DMSO-d.sub.6) .delta. = 11.59 (s, 1
H), 519 6-methyl-2-oxo-1,2- 7.74 (d, J = 7.6 Hz, 1 H), 7.71-7.66
(m, 1 dihydropyridin-3- H), 7.61 (d, J = 7.8 Hz, 1 H), 7.13-7.01
(m, yl)methyl)-2-methyl-1- 2 H), 6.15 (s, 1 H), 4.32 (d, J = 4.9
Hz, 2 H), (1-(1-(2,2,2- 4.22-4.12 (m, 1 H), 3.84 (s, 3 H), 3.15-
trifluoroethyl)piperidin- 2.95 (m, 3 H), 2.75-2.66 (m, 1 H), 2.60
(s, 4-yl)ethyl)-1H-indole-3- 3 H), 2.39-2.31 (m, 1 H), 2.20 (s, 3
H), carboxamide 2.05-1.98 (m, 1 H), 1.92-1.84 (m, 1 H), 1.56-1.46
(m, 4 H), 1.42-1.32 (m, 1 H), 1.11-1.01 (m, 1 H), 0.69-0.62 (m, 1
H) 441 (R or S)-1-(1-(1-(2,2- (400 MHz, DMSO-d.sub.6) .delta. =
11.59 (s, 1 H), 485 difluoroethyl)piperidin- 7.73 (d, J = 7.8 Hz, 1
H), 7.65-7.55 (m, 2 4-yl)ethyl)-N-((4,6- H), 7.12-7.00 (m, 2 H),
6.22-5.90 (m, 1 dimethyl-2-oxo-1,2- H), 5.89 (s, 1 H), 4.36-4.25
(m, 2 H), 4.20- dihydropyridin-3- 4.09 (m, 1 H), 3.01-2.93 (m, 1
H), 2.72- yl)methyl)-2-methyl-1H- 2.59 (m, 3 H), 2.58 (s, 3 H),
2.26 (s, 3 H), indole-3-carboxamide 2.21-2.13 (m, 2 H), 2.12 (s, 3
H), 1.92- 1.79 (m, 2 H), 1.53 (s, 4 H), 1.41-1.29 (m, 1 H),
1.10-0.97 (m, 1 H), 0.70-0.59 (m, 1 H)
Example 4
Synthesis of (R or
S)--N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-
-1-(1-(1-(pyrimidin-2-yl)piperidin-4-yl)ethyl)-1H-indole-3-carboxamide
(Compound 361)
##STR00036## To a re-sealable vial was added 2-chloropyrimidine
(185 mg, 1.611 mmol), (R or
S)--N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-
-1-(1-(piperidin-4-yl)ethyl)-1H-indole-3-carboxamide hydrochloride
(508 mg, 1.074 mmol), and EtOH (8 mL). To this solution was added
Et.sub.3N (449 .mu.l, 3.22 mmol). The vial was sealed and heated to
100.degree. C. overnight. The solution was allowed to cool to room
temperature and concentrated in vacuo. The crude residue was
purified via silica gel chromatography (hexanes: (3:2 DCM:IPA)) to
afford the title compound as a solid (357 mg, 0.694 mmol, 64.6%
yield). LCMS 515 (M+1).sup.+; .sup.1H NMR (DMSO-d6, 400 MHz)
.delta. 11.60 (s, 1H), 8.30 (d, J=4.7 Hz, 2H), 7.76 (d, J=7.6 Hz,
1H), 7.73-7.64 (m, 2H), 7.14-7.01 (m, 2H), 6.55 (t, J=4.7 Hz, 1H),
6.15 (s, 1H), 4.84-4.75 (m, 1H), 4.57-4.47 (m, 1H), 4.33 (d, J=4.2
Hz, 2H), 4.22-4.11 (m, 1H), 3.84 (s, 3H), 2.92-2.81 (m, 1H),
2.63-2.52 (m, 4H), 2.20 (s, 3H), 2.05-1.94 (m, 1H), 1.61-1.49 (m,
4H), 1.34-1.21 (m, 1H), 1.04-0.91 (m, 1H), 0.83-0.75 (m, 1H).
The compound shown in the following table was prepared according to
the general procedure outlined in this Example using the
appropriate starting materials. The structure of the compound is
shown in FIG. 1.
TABLE-US-00013 Compound Number Name .sup.1H NMR m/z 373 (R or
S)-N-((4- (400 MHz, DMSO-d.sub.6) .delta. 11.60 (br. s., 1 H), 8.06
514 methoxy-6-methyl- (d, J =3.6 Hz, 1 H), 7.82-7.62 (m, 3 H),
7.51- 2-oxo-1,2- 7.39 (m, 1 H), 7.17-6.98 (m, 2 H), 6.75 (d, J =
dihydropyridin-3- 8.5 Hz, 1 H), 6.61-6.49 (m, 1 H), 6.15 (s, 1 H),
yl)methyl)-2- 4.49-4.38 (m, 1 H), 4.33 (d, J = 3.8 Hz, 2 H),
methyl-1-(1-(1- 4.24-4.03 (m, 2 H), 3.85 (s, 3 H), 2.90-2.70
(pyridin-2- (m, 2 H), 2.58 (s, 3 H), 2.20 (s, 3 H), 2.06-1.91
yl)piperidin-4- (m, 1 H), 1.63-1.47 (m, 4 H), 1.40-1.27 (m, 1
yl)ethyl)-1H- H), 1.07-0.94 (m, 1 H), 0.82-0.72 (m, 1 H) indole-3-
carboxamide
Example 5
Synthesis of (R or
S)-1-(1-(1-(2-hydroxyethyl)piperidin-4-yl)ethyl)-N-((4-methoxy-6-methyl-2-
-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-1H-indole-3-carboxamide
(Compound 347)
##STR00037## To a sealed tube charged with a magnetic stir bar was
added (R or
S)--N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-
-1-(1-(piperidin-4-yl)ethyl)-1H-indole-3-carboxamide (0.1 g, 0.229
mmol) was added DCM (3 mL) and the reaction cooled to 0.degree. C.
To the cooled reaction mixture was added oxirane which was
condensed into the reaction vial (.about.1 mL). The reaction was
allowed to stir to rt over 4 h and was then conc. in vacuo to
afford the crude material which was purified via silica gel
chromatography (12 g) using ethyl acetate/MeOH (4:1) as eluent to
afford the title compound as a white solid (50 mg). LCMS 481
(M+1).sup.+; .sup.1H NMR (DMSO-d6, 400 MHz) .delta.) .delta. 11.58
(s, 1H), 7.77-7.65 (m, 2H), 7.59 (d, J=7.8 Hz, 1H), 7.11-6.99 (m,
2H), 6.14 (s, 1H), 4.54-4.44 (m, 1H), 4.31 (d, J=5.1 Hz, 3H), 4.13
(dd, J=7.1, 10.3 Hz, 1H), 3.83 (s, 3H), 3.42 (q, J=6.0 Hz, 2H),
2.93 (br. s., 1H), 2.71-2.56 (m, 4H), 2.31 (br. s., 2H), 2.19 (s,
3H), 2.03-1.83 (m, 2H), 1.64 (br. s., 1H), 1.53 (d, J=6.9 Hz, 3H),
1.32 (d, J=11.1 Hz, 1H), 1.02 (d, J=10.3 Hz, 1H), 0.65 (d, J=11.8
Hz, 1H).
The compounds shown in the following table were prepared according
to the general procedure outlined in this Example using the
appropriate starting materials. The structures of the compounds are
shown in FIG. 1.
TABLE-US-00014 Compound Number Name .sup.1H NMR m/z 352 (R or
S)-1-(1-(1-(2- NMR (400 MHz, DMSO-d6) .delta. 11.58 (br. s., 1 509
hydroxy-2- H), 7.76-7.65 (m, 2 H), 7.58 (d, J = 7.8 Hz, 1
methylpropyl)piperidin- H), 7.10-6.99 (m, 2 H), 6.14 (s, 1 H), 4.31
(d, 4-yl)ethyl)-N-((4- J = 4.9 Hz, 2 H), 4.14 (br. s., 1 H), 3.94
(s, 1 methoxy-6-methyl-2- H), 3.83 (s, 3 H), 3.56 (s, 2 H), 3.01
(d, J = oxo-1,2- 11.4 Hz, 1 H), 2.73-2.64 (m, 1 H), 2.59 (s, 3
dihydropyridin-3- H), 2.19 (s, 3 H), 2.16-2.03 (m, 2 H), 1.52 (d,
yl)methyl)-2-methyl- J = 6.9 Hz, 4 H), 1.34 (br. s., 2 H), 1.02 (d,
J = 1H-indole-3- 4.5 Hz, 7 H), 0.66-0.58 (m, 1 H) carboxamide 369
(R or S)-N-((4,6- (400 MHz, DMSO-d6) .delta. 11.59 (br. s., 1 H),
493 dimethyl-2-oxo-1,2- 7.77-7.69 (m, 1 H), 7.60 (br. s., 2 H),
7.06 dihydropyridin-3- (br. s., 2 H), 5.89 (s, 1 H), 4.31 (t, J =
5.7 Hz, 2 yl)methyl)-1-(1-(1-(2- H), 4.20-4.09 (m, 1 H), 4.00-3.92
(m, 1 H), hydroxy-2- 3.58-3.55 (m, 2 H), 3.19-3.10 (m, 1 H), 3.07-
methylpropyl)piperidin- 2.95 (m, 1 H), 2.74-2.63 (m, 1 H), 2.59
(br. 4-yl)ethyl)-2-methyl- s., 3 H), 2.27 (s, 3 H), 2.12 (s, 3 H),
1.89-1.72 1H-indole-3- (m, 1 H), 1.54 (br. s., 4 H), 1.30-1.14 (m,
2 carboxamide H), 1.03 (br. s., 6 H), 0.84-0.57 (m, 2 H)
Example 6
Synthesis of (R or
S)--N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-
-6-phenyl-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-indole-3-carboxamide
(Compound 374)
##STR00038## A 25 mL reaction tube was charged with a magnetic stir
bar, phenyl boronic acid (72.6 mg, 0.596 mmol), K.sub.3PO.sub.4
(103 mg, 0.447 mmol), X-Phos pre-catalyst
(Chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2-
-aminoethyl)phenyl)]palladium(II)) (4.92 mg, 5.96 .mu.mol), and the
vial was sealed. The vial was evacuated/backfilled with nitrogen
(3.times.) before the addition of methyl
6-chloro-2-methyl-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-indole-3-carbo-
xylate (Compound 314) (100 mg, 0.298 mmol) as a solution in
1,4-dioxane (1 mL). The vial was then heated to 100.degree. C.
overnight with stirring. The vial was then allowed to cool to room
temperature and the reaction concentrated in vacuo. The crude
residue was purified via SiO.sub.2 chromatography (10 g) using an
eluent of ethyl acetate/hexanes (4:1) the title compound as a white
solid (106 mg, 0.281 mmol, 94% yield).). LCMS 514 (M+1).sup.+;
.sup.1H NMR .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.=11.59 (s,
1H), 7.98-7.84 (m, 2H), 7.75-7.67 (m, 3H), 7.47 (t, J=7.8 Hz, 2H),
7.39 (d, J=8.5 Hz, 1H), 7.35-7.27 (m, 1H), 6.15 (s, 1H), 4.35 (d,
J=4.9 Hz, 2H), 4.25-4.12 (m, 1H), 3.93 (d, J=8.5 Hz, 1H), 3.86-3.77
(m, 3H), 3.67 (d, J=8.5 Hz, 1H), 3.39-3.32 (m, 1H), 3.10-3.00 (m,
1H), 2.62 (s, 3H), 2.21 (s, 3H), 1.85 (d, J=10.0 Hz, 1H), 1.63-1.49
(m, 4H), 1.45-1.33 (m, 1H), 1.20-0.99 (m, 1H), 0.66 (d, J=12.0 Hz,
1H).
Example 7
Synthesis of (R or
S)-2-(4-(1-(3-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methylca-
rbamoyl)-2-methyl-1H-indol-1-yl)ethyl)piperidin-1-yl) acetic acid
(Compound 364)
##STR00039## To a round bottomed flask was charged with a magnetic
stir bar was added (R or
S)-ethyl-2-(4-(1-(3-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)me-
thylcarbamoyl)-2-methyl-1H-indol-1-yl)ethyl)piperidin-1-yl)acetate
(Compound 363) (69 mg, 0.132 mmol), THF (1.5 mL), MeOH (1.5 mL),
and water (0.75 mL). To this solution was added lithium hydroxide
monohydrate (5.54 mg, 0.132 mmol) and the reaction stirred at room
temperature for 1 h. The organics were removed under reduced
pressure and the resulting aqueous solution purified via reverse
phase-HPLC (water/MeCN) 0.fwdarw.95% to afford the title compound
(66 mg, 0.108 mmol, 82% yield). LCMS 514 (M+1).sup.+ 1H NMR (400
MHz, DMSO-d.sub.6) .delta.=11.67 (s, 1H), 9.65 (s, 1H), 7.84-7.68
(m, 2H), 7.63 (d, J=7.4 Hz, 1H), 7.14-7.03 (m, 2H), 6.18 (s, 1H),
4.33 (d, J=3.6 Hz, 2H), 4.27-4.15 (m, 1H), 4.04 (br. s., 2H), 3.85
(s, 3H), 3.57 (s, 1H), 3.35-3.23 (m, 1H), 3.14-2.99 (m, 1H),
2.86-2.74 (m, 1H), 2.62 (s, 3H), 2.21 (s, 3H), 2.18-2.08 (m, 1H),
1.75 (s, 1H), 1.60-1.49 (m, 4H), 1.46-1.33 (m, 1H), 0.92-0.81 (m,
1H).
Example 8
Synthesis of (R or S)-methyl
2-methyl-6-(pyridin-3-yl)-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-indole-
-3-carboxylate
This intermediate was used as an alternate starting material in
Step 7 set forth in Example 1 for the synthesis of other compounds
of the invention.
(R or S)-methyl
2-methyl-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-6-(4,4,5,5-tetramethyl-1,3-
,2-dioxaborolan-2-yl)-1H-indole-3-carboxylate
##STR00040## To a round bottomed flask was added Pd(OAc).sub.2
(10.03 mg, 0.045 mmol), potassium acetate (219 mg, 2.233 mmol),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (567
mg, 2.233 mmol), and
2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (XPhos) (85
mg, 0.179 mmol), and the vial was sealed. To this vessel was added
(R or S)-methyl
6-chloro-2-methyl-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-indole-3-carbo-
xylate (Step 6) (500 mg, 1.489 mmol) dissolved in dioxane (3.4 mL)
and the reaction evacuated/back-filled with N.sub.2 (3.times.)
before heating to 100.degree. C. overnight. The reaction was then
allowed to cool to rt and was diluted with EtOAc. The reaction was
filtered through diatomaceous earth and the filtrate concentrated
to afford the title compound which was used in subsequent reactions
without further purification. LCMS 428 (M+1).sup.+.
(R or S)-methyl
2-methyl-6-(pyridin-3-yl)-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-indole-
-3-carboxylate
##STR00041## To a re-sealable vial was added K.sub.2CO.sub.3 (206
mg, 1.488 mmol), PdCl.sub.2(dppf)-CH.sub.2Cl.sub.2 adduct (60.8 mg,
0.074 mmol), and the vial was sealed. This vial was
evacuated/backfilled with N.sub.2 (3.times.) before addition of (R
or S)-methyl
2-methyl-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-6-(4,4,5,5-tetramethyl-1,3-
,2-dioxaborolan-2-yl)-1H-indole-3-carboxylate (318 mg, 0.744 mmol)
dissolved in 1,4-dioxane (4 mL), 3-bromopyridine (71.7 .mu.l, 0.744
mmol), and water (400 .mu.L). The reaction was evacuated/backfilled
with N.sub.2 (3.times.) before heating to 100.degree. C. The
solution was cooled to room temperature and diluted with EtOAc. The
solution was filtered and concentrated in vacuo. The crude residue
was purified via silica gel chromatography (10 g, EtOAc/hex (1:1))
to afford the title compound (101 mg, 0.267 mmol, 35.9% yield).
LCMS 379 (M+1).sup.+.
Example 9
Other Alkyl Carboxylate Intermediates
The following alkyl carboxylate intermediates were synthesized in
an analogous manner to that set forth in Step 2 of Example 1, using
an appropriate starting material and reactant.
TABLE-US-00015 Name Structure m/z (.+-.)-ethyl 5- fluoro-1-(1-
methoxypropan- 2-yl)-2-methyl- 1H-indole-3- carboxylate
##STR00042## 294 (.+-.)-ethyl 6- fluoro-1-(1- methoxypropan-
2-yl)-2-methyl- 1H-indole-3- carboxylate ##STR00043## 294
(.+-.)-ethyl 1-(1- methoxypropan- 2-yl)-2-methyl- 1H-indole-3-
carboxylate ##STR00044## 276 (.+-.)-tert-butyl 1-(1- methoxypropan-
2-yl)-2-methyl- 1H-pyrrolo[2,3- b]pyridine-3- carboxylate
##STR00045## 305 (.+-.)-tert-butyl 1- (1-ethoxypropan-
2-yl)-2-methyl- 1H-pyrrolo[2,3- b]pyridine-3- carboxylate
##STR00046## 319 tert-butyl 1-(3- methoxybutan-2- yl)-2-methyl-
1H-pyrrolo[2,3- b]pyridine-3- carboxylate ##STR00047## 319 ethyl
1-(3- methoxybutan- 2-yl)-2- methyl-6- (methylsulfonyl)-
1H-indole-3- carboxylate ##STR00048## 368 (.+-.)-ethyl 1-(3-
methoxypentan- 2-yl)-2-methyl- 1H-indole-3- carboxylate
##STR00049## 304
Example 10
Other Compounds of the Invention Produced from Carboxylic Acid
Intermediates
The following compounds were synthesized in an analogous manner to
that set forth in Step 4 of Example 1, using an appropriate
starting material. Structures of these compounds are set forth in
FIG. 1.
TABLE-US-00016 Compound Name .sup.1H NMR m/z 304 (.+-.)-1-(1-(4,4-
(CDCl.sub.3 400 MHz) .delta. 12.63-12.64 (d, J = 3.2 427
difluorocyclohexyl)ethyl)- Hz, 1H), 7.84 (s, 1H), 7.49 (s, 1H),
7.42-7.40 N-((4-methoxy-6-methyl- (d, J = 9.2 Hz, 1H), 7.06-7.00
(m, 2H), 5.90- 2-oxo-1,2-dihydropyridin- 5.89 (d, J = 3.6 Hz 1H),
4.66-4.62 (t, J = 14 3-yl)methyl)-2-methyl- Hz, 2H), 4.11-4.08 (m,
1H), 3.88-3.87 (d, 1H-indole-3-carboxamide J = 3.6 Hz, 3H),
2.99-2.76 (m, 3H), 2.36 (s, 1H), 2.25 (s, 3H), 2.17-2.16 (d, J =
3.2 Hz, 2H), 2.08-2.05 (m, 2H), 1.84-1.70 (m, 2H), 1.61 (s, 1H),
1.51-1.47 (m, 2H) 230 (.+-.)-5-fluoro-N-((4- (400 MHz, CD.sub.3OD)
.delta. 7.59-7.55 (m, 1H), 416 methoxy-6-methyl-2-oxo- 7.42-7.39
(m, 1H), 6.95-6.90 (m, 2H), 4.57 1,2-dihydropyridin-3- (s, 2H),
4.12 (s, 3H), 3.99-3.94 (m, 1H), yl)methyl)-1-(1- 3.72-3.65 (m,
1H), 3.19 (s, 3H), 2.64 (s, 3H), methoxypropan-2-yl)-2- 2.54 (s,
3H), 1.59-1.57 (d, 3H) methyl-1H-indole-3- carboxamide 231
(.+-.)-6-fluoro-N-((4- (400 MHz, CD.sub.3OD) .delta. 7.70-7.66 (m,
1H), 416 methoxy-6-methyl-2-oxo- 7.36-7.33 (m, 1H), 6.94-6.89 (m,
2H), 4.56 1,2-dihydropyridin-3- (s, 2H), 4.11 (s, 3H), 3.97-3.92
(m, 1H), yl)methyl)-1-(1- 3.71-3.67 (m, 1H), 3.20 (s, 3H), 2.62 (s,
3H), methoxypropan-2-yl)-2- 2.53 (s, 3H), 1.58-1.56 (d, 3H)
methyl-1H-indole-3- carboxamide 218 (.+-.)-N-((4-methoxy-6- (400
MHz, CD.sub.3OD) .delta. 7.69 (d, J = 7.2 398 methyl-2-oxo-1,2- Hz,
1H), 7.53 (d, J = 7.6 Hz, 1H), 7.12 (m, dihydropyridin-3- 2H), 6.26
(s, 1H), 4.80 (m, 1H), 4.52 (s, 2H), yl)methyl)-1-(1- 3.99 (m, 4H),
3.75 (m, 1H), 3.20 (s, 3H), 2.62 methoxypropan-2-yl)-2- (s, 3H),
2.31 (s, 3H), 1.59 (d, J = 7.2 Hz, 3H) methyl-1H-indole-3-
carboxamide 183 (.+-.)-N-((4,6-dimethyl-2- (400 MHz, CD.sub.3OD)
.delta. 7.74 (m, 1H), 7.57 382 oxo-1,2-dihydropyridin-3- (d, J =
7.6 Hz, 1H), 7.15 (m, 2H), 6.14 yl)methyl)-1-(1- (s, 1H), 4.86 (m,
1H), 4.55 (s, 2H), 4.02 (m, methoxypropan-2-yl)-2- 1H), 3.77 (m,
1H), 3.22 (s, 3H), 2.65 (s, 3H), methyl-1H-indole-3- 2.43 (s, 3H),
2.26 (s, 3H), 1.62 (d, J = 7.2 Hz, carboxamide 3H) 204
(.+-.)-N-((4-methoxy-6- (400 MHz, CDCl.sub.3) .delta. 13.23 (s,
1H), 8.16- 399 methyl-2-oxo-1,2- 8.17 (m, 1H), 8.11-8.13 (m, 1H),
7.57-7.60 dihydropyridin-3- (t, J = 5.2 Hz, 1H), 6.93-6.96 (m, 1H),
5.92 yl)methyl)-1-(1- (s, 1H), 4.82-4.83 (d, J = 2.4 Hz, 1H), 4.65-
methoxypropan-2-yl)-2- 4.66 (d, J = 6.4 Hz, 2H), 3.89 (s, 3H),
3.81- methyl-1H-pyrrolo[2,3- 3.85 (m, 1H), 3.22 (s, 3H), 2.79 (s,
3H), b]pyridine-3-carboxamide 2.17 (s, 3H), 1.64-1.66 (d, J = 8.0
Hz, 3H) 211 (.+-.)-1-(1- 379 cyclopropylethyl)-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-1H-
pyrrolo[2,3-b]pyridine-3- carboxamide 212
(.+-.)-1-(1-ethoxypropan-2- (400 MHz, CDCl.sub.3) .delta.
8.173-8.189 (m, 1H), 413 yl)-N-((4-methoxy-6- 8.13-8.153 (m, 1H),
7.563 (s, 1H), 6.977- methyl-2-oxo-1,2- 7.008 (m, 1H), 5.938 (s,
1H), 4.652-4.667 dihydropyridin-3- (d, 2H), 4.177 (s, 1H),
3.309-3.454 (m, 2H), yl)methyl)-2-methyl-1H- 3.94-3.98 (m, 1H),
2.806 (s, 3H), 2.212 (s, pyrrolo[2,3-b]pyridine-3- 3H), 1.665-1.682
(d, 3H), 1.044 (t, 3H) carboxamide 235 (.+-.)-N-((4-ethoxy-6- (400
MHz, CDCl.sub.3) .delta. 413 methyl-2-oxo-1,2- 12.5 (s, 1H),
8.11-8.18 (m, 2H), 7.60 (s, dihydropyridin-3- 1H), 6.95-6.98 (m,
1H), 5.90 (s, 1H), 5.96 yl)methyl)-1-(1- (s, 1H), 4.83 (s, 1H),
4.10-4.21 (m, 3H), methoxypropan-2-yl)-2- 3.82-3.83 (m, 1H), 3.23
(s, 3H), 3.79 (s, methyl-1H-pyrrolo[2,3- 3H), 2.15 (s, 3H),
1.65-1.66 (d, J = 6.8 Hz, b]pyridine-3-carboxamide 6H), 1.44-1.47
(t, J = 7.2 Hz, 3H). 241 N-((4,6-dimethyl-2-oxo- (400 MHz,
CD.sub.3OD): .delta. 8.67-8.65 (d, 1H), 397 1,2-dihydropyridin-3-
8.45-8.44 (d, 1H), 7.59-7.55 (m, 1H), 6.70 yl)methyl)-1-(3- (s,
1H), 4.79 (s, 1H), 4.61 (s, 2H), 4.07 (s, methoxybutan-2-yl)-2-
1H), 3.32 (s, 3H), 2.75 (s, 3H), 2.56 (s, 3H),
methyl-1H-pyrrolo[2,3- 2.42 (s, 3H), 1.68-1.66 (d, 3H), 1.16-1.15
(d, b]pyridine-3-carboxamide 3H). 280 (.+-.)-N-((6-ethyl-4- 1H NMR
(400 MHz, CD.sub.3OD) .delta. 8.25-8.29 413 methoxy-2-oxo-1,2- (m,
2H). .delta. 7.28-7.31 (m, 1H). 6.89 (s, 1H), dihydropyridin-3-
4.93-4.95 (br, 1H), 4.58 (s, 2H), 4.2-4.25 yl)methyl)-1-(1- (m,
1H), 4.13 (s, 3H), 3.77-3.81 (m, 1H), methoxypropan-2-yl)-2- 3.24
(s, 3H), 2.79-2.84 (q, 1H), 2.72 (s, 3H), methyl-1H-pyrrolo[2,3-
1.66-1.68 (d, J = 7.2 Hz, 3H) 1.32-1.36 (t,
b]pyridine-3-carboxamide 3H) 288 (R or S)-N-((4-Methoxy- (400 MHz,
d6-DMSO) .delta. 11.57-11.65 (m, 439 6-methyl-2-oxo-1,2- 1H),
8.18-8.23 (m, 1H), 8.07-8.12 (m, dihydropyridin-3- 1H), 7.83-7.91
(m, 1H), 7.07-7.15 (m, yl)methyl)-2-methyl-1-(1- 1H), 6.15 (s, 1H),
4.31 (d, J = 4.46 Hz, 1H), (tetrahydro-2H-pyran-4- 4.04-4.20 (m,
1H), 3.88-3.97 (m, 1H), yl)ethyl)-1H-pyrrolo[2,3- 3.84 (s, 3H),
3.59-3.70 (m, 1H), 2.97- b]pyridine-3-carboxamide 3.10 (m, 1H),
2.79-2.93 (m, 1H), 2.67 (br. S., 3H), 2.20 (s, 3H), 1.78-1.88 (m,
1H), 1.53-1.68 (m, 3H), 1.28-1.41 (m, 2H), 0.97-1.13 (m, 2H),
0.56-0.68 (m, 1H) 306 (R or S)-N-((4,6-dimethyl- (400 MHz,
DMSO-d.sub.6) .delta. = 11.73-11.56 (m, 423
2-oxo-1,2-dihydropyridin- 1 H), 8.19 (d, J = 3.1 Hz, 1 H), 8.06
(dd, J = 3-yl)methyl)-2-methyl-1- 1.4, 7.9 Hz, 1 H), 7.82 (br. S.,
1 H), 7.10 (1-(tetrahydro-2H-pyran- (dd, J = 4.7, 7.8 Hz, 1 H),
5.91 (s, 1 H), 4.30 4-yl)ethyl)-1H-indole-3- (br. S., 2 H),
4.19-4.02 (m, 1 H), 3.90 (d, J = carboxamide 8.5 Hz, 1 H), 3.63 (d,
J = 7.8 Hz, 1 H), 3.29 (s, 1 H), 3.06 (s, 1 H), 2.92-2.74 (m, 1 H),
2.64 (br. S., 3 H), 2.25 (s, 3 H), 2.11 (s, 3 H), 1.80 (br. S., 1
H), 1.59 (br. S., 3 H), 1.41-1.24 (m, 1 H), 1.09 (s, 2 H), 0.67-
0.52 (m, 1 H) 277 (.+-.)-1-(3-methoxy-3- (400 MHz, DMSO-d.sub.6)
.delta. = 12.01-11.82 (m, 426 methylbutan-2-yl)-N-((4- 1 H),
7.91-7.82 (m, 2 H), 7.71-7.64 (m, 1 methoxy-6-methyl-2-oxo- H),
7.06-6.96 (m, 2 H), 6.25 (s, 1 H), 4.43 1,2-dihydropyridin-3- (q, J
= 7.1 Hz, 1 H), 4.33 (br. S., 2 H), 3.86 yl)methyl)-2-methyl-1H-
(s, 3 H), 3.14-3.09 (m, 3 H), 2.61 (s, 3 H), indole-3-carboxamide
2.23 (s, 3 H), 1.58-1.52 (m, 3 H), 1.27 (s, 3 H), 0.88 (s, 3 H) 275
(.+-.)-N-((4,6-dimethyl-2- 410 oxo-1,2-dihydropyridin-3-
yl)methyl)-1-(3- methoxypentan-2-yl)-2- methyl-1H-indole-3-
carboxamide 294 (.+-.)-N-((4-methoxy-6- (CDCl.sub.3, 400 MHz)
.delta. 7.85 (t, J = 6.4 Hz, 412 methyl-2-oxo-1,2- 1H), 7.45 (s,
2H), 7.08-7.03 (m, 2H), 5.93 dihydropyridin-3- (s, 1H), 4.71-4.61
(m, 2H), 4.36 (s, 1H), yl)methyl)-1-(3- 3.90 (s, 4H), 2.95 (s, 3H),
2.75 (s, 3H), 2.17 methoxybutan-2-yl)-2- (s, 3H), 1.57 (d, J = 7.2
Hz, 3H), 1.23 (d, methyl-1H-indole-3- J =6.0 Hz, 3H) carboxamide
290 (.+-.)-1-(3-ethoxybutan-2- (400 MHz, DMSO-d.sub.6) .delta. =
11.60 (br. s., 1 426 yl)-N-((4-methoxy-6- H), 7.72 (d, J = 7.6 Hz,
1 H), 7.67 (d, J = methyl-2-oxo-1,2- 5.1 Hz, 2 H), 7.09-6.98 (m, 2
H), 6.14 (s, 1 dihydropyridin-3- H), 4.41-4.35 (m, 1 H), 4.32 (d, J
= 4.9 Hz, yl)methyl)-2-methyl-1H- 2 H), 4.03-3.93 (m, 1 H), 3.83
(s, 3 H), indole-3-carboxamide 3.25 (d, J = 9.4 Hz, 1 H), 2.82-2.72
(m, 1 H), 2.62 (br. s., 3 H), 2.19 (s, 3 H), 1.52 (d, J = 7.1 Hz, 3
H), 1.15 (d, J = 6.0 Hz, 3 H), 0.68 (t, J = 6.9 Hz, 3 H) 293
N-((4-methoxy-6-methyl- (400 MHz, CDCl.sub.3): .delta. 8.19-8.13
(m, 2H), 427 2-oxo-1,2-dihydropyridin- 7.57-7.55 (t, 1H), 6.99-6.96
(m, 1H), 5.94 3-yl) methyl)-1-(3- (s, 1H), 4.67-4.65 (m, 2H), 4.40
(m, 1H), methoxypentan-2-yl)-2- 4.16 (m, 1H), 3.16 (s, 3H), 2.80
(s, 3H), methyl-1H-pyrrolo [2,3- 2.77 (s, 3H), 2.20 (s, 3H),
1.87-1.81 (m, b] pyridine-3-carboxamide 1H), 1.67-1.65 (m, 3H),
1.53-1.45 (m, 3H), 1.02-0.99 (m, 3H) 299 N-((4-methoxy-6-methyl-
(400 MHz, CDCl.sub.3) .delta. 7.87-7.86 (d, 1H), 425
2-oxo-1,2-dihydropyridin- 7.52-7.45 (m, 2H), 7.10-7.02 (m, 2H),
4.72- 3-yl) methyl)-1-(3- 4.64 (dd, 2H), 4.45-4.42 (s 1H), 3.9 (s,
3H), methoxypentan-2-yl)-2- 3.73 (s, 1H), 2.8-2.7 (d, 6H), 2.17 (s,
3H), methyl-1H-indole-3- 1.80-1.75 (m, 1H), 1.58 (s, 3H), 1.25
carboxamide (m, 1H), 1.03-0.99 (t, 3H)
Example 11
Synthesis of Methyl
1-(1-(1,4-dioxan-2-yl)ethyl)-2-methyl-1H-indole-3-carboxylate
The title compound was used as an alternate alkyl carboxylate
starting material in Step 3 of Example 1.
Step 1: 1-(1,4-dioxan-2-yl)ethanone
##STR00050## To a solution of benzoic peroxide (20 g, 141 mmol) in
200 mL 1,4-dioxane at room temperature under nitrogen atmosphere
was added biacetyl (24.3 g, 282 mmol). After the addition, the
mixture was heated to reflux and stirred for 24 hours. The reaction
mixture was cooled to 0.degree. C. The pH was adjusted to around 9
by progressively adding 2N sodium hydroxide below 0.degree. C.,
extracted with 2-methoxy-2-methylpropane (10 mL.times.3), and
concentrated to give 1-(1,4-dioxan-2-yl)ethanone (13 g, 36%) as a
yellow oil which was used directly in the next step without
purification.
Step 2: 1-(1,4-dioxan-2-yl)ethanamine
##STR00051## To a solution of 1-(1,4-dioxan-2-yl)ethanone (12 g,
92.2 mmol) in 1,2-dichloroethane (100 mL) was added
(4-methoxyphenyl)methanamine (25 g, 184.4 mmol) at room
temperature. The mixture was allowed to stir for 3 hours, and then
sodium triacetoxyborohydride (39 g, 184.4 mmol) was added. The
resulting mixture was allowed to stir for 48 hours at room
temperature. The reaction mixture was quenched by adding water,
extracted with dichloromethane (100 mL.times.3). The combined
organic phase was dried by anhydrous sodium sulphate, and then
filtered. The filtrate was concentrated and purified by column
chromatograph on silica gel (elute: dichloromethane/methanol
100:1.fwdarw.50:1.fwdarw.20:1) to give
1-(1,4-dioxan-2-yl)-N-(4-methoxybenzyl)ethanamine (16.4 g, 71%) as
a yellow solid. LCMS (M+H.sup.+) m/z: calcd. 251.15. found 251.9.
To a solution of 1-(1,4-dioxan-2-yl)-N-(4-methoxybenzyl)ethanamine
(5 g, 19.9 mmol) in anhydrous methanol (100 mL) was added palladium
10% on carbon (240 mg, 2 mmol), then purged with hydrogen (30 psi),
the mixture was allowed to stir overnight at room temperature. The
reaction mixture was filtered, and the filtrate was concentrated to
afford the title compound (2.5 g, 96%) as a brown solid.
The amine intermediates shown in the following table were prepared
according to the general procedure outlined above using the
appropriate starting materials and modifications.
TABLE-US-00017 Name Structure m/z tert-butyl 3-(1-
aminoethyl)piperidine- 1-carboxylate ##STR00052## 228
(.+-.)-1-(4,4- difluorocyclohexyl) ethanamine ##STR00053## 164
(.+-.)-1-(1- (methylsulfonyl)azetidin- 3-yl)ethanamine ##STR00054##
179 (.+-.)-tert-butyl 4-(4-(1- aminoethyl)47yridine-
2-yl)piperazine-1- carboxylate ##STR00055## 307
Step 3: (E)-methyl
3-((1-(1,4-dioxan-2-yl)ethyl)imino)-2-(2-bromophenyl)butanoate
##STR00056## To a solution of 1-(1,4-dioxan-2-yl)ethanamine (2.5 g,
19 mmol) in methanol (100 mL) was added methyl
2-(2-bromophenyl)-3-oxobutanoate (5.4 g, 20 mmol) and acetic acid
(1.8 g, 30 mmol). The resulting reaction system was warm to reflux
and allowed to stir overnight. The reaction mixture was
concentrated and purified by column chromatographed on silica gel
(eluted: dichloromethane/methanol 50:1.fwdarw.20:1.fwdarw.5:1) the
title compound (1 g, 14%) as a brown solid. LCMS (M+H.sup.+) m/z:
calcd. 383.07. found 384.9.
The imino-bromo intermediates shown in the following table were
prepared according to the general procedure outlined above using
the appropriate starting materials (e.g., one of the amines set
forth in the table in Step 2 of this example) and
modifications.
TABLE-US-00018 Name Structure m/z (E)-tert-butyl 3-(1-((3-(2-
bromophenyl)- 4-methoxy-4- oxobutan-2- ylidene)amino)
ethyl)piperidine- 1-carboxylate ##STR00057## 482 (.+-.)-(E)-methyl
2-(2-bromophenyl)- 3-((1-(4,4- difluorocyclohexyl) ethyl)imino)
butanoate ##STR00058## 417 (E)-tert-butyl 4-((3-(2- bromophenyl)-
4-methoxy-4- oxobutan-2- ylidene)amino) piperidine-1- carboxylate
##STR00059## 454 (Z)-methyl 2-(2- bromophenyl)-3- (quinolin-5-
ylamino)but-2- enoate ##STR00060## 398 (E)-methyl 2-(2-
bromophenyl)-3- (cyclopentylimino) butanoate ##STR00061## 339
(E)-methyl 2-(2- bromophenyl)-3- ((6- methylquinolin-5-
yl)imino)butanoate ##STR00062## 412 (.+-.)-(E)-methyl 2-
(2-bromophenyl)- 3-((1-(1- (methylsulfonyl) azetidin-3-
yl)ethyl)imino) butanoate ##STR00063## 432 (.+-.)-(E)-tert-butyl
4-(4-((3-(2- bromophenyl)-4- methoxy-4- oxobutan- 2-ylidene)
amino)pyridine- 2-yl)piperazine- 1-carboxylate ##STR00064## 559
(E)-methyl 2-(2- bromophenyl)-3- ((2,5- dimethylphenyl)
amino)but-2- enoate ##STR00065## 375 (E)-methyl 2-(2-
bromophenyl)-3- ((2,3- dimethylphenyl) amino)but-2- enoate
##STR00066## 375 (E)-methyl 2-(2- bromophenyl)-3- (quinolin-6-
ylimino)butanoate ##STR00067## 398
Step 4: Methyl
1-(1-(1,4-dioxan-2-yl)ethyl)-2-methyl-1H-indole-3-carboxylate
##STR00068## To a solution of (E)-methyl
3-((1-(1,4-dioxan-2-yl)ethyl)imino)-2-(2-bromophenyl)butanoate (400
mg, 1.1 mmol) in dioxane (3 mL) was added
Chloro[2-(dicyclohexylphosphino)-3,6-dimethoxy-2',4',6'-triisopropylbiphe-
nyl][2-(2-aminoethyl)phenyl]Pd(II) (160 mg, 0.2 mmol),
2-Dicyclohexyphosphino-2',6'-diisopropoxybiphenyl (93 mg, 0.2 mmol)
and sodium tert-butoxide (192 mg, 2 mmol). The resulting reaction
mixture was heated to 120.degree. C. with stirring for 30 mins in a
microwave. The reaction mixture was quenched by adding water and
was extracted with ethyl acetate (25 mL.times.3). The combined
organic phase was dried by anhydrous sodium sulphate, and then
filtered. The filtrate was concentrated and purified by column
chromatograph on silica gel (eluted: petrol ether/acetic ester
10:1.fwdarw.5:1.fwdarw.2:1) to afford the title compound (282 mg,
89%) as yellow solid. LCMS (M+H.sup.+) m/z: calcd. 303.15. found
303.9.
The compound shown in the following table was prepared according to
the general procedure outlined above using the appropriate starting
materials (e.g., one of the imino-bromo intermediates shown in the
table in Step 3 of this example) and modifications.
TABLE-US-00019 Name Structure m/z (.+-.)-methyl 1-(1-(4,4-
difluorocyclohexyl) ethyl)- 2-methyl- 1H-indole-3- carboxylate
##STR00069## 336
These alkyl carboxylates were also used as starting material in
Step 3 of Example 1 in the synthesis of certain compounds of the
invention.
Example 12
Chiral Separation of Compound 219 to Afford Compounds 223 and
224
N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(1-methoxy-
propan-2-yl)-2-methyl-1H-indole-3-carboxamide (200 mg) (Compound
219) was subjected to chiral chromatography via supercritical fluid
chromatography (SFC) (A: C.sub.2H.sub.5OH, B: NH.sub.3--H.sub.2O.
A:B=55:45 AD column) to afford the separate enantiomers 223 (peak
1) and 224 (Peak 2) (60 mg each) LCMS 398 (M+1).sup.+ 1H NMR (400
MHz, CD.sub.3OD) .delta. 7.69 (d, J=7.2 Hz, 1H), 7.53 (d, J=7.6 Hz,
1H), 7.12 (m, 2H), 6.26 (s, 1H), 4.80 (m, 1H), 4.52 (s, 2H), 3.99
(m, 4H), 3.75 (m, 1H), 3.20 (s, 3H), 2.62 (s, 3H), 2.31 (s, 3H),
1.59 (d, J=7.2 Hz, 3H). The optical rotation of each enantiomer was
not determined.
The compounds shown in the following table were prepared according
to the general chiral chromatography procedure outlined above. The
optical rotation of the separated enantiomers was not determined,
but the elution peak ("Peak 1" or "Peak 2") is indicated.
Structures of each compound are shown in FIG. 1.
TABLE-US-00020 Com- pound Name .sup.1H NMR m/z 217 (R or
S)-N-((4,6-dimethyl-2- (400 MHz, CD.sub.3OD) .delta. 7.74 (m, 1H),
382 oxo-1,2-dihydropyridin-3- 7.57 (d, J = 7.6 Hz, 1H), 7.15 (m,
2H), yl)methyl)-1-(1- 6.14 (s, 1H), 4.86 (m, 1H), 4.55 (s, 2H),
methoxypropan-2-yl)-2- 4.02 (m, 1H), 3.77 (m, 1H), 3.22 (s,
methyl-1H-indole-3- 3H), 2.65 (s, 3H), 2.43 (s, 3H), 2.26 (s,
carboxamide-PEAK 1 3H), 1.62 (d, J = 7.2 Hz, 3H) 218 (R or
S)-N-((4,6-dimethyl-2- (400 MHz, CD.sub.3OD) 6 7.74 (m, 1H), 382
oxo-1,2-dihydropyridin-3- 7.57 (d, J = 7.6 Hz, 1H), 7.15 (m, 2H),
yl)methyl)-1-(1- 6.14 (s, 1H), 4.86 (m, 1H), 4.55 (s, 2H),
methoxypropan-2-yl)-2- 4.02 (m, 1H), 3.77 (m, 1H), 3.22 (s, 3H),
methyl-1H-indole-3- 2.65 (s, 3H), 2.43 (s, 3H), 2.26 (s, 3H),
carboxamide-PEAK 2 1.62 (d, J = 7.2 Hz, 3H) 252 (R or
S)-(.+-.)-1-(1- NMR (400 MHz, CDCl.sub.3): 379
cyclopropylethyl)-N-((4,6- .delta. 8.32-8.34 (d, 1H), 8.18-8.2 (d,
1H), dimethyl-2-oxo-1,2- 7.27-7.30 (m, 1H), 6.70 (s, 1H), 4.47 (s,
dihydropyridin-3-yl)methyl)- 2H), 3.94-3.95 (d, 1H), 2.61 (s, 3H),
2-methyl-1H-pyrrolo[2,3- 2.43 (s, 3H), 2.29-2.30 (s, 3H), 1.57-
b]pyridine-3-carboxamide 1.59 (d, 3H), 0.63-0.64 (t, 1H), 0.27-
PEAK 1 0.64 (m, 2H), 0.02-0.04 (t, 1H) 253 (R or S)-(.+-.)-1-(1-
NMR (400 MHz, CDCl.sub.3): 379 cyclopropylethyl)-N-((4,6- .delta.
8.32-8.34 (d, 1H), 8.18-8.2 (d, 1H), dimethyl-2-oxo-1,2- 7.27-7.30
(m, 1H), 6.70 (s, 1H), 4.47 (s, dihydropyridin-3-yl)methyl)- 2H),
3.94-3.95 (d, 1H), 2.61 (s, 3H), 2-methyl-1H-pyrrolo]2,3- 2.43 (s,
3H), 2.29-2.30 (s, 3H), 1.57- b]pyridine-3-carboxamide 1.59 (d,
3H), 0.63-0.64 (t, 1H), 0.27- PEAK 2 0.64 (m, 2H), 0.02-0.04 (t,
1H) 256 (R or S)-N-((4-methoxy-6- (400 MHz, CDCl.sub.3) .delta.
13.23 (s, 1H), 399 methyl-2-oxo-1,2- 8.16-8.17 (m, 1H), 8.11-8.13
(m, 1H), dihydropyridin-3-yl)methyl)- 7.57-7.60 (t, J = 5.2 Hz,
1H), 6.93-6.96 1-(1-methoxypropan-2-yl)-2- (m, 1H), 5.92 (s, 1H),
4.82-4.83 (d, J = methyl-1H-pyrrolo[2,3- 2.4 Hz, 1H), 4.65-4.66 (d,
J = 6.4 Hz, b]pyridine-3-carboxamide 2H), 3.89 (s, 3H), 3.81-3.85
(m, 1H), PEAK 1 3.22 (s, 3H), 2.79 (s, 3H), 2.17 (s, 3H), 1.64-1.66
(d, J = 8.0 Hz, 3H) 257 (R or S)-N-((4-methoxy-6- (400 MHz,
CDCl.sub.3) .delta. 13.23 (s, 1H), 399 methyl-2-oxo-1,2 8.16-8.17
(m, 1H), 8.11-8.13 (m, 1H), dihydropyridin-3-yl)methyl)- 7.57-7.60
(t, J = 5.2 Hz, 1H), 6.93-6.96 1-(1-methoxypropan-2-yl)-2- (m, 1H),
5.92 (s, 1H), 4.82-4.83 (d, J = methyl-1H-pyrrolo[2,3- 2.4 Hz, 1H),
4.65-4.66 (d, J = 6.4 Hz, b]pyridine-3-carboxamide 2H), 3.89 (s,
3H), 3.81-3.85 (m, 1H), PEAK 2 3.22 (s, 3H), 2.79 (s, 3H), 2.17 (s,
3H), 1.64-1.66 (d, J = 8.0 Hz, 3H) 307 Trans-(R or S, R or
S)-N-((4- (CDCl.sub.3, 400 MHz) .delta. 7.85 (t, J = 6.4 412
methoxy-6-methyl-2-oxo-1,2- Hz, 1H), 7.45 (s, 2H), 7.08-7.03 (m,
dihydropyridin-3-yl)methyl)- 2H), 5.93 (s, 1H), 4.71-4.61 (m, 2H),
1-(3-methoxybutan-2-yl)-2- 4.36 (s, 1H), 3.90 (s, 4H), 2.95 (s,
3H), methyl-1H-indole-3- 2.75 (s, 3H), 2.17 (s, 3H), 1.57 (d, J =
carboxamide 7.2 Hz, 3H), 1.23 (d, J = 6.0 Hz, 3H) PEAK 1 308
Trans-(R or S, R or S)-N-((4- (CDCl.sub.3, 400 MHz) .delta. 7.85
(t, J = 6.4 412 methoxy-6-methyl-2-oxo-1,2- Hz, 1H), 7.45 (s, 2H),
7.08-7.03 (m, dihydropyridin-3-yl)methyl)- 2H), 5.93 (s, 1H),
4.71-4.61 (m, 2H), 1-(3-methoxybutan-2-yl)-2- 4.36 (s, 1H), 3.90
(s, 4H), 2.95 (s, 3H), methyl-1H-indole-3- 2.75 (s, 3H), 2.17 (s,
3H), 1.57 (d, J = carboxamide 7.2 Hz, 3H), 1.23 (d, J = 6.0 Hz, 3H)
PEAK 2
Example 1
Synthesis of tert-butyl
1-(2,3-dihydro-1H-inden-1-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carbox-
ylate
The title compound as starting material in Step 3 of Example 36 in
the synthesis of certain compounds of the invention.
Tert-butyl 2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate
##STR00070## To a 500 mL round-bottom flask that contains
N-acetyl-N-(3-bromopyridin-2-yl)acetamide (14.815 g, 57.6 mmol),
was added copper(I) iodide (1.098 g, 5.76 mmol), L-proline (1.327
g, 11.53 mmol), cesium carbonate (28.2 g, 86 mmol), then t-butyl
acetoacetate (11.47 ml, 69.2 mmol) and dioxane (100 mL). The
reaction was vac/purged with N.sub.2 3.times. then fitted with a
septum and a N.sub.2 inlet and heated overnight at 70.degree. C.
The inorganic solids were removed by filtration over celite and the
cake was washed with 100 mL EtOAc. This solution was concentrated
and the residue was partitioned between 250 mL brine and 250 mL
EtOAc. The aq. Layer was further extracted with EtOAc (2.times.250
mL) and the combined organic layer was dried over Na.sub.2SO.sub.4,
filtered, concentrated and purified by CC using 1:1 EtOAc:Hex as
eluent to provide (2.7 g, 20.2%) of tert-butyl
2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate. LRMS (M+H.sup.+)
m/z: calc'd 233.28. found 233.1.
Tert-butyl
1-(2,3-dihydro-1H-inden-1-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridin-
e-3-carboxylate
##STR00071## A solution of ethyl tert-butyl
2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate (100 mg, 0.74
mmol), 2,3-dihydro-1H-inden-1-ol (176 mg, 0.74 mmol), PPh.sub.3
(195 mg, 1.49 mmol) was stirred in dry THF (10 mL) at 0.degree. C.
under a nitrogen atmosphere. To this mixture was added drop-wise
DIAD (150 mg, 1.48 mmol) over a period of 5 min, and the reaction
was stirred at room temperature for 16 hours. The mixture was
washed with brine, dried and concentrated to afford the crude
product. The crude product was purified by silca gel chromatography
(petroleum ether/ethyl acetate=5:1) to afford the
tert-butyl-1-(2,3-dihydro-1H-inden-1-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridi-
ne-3-carboxylate (150 mg, 60%).
The compound shown in the following table was prepared according to
the general procedure outlined above using the appropriate starting
materials and modifications.
TABLE-US-00021 Name Structure m/z (.+-.)-tert-butyl 1-
(1-cyclopropylethyl)- 2-methyl-1H-pyrrolo [2,3-b]pyridine-3-
carboxylate ##STR00072## 301
Each of the above alkyl carboxylates was used as starting material
in Step 3 of Example 1 in the synthesis of certain compounds of the
invention.
Example 13
Synthesis of isolated
N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-((2R
or 2S, 3R or
3S)-3-methoxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-c-
arboxamide diastereomers (Compounds 261, 266, 267 and 302)
Step 1: Tert-butyl
2-methyl-1-(3-oxobutan-2-yl)-1H-pyrrolo[2,3-b]pyridine-3-carboxylate
##STR00073## To a solution of tert-butyl
2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate (5.0 g, 21.53
mmol) in CH.sub.3CN (50 mL) was added Cs.sub.2CO.sub.3 (21.0 g,
64.58 mmol), potassium iodide (3.57 g, 21.53 mmol). The mixture was
stirred at 27.degree. C. for 30 minutes. Then 3-chlorobutan-2-one
(2.75 g, 25.83 mmol) was added and the mixture was stirred at
70.degree. C. for 12 hours. The mixture was filtered and the
filtrate was concentrated. The residue was purified by column
(Elute: Petroleum ether:Ethyl acetate=50:1) to give tert-butyl
2-methyl-1-(3-oxobutan-2-yl)-1H-pyrrolo[2,3-b]pyridine-3-carboxylate
as a yellow-green oil. (3.23 g, yield 50%) LCMS (M+H.sup.+) m/z:
calcd 303.37. found 302.9. 1H NMR (400 MHz, CDCl.sub.3): .delta.
8.32-8.30 (m, 1H), 8.25-8.23 (m, 1H), 7.17-7.14 (m, 1H), 5.50-5.44
(m, 1H), 2.71 (s, 3H), 1.96 (s, 3H), 1.65-1.67 (d, 3H), 1.64 (s,
9H).
Step 2: Tert-butyl
1-(3-hydroxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate
##STR00074## To the solution of tert-butyl
2-methyl-1-(3-oxobutan-2-yl)-1H-pyrrolo[2,3-b]pyridine-3-carboxylate
(3.1 g, 10.25 mmol) in methanol (30 mL) was added sodium
borohydride (0.30 g, 8.2 mmol) at 0.degree. C. After 30 minutes,
another batch of sodium borohydride (0.30 g, 8.2 mmol) was added at
0.degree. C. After the reaction completed about 2 h later, water
(30 ml) was added dropwise very carefully to quench the reaction.
The mixture was extracted with CH.sub.2Cl.sub.2. The extraction was
dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum
to give tert-butyl
1-(3-hydroxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate
as a yellow solid. (3.0 g, yield 96%) LCMS (M+H.sup.+) m/z: calcd
305.38. found 304.9. 1H NMR (400 MHz, CDCl.sub.3): .delta.
8.31-8.29 (m, 1H), 8.13-8.12 (m, 1H), 7.11-7.07 (m, 1H), 4.46-4.43
(m, 1H), 4.12 (m, 1H), 2.73 (s, 3H), 1.58 (s, 9H), 1.51-1.49 (d,
3H), 0.92-0.91 (d, 3H).
Step 3:
Tert-butyl-1-(3-methoxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyrid-
ine-3-carboxylate
##STR00075## To dry THF (20 mL) was added NaH (60% in mineral oil,
2.37 g, 59.14 mmol). Then the mixture was stirred at 27.degree. C.
for 20 minutes, then tert-butyl
1-(3-hydroxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate
(3.0 g, 9.86 mmol) was added. The mixture was stirred at 27.degree.
C. for 1 hour, then added by CH.sub.3I (13.99 g, 98.6 mmol). The
mixture was stirred for 12 hours at 27.degree. C. and then cooled
to 0.degree. C. Sat. NH.sub.4Cl was added and extracted with
CH.sub.2Cl.sub.2. The extraction was dried over sodium sulfate,
filtered and concentrated to give
tert-butyl-1-(3-methoxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridin-
e-3-carboxylate as a yellow oil. (3.2 g, yield 100%) LCMS
(M+H.sup.+) m/z: calcd. 319.41. found 318.9.
Step 4:
1-(3-methoxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carbo-
xylic acid
##STR00076## To the pre-cooled solution of tert-butyl
1-(3-methoxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate
(3.0 g, 9.42 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added
trifluoroacetic acid (20 mL) dropwise. The solution was stirred at
27.degree. C. for 1.5 hours. The solvent was removed under vacuum
at 27.degree. C. The residue was used for next step without
purified. LCMS (M+H+) m/z: calcd 263.30. found 262.9.
Step 5:
N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(3--
methoxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide
##STR00077## To a solution of
1-(3-methoxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylic
acid (2.4 g, 9.15 mmol) in DMF (30 mL) was added TEA (4.2 g, 41.50
mmol), 3-(aminomethyl)-4-methoxy-6-methylpyridin-2(1H)-one
hydrochloride (2.1 g, 12.81 mmol) After stirred for 10 minutes at
27.degree. C., the mixture was cooled and added HATU (5.56 g, 14.64
mmol). The mixture was stirred at 27.degree. C. for 72 hours and
30% of S.M. remained. Then the mixture was heated at 80.degree. C.
for 5 hours. The solution was diluted with brine (100 mL) and
extracted with CH.sub.2Cl.sub.2 (100 mL*3). The extractions were
combined and dried over Na.sub.2SO.sub.4. The solvent was
evaporated under vacuum and the residue was purified by flash
column (Eluent: dichloromethane:methanol=95:5) to give
N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(3-methoxy-
butan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide. (3.6
g, yield 95%)
Step 6: Separation of
N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(3-methoxy-
butan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide:
Isomers (Compounds 261, 266, 267, and 302)
##STR00078## ##STR00079## The mixture or isomers from step 5,
N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(3-methoxy-
butan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide was
purified by prep-HPLC (Condition: Column: SHIMADZU LC-8A, 250*50
mm*10 um; Mobile phase A: water with 0.2% formic acid; Mobile phase
B: MeCN; column temperature: 30.degree. C.; Gradient: B in A
10-50%) to give a major isomer pair (Compound 261 and Compound 266
combined) (1.0 g, purity 98.8%) and a minor isomer pair (Compound
267 and Compound 302 combined) (180 mg, purity 63%). The resulting
isomer pairs were individually separated by SFC (Condition: Column:
Chiral-pak AD 250*30 mm*5 um; Mobile phase A: Supercritical
CO.sub.2; Mobile phase B: IPA+NH.sub.3.H.sub.2O; Gradient: B/A:
75:25) to give the following individual single compounds:
Compound 261,
N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-((2R
or 2S, 3R or
3S)-3-methoxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-c-
arboxamide (Major Isomer Pair; Peak 1): .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 8.173-8.157 (m, 1H), 8.140-8.116 (m, 1H),
7.582-7.555 (m, 1H), 6.968-6.936 (m, 1H), 5.927 (s, 1H),
4.707-4.609 (m, 2H), 4.348 (s, 1H), 3.892 (s, 3H), 2.869 (s, 3H),
2.788 (s, 3H), 2.173 (s, 3H), 1.644-1.627 (d, 3H), 1.263-1.249 (d,
3H).
Compound 266,
N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-((2R
or 2S, 3R or
3S)-3-methoxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-c-
arboxamide (Major Isomer Pair; Peak 2): .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 8.179-8.163 (m, 1H), 8.143-8.120 (m, 1H),
7.558-7.531 (m, 1H), 6.986-6.954 (m, 1H), 5.931 (s, 1H),
4.702-4.605 (m, 2H), 3.897 (s, 3H), 2.892 (s, 3H), 2.789 (s, 3H),
2.189 (s, 3H), 1.647-1.629 (d, 3H), 1.267-1.252 (d, 3H).
Compound 267,
N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-((2R
or 2S, 3R or
3S)-3-methoxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-c-
arboxamide (Minor Isomer Pair; Peak 1): .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 8.174-8.162 (d, 1H), 8.111-8.094 (d, 1H),
7.551-7.526 (m, 1H), 6.993-6.961 (m, 1H), 5.935 (s, 1H),
4.683-4.579 (m, 2H), 3.887 (s, 3H), 3.442 (s, 3H), 2.753 (s, 3H),
2.194 (s, 3H), 1.695-1.678 (d, 3H), 0.781-0.768 (d, 3H).
Compound 302,
N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-((2R
or 2S, 3R or
3S)-3-methoxybutan-2-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridine-3-c-
arboxamide (Minor Isomer Pair; Peak 2): .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 8.177-8.166 (d, 1H), 8.122-8.104 (d, 1H),
7.587-7.562 (m, 1H), 6.984-6.952 (m, 1H), 5.933 (s, 1H),
4.698-4.591 (m, 2H), 4.426 (s, 2H), 3.983 (s, 3H), 3.448 (s, 3H),
2.764 (s, 3H), 2.180 (s, 3H), 1.701-1.684 (d, 3H), 0.786-0.772 (d,
3H).
Example 14
Synthesis of
(.+-.)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-1--
(1-phenylethyl)-1H-pyrrolo[2,3-c]pyridine-3-carboxamide
Step 1: 1-(3-methoxyphenyl)ethanol
##STR00080## To a stirred solution of 3-Amino-4-picoline (7 g, 64.8
mmol) in anhydrous THF (200 mL), sec-BuLi (150 mL, 1.3M in
cyclohexane, 194 mmol) was added dropwise over 20 minutes at
-78.degree. C. The solution was warmed to room temperature and
stirred at 3 hours. Ethyl acetate (2.3 g, 25.9 mmol) was added
dropwise into the reaction at -78.degree. C. and the mixture was
stirred at the same temperature for 2 hours. Methanol (50 mL) was
added dropwise into the reaction over 10 minutes. The mixture was
warmed to room temperature and stirred for 1 hour. A half-saturated
NH4Cl (250 mL) was added. The mixture was extracted with EA. The
combined organic layers were washed with brine, dried and
concentrated to afford the crude product. The crude product was
purified by silica gel chromatography (petroleum ether/ethyl
acetate=10:1) to afford 2-methyl-1H-pyrrolo[2,3-c]pyridine (2.5 g,
73.5%).
Step 2:
2,2,2-trichloro-1-(2-methyl-1H-pyrrolo[2,3-c]pyridin-3-yl)ethanone
##STR00081## To a stirred solution of
2-methyl-1H-pyrrolo[2,3-c]pyridine (2.5 g, 18.9 mmol) and aluminum
chloride (5 g, 37.8 mmol) in DCM (100 mL), trichloroacetylchloride
(4.1 g, 22.7 mmol) was added dropwise into the reaction over 0.5
hours at room temperature. After stirring 2 hours, the reaction was
cooled to 0.degree. C. and was quenched with water (100 mL). The
resulting precipitate was isolated by filtration to afford
2,2,2-trichloro-1-(2-methyl-1H-pyrrolo[2,3-c]pyridin-3-yl)ethanone
which was used for next step without further purification. Assumed
100% yield. (5.24 g).
Step 3: Methyl 2-methyl-1H-pyrrolo[2,3-c]pyridine-3-carboxylate
##STR00082## A mixture of
2,2,2-trichloro-1-(2-methyl-1H-pyrrolo[2,3-c]pyridin-3-yl)ethanone
(5.24 g, 18.9 mmol) and KOH (1.2 g, 20.9 mmol) in MeOH (100 mL) was
stirred at room temperature for 16 hour. The reaction mixture was
concentrated to remove MeOH, the residue was partitioned between EA
and Water. The organic layer was washed with brine, dried and
concentrated to afford methyl
2-methyl-1H-pyrrolo[2,3-c]pyridine-3-carboxylate (3 g, 83%).
Step 4: Methyl methyl
2-methyl-1-(1-phenylethyl)-1H-pyrrolo[2,3-c]pyridine-3-carboxylate
##STR00083## A mixture of methyl
2-methyl-1H-pyrrolo[2,3-c]pyridine-3-carboxylate (550 mg, 2.89
mmol) and sodium hydride (200 mg, 4.34 mmol) in
N,N-dimethylformamide (3.0 mL) was stirred at room temperature for
0.5 hour, and then (1-bromoethyl)benzene (589 mg, 3.18 mmol) was
added. The mixture was stirred at room temperature for 3 hours. The
reaction mixture was poured into saturated NH.sub.4Cl and extracted
with ethyl acetate. Organic layers were combined and concentrated
to give a residue. The residue was purified by chromatography
(petroleum ether/ethyl acetate=5:1) to give methyl
2-methyl-1-(1-phenylethyl)-1H-pyrrolo[2,3-c]pyridine-3-carboxylate
(800 mg, 94%).
Step 5:
2-methyl-1-(1-phenylethyl)-1H-pyrrolo[2,3-c]pyridine-3-carboxylic
acid
##STR00084## To a mixture of methyl
2-methyl-1-(1-phenylethyl)-1H-pyrrolo[2,3-c]pyridine-3-carboxylate
(800 mg, 2.72 mmol) and KOH (1.5 g, 27.2 mmol) in (15 mL) and water
(5 mL) was refluxed for 2 hours. The mixture was adjust PH to 2 by
10% HCl and extracted with EA. The combined organic layers were
washed with brine, dried and concentrated to afford the crude
product. The crude product was used into the next step without more
purification. 100% yield. (760 mg).
Step 6:
(.+-.)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-me-
thyl-1-(1-phenylethyl)-1H-pyrrolo[2,3-c]pyridine-3-carboxamide
(Compound 203)
##STR00085## A mixture of
2-methyl-1-(1-phenylethyl)-1H-pyrrolo[2,3-c]pyridine-3-carboxylic
acid (280 mg, 1.0 mmol) was added HATU (456 mg, 1.2 mmol), TEA (1
g, 10 mmol) and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (182
mg, 1.2 mmol) in anhydrous dichloromethane (30 mL) was stirred at
room temperature for 16 hours. To the reaction mixture was added
water (10 mL), extracted with dichloromethane (30 mL.times.2). The
organic layers were combined and concentrated to give a residue.
The residue was rereystallized from MeCN to afford compound
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-1-(1-phen-
ylethyl)-1H-pyrrolo[2,3-c]pyridine-3-carboxamide as an off-white
solid (80 mg, 21.6%). LRMS (M+H.sup.+) m/z: calcd 414.21. found
414. .sup.1H NMR (400 MHz, Methanol-d4) .delta.: 8.84 (s, 1H), 8.16
(d, J=7.6 Hz, 1H), 8.03 (d, J=6.8 Hz, 1H), 7.44-7.37 (m, 5H), 6.09
(s, 1H), 6.01-5.99 (m, 1H), 4.49 (s, 2H), 2.73 (s, 3H), 2.38 (s,
3H), 2.22 (s, 3H), 2.06 (d, J=7.2 Hz, 3H).
The compounds shown in the following table were prepared according
to the general procedure outlined in this example using the
appropriate starting materials and modifications. Structures are
shown in FIG. 1.
TABLE-US-00022 Compound Name NMR m/z 240 (.+-.)-N-((4,6-dimethyl-2-
(400 MHz, CHLOROFORM-d) .delta. ppm 1.63 383
oxo-1,2-dihydropyridin- (br. s., 3 H) 2.21 (s, 3 H) 2.41 (s, 3 H)
2.73 (s, 3-yl)methyl)-1-(1- 3 H) 3.24 (s, 3 H) 3.72 (dd, J = 9.81,
5.40 Hz, methoxypropan-2-yl)-2- 1 H) 3.80-3.88 (m, 1 H) 4.60 (d, J
= 5.95 Hz, methyl-1H-pyrrolo[2,3- 2 H) 4.71 (dd, J = 13.23, 7.06
Hz, 1 H) 5.92 (s, c]pyridine-3- 1 H) 7.31 (d, J = 5.73 Hz, 1 H)
7.38 (br. s., 1 carboxamide H) 8.26 (d, J = 5.29 Hz, 1 H) 9.09 (br.
s., 1 H) 11.07 (br. s., 1 H) 243 (.+-.)-N-((4-methoxy-6- (400 MHz,
CHLOROFORM-d) .delta. ppm 1.62 431 methyl-2-oxo-1,2- (br. s., 3 H)
2.26 (s, 3 H) 2.75 (s, 3 H) 3.25 (s, dihydropyridin-3- 3 H) 3.72
(dd, J = 9.81, 5.40 Hz, 1 H) 3.80- yl)methyl)-2-methyl-1- 3.87 (m,
1 H) 3.90 (s, 3 H) 4.65 (d, J = 5.29 (1-phenylethyl)-1H- Hz, 2 H)
4.71 (dd, J = 13.78, 6.95 Hz, 1 H) pyrrolo[2,3-c]pyridine- 5.93 (s,
1 H) 7.32 (br. s., 1 H) 7.50 (br. s., 1 3-carboxamide H) 8.25 (br.
s., 1 H) 9.11 (br. s., 1 H)
Example 15
General Procedures for Synthesizing Other Compounds of the
Invention
General Procedure A: Indole Alkyation
##STR00086## To a cooled (0.degree. C.) solution of NH indole ester
(1 equivalent) in N,N-dimethylformamide (volume to make
concentration 0.4M) was added sodium hydride (60% w/w, 1.1
equivalents relative to indole). The resultant mixture was stirred
for 15 minutes. Then RX (2 equivalents) was added and the reaction
was allowed to warm to room temperature. The reaction was
maintained at ambient temperature for 12 hours. The reaction
mixture was poured into saturated ammonium chloride solution (100
mL) with stirring. The mixture was extracted with ethyl acetate
(200 mL.times.2) and the combined organic phase was washed with
brine, dried over magnesium sulfate, filtered, and concentrated to
give crude product which was purified by column chromatography
(silica gel, petroleum ether/ethyl acetate=20:1) to afford the
desired alkylated Indole ester product.
General Procedure B: Saponification of Alkylated Indole Ester
##STR00087## To a solution of alkylated Indole ester (1 equivalent)
in tetrahydrofuran:methanol:water (2.5:5:1, volume to make
concentration 0.05M) was added lithium hydroxide (4 equivalents).
The resultant reaction mixture was stirred at 60.degree. C. for 48
hours. The mixture was concentrated in vacuo. Then the residue was
diluted with water (40 mL) and slowly acidified with 1N hydrogen
chloride to pH=4-5. The mixture was extracted with ethyl acetate
(100 mL.times.3). The combined organic layers were washed with
brine, dried over magnesium sulfate, filtered and concentrated to
give crude indole acid, which was used in the subsequent step
without additional purification.
General Procedure C: Amide Bond Formation
##STR00088## To a solution of Indole acid (1 equivalent) in
dichloromethane (volume to make concentration 0.05M) were added
1-hydroxybenzotriazole (1.5 equivalents),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5
equiv.) and triethylamine (3 equiv.). The resultant mixture was
stirred at room temperature for 30 minutes. Then Pyridone amine
(1.2 equiv.) was added and the resultant mixture was stirred at
room temperature for 16 hours. Water (50 mL) was added to the
mixture. The mixture was extracted with dichloromethane (100
mL.times.2). The organic layer was concentrated in vacuo to provide
crude product which was purified by column chromatography (silica
gel, dichloromethane/methanol=20:1) to afford the target
compound.
General Procedure D: Chiral Chromatography
Separation of chiral compounds was accomplished via normal phase
HPLC or SFC (supercritical carbon dioxide fluid chromatography).
Separated compounds were typically>95% ee. The absolute
configuration of chiral centers was not determined.
General Procedure L: Sulphonylation
##STR00089## To a solution of Chiral amine (1 equiv.) in
dichloromethane (volume to make concentration 0.1M) was added
triethylamine (4 equiv.) at 18.degree. C. under N.sub.2. The
reaction was cooled to 0.degree. C. and methanesulfonyl chloride
(1.5 equiv.) was added. The reaction was stirred at 0.degree. C.
for 1 h. Then the mixture was concentrated in vacuo and methanol
and potassium carbonate were added and the reaction was stirred for
another 1 h. The mixture was filtered and the crude product was
purified by preparative-HPLC.
The table below lists compounds of the invention and which of the
above general methods was used in their synthesis. Structures of
these compounds are set forth in FIG. 1.
TABLE-US-00023 General Com- Methods pound Used and Notes Name NMR
data m/z 370 General .sup.1H NMR (CDCl3, 400 MHz) 517 procedure L
.delta. 12.12 (s, 1H), 9.20 (s, 1H), on (.+-.)-N-((4- 8.78 (s, 1H),
7.68 (s, 1H), methoxy-6- 5.97 (s, 1H), 4.70-4.58 (m, methyl-2-oxo-
2H), 3.91 (s, 4H), 3.62 (d, J = 1,2-dihydro- 12.0 Hz, 1H),
2.76-2.67 (m, pyridin-3- 7H), 2.45 (dd, J1 = 2.0 Hz, J, =
yl)methyl)-6- 11.6 Hz, 1H), 2.32 (s, 3H), methyl-7-(1- 2.10 (t, J =
12.0 Hz, 1H), (piperidin-4- 1.67 (d, J = 6.8 Hz, 4H), yl)ethyl)-7H-
1.48-1.40 (m, 1H), 1.37-1.29 pyrrolo[2,3- (m, 1H), 1.27-1.19 (m,
1H), d]pyrimidine- 0.9-0.78 (m, 1H). 5-carboxamide
Example 16
IC.sub.50 Measurements for Inhibitors Using EZH2
EZH2 Assay:
Assays were carried out by mixing rPRC2 together with biotinylated
oligonucleosome substrates in the presence of the radio-labeled
enzyme co-factor, S-adenosyl-L-methionine (.sup.3H SAM) (Perkin
Elmer) and monitoring the enzymatically mediated transfer of
tritiated methyl groups from .sup.3H SAM to histone lysine
residues. The amount of resulting tritiated methyl histone product
was measured by first capturing the biotinylated oligonucleosomes
in streptavidin (SAV) coated FlashPlates (Perkin Elmer), followed
by a wash step to remove un-reacted .sup.3H SAM, and then counting
on a TopCount NXT 384 well plate scintillation counter (Perkin
Elmer). The final assay conditions for EZH2 were as follows: 50 mM
Tris Buffer pH 8.5, 1 mM DTT, 69 .mu.M Brij-35 detergent, 5.0 mM
MgCl.sub.2, 0.1 mg/mL BSA, 0.2 .mu.M .sup.3H SAM, 0.2 .mu.M
biotinylated oligonucleosomes, 3.6 .mu.M H3K27me3 peptide and 2 nM
EZH2.
Compound IC.sub.50 measurements were obtained as follows: Compounds
were first dissolved in 100% DMSO as 10 mM stock solutions. Ten
point dose response curves were generated by dispensing varying
amounts of the 10 mM compound solution in 10 wells of the 384 well
plate (Echo; Labcyte), pure DMSO was then used to backfill the
wells to insure all wells have the same amount of DMSO. A 12.5
.mu.L volume of the HMT enzyme, H3K27me3 peptide and
oligonucleosome substrate in assay buffer was added to each well of
the assay plate using a Multidrop Combi (Thermo-Fisher). Compounds
were pre-incubated with the enzyme for 20 min, followed by
initiation of the methyltransferase reaction by addition of 12.5
.mu.L of .sup.3H SAM in assay buffer (final volume=25 .mu.L). The
final concentrations of compounds ranged from a top default
concentration of 80 .mu.M down to 0.16 .mu.M in ten 2-fold dilution
steps. Reactions were carried out for 60 minutes and quenched with
20 .mu.L per well of 1.96 mM SAH, 50 mM Tris pH 8.5, 200 mM EDTA.
Stopped reactions were transferred to SAV coated Flash-plates
(Perkin Elmer), incubated for 120 min, washed with a plate washer,
and then read on the TopCount NXT (1.0 min/well) to measure the
amount of methyl histone product formed during the reaction. The
amount of methyl histone product was compared with the amount of
product formed in the 0% and 100% inhibition control wells allowing
the calculation of % Inhibition in the presence of the individual
compounds at various concentrations. IC.sub.50's were computed
using a 4 parameter fit non-linear curve fitting software package
(XLFIT, part of the database package, ActivityBase (IDBS)) where
the four parameters were IC.sub.50, Hill slope, pre-transitional
baseline (0% INH), and post-transitional baseline (100% INH); with
the latter two parameters being fixed to zero and 100%,
respectively, by default.
Assay for Y641N EZH2 was performed as above using reconstituted
H3K27Me2 oligonucleosomes as substrate.
Table 2 shows the activity of selected compounds of this invention
in the EZH2 and Y641N EZH2 activity inhibition assay. IC.sub.50
values are reported as follows: "A" indicates an IC.sub.50 value of
less than 100 nM; "B" indicates an IC.sub.50 value of 100 nM to 1
.mu.M; "C" indicates an IC.sub.50 value of greater than 1 .mu.M and
less than 10 .mu.M for each enzyme; "D" indicates an IC.sub.50
value of greater than 10 .mu.M for each enzyme; and "*(X .mu.M)"
indicates that no inhibition was observed at the highest
concentration (i.e., X .mu.M) of compound tested.
TABLE-US-00024 TABLE 2 IC50 Values for Compounds of Formula I
against EZH2 and Y641N EZH2 Mutant Enzymes. Compound Y641N No. EZH2
IC.sub.50 EZH2 IC.sub.50 183 A A 204 A B 211 A B 212 A B 217 B B
218 A A 219 A A 223 A B 224 A A 229 C D 230 A B 231 A B 234 C D 235
B C 236 *(0.5 .mu.M) *(10 .mu.M) 240 A B 241 A B 243 A B 252 A B
253 A B 256 A B 257 B C 261 A A 266 B B 267 A B 273 A A 275 A A 277
A A 280 B C 284 A B 288 A A 290 A B 293 A B 294 A A 298 A A 299 A A
300 A A 302 B C 304 A A 306 A A 307 A A 308 B B 310 A A 313 A A 314
A A 316 A A 317 A A 321 A A 327 A A 335 A A 336 A A 337 A A 341 A A
342 A A 343 A A 344 A A 345 A A 346 A A 347 A A 352 A A 355 A A 356
A A 357 A A 358 A A 359 A A 360 A A 362 A A 363 A A 364 A B 365 A A
366 A A 367 A A 368 A A 369 A A 370 A A 373 A A 375 A A 376 A A 377
A A
Example 17
EC50 Measurements for Inhibitors in HeLa Cell Assays
H3K27me3 MSD Hela Assay.
Trypsinized HeLa cells were counted and diluted in 10% DMEM (Life
Technologies, Cat. #10569) to 5000 cells/75 .mu.L. Seventy-five
.mu.L of cells were place in each well of a 96-well flat-bottomed
plate and incubated at 37.degree. C. for 4 hours. Twenty-five .mu.L
of test compound (at various concentrations) was added to the cells
and incubation continued at 37.degree. C. for 96 hours. Media was
then removed and the cells rinsed once with ice cold PBS. Forty
.mu.L of ice-cold MSD Buffer AT (10 mM HEPES, pH 7.9, 5 mM
MgCl.sub.2, 0.25M sucrose, Benzonase (1:10000), 1% Triton X-100
supplemented with fresh 1.times. Protease Inhibitor cocktail and 1
mM 4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF))
was added to each well and the plates placed on ice for 30 minutes.
Ten .mu.L of 5M NaCl was then added to each well and incubation on
ice continued for another 15 minutes. The material in each well was
suspended pipetting up and down and then transferred to a new 96
well plate. The emptied wells were rinsed with 150 uL ice-cold 20
mM Tris pH 7.5, 1 mM EDTA, 1 mM EGTA, supplemented with fresh
1.times. Protease Inhibitor cocktail and 1 mM AEBSF ("NO salt NO
detergent buffer) and transferred to the respective wells in the
new plate. Three hundred .mu.L of NO Salt NO detergent buffer was
then added to each well of lysates and the plates frozen at
-80.degree. C.
On the same day, an appropriate number of MSD standard bind 96-well
plates were coated with 30 .mu.L/well of total H3 capture antibody
(Millipore, Cat # MAB3422) at 1 .mu.g/mL concentration in PBS. The
antibody solution was evenly distributed first by tapping gently on
the sides of the plates and then by shaking the plates for a few
minutes at 1000 rpm. Antibody coated plates were stored at
4.degree. C. overnight.
The next day the lysates are thawed to RT. The antibody coated MSD
plates are washed 3.times. with TBS-T (Tris-buffered saline (Fisher
Scientific, Cat #BP2471-1)+0.2% Tween-20). One-hundred fifty .mu.L
of 5% Blocker A in TBS-T is added to each well. The wells are
covered and shaken on a shaker at RT for one hour. The Blocker A
step is repeated a second time. After removing the blocker, 25
.mu.L of cell lysate is transferred into each antibody coated well.
The plates are shaken for 2 hours at RT, the lysate removed and the
plates again washed with Blocker A in TBS-T. Twenty-five .mu.L of
appropriate freshly prepared antibody mix (including both primary
and secondary antibodies) is added to each well and the plates
shaken for 1 hour at RT. The antibody mix used was one (or both) of
those indicated in the table below:
TABLE-US-00025 Concen- Anti-rabbit 1% tration Primary detection
blocker Ab (.mu.g/mL) Ab (.mu.L) Ab (.mu.L) A (.mu.L) H3K27me3 33
37.88 5.00 5000 H3 12 52.08 5.00 5000
Both H3 antibodies were obtained from Cell Signalling (Cat #s 4499
and 9733). The goat anti-rabbit antibody was obtained from
Meso-Scale Discovery (Cat #R32AB-1).
The antibody mix was then removed and the wells washed with Blocker
A. One hundred-fifty .mu.L of freshly prepared 1.times.MSD Read
Buffer (Meso-Scale Discovery; Cat #R927C-2) was then added to each
well and the plates read on a MSD Sector 2400 Plate Reader.
Data was analyzed using Assay Assistant (Constellation
Pharmaceuticals In-house product) and Activity Base (IDBS Ltd,
Surrey, UK) template. Data files were imported to Assay Assistant
and assay conditions were specified. A unique Analysis ID was
created and the data files exported to Activity Base. An analysis
template was created on Activity Base to measure dose-dependent
inhibition of H3K27me3 mark and cell viability respectively.
Readout of DMSO wells were used to normalize the data. Resulting
curves were fitted using Activity base software Model 205 (IDBS
Ltd, Surrey, UK). The data was checked for quality, validated and
integrated in excel format using SARview (IDBS Ltd, Surrey,
UK).
H3K27me3 Alpha Hela Assay (AlphaLISA).
Ten different doses of each test compound (in a series of 3-fold
dilutions) were plated in duplicate 384-well tissue culture treated
plates (Catalog #781080; Greiner Bio One; Monroe, N.C.). Hela cells
grown in culture were trypsinized and counted using a Countess.RTM.
cell counter (Catalog #C10281; Life Technologies, Grand Island,
N.Y.). Cell were diluted to 67,000 cells per mL in 10% DMEM
(Catalog #10569-010 Life Technologies, Grand Island, N.Y.) and 15
.mu.L (1,000 cells) were plated into each well using the Biotek
MicroFlo.TM. Select Dispenser (BioTek Instruments, Inc. Vermont,
USA),) of the 384-well plate. Plates were incubated at 37.degree.
C./5% CO.sub.2 for 72 hrs. One of the duplicate plates was
processed for HeLa assay and the other for viability.
To the plate processed for AlphaLISA was added 5 .mu.L per well
Cell-Histone Lysis buffer (1.times.) (Catalog #AL009F1 Perkin
Elmer; Waltham, Mass.) and the plate was incubated at RT for 30
minutes on a plate shaker with low speed (Model #4625-Q Thermo
Scientific; Waltham, Mass.). Then, 10 .mu.L per well Histone
Extraction buffer (catalog #AL009F2; Perkin Elmer; Waltham, Mass.)
was added and the plate further incubated at RT for 20 min on plate
shaker with low speed. To each well was then added 10 .mu.L per
well of a 5.times. mix of anti-K27me3 acceptor beads plus
Biotinylated anti-Histone H3 (C-ter) Antibody (diluted to 3 nM
final) (Catalog #AL118 Perkin Elmer; Waltham, Mass.). Dilution of
the acceptor beads and then anti-Histone H3 was with 1.times.
Histone Detection buffer (Catalog #AL009F3 Perkin Elmer; Waltham,
Mass.) which was produced diluted from the 10.times. stock
provided. The plate was sealed with an aluminum plate sealer and
incubated at 23.degree. C. for 60 min. We then added 10 .mu.L
5.times. solution of Streptavidin Donor beads (Catalog #6760002
Perkin Elmer; Waltham, Mass.) (20 .mu.g/mL final in 1.times.
Histone Detection Buffer), sealed the plate with Aluminum plate
sealer and incubated at 23.degree. C. for 30 min. The plates were
then read using an EnVision-Alpha Reader (model #2104 Perkin Elmer;
Waltham, Mass.).
Cell viability was assayed by adding 15 .mu.L of Cell Titer Glo
((Catalog #G7571 Promega Madison, Wis.) to each well with cells
with media. The plates were incubated foat RT for 15-20 minutes on
a plate shaker at low speed. The plates were then read using an
EnVision-Alpha Reader (model #2104 Perkin Elmer; Waltham,
Mass.).
Data from both assays was analyzed using Assay Assistant
(Constellation Pharmaceuticals In-house product) and Activity Base
(IDBS Ltd, Surrey, UK) template. Data files were imported to Assay
Assistant and assay conditions were specified. A unique Analysis ID
was created and the data files exported to Activity Base. An
analysis template was created on Activity Base to measure
dose-dependent inhibition of H3K27me3 mark and cell viability
respectively. Readout of DMSO wells were used to normalize the
data. Resulting curves were fitted using Activity base software
Model 205 (IDBS Ltd, Surrey, UK). The data was checked for quality,
validated and integrated in excel format using SARview (IDBS Ltd,
Surrey, UK).
Table 3 shows the activity of selected compounds of this invention
in the two different HeLa cell assays described above. EC.sub.50
values are reported as follows: "A" indicates an EC.sub.50 value of
less than 400 nM; "B" indicates an EC.sub.50 value of 400 nM to 2
.mu.M; "C" indicates an EC.sub.50 value of greater than 2 .mu.M and
less than 10 .mu.M for each enzyme; "D" indicates an EC.sub.50
value of greater than 10 .mu.M for each enzyme; and "*(X .mu.M)"
indicates that no inhibition was observed at the highest
concentration (i.e., X .mu.M) of compound tested.
TABLE-US-00026 TABLE 3 Ec50 Values for Selected Compounds of the
Invention In Hela Cells Expressing H3k27 Mutant EZH2. H3K27me3
H3K27me3 _Alpha_ _MSD_ Compound HeLa HeLa_ No. (EC50) (EC50) 204 B
211 B 212 B 218 A 219 B 224 A A 230 B 240 C 241 B 243 C 253 A 256 B
261 A A 273 A 284 B 288 A B 294 A A 298 A A 300 A A 304 A A 310 A A
313 A A 314 A 315 D 316 B 317 A 321 A 327 A 335 A 336 A 337 A 341 A
342 A 343 B 344 A 345 A 346 A 347 B 352 B 355 A 356 A 357 A 358 B
359 B 360 C 362 A 363 A 364 *(3.33 .mu.M) 365 A 366 B 367 A 368 A
369 A 370 *(3.33 .mu.M) 373 A 374 A 375 A 376 NaN 377 B
Example 18
Tumor Growth Inhibition Analysis
The anti-tumor efficacy of Compound 362 and 365 in the subcutaneous
Karpas422 human lymphoma xenograft model in female CB-17 SCID mice
was as follows.
Animals
Species: Mus Musculus
Strain: CB-17 SCID mice
Age: 6-8 weeks
Sex: female
Body weight: 18-22 g
Number of animals: 50 mice plus spare
Animal supplier: Shanghai SLAC Laboratory Animal Co., LTD.
Cell Culture
The Karpas422 tumor cells were maintained in vitro as suspension
culture in RPMI1640 medium supplemented with 10% heat inactivated
fetal calf serum at 37.degree. C. in an atmosphere of 5% CO.sub.2
in air. The tumor cells were routinely subcultured twice weekly.
The cells growing in an exponential growth phase were harvested and
counted for tumor inoculation.
Tumor Inoculation
Each mouse was inoculated subcutaneously at the right flank with
the Karpas422 tumor cells (5.times.10.sup.6) in 0.2 ml of PBS with
Matrigel (1:1) for tumor development. Day 23 after tumor
inoculation was as day 0 after the start of treatment when the
average tumor size reached approximately 300 mm.sup.3. Each group
consisted of 10 tumor-bearing mice.
Tumor Measurements
Tumor size was measured three times weekly in two dimensions using
a caliper, and the volume was expressed in mm.sup.3 using the
formula: V=0.536a.times.b.sup.2 where a and b are the long and
short diameters of the tumor, respectively. The tumor size was then
used for calculations of T/C values. The T/C value (in percent) is
an indication of antitumor effectiveness; T and C are the mean
volumes of the treated and control groups, respectively, on a given
day. TGI was calculated for each group using the formula: TGI
(%)[1-(Ti-T0)/(Vi-V0)].times.100; Ti is the average tumor volume of
a treatment group on a given day, T0 is the average tumor volume of
the treatment group on the day of treatment start, Vi is the
average tumor volume of the vehicle control group on the same day
with Ti, and V0 is the average tumor volume of the vehicle group on
the day of treatment start.
Experimental Endpoint and Sample Collection
1). Plasma, tumor and muscle in EPZ-6438 group were collected on
day 16 after the start of treatment at 6 h after dosing. Plasma,
tumor and muscle in vehicle, CPI-524369, CPI-524416 and CPI-591780
groups were collected on day 25 after the start of treatment at 1 h
after dosing. 2). All the blood was taken from each animal with
EDTA-K2 as anticoagulant. Plasma was divided into two parts. The
first part was for PK analysis; the second part was frozen for
backup. 3). Tumor was divided into three parts. The first part was
snap-frozen for PK; the second part was snap-frozen for PD
analysis; the third part was frozen for backup. 4). Muscle was
divided into two parts. The first part was snap-frozen for PK; the
second part was frozen for backup.
Tumor Growth Inhibition Analysis
TABLE-US-00027 TABLE 4 Tumor growth inhibition calculation for
Compounds 362 and 365 in the karpass422 xenograft model calculated
based on tumor volume measurements on day 25 or day 16 after the
start of treatment Tumor Size (mm3).sup.a T/C.sup.b TGI Treatment
on day 25 (%) (%) Significance.sup.d Vehicle 1704 .+-. 123 -- -- --
Compound 362 385 .+-. 66 22.59 92.75 *** (160 mg/kg) Compound 365
319 .+-. 67 18.72 97.25 *** (160 mg/kg) Note: .sup.aMean .+-. SEM.
.sup.bTumor Growth Inhibition is calculated by dividing the group
average tumor volume for the treated group by the group average
tumor volume for the control group (T/C). For a test article to be
considered to have anti-tumor activity, T/C must be 0.5 or less.
.sup.dStatistically significant difference (one-way ANOVA), vs
vehicle: *** p < 0.001.
* * * * *
References