U.S. patent application number 15/999664 was filed with the patent office on 2020-08-27 for tetracyclic pyridone compounds as antivirals.
The applicant listed for this patent is Novartis AG. Invention is credited to Jiping FU, Xianming JIN, Patrick LEE, Peichao LU, Joseph Michael YOUNG.
Application Number | 20200270265 15/999664 |
Document ID | / |
Family ID | 1000004841253 |
Filed Date | 2020-08-27 |
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United States Patent
Application |
20200270265 |
Kind Code |
A1 |
FU; Jiping ; et al. |
August 27, 2020 |
TETRACYCLIC PYRIDONE COMPOUNDS AS ANTIVIRALS
Abstract
The invention provides compounds of Formula (I) ##STR00001## as
described herein, along with pharmaceutically acceptable salts,
pharmaceutical compositions containing such compounds, and methods
to use these compounds, salts and compositions for treating viral
infections, particularly infections caused by hepatitis B virus,
and reducing the occurrence of serious conditions associated with
HBV.
Inventors: |
FU; Jiping; (Danville,
CA) ; JIN; Xianming; (San Ramon, CA) ; LEE;
Patrick; (Walnut Creek, CA) ; LU; Peichao;
(Cambridge, MA) ; YOUNG; Joseph Michael; (Pleasant
Hill, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Novartis AG |
Basel |
|
CH |
|
|
Family ID: |
1000004841253 |
Appl. No.: |
15/999664 |
Filed: |
February 16, 2017 |
PCT Filed: |
February 16, 2017 |
PCT NO: |
PCT/EP2017/053568 |
371 Date: |
August 20, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62297590 |
Feb 19, 2016 |
|
|
|
62434658 |
Dec 15, 2016 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 491/147 20130101;
C07D 471/04 20130101; C07D 519/00 20130101; A61P 31/20
20180101 |
International
Class: |
C07D 491/147 20060101
C07D491/147; C07D 471/04 20060101 C07D471/04; C07D 519/00 20060101
C07D519/00; A61P 31/20 20060101 A61P031/20 |
Claims
1. A compound of formula (I): ##STR00103## wherein: R.sup.1 is H,
halo, or C.sub.1-C.sub.3 alkyl; R.sup.2 is H, halo, CN,
C.sub.1-C.sub.3 alkyl, C.sub.1-C.sub.3 haloalkyl, or
C.sub.1-C.sub.3 alkoxy; R.sup.3 is OH, halo, CN, C.sub.1-C.sub.3
alkyl, C.sub.3-C.sub.6 cycloalkyl, C.sub.1-C.sub.3 haloalkyl,
C.sub.1-C.sub.3 alkoxy, or C.sub.1-C.sub.3 haloalkoxy; R.sup.4 is
selected from R.sup.11, --OR.sup.11, --SR.sup.1, and --NRR.sup.11;
R.sup.11 is C.sub.1-C.sub.4 alkyl, C.sub.3-C.sub.6 cycloalkyl,
oxetanyl, tetrahydrofuranyl, or tetrahydropyranyl, each of which is
optionally substituted with up to three groups selected from halo,
CN, --OR, C.sub.1-C.sub.3 haloalkoxy, --NR.sub.2, and a 4-7
membered heterocyclic group containing one or two heteroatoms
selected from N, O and S as ring members that is optionally
substituted with one or two groups selected from halo, oxo, CN, R,
--OR, and --NR.sub.2; R is independently selected at each
occurrence from H and C.sub.1-C.sub.3 alkyl optionally substituted
with one to three groups selected from halo, --OH, C.sub.1-C.sub.3
alkoxy, oxo, CN, --NH.sub.2, --NH(C.sub.1-C.sub.3 alkyl),
--N(C.sub.1-C.sub.3 alkyl).sub.2, and cyclopropyl; and two R groups
directly attached to the same atom, which may be C or N, can
optionally be taken together to form a 3-6 membered ring that can
optionally contain an added heteroatom selected from N, O and S as
a ring member, and can be substituted by up to two groups selected
from --OH, oxo, C.sub.1-C.sub.3 alkyl, and C.sub.1-C.sub.3 alkoxy;
R.sup.5 is H, halo, CN, C.sub.1-C.sub.3 alkyl, or C.sub.1-C.sub.3
haloalkyl; R.sup.6 is H, halo, C.sub.1-C.sub.3 alkoxy, or
C.sub.1-C.sub.6 alkyl; R.sup.7 is II, halo, C.sub.1-C.sub.3 alkoxy,
or C.sub.1-C.sub.6 alkyl; R.sup.8 is H or C.sub.1-C.sub.6 alkyl;
R.sup.9 taken together with one group selected from R.sup.6,
R.sup.7 and R.sup.8 forms a 3-7 membered cycloalkyl ring or a 3-7
membered heterocyclic ring containing N, O or S as a ring member;
wherein the cycloalkyl or heterocyclic ring is optionally
substituted with up to three groups selected from R, --OR,
--NR.sub.2, halo, CN, COOR, CONR.sub.2, and oxo; W is
--COOR.sup.10, --C(O)NH--SO.sub.2R, --C(O)NH--SO.sub.2NR.sub.2,
5-tetrazolyl, or 1,2,4-oxadiazol-3-yl-5(4H)-one; R.sup.10 is H or
C.sub.1-C.sub.6 alkyl that is optionally substituted with one or
two groups selected from halo, --OR, oxo, CN, --NR.sub.2, COOR, and
CONR.sub.2; or a pharmaceutically acceptable salt thereof.
2. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein R.sup.1 is H.
3. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein R.sup.2 is H or halo.
4. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein R.sup.3 is C.sub.1-C.sub.3 alkoxy
or halo.
5. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein R.sup.4 is --OR.sup.11.
6. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein R.sup.5 is H or halo.
7. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, which is of the formula: ##STR00104##
wherein R.sup.9 taken together with R.sup.7 forms a 3-7 membered
cycloalkyl ring or a 3-7 membered heterocyclic ring containing N, O
or S as a ring member; wherein the cycloalkyl or heterocyclic ring
is optionally substituted with up to three groups selected from R,
--OR, --NR.sub.2, halo, CN, COOR, CONR.sub.2, and oxo; or a
pharmaceutically acceptable salt thereof.
8. The compound according to claim 1, which is of the formula:
##STR00105## wherein R.sup.9 taken together with R.sup.8 forms a
3-7 membered cycloalkyl ring or a 3-7 membered heterocyclic ring
containing N, O or S as a ring member; wherein the cycloalkyl or
heterocyclic ring is optionally substituted with up to three groups
selected from R, --OR, --NR.sub.2, halo, CN, COOR, CONR.sub.2, and
oxo; or a pharmaceutically acceptable salt thereof.
9. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein R.sup.11 is C.sub.1-C.sub.4 alkyl,
optionally substituted with up to two groups selected from halo,
CN, --OR, C.sub.1-C.sub.3 haloalkoxy, and a 4-7 membered
heterocyclic group containing one or two heteroatoms selected from
N, O and S as ring members that is optionally substituted with one
or two groups selected from halo, oxo, CN, R, --OR, and
--NR.sub.2.
10. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein the R.sup.11 is selected from
--CH.sub.2CH.sub.2OMe, --CH.sub.2CH.sub.2CH.sub.2OMe,
--CH.sub.2--OEt, --CH.sub.2CH.sub.2-Q, and
--CH.sub.2CH.sub.2CH.sub.2-Q, where Q is selected from
##STR00106##
11. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein R.sup.9 taken together with one
group selected from R.sup.6, R.sup.7 and R.sup.8 forms a 4-6
membered cycloalkyl ring or a 5-6 membered heterocyclic ring
containing N, O or S as a ring member; wherein the cycloalkyl or
heterocyclic ring is optionally substituted with up to three groups
selected from R, --OR, --NR.sub.2, halo, CN, COOR, CONR.sub.2, and
oxo.
12. The compound according to claim 1, which is selected from:
##STR00107## ##STR00108## ##STR00109## ##STR00110## and the
enantiomers of these compounds; or a pharmaceutically acceptable
salt thereof.
13. The compound of claim 1, which is selected from: ##STR00111##
##STR00112## ##STR00113## or a pharmaceutically acceptable salt
thereof.
14. The compound of claim 1, wherein R.sup.1 is F.
15. A pharmaceutical composition, comprising a compound of claim 1
admixed with at least one pharmaceutically acceptable carrier.
16. A method to treat a hepatitis B infection, which comprises
administering to a patient having a hepatitis B infection a
compound of claim 1 or a pharmaceutical composition of claim
15.
17. The method of claim 16, wherein the compound of claim 1 or the
pharmaceutical composition of claim 15 is used in combination with
an additional therapeutic agent selected from an interferon or
peginterferon, an HBV polymerase inhibitor, a viral entry
inhibitor, a viral maturation inhibitor, a capsid assembly
inhibitor, an HBV core modulator, a reverse transcriptase
inhibitor, a TLR-agonist, or an immunomodulator.
18. A method to inhibit replication of hepatitis B virus, which
comprises contacting the hepatitis B virus, either in vitro or in
vivo, with a compound according to claim 1.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to novel tetracyclic pyridone
compounds that are inhibitors of hepatitis virus replication, and
are thus useful to treat viral infections, and particularly
hepatitis B virus (HBV). The invention provides novel tetracyclic
pyridone compounds as disclosed herein, pharmaceutical compositions
containing such compounds, and methods of using these compounds and
compositions in the treatment and prevention of HBV infections.
BACKGROUND
[0002] Globally, over 240 million people are chronically infected
with hepatitis B virus (HBV), and more than 2 million reside in the
United States alone. Of those chronically infected patients, up to
40 percent will eventually develop complications of liver failure
from cirrhosis or development of hepatocellular carcinoma (HCC).
Hepatitis B virus (HBV) belongs to the family of Hepadnaviridae, a
group of small hepatotropic DNA viruses that replicate through the
reverse transcription of an RNA intermediate. The 3.2-kb HBV genome
in viral particles is in a circular, partially doublestranded DNA
conformation (relaxed circular DNA or rcDNA). The HBV genome
consists of four overlapping open reading frames (ORF), which
encode for the core, polymerase (Pol), envelope, and X proteins.
rcDNA is transcriptionally inert and must be converted into
covalently closed circular DNA (cccDNA) in the nucleus of infected
cells before viral RNAs can be transcribed. cccDNA is the only
template for HBV transcription and, because HBV RNA templates
genomic reverse transcription, its persistence is required for
persistent infection.
[0003] The envelope of HBV comprises a mixture of surface antigen
proteins (HBsAg). The HBsAg coat is a mixture of three overlapping
proteins: all three share a common region, which corresponds to the
smallest of the three proteins (SHBsAg). The mixture consists
mostly of SHBsAg, but also includes Medium HBsAg, which comprises
SHBsAg plus an additional polypeptide segment, and Large HBsAg,
which comprises M HBsAg plus another added polypeptide segment. In
addition to forming the infectious virion particle, the S, M and L
HBsAg proteins also assemble into a subviral particle knows as the
22-nm particle, which is not infectious but contains the same
proteins that envelope the infectious virus particles. Indeed,
these subviral, non-infectious particles have been used as a
vaccine, since they contain the same antigenic surface proteins
that envelope the infectious HBV virion and thus elicit antibodies
that recognize the infectious agent. Interestingly, these subviral
particles greatly outnumber infectious virions, and are believed to
protect the infectious virions from the immune system of the
infected host. By sheer numbers, they may act as decoys,
distracting immune responses from the infectious virus particles,
but in addition they are reported to suppress the function of
immune cells (monocytes, dendritic cells and natural killer cells)
and may thus impair the immune response to HBV. Because these
subviral particles protect infectious HBV from the host immune
system, reducing the level of subviral particles has been
recognized as a viable therapeutic approach. See, e.g.,
WO2015/113990.
[0004] One of the key diagnostic symptoms of chronic HBV is the
high serum levels of the hepatitis B surface antigen (HBsAg).
Clinical data in recent years suggest that sustained virologic
response is often associated with on-treatment HBsAg decline during
the early phase of the treatment as early as week 8, while
sustained exposure to HBsAg and other viral antigens may lead to
HBV-specific immune-tolerance. Chronic HB patients who experienced
larger and faster decreases in serum HBsAg levels achieved
significantly higher rate (.about.40%) of sustained virologic
response as defined by sustained viral control post treatment.
[0005] Current treatment options for HBV include interferon
therapies and nucleoside/nucleotide inhibitors of the viral DNA
polymerase, such as entecavir and tenofovir. These focus on
reduction in the level of viremia and toleration of hepatic
dysfunction, and may have adverse side-effects and also select for
drug-resistant virus variants during long term therapy. More
importantly, these therapies cannot eradicate the intrahepatic HBV
cccDNA pool in chronic hepatitis B patients or limit the
transcription of HBsAg from the pre-existing cccDNA, nor do they
affect the secretion of synthesized HBsAg into patients' blood to
counteract the host innate immune response. As a result, these HBV
treatments are in most cases life-long therapy and discontinuation
often leads to virological relapse. Some compounds have been
reported to reduce serum HBsAg levels but so far have not resulted
in new approved therapeutic agents. See for example WO2015/113990,
WO2015/173164, WO2016/023877, WO2016/071215, and WO2016/128335.
[0006] Accordingly, there remains a need for more effective
treatments for HBV, especially for treating chronic HBV infections.
The invention provides compounds that are believed to operate by
suppression of the secretion of the 22 nm subviral particles
containing HBsAg. These compounds are useful to treat HBV
infections and to reduce the incidence of serious liver disorders
caused by HBV infections. They also exhibit improved properties
relative to prior art compounds having similar biological activity,
such as improved solubility in buffered aqueous systems and lower
predicted propensity for certain adverse effects.
SUMMARY
[0007] The present invention provides novel compounds that inhibit
secretion of HBsAg from cells infected with hepatitis B virus and
thereby reduce viral load and viral replication in patients having
chronic HBV infection. Some of the compounds of the invention, in
addition to being highly effective at suppression of HBsAg levels,
also exhibit improved safety relative to similar compounds known in
the art, such as reduced inhibition of sodium ion channels that can
be indicative of potential cardiotoxicity, reduced drug-drug
interactions, and lower risk of time-dependent cytochrome (CYP)
inhibition. Thus the compounds of the invention are suitable for
treatment of patients with HBV. The invention also provides
pharmaceutical compositions containing the novel compounds as well
as methods to use the compounds and compositions to inhibit
hepatitis B virus replication, and to treat disease conditions
associated with or caused by HBV. Further objects of this invention
are described in the following description and the examples.
[0008] In one aspect, the invention provides compounds of Formula
(I):
##STR00002##
wherein:
[0009] R.sup.1 is H, halo, or C.sub.1-C.sub.3 alkyl;
[0010] R.sup.2 is H, halo, CN, C.sub.1-C.sub.3 alkyl or
C.sub.1-C.sub.3 haloalkyl, C.sub.1-C.sub.3 alkoxy;
[0011] R.sup.3 is OH, halo, CN, C.sub.1-C.sub.3 alkyl,
C.sub.3-C.sub.6 cycloalkyl, C.sub.1-C.sub.3 haloalkyl,
C.sub.1-C.sub.3 alkoxy, or C.sub.1-C.sub.3 haloalkoxy;
[0012] R.sup.4 is selected from R.sup.1, --OR.sup.11, --SR.sup.11,
and --NRR.sup.11;
[0013] R.sup.11 is C.sub.1-C.sub.4 alkyl, C.sub.3-C.sub.6
cycloalkyl, oxetanyl, tetrahydrofuranyl, or tetrahydropyranyl, each
of which is optionally substituted with up to three groups selected
from halo, CN, --OR, C.sub.1-C.sub.3 haloalkoxy, and a 4-7 membered
heterocyclic group containing one or two heteroatoms selected from
N, O and S as ring members that is optionally substituted with one
or two groups selected from halo, oxo, CN, R, --OR, and
--NR.sub.2;
[0014] R is independently selected at each occurrence from H and
C.sub.1-C.sub.3 alkyl optionally substituted with one to three
groups selected from halo, --OH, C.sub.1-C.sub.3 alkoxy, oxo, CN,
--NH.sub.2, --NH(C.sub.1-C.sub.3 alkyl), --N(C.sub.1-C.sub.3
alkyl).sub.2, and cyclopropyl; [0015] and two R groups directly
attached to a single atom can optionally be taken together to form
a 3-6 membered ring that can optionally contain a heteroatom
selected from N, O and S as a ring member, and can be substituted
by up to two groups selected from --OH, oxo, C.sub.1-C.sub.3 alkyl,
and C.sub.1-C.sub.3 alkoxy;
[0016] R.sup.5 is H, halo, CN, C.sub.1-C.sub.3 alkyl, or
C.sub.1-C.sub.3 haloalkyl;
[0017] R.sup.6 is H, halo, C.sub.1-C.sub.3 alkoxy, or
C.sub.1-C.sub.6 alkyl;
[0018] R.sup.7 is H, halo, C.sub.1-C.sub.3 alkoxy, or
C.sub.1-C.sub.6 alkyl;
[0019] R.sup.8 is H or C.sub.1-C.sub.6 alkyl;
[0020] R.sup.9 taken together with one group selected from R.sup.6,
R.sup.7 and R.sup.8 forms a 3-7 membered cycloalkyl ring or a 3-7
membered heterocyclic ring containing N, O or S as a ring member;
wherein the cycloalkyl or heterocyclic ring is optionally
substituted with up to three groups selected from R, --OR,
--NR.sub.2, halo, CN, COOR, CONR.sub.2, and oxo;
[0021] W is --COOR.sup.10, --C(O)NH--SO.sub.2R,
--C(O)NH--SO.sub.2NR.sub.2, 5-tetrazolyl, or
1,2,4-oxadiazol-3-yl-5(4H)-one;
[0022] R.sup.10 is H or C.sub.1-C.sub.6 alkyl that is optionally
substituted with one or two groups selected from halo, --OR, oxo,
CN, --NR.sub.2, COOR, and CONR.sub.2;
[0023] or a pharmaceutically acceptable salt thereof.
DETAILED DESCRIPTION
[0024] For purposes of interpreting this specification, the
following definitions will apply, and whenever appropriate, terms
used in the singular will also include the plural.
[0025] Terms used in the specification have the following meanings
unless the context clearly indicates otherwise:
[0026] As used herein, the term "subject" refers to an animal. In
certain aspects, the animal is a mammal. A subject also refers to
for example, primates (e.g., humans), cows, sheep, goats, horses,
dogs, cats, rabbits, rats, mice, fish, birds and the like. In
certain embodiments, the subject is a human. A "patient" as used
herein refers to a human subject.
[0027] As used herein, the term "inhibition" or "inhibiting" refers
to the reduction or suppression of a given condition, symptom, or
disorder, or disease, or a significant decrease in the baseline
activity of a biological activity or process.
[0028] As used herein, the term "treating" or "treatment" of any
disease or disorder refers in one embodiment, to ameliorating the
disease or disorder (i.e., slowing or arresting or reducing the
development of the disease or at least one of the clinical symptoms
thereof). In another embodiment "treating" or "treatment" refers to
alleviating or ameliorating at least one physical parameter
including those which may not be discernible by the patient. In yet
another embodiment, "treating" or "treatment" refers to modulating
the disease or disorder, either physically, (e.g., stabilization of
a discernible symptom), physiologically, (e.g., stabilization of a
physical parameter), or both. In yet another embodiment, "treating"
or "treatment" refers to preventing or delaying the onset or
development or progression of the disease or disorder.
[0029] As used herein, the term "a," "an," "the" and similar terms
used in the context of the present invention (especially in the
context of the claims) are to be construed to cover both the
singular and plural unless otherwise indicated herein or clearly
contradicted by the context.
[0030] All methods described herein can be performed in any
suitable order unless otherwise indicated herein or otherwise
clearly contradicted by context. The use of any and all examples,
or exemplary language (e.g. "such as") provided herein is intended
merely to better illuminate the invention and does not pose a
limitation on the scope of the invention otherwise claimed.
[0031] "Optionally substituted" means the group referred to can be
substituted at one or more positions by any one or any combination
of the radicals listed thereafter. The number, placement and
selection of substituents is understood to encompass only those
substitutions that a skilled chemist would expect to be reasonably
stable; thus `oxo` would not be a substituent on an aryl or
heteroaryl ring, for example, and a single carbon atom would not
have three hydroxy or amino substituents. Unless otherwise
specified, optional substituents are typically up to four groups
selected from halo, oxo, CN, amino, hydroxy, --C.sub.1-3 alkyl,
--OR*, --NR*.sub.2, --SR*, --SO.sub.2R*, --COOR*, and
--CONR*.sub.2, where each R* is independently H or C.sub.1-3
alkyl.
[0032] "Aryl" as used herein refers to a phenyl or naphthyl group
unless otherwise specified. Aryl groups unless otherwise specified
may be optionally substituted with up to four groups selected from
halo, CN, amino, hydroxy, C.sub.1-3 alkyl, --OR*, --NR*.sub.2,
--SR*, --SO.sub.2R*, --COOR*, and --CONR*.sub.2, where each R* is
independently H or C.sub.1-3 alkyl.
[0033] "Halo" or "halogen", as used herein, may be fluorine,
chlorine, bromine or iodine.
[0034] "C.sub.1-6 alkyl" or "C.sub.1-C.sub.6 alkyl", as used
herein, denotes straight chain or branched alkyl having 1-6 carbon
atoms. If a different number of carbon atoms is specified, such as
C.sub.4 or C.sub.3, then the definition is to be amended
accordingly, such as "C.sub.1-4 alkyl" will represent methyl,
ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and
tert-butyl.
[0035] "C.sub.1-6 alkylene" or "C.sub.1-C.sub.6 alkylene", as used
herein, denotes straight chain or branched alkyl having 1-6 carbon
atoms and two open valences for connection to two other groups. If
a different number of carbon atoms is specified, such as C.sub.4 or
C.sub.3, then the definition is to be amended accordingly, such as
"C.sub.1-4 alkylene" will represent methylene (--CH.sub.2--),
ethylene (--CH.sub.2CH.sub.2--), straight chain or branched
propylene (--CH.sub.2CH.sub.2CH.sub.2-- or
--CH.sub.2--CHMe-CH.sub.2--), and the like.
[0036] "C.sub.1-6 alkoxy", as used herein, denotes straight chain
or branched alkoxy (--O-Alkyl) having 1-6 carbon atoms. If a
different number of carbon atoms is specified, such as C.sub.4 or
C.sub.3, then the definition is to be amended accordingly, such as
"C.sub.1-4 alkoxy" will represent methoxy, ethoxy, propoxy,
isopropoxy, butoxy, isobutoxy, sec-butoxy and tert-butoxy.
[0037] "C.sub.1-4 Haloalkyl" or "C.sub.1-C.sub.4 haloalkyl" as used
herein, denotes straight chain or branched alkyl having 1-4 carbon
atoms wherein at least one hydrogen has been replaced with a
halogen. The number of halogen replacements can be from one up to
the number of hydrogen atoms on the unsubstituted alkyl group. If a
different number of carbon atoms is specified, such as C.sub.6 or
C.sub.3, then the definition is to be amended accordingly. Thus
"C.sub.1-4 haloalkyl" will represent methyl, ethyl, propyl,
isopropyl, butyl, isobutyl, sec-butyl and tert-butyl that have at
least one hydrogen substituted with halogen, such as where the
halogen is fluorine: CF.sub.3CF.sub.2--, (CF.sub.3).sub.2CH--,
CH.sub.3--CF.sub.2--, CF.sub.3CF.sub.2--, CF.sub.3, CF.sub.2H--,
CF.sub.3CF.sub.2CH(CF.sub.3)-- or
CF.sub.3CF.sub.2CF.sub.2CF.sub.2--.
[0038] "C.sub.3-8 cycloalkyl" as used herein refers to a saturated
monocyclic hydrocarbon ring of 3 to 8 carbon atoms. Examples of
such groups include cyclopropyl, cyclobutyl, cyclopentyl and
cyclohexyl. If a different number of carbon atoms is specified,
such as C.sub.3-C.sub.6, then the definition is to be amended
accordingly.
[0039] "4- to 8-Membered heterocyclyl", "5- to 6-membered
heterocyclyl", "3- to 10-membered heterocyclyl", "3- to 14-membered
heterocyclyl", "4- to 14-membered heterocyclyl" and "5- to
14-membered heterocyclyl", refers, respectively, to 4- to
8-membered, 5- to 6-membered, 3- to 10-membered, 3- to 14-membered,
4- to 14-membered and 5- to 14-membered heterocyclic rings; unless
otherwise specified, such rings contain 1 to 7, 1 to 5, or 1 to 3
heteroatoms selected from the group consisting of nitrogen, oxygen
and sulfur as ring members, and the rings may be saturated, or
partially saturated but not aromatic. The heterocyclic group can be
attached to another group at a nitrogen or a carbon atom. The term
"heterocyclyl" includes single ring groups, fused ring groups and
bridged groups. Examples of such heterocyclyl include, but are not
limited to pyrrolidine, piperidine, piperazine, pyrrolidinone,
morpholine, tetrahydrofuran, tetrahydrothiophene,
tetrahydrothiopyran, tetrahydropyran, 1,4-dioxane, 1,4-oxathiane,
8-aza-bicyclo[3.2.1]octane, 3,8-diazabicyclo[3.2.1]octane,
3-Oxa-8-aza-bicyclo[3.2.1]octane, 8-Oxa-3-aza-bicyclo[3.2.1]octane,
2-Oxa-5-aza-bicyclo[2.2.1]heptane, 2,5-Diazabicyclo[2.2.1]heptane,
azetidine, ethylenedioxo, oxetane or thiazole. In certain
embodiments, if not otherwise specified, heterocyclic groups have
1-2 heteroatoms selected from N, O and S as ring members, and 4-7
ring atoms, and are optionally substituted with up to four groups
selected from halo, oxo, CN, amino, hydroxy, C.sub.1-3 alkyl,
--OR*, --NR*.sub.2, --SR*, --SO.sub.2R*, --COOR*, and
--CONR*.sub.2, where each R* is independently H or C.sub.1-3 alkyl.
In particular, heterocyclic groups containing a sulfur atom are
optionally substituted with one or two oxo groups on the
sulfur.
[0040] "Heteroaryl" is a completely unsaturated (aromatic) ring.
The term "heteroaryl" refers to a 5-14 membered monocyclic- or
bicyclic- or tricyclic-aromatic ring system, having 1 to 8
heteroatoms selected from N, O or S. Typically, the heteroaryl is a
5-10 membered ring or ring system (e.g., 5-7 membered monocyclic
group or an 8-10 membered bicyclic group), often a 5-6 membered
ring containing up to four heteroatoms selected from N, O and S,
though often a heteroaryl ring contains no more than one divalent O
or S in the ring. Typical heteroaryl groups include furan,
isothiazole, thiadiazole, oxadiazole, indazole, indole, quinoline,
2- or 3-thienyl, 2- or 3-furyl, 2- or 3-pyrrolyl, 2-, 4-, or
5-imidazolyl, 3-, 4-, or 5-pyrazolyl, 2-, 4-, or 5-thiazolyl, 3-,
4-, or 5-isothiazolyl, 2-, 4-, or 5-oxazolyl, 3-, 4-, or
5-isoxazolyl, 3- or 5-(1,2,4-triazolyl), 4- or 5-(1,2,3-triazolyl),
tetrazolyl, triazine, pyrimidine, 2-, 3-, or 4-pyridyl, 3- or
4-pyridazinyl, 3-, 4-, or 5-pyrazinyl, 2-pyrazinyl, and 2-, 4-, or
5-pyrimidinyl. Heteroaryl groups are and are optionally substituted
with up to four groups selected from halo, CN, amino, hydroxy,
C.sub.1-3 alkyl, --OR*, --NR*.sub.2, --SR*, --SO.sub.2R*, --COOR*,
and --CONR*.sub.2, where each R* is independently H or C.sub.1-3
alkyl.
[0041] The term "hydroxy" or "hydroxyl" refers to the group
--OH.
[0042] Various embodiments of the invention are described herein.
It will be recognized that features specified in each embodiment
may be combined with other specified features to provide further
embodiments. The following enumerated embodiments are
representative of the invention:
[0043] 1. A compound of formula (I):
##STR00003##
wherein:
[0044] R.sup.1 is H, halo, or C.sub.1-C.sub.3 alkyl;
[0045] R.sup.2 is H, halo, CN, C.sub.1-C.sub.3 alkyl or
C.sub.1-C.sub.3 haloalkyl, C.sub.1-C.sub.3 alkoxy;
[0046] R.sup.3 is OH, halo, CN, C.sub.1-C.sub.3 alkyl,
C.sub.3-C.sub.6 cycloalkyl, C.sub.1-C.sub.3 haloalkyl, CO.sub.1-3
alkoxy, or C.sub.1-C.sub.3 haloalkoxy;
[0047] R.sup.4 is selected from R.sup.1, --OR.sup.11, --SR.sup.11,
and --NRR.sup.11; R.sup.11 is C.sub.1-C.sub.4 alkyl,
C.sub.3-C.sub.6 cycloalkyl, oxetanyl, tetrahydrofuranyl, or
tetrahydropyranyl, each of which is optionally substituted with up
to three groups selected from halo, CN, --OR, C.sub.1-C.sub.3
haloalkoxy, --NR.sub.2, and a 4-7 membered heterocyclic group
containing one or two heteroatoms selected from N, O and S as ring
members that is optionally substituted with one or two groups
selected from halo, oxo, CN, R, --OR, and --NR.sub.2;
[0048] R is independently selected at each occurrence from H and
C.sub.1-C.sub.3 alkyl optionally substituted with one to three
groups selected from halo, --OH, C.sub.1-C.sub.3 alkoxy, oxo, CN,
--NH.sub.2, --NH(C.sub.1-C.sub.3 alkyl), --N(C.sub.1-C.sub.3
alkyl).sub.2, and cyclopropyl; [0049] and two R groups directly
attached to the same atom, which may be C or N, can optionally be
taken together to form a 3-6 membered ring that can optionally
contain an added heteroatom selected from N, O and S as a ring
member, and can be substituted by up to two groups selected from
--OH, oxo, C.sub.1-C.sub.3 alkyl, and C.sub.1-C.sub.3 alkoxy;
[0050] R.sup.5 is H, halo, CN, C.sub.1-C.sub.3 alkyl, or
C.sub.1-C.sub.3 haloalkyl;
[0051] R.sup.6 is H, halo, C.sub.1-C.sub.3 alkoxy, or
C.sub.1-C.sub.6 alkyl;
[0052] R.sup.7 is H, halo, C.sub.1-C.sub.3 alkoxy, or
C.sub.1-C.sub.6 alkyl;
[0053] R.sup.8 is H or C.sub.1-C.sub.6 alkyl;
[0054] R.sup.9 taken together with one group selected from R.sup.6,
R.sup.7 and R.sup.8 forms a 3-7 membered cycloalkyl ring or a 3-7
membered heterocyclic ring containing N, O or S as a ring member;
wherein the cycloalkyl or heterocyclic ring is optionally
substituted with up to three groups selected from R, --OR,
--NR.sub.2, halo, CN, COOR, CONR.sub.2, and oxo;
[0055] W is --COOR.sup.10, --C(O)NH--SO.sub.2R,
--C(O)NH--SO.sub.2NR.sub.2, 5-tetrazolyl, or
1,2,4-oxadiazol-3-yl-5(4H)-one;
[0056] R.sup.10 is H or C.sub.1-C.sub.6 alkyl that is optionally
substituted with one or two groups selected from halo, --OR, oxo,
CN, --NR.sub.2, COOR, and CONR.sub.2;
[0057] or a pharmaceutically acceptable salt thereof.
[0058] A preferred option for W in embodiment 1 is --COOH.
[0059] 2. A compound according to embodiment 1 or a
pharmaceutically acceptable salt thereof, wherein R.sup.1 is H.
Alternatively, a compound of embodiment 1 wherein R.sup.1 is F or
Cl.
[0060] 3. A compound according to embodiment 1 or embodiment 2 or a
pharmaceutically acceptable salt thereof, wherein R.sup.2 is H or
halo.
[0061] 4. A compound according to any one of embodiments 1 to 3 or
a pharmaceutically acceptable salt thereof, wherein R.sup.3 is
C.sub.1-C.sub.3 alkoxy or halo.
[0062] 5. A compound according to any of the preceding embodiments
or a pharmaceutically acceptable salt thereof, wherein R.sup.4 is
--OR.sup.11.
[0063] 6. A compound according to any of the preceding embodiments
or a pharmaceutically acceptable salt thereof, wherein R.sup.5 is H
or halo.
[0064] 7. A compound according to any of the preceding embodiments
or a pharmaceutically acceptable salt thereof, which is of the
formula:
##STR00004##
[0065] wherein R.sup.9 taken together with R.sup.7 forms a 3-7
membered cycloalkyl ring or a 3-7 membered heterocyclic ring
containing N, O or S as a ring member; wherein the cycloalkyl or
heterocyclic ring is optionally substituted with up to three groups
selected from R, --OR, --NR.sub.2, halo, CN, COOR, CONR.sub.2, and
oxo; or a pharmaceutically acceptable salt thereof.
[0066] In preferred compounds of this embodiment, the ring formed
by R.sup.9 and R.sup.7 taken together is cis-fused onto the
tricyclic core. In certain compounds of this embodiment, R.sup.8 is
H. Compounds of special interest in this embodiment include
compounds with this absolute stereochemistry:
##STR00005##
[0067] 8. A compound according to any of embodiments 1-6, which is
of the formula:
##STR00006##
[0068] wherein R.sup.9 taken together with R.sup.8 forms a 3-7
membered cycloalkyl ring or a 3-7 membered heterocyclic ring
containing N, O or S as a ring member; wherein the cycloalkyl or
heterocyclic ring is optionally substituted with up to three groups
selected from R, --OR, --NR.sub.2, halo, CN, COOR, CONR.sub.2, and
oxo; or a pharmaceutically acceptable salt thereof.
[0069] 9. A compound according to any of the preceding embodiments
or a pharmaceutically acceptable salt thereof, wherein R.sup.11 is
C.sub.1-C.sub.4 alkyl, optionally substituted with up to two groups
selected from halo, CN, --OR, C.sub.1-C.sub.3 haloalkoxy, and a 4-7
membered heterocyclic group containing one or two heteroatoms
selected from N, O and S as ring members that is optionally
substituted with one or two groups selected from halo, oxo, CN, R,
--OR, and --NR.sub.2.
[0070] 10. A compound according to any of embodiments 1-9 or a
pharmaceutically acceptable salt thereof, wherein the R.sup.11 is
selected from --CH.sub.2CH.sub.2OMe, --CH.sub.2CH.sub.2CH.sub.2OMe,
--CH.sub.2--OEt, --CH.sub.2CH.sub.2-Q, and
--CH.sub.2CH.sub.2CH.sub.2-Q, where Q is selected from
##STR00007##
[0071] 11. A compound according to any of the preceding embodiments
or a pharmaceutically acceptable salt thereof, wherein
[0072] R.sup.9 taken together with one group selected from R.sup.6,
R.sup.7 and R.sup.8 forms a 4-6 membered cycloalkyl ring or a 5-6
membered heterocyclic ring containing N, O or S as a ring member;
wherein the cycloalkyl or heterocyclic ring is optionally
substituted with up to three groups selected from R, --OR,
--NR.sub.2, halo, CN, COOR, CONR.sub.2, and oxo.
[0073] 12. The compound according to embodiment 1, which is
selected from:
##STR00008## ##STR00009## ##STR00010## ##STR00011##
and the enantiomers of these compounds; or a pharmaceutically
acceptable salt thereof. Additional compounds of embodiment 1
include the following, and their pharmaceutically acceptable
salts:
##STR00012## ##STR00013##
[0074] 13. The compound of any of the Examples, or a
pharmaceutically acceptable salt thereof. Specific compounds of
this embodiment include any or all of the following:
##STR00014## ##STR00015## ##STR00016##
and the pharmaceutically acceptable salts of these.
[0075] 14. A pharmaceutical composition, comprising a compound of
any of the preceding embodiments admixed with at least one
pharmaceutically acceptable carrier.
[0076] 15. A method to treat a hepatitis B infection, which
comprises administering to a patient having a hepatitis B infection
a compound of any of embodiments 1-13 or a pharmaceutical
composition of embodiment 14.
[0077] 16. The method of embodiment 15, wherein the compound of any
one of claims 1-13 or the pharmaceutical composition of claim 14 is
used in combination with an additional therapeutic agent selected
from an interferon or peginterferon, an HBV polymerase inhibitor, a
viral entry inhibitor, a viral maturation inhibitor, a capsid
assembly inhibitor, an HBV core modulator, a reverse transcriptase
inhibitor, a TLR-agonist, or an immunomodulator.
[0078] 17. A method to inhibit replication of hepatitis B virus,
which comprises contacting the hepatitis B virus, either in vitro
or in vivo, with a compound according to any one of embodiments
1-13.
[0079] 18. A compound according to any one of embodiments 1-11,
wherein R.sup.1 is F.
[0080] Another embodiment of the invention provides a compound as
described above, or a pharmaceutically acceptable salt thereof, as
a medicament.
[0081] Also within the scope of this invention is the use of a
compound of formula (I), or a pharmaceutically acceptable salt
thereof, for the manufacture of a medicament for the treatment or
prevention of a viral disease and/or infection in a human being,
including HBV.
[0082] Included within the scope of this invention are
pharmaceutical compositions comprising a compound of Formula (I),
or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier, and optionally further
including an additional pharmaceutically acceptable carrier or
excipient.
[0083] According to a further aspect of this embodiment the
pharmaceutical composition according to this invention further
comprises a therapeutically effective amount of at least one other
antiviral agent.
[0084] The invention also provides the use of a pharmaceutical
composition as described hereinabove for the treatment of a HBV
infection in a human being having or at risk of having the
infection.
[0085] The invention also provides the use of a pharmaceutical
composition as described hereinabove for the treatment of HBV
infection in a human being having or at risk of having the
disease.
[0086] Another aspect of the invention involves a method of
treating or preventing a hepatitis B viral disease and/or infection
in a human being by administering to the human being an antivirally
effective amount of a compound of the invention, a pharmaceutically
acceptable salt thereof, or a composition as described above, alone
or in combination with at least one other antiviral agent,
administered together or separately.
[0087] An additional aspect of this invention refers to an article
of manufacture comprising a composition effective to treat a
hepatitis B viral disease and/or infection; and packaging material
comprising a label which indicates that the composition can be used
to treat disease and/or infection by a hepatitis B virus; wherein
the composition comprises a compound of formula (I) according to
this invention or a pharmaceutically acceptable salt thereof.
[0088] Still another aspect of this invention relates to a method
of inhibiting the replication of HBV, comprising exposing the virus
to an effective amount of the compound of formula (I), or a salt
thereof, under conditions where replication of the virus is
inhibited. This method can be practiced in vitro or in vivo.
[0089] Further included in the scope of the invention is the use of
a compound of formula (I), or a salt thereof, to inhibit the
replication of HBV.
[0090] In some embodiments, the compound of Formula (I) is
co-administered with or used in combination with at least one
additional therapeutic agent selected from: an interferon or
peginterferon, an HBV polymerase inhibitor, a viral entry
inhibitor, a viral maturation inhibitor, a capsid assembly
inhibitor, an HBV core modulator, a reverse transcriptase
inhibitor, a TLR-agonist, or an immunomodulator. Some particular
therapeutic agents that may be used in combination with the
compounds of the invention include immunomodulators described
herein, interferon alfa 2a, interferon alfa-2b, pegylated
interferon alfa-2a, pegylated interferon alfa-2b, TLR-7 and TLR-9
agonists, entecavir, tenofovir, cidofovir, telbivudine, didanosine,
zalcitabine, stavudine, lamivudine, abacavir, emtricitabine,
apricitabine, atevirapine, ribavirin, acyclovir, famciclovir,
valacyclovir, ganciclovir, adefovir, efavirenz, nevirapine,
delavirdine, and etravirine. Suitable core modulators are disclosed
in WO2013/096744; suitable HBV capsid inhibitors are described in
US2015/0252057.
[0091] These additional agents may be combined with the compounds
of this invention to create a single pharmaceutical dosage form.
Alternatively these additional agents may be separately
administered to the patient as part of a multiple dosage form, for
example, using a kit. Such additional agents may be administered to
the patient prior to, concurrently with, or following the
administration of a compound of the invention, or a
pharmaceutically acceptable salt thereof. Alternatively, these
additional therapeutic agents may be administered separately from
and optionally by different routes of administration and on
different dosing schedules from the compound of the invention,
provided the compound of the invention and the additional
therapeutic agent are used concurrently for treatment of an HBV
infection or a disorder caused or complicated by an HBV
infection.
[0092] The dose range of the compounds of the invention applicable
per day is usually from 0.01 to 100 mg/kg of body weight,
preferably from 0.1 to 50 mg/kg of body weight. In some
embodiments, the total daily dosage is between 1 and 25 mg, and may
be administered in a single dose or in divided doses at different
times to maintain a suitable plasma concentration. Each dosage unit
may conveniently contain from 5% to 95% active compound (w/w).
Preferably such preparations contain from 20% to 80% active
compound, which is typically admixed with one or more
pharmaceutically acceptable carriers or excipients.
[0093] The actual pharmaceutically effective amount or therapeutic
dosage will of course depend on factors known by those skilled in
the art such as age and weight of the patient, route of
administration and severity of disease. In any case the combination
will be administered at dosages and in a manner which allows a
pharmaceutically effective amount to be delivered based upon
patient's unique condition.
[0094] When the composition of this invention comprises a
combination of a compound of the invention and one or more
additional therapeutic or prophylactic agent, both the compound and
the additional agent may be used at lower dosages than would be
used typically for the individual compound when used as a
single-agent treatment. Thus in some embodiments, each component
may be present at dosage levels of between about 10 to 100%, and
more preferably between about 10 and 80% of the dosage normally
administered in a monotherapy regimen.
[0095] It is anticipated that the compounds of the invention may be
used in combination with other therapeutic agents, just as
combinations of therapeutic agents are currently used for the
treatment of hepatitis C virus (HCV) infections. Thus a compound of
the invention may be used in combination with a different anti-HBV
therapeutic agent such as a nucleoside or an immunomodulatory
agent. These combination therapies provide complementary mechanisms
to suppress HBV and thus their use in combination should enhance
efficacy and also reduce the frequency of resistance
development.
[0096] Antiviral agents contemplated for use in such combination
therapy include agents (compounds or biologicals) that are
effective to inhibit the formation and/or replication of a virus in
a human being, including but not limited to agents that interfere
with either host or viral mechanisms necessary for the formation
and/or replication of a virus in a human being. Such agents can be
selected from entecavir, tenofovir, cidofovir, telbivudine,
didanosine, zalcitabine, stavudine, lamivudine, abacavir,
emtricitabine, apricitabine, atevirapine, ribavirin, acyclovir,
famciclovir, valacyclovir, ganciclovir, adefovir, efavirenz,
nevirapine, delavirdine, and etravirine, and immunomodulators
described herein including interferons and pegylated interferons,
TLR-7 agonists, and TLR-9 agonists. Current HBV treatments
including immunomodulatory agents, such as interferon-.alpha. and
pegylated interferon-.alpha., and oral nucleoside/nucleotide
analogues (NAs), including lamivudine, adefovir, telbivudine,
entecavir and tenofovir, are known to suppress but not eliminate
HBV. J. Antimicrob. Chemother. 2011, vol. 66(12), 2715-25, and thus
those therapeutics may be used in combination with a compound of
the invention.
[0097] Many compounds of the invention contain one or more chiral
centers. These compounds may be made and used as single isomers or
as mixtures of isomers. Methods for separating the isomers,
including diastereomers and enantiomers, are known in the art, and
examples of suitable methods are described herein. In certain
embodiments, the compounds of the invention are used as a single
substantially pure isomer, meaning at least 90% of a sample of the
compound is the specified isomer and less than 10% of the sample is
any other isomer or mixture of isomers. Preferably, at least 95% of
the sample is a single isomer. Selection of a suitable isomer is
within the ordinary level of skill, as one isomer will typically be
more active in the in vivo or in vitro assay described herein for
measuring HBV activity, and will be the preferred isomer. Where in
vitro activity differences between isomers are relatively small,
e.g. less than about a factor of 4, a preferred isomer may be
selected based on activity level against viral replication in cell
culture, using methods such as those described herein: the isomer
having a lower MIC (minimum inhibitory concentration) or EC-50 is
preferred.
[0098] The compounds of the invention may be synthesized by the
general synthetic routes illustrated below, specific examples of
which are described in more detail in the Examples.
[0099] The term "an optical isomer" or "a stereoisomer" refers to
any of the various stereoisomeric configurations which may exist
for a given compound of the present invention and includes
geometric isomers. It is understood that a substituent may be
attached at a chiral center of a carbon atom. The term "chiral"
refers to molecules which have the property of
non-superimposability on their mirror image partner, while the term
"achiral" refers to molecules which are superimposable on their
mirror image partner. Therefore, the invention includes
enantiomers, diastereomers or racemates of the compound.
"Enantiomers" are a pair of stereoisomers that are
non-superimposable mirror images of each other. A 1:1 mixture of a
pair of enantiomers is a "racemic" mixture. The term is used to
designate a racemic mixture where appropriate. "Diastereoisomers"
are stereoisomers that have at least two asymmetric atoms, but
which are not mirror-images of each other. The absolute
stereochemistry is specified according to the Cahn-Ingold-Prelog
R-S system. When a compound is a pure enantiomer the
stereochemistry at each chiral carbon may be specified by either R
or S. Resolved compounds whose absolute configuration is unknown
can be designated (+) or (-) depending on the direction (dextro- or
levorotatory) which they rotate plane polarized light at the
wavelength of the sodium D line. Certain compounds described herein
contain one or more asymmetric centers or axes and may thus give
rise to enantiomers, diastereomers, and other stereoisomeric forms
that may be defined, in terms of absolute stereochemistry, as (R)-
or (S)-.
[0100] Depending on the choice of the starting materials and
procedures, the compounds can be present in the form of one of the
possible isomers or as mixtures thereof, for example as pure
optical isomers, or as isomer mixtures, such as racemates and
diastereoisomer mixtures, depending on the number of asymmetric
carbon atoms. The present invention is meant to include all such
possible stereoisomers, including racemic mixtures, diasteriomeric
mixtures and optically pure forms. Optically active (R)- and
(S)-isomers may be prepared using chiral synthons or chiral
reagents, or resolved using conventional techniques. If the
compound contains a double bond, the substituent may be E or Z
configuration. If the compound contains a disubstituted cycloalkyl,
the cycloalkyl substituent may have a cis- or trans-configuration.
All tautomeric forms are also intended to be included.
[0101] Any resulting mixtures of isomers can be separated on the
basis of the physicochemical differences of the constituents, into
the pure or substantially pure geometric or optical isomers or
diastereomers, for example, by chromatography and/or fractional
crystallization.
[0102] Any resulting racemates of final products or intermediates
can be resolved into the optical antipodes by known methods, e.g.,
by separation of the diastereomeric salts thereof, obtained with an
optically active acid or base, and liberating the optically active
acidic or basic compound. In particular, a basic moiety may thus be
employed to resolve the compounds of the present invention into
their optical antipodes, e.g., by fractional crystallization of a
salt formed with an optically active acid, e.g., tartaric acid,
dibenzoyl tartaric acid, diacetyl tartaric acid, di-O,O'-p-toluoyl
tartaric acid, mandelic acid, malic acid or camphor-10-sulfonic
acid. Racemic products can also be resolved by chiral
chromatography, e.g., high pressure liquid chromatography (HPLC)
using a chiral adsorbent.
[0103] Furthermore, the compounds of the present invention,
including their salts, can also be obtained in the form of their
hydrates, or include other solvents used for their crystallization.
The compounds of the present invention may inherently or by design
form solvates with pharmaceutically acceptable solvents (including
water); therefore, it is intended that the invention embrace both
solvated and unsolvated forms. The term "solvate" refers to a
molecular complex of a compound of the present invention (including
pharmaceutically acceptable salts thereof) with one or more solvent
molecules. Such solvent molecules are those commonly used in the
pharmaceutical art, which are known to be innocuous to the
recipient, e.g., water, ethanol, and the like. The term "hydrate"
refers to the complex where the solvent molecule is water.
[0104] The compounds of the present invention, including salts,
hydrates and solvates thereof, may inherently or by design form
polymorphs.
[0105] As used herein, the terms "salt" or "salts" refers to an
acid addition or base addition salt of a compound of the present
invention. "Salts" include in particular "pharmaceutically
acceptable salts". The term "pharmaceutically acceptable salts"
refers to salts that retain the biological effectiveness and
properties of the compounds of this invention and, which typically
are not biologically or otherwise undesirable. In many cases, the
compounds of the present invention are capable of forming acid
and/or base salts by virtue of the presence of amino and/or
carboxyl groups or groups similar thereto.
[0106] Pharmaceutically acceptable acid addition salts can be
formed with inorganic acids and organic acids, e.g., acetate,
aspartate, benzoate, besylate, bromide/hydrobromide,
bicarbonate/carbonate, bisulfate/sulfate, camphorsulfonate,
chloride/hydrochloride, chlortheophyllonate, citrate,
ethandisulfonate, fumarate, gluceptate, gluconate, glucuronate,
hippurate, hydroiodide/iodide, isethionate, lactate, lactobionate,
laurylsulfate, malate, maleate, malonate, mandelate, mesylate,
methylsulphate, naphthoate, napsylate, nicotinate, nitrate,
octadecanoate, oleate, oxalate, palmitate, pamoate,
phosphate/hydrogen phosphate/dihydrogen phosphate,
polygalacturonate, propionate, stearate, succinate,
sulfosalicylate, tartrate, tosylate and trifluoroacetate salts.
[0107] Inorganic acids from which salts can be derived include, for
example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric
acid, phosphoric acid, and the like.
[0108] Organic acids from which salts can be derived include, for
example, acetic acid, propionic acid, glycolic acid, oxalic acid,
maleic acid, malonic acid, succinic acid, fumaric acid, tartaric
acid, citric acid, benzoic acid, mandelic acid, methanesulfonic
acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic
acid, and the like. Pharmaceutically acceptable base addition salts
can be formed with inorganic and organic bases.
[0109] Inorganic bases from which salts can be derived include, for
example, ammonium salts and metals from columns I to XII of the
periodic table. In certain embodiments, the salts are derived from
sodium, potassium, ammonium, calcium, magnesium, iron, silver,
zinc, and copper; particularly suitable salts include ammonium,
potassium, sodium, calcium and magnesium salts.
[0110] Organic bases from which salts can be derived include, for
example, primary, secondary, and tertiary amines, substituted
amines including naturally occurring substituted amines, cyclic
amines, basic ion exchange resins, and the like. Certain organic
amines include isopropylamine, benzathine, chlorinate,
diethanolamine, diethylamine, lysine, meglumine, piperazine and
tromethamine.
[0111] The pharmaceutically acceptable salts of the present
invention can be synthesized from a basic or acidic moiety, by
conventional chemical methods. Generally, such salts can be
prepared by reacting free acid forms of these compounds with a
stoichiometric amount of the appropriate base (such as Na, Ca, Mg,
or K hydroxide, carbonate, bicarbonate or the like), or by reacting
free base forms of these compounds with a stoichiometric amount of
the appropriate acid. Such reactions are typically carried out in
water or in an organic solvent, or in a mixture of the two.
Generally, use of non-aqueous media like ether, ethyl acetate,
ethanol, isopropanol, or acetonitrile is desirable, where
practicable. Lists of additional suitable salts can be found, e.g.,
in "Remington's Pharmaceutical Sciences", 20th ed., Mack Publishing
Company, Easton, Pa., (1985); and in "Handbook of Pharmaceutical
Salts: Properties, Selection, and Use" by Stahl and Wermuth
(Wiley-VCH, Weinheim, Germany, 2002).
[0112] Any formula given herein is intended to represent unlabeled
forms as well as isotopically labeled forms of the compounds of the
present invention having up to three atoms with non-natural isotope
distributions, e.g., sites that are enriched in deuterium or
.sup.13C or .sup.15N. Isotopically labeled compounds have
structures depicted by the formulas given herein except that one or
more atoms are replaced by an atom having a selected atomic mass or
mass number other than the natural-abundance mass distribution.
Examples of isotopes that can be usefully over-incorporated into
compounds of the invention include isotopes of hydrogen, carbon,
nitrogen, oxygen, phosphorous, fluorine, and chlorine, such as
.sup.2H, .sup.3H, .sup.11C, .sup.13C, .sup.14O, .sup.15N, .sup.18F
.sup.31P, .sup.32P, .sup.35S, .sup.36Cl, .sup.125I respectively.
The invention includes various isotopically labeled compounds of
the present invention, for example those into which radioactive
isotopes, such as .sup.3H and .sup.14C, or those in which
non-radioactive isotopes, such as .sup.2H and .sup.13C are present
at levels substantially above normal isotope distribution. Such
isotopically labelled compounds are useful in metabolic studies
(with .sup.14C, for example), reaction kinetic studies (with, for
example .sup.2H or .sup.3H), detection or imaging techniques, such
as positron emission tomography (PET) or single-photon emission
computed tomography (SPECT) including drug or substrate tissue
distribution assays, or in radioactive treatment of patients. In
particular, an .sup.18F labeled compound of the present invention
may be particularly desirable for PET or SPECT studies.
Isotopically-labeled compounds of the present invention can
generally be prepared by conventional techniques known to those
skilled in the art or by processes analogous to those described in
the accompanying Examples and Preparations using an appropriate
isotopically-labeled reagent in place of the non-labeled reagent
typically employed. Labeled samples may be useful with quite low
isotope incorporation, such as where a radiolabel is used to detect
trace amounts of the compound.
[0113] Further, more extensive substitution with heavier isotopes,
particularly deuterium (i.e., .sup.2H or D), may afford certain
therapeutic advantages resulting from greater metabolic stability,
for example increased in vivo half-life or reduced dosage
requirements or an improvement in therapeutic index. It is
understood that deuterium in this context is regarded as a
substituent of a compound of the present invention, and typically a
sample of a compound having deuterium as a substituent has at least
50% deuterium incorporation at the labeled position(s). The
concentration of such a heavier isotope, specifically deuterium,
may be defined by the isotopic enrichment factor. The term
"isotopic enrichment factor" as used herein means the ratio between
the isotopic abundance and the natural abundance of a specified
isotope. If a substituent in a compound of this invention is
denoted deuterium, such compound has an isotopic enrichment factor
for each designated deuterium atom of at least 3500 (52.5%
deuterium incorporation at each designated deuterium atom), at
least 4000 (60% deuterium incorporation), at least 4500 (67.5%
deuterium incorporation), at least 5000 (75% deuterium
incorporation), at least 5500 (82.5% deuterium incorporation), at
least 6000 (90% deuterium incorporation), at least 6333.3 (95%
deuterium incorporation), at least 6466.7 (97% deuterium
incorporation), at least 6600 (99% deuterium incorporation), or at
least 6633.3 (99.5% deuterium incorporation).
[0114] Pharmaceutically acceptable solvates in accordance with the
invention include those wherein the solvent of crystallization may
be isotopically substituted, e.g. D.sub.2O, d.sup.6-acetone,
d.sup.6-DMSO.
[0115] Compounds of the present invention that contain groups
capable of acting as donors and/or acceptors for hydrogen bonds may
be capable of forming co-crystals with suitable co-crystal formers.
These co-crystals may be prepared from compounds of the present
invention by known co-crystal forming procedures. Such procedures
include grinding, heating, co-subliming, co-melting, or contacting
in solution compounds of the present invention with the co-crystal
former under crystallization conditions and isolating co-crystals
thereby formed. Suitable co-crystal formers include those described
in WO 2004/078163. Hence the invention further provides co-crystals
comprising a compound of the present invention.
Methods of Use
[0116] All methods described herein can be performed in any
suitable order unless otherwise indicated herein or otherwise
clearly contradicted by context. The use of any and all examples,
or exemplary language (e.g. "such as") provided herein is intended
merely to better illuminate the invention and does not pose a
limitation on the scope of the invention otherwise claimed.
[0117] The compounds of the invention can be administered by known
methods, including oral, parenteral, inhalation, and the like. In
certain embodiments, the compound of the invention is administered
orally, as a pill, lozenge, troche, capsule, solution, or
suspension. In other embodiments, a compound of the invention is
administered by injection or infusion. Infusion is typically
performed intravenously, often over a period of time between about
15 minutes and 4 hours. In other embodiments, a compound of the
invention is administered intranasally or by inhalation; inhalation
methods are particularly useful for treatment of respiratory
infections. Compounds of the present invention exhibit oral
bioavailability, so oral administration is sometimes preferred.
[0118] In certain embodiments of the present invention, a compound
of the present invention is used in combination with a second
antiviral agent, such as those named herein.
[0119] By the term "combination", is meant either a fixed
combination in one dosage unit form, as separate dosage forms
suitable for use together either simultaneously or sequentially, or
as a kit of parts for the combined administration where a compound
of the present invention and a combination partner may be
administered independently at the same time or separately within
time intervals that especially allow that the combination partners
show a cooperative, e.g., synergistic, effect, or any combination
thereof.
[0120] The second antiviral agent may be administered in
combination with the compounds of the present inventions wherein
the second antiviral agent is administered prior to,
simultaneously, or after the compound or compounds of the present
invention. When simultaneous administration of a compound of the
invention with a second agent is desired and the route of
administration is the same, then a compound of the invention may be
formulated with a second agent into the same dosage form. An
example of a dosage form containing a compound of the invention and
a second agent is a tablet or a capsule.
[0121] In some embodiments, a combination of a compound of the
invention and a second antiviral agent may provide synergistic
activity. The compound of the invention and second antiviral agent
may be administered together, separate but simultaneously, or
sequentially.
[0122] An "effective amount" of a compound is that amount necessary
or sufficient to treat or prevent a viral infection and/or a
disease or condition described herein. In an example, an effective
amount of a compound of Formula I is an amount sufficient to treat
viral infection in a subject. In another example, an effective
amount is an amount sufficient to treat HBV in a subject in need of
such treatment. The effective amount can vary depending on such
factors as the size and weight of the subject, the type of illness,
or the particular compound of the invention. For example, the
choice of the compound of the invention can affect what constitutes
an "effective amount." One of ordinary skill in the art would be
able to study the factors contained herein and make the
determination regarding the effective amount of the compounds of
the invention without undue experimentation.
[0123] The regimen of administration can affect what constitutes an
effective amount. The compound of the invention can be administered
to the subject either prior to or after the onset of a viral
infection. Further, several divided dosages, as well as staggered
dosages, can be administered daily or sequentially, or the dose can
be continuously infused, or can be a bolus injection. Further, the
dosages of the compound(s) of the invention can be proportionally
increased or decreased as indicated by the exigencies of the
therapeutic or prophylactic situation.
[0124] Compounds of the invention may be used in the treatment of
states, disorders or diseases as described herein, or for the
manufacture of pharmaceutical compositions for use in the treatment
of these diseases. The invention provides methods of use of
compounds of the present invention in the treatment of these
diseases or for preparation of pharmaceutical compositions having
compounds of the present invention for the treatment of these
diseases.
[0125] The language "pharmaceutical composition" includes
preparations suitable for administration to mammals, e.g., humans.
When the compounds of the present invention are administered as
pharmaceuticals to mammals, e.g., humans, they can be given per se
or as a pharmaceutical composition containing, for example, 0.1 to
99.5% (more preferably, 0.5 to 90%) of at least one compound of
Formula (I) or any subgenus thereof as active ingredient in
combination with a pharmaceutically acceptable carrier, or
optionally two or more pharmaceutically acceptable carriers.
[0126] The phrase "pharmaceutically acceptable carrier" is art
recognized and includes a pharmaceutically acceptable material,
composition or vehicle, suitable for administering compounds of the
present invention to mammals. The carriers include liquid or solid
filler, diluent, excipient, solvent or encapsulating material,
involved in carrying or transporting the subject agent from one
organ, or portion of the body, to another organ, or portion of the
body. Each carrier must be "acceptable" in the sense of being
compatible with the other ingredients of the formulation and not
injurious to the patient. Some examples of materials which can
serve as pharmaceutically acceptable carriers include: sugars, such
as lactose, glucose and sucrose; starches, such as corn starch and
potato starch; cellulose, and its derivatives, such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa
butter and suppository waxes; oils, such as peanut oil, cottonseed
oil, safflower oil, sesame oil, olive oil, corn oil and soybean
oil; glycols, such as propylene glycol; polyols, such as glycerin,
sorbitol, mannitol and polyethylene glycol; esters, such as ethyl
oleate and ethyl laurate; agar; buffering agents, such as magnesium
hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water;
isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer
solutions; and other non-toxic compatible substances employed in
pharmaceutical formulations. Typically, pharmaceutically acceptable
carriers are sterilized and/or substantially pyrogen-free.
[0127] Wetting agents, emulsifiers and lubricants, such as sodium
lauryl sulfate and magnesium stearate, as well as coloring agents,
release agents, coating agents, sweetening, flavoring and perfuming
agents, preservatives and antioxidants can also be present in the
compositions.
[0128] Examples of pharmaceutically acceptable antioxidants
include: water soluble antioxidants, such as ascorbic acid,
cysteine hydrochloride, sodium bisulfate, sodium metabisulfite,
sodium sulfite and the like; oil-soluble antioxidants, such as
ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated
hydroxytoluene (BHT), lecithin, propyl gallate, .alpha.-tocopherol,
and the like; and metal chelating agents, such as citric acid,
ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid,
phosphoric acid, and the like.
[0129] Formulations of the present invention include those suitable
for oral, nasal, inhalation, topical, transdermal, buccal,
sublingual, rectal, vaginal and/or parenteral administration. The
formulations may conveniently be presented in unit dosage form and
may be prepared by any methods well known in the art of pharmacy.
The amount of active ingredient that can be combined with a carrier
material to produce a single dosage form will generally be that
amount of the compound that produces a therapeutic effect.
Generally, out of one hundred percent, this amount will range from
about 1 percent to about ninety-nine percent of active ingredient,
preferably from about 5 percent to about 70 percent, most
preferably from about 10 percent to about 30 percent.
[0130] Methods of preparing these formulations or compositions
include the step of bringing into association a compound of the
present invention with the carrier and, optionally, one or more
accessory ingredients. In general, the formulations are prepared by
uniformly and intimately bringing into association a compound of
the present invention with liquid carriers, or finely divided solid
carriers, or both, and then, if necessary, shaping the product.
[0131] Formulations of the invention suitable for oral
administration may be in the form of capsules, cachets, pills,
tablets, lozenges (using a flavored base, for example, usually
sucrose and acacia or tragacanth), powders, granules, or as a
solution or a suspension in an aqueous or non-aqueous liquid, or as
an oil-in-water or water-in-oil liquid emulsion, or as an elixir or
syrup, or as pastilles (using an inert base, such as gelatin and
glycerin, or sucrose and acacia) and/or as mouth washes and the
like, each containing a predetermined amount of a compound of the
present invention as an active ingredient. A compound of the
present invention may also be administered as a bolus, electuary or
paste.
[0132] In solid dosage forms of the invention for oral
administration (capsules, tablets, pills, dragees, powders,
granules and the like), the active ingredient is mixed with one or
more pharmaceutically acceptable carriers, such as sodium citrate
or dicalcium phosphate, and/or any of the following: fillers or
extenders, such as starches, lactose, sucrose, glucose, mannitol,
and/or silicic acid; binders, such as, for example,
carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,
sucrose and/or acacia; humectants, such as glycerol; disintegrating
agents, such as agar-agar, calcium carbonate, potato or tapioca
starch, alginic acid, certain silicates, and sodium carbonate;
solution retarding agents, such as paraffin; absorption
accelerators, such as quaternary ammonium compounds; wetting
agents, such as, for example, cetyl alcohol and glycerol
monostearate; absorbents, such as kaolin and bentonite clay;
lubricants, such a talc, calcium stearate, magnesium stearate,
solid polyethylene glycols, sodium lauryl sulfate, and mixtures
thereof; and coloring agents. In the case of capsules, tablets and
pills, the pharmaceutical compositions may also comprise buffering
agents. Solid compositions of a similar type may also be employed
as fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugars, as well as high molecular
weight polyethylene glycols and the like.
[0133] A tablet may be made by compression or molding, optionally
with one or more accessory ingredients. Compressed tablets may be
prepared using binder (for example, gelatin or hydroxypropylmethyl
cellulose), lubricant, inert diluent, preservative, disintegrant
(for example, sodium starch glycolate or cross-linked sodium
carboxymethyl cellulose), surface-active or dispersing agent.
Molded tablets may be made by molding in a suitable machine a
mixture of the powdered compound moistened with an inert liquid
diluent.
[0134] The tablets, and other solid dosage forms of the
pharmaceutical compositions of the present invention, such as
dragees, capsules, pills and granules, may optionally be scored or
prepared with coatings and shells, such as enteric coatings and
other coatings well known in the pharmaceutical-formulating art.
They may also be formulated so as to provide slow or controlled
release of the active ingredient therein using, for example,
hydroxypropylmethyl cellulose in varying proportions to provide the
desired release profile, other polymer matrices, liposomes and/or
microspheres. They may be sterilized by, for example, filtration
through a bacteria-retaining filter, or by incorporating
sterilizing agents in the form of sterile solid compositions that
can be dissolved in sterile water, or some other sterile injectable
medium immediately before use. These compositions may also
optionally contain opacifying agents and may be of a composition
that they release the active ingredient(s) only, or preferentially,
in a certain portion of the gastrointestinal tract, optionally, in
a delayed manner. Examples of embedding compositions that can be
used include polymeric substances and waxes. The active ingredient
can also be in micro-encapsulated form, if appropriate, with one or
more of the above-described excipients.
[0135] Liquid dosage forms for oral administration of the compounds
of the invention include pharmaceutically acceptable emulsions,
microemulsions, solutions, suspensions, syrups and elixirs. In
addition to the active ingredient, the liquid dosage forms may
contain inert diluent commonly used in the art, such as, for
example, water or other solvents, solubilizing agents and
emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, oils (in particular,
cottonseed, groundnut, corn, germ, olive, castor and sesame oils),
glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty
acid esters of sorbitan, and mixtures thereof.
[0136] Besides inert diluents, the oral compositions can also
include adjuvants such as wetting agents, emulsifying and
suspending agents, sweetening, flavoring, coloring, perfuming and
preservative agents.
[0137] Suspensions, in addition to the active compounds, may
contain suspending agents as, for example, ethoxylated isostearyl
alcohols, polyoxyethylene sorbitol and sorbitan esters,
microcrystalline cellulose, aluminum metahydroxide, bentonite,
agar-agar and tragacanth, and mixtures thereof.
[0138] Formulations of the pharmaceutical compositions of the
invention for rectal or vaginal administration may be presented as
a suppository, which may be prepared by mixing one or more
compounds of the invention with one or more suitable nonirritating
excipients or carriers comprising, for example, cocoa butter,
polyethylene glycol, a suppository wax or a salicylate, and which
is solid at room temperature, but liquid at body temperature and,
therefore, will melt in the rectum or vaginal cavity and release
the active compound.
[0139] Formulations of the present invention which are suitable for
vaginal administration also include pessaries, tampons, creams,
gels, pastes, foams or spray formulations containing such carriers
as are known in the art to be appropriate.
[0140] Dosage forms for the topical or transdermal administration
of a compound of this invention include powders, sprays, ointments,
pastes, creams, lotions, gels, solutions, patches and inhalants.
The active compound may be mixed under sterile conditions with a
pharmaceutically acceptable carrier, and with any preservatives,
buffers, or propellants that may be required.
[0141] The ointments, pastes, creams and gels may contain, in
addition to an active compound of this invention, excipients, such
as animal and vegetable fats, oils, waxes, paraffins, starch,
tragacanth, cellulose derivatives, polyethylene glycols, silicones,
bentonites, silicic acid, talc and zinc oxide, or mixtures
thereof.
[0142] Powders and sprays can contain, in addition to a compound of
this invention, excipients such as lactose, talc, silicic acid,
aluminum hydroxide, calcium silicates and polyamide powder, or
mixtures of these substances. Sprays can additionally contain
customary propellants, such as chlorofluorohydrocarbons and
volatile unsubstituted hydrocarbons, such as butane and
propane.
[0143] Transdermal patches have the added advantage of providing
controlled delivery of a compound of the present invention to the
body. Such dosage forms can be made by dissolving or dispersing the
compound in the proper medium. Absorption enhancers can also be
used to increase the flux of the compound across the skin. The rate
of such flux can be controlled by either providing a rate
controlling membrane or dispersing the active compound in a polymer
matrix or gel.
[0144] Ophthalmic formulations, eye ointments, powders, solutions
and the like, are also contemplated as being within the scope of
this invention.
[0145] Pharmaceutical compositions of this invention suitable for
parenteral administration may comprise one or more compounds of the
invention in combination with one or more pharmaceutically
acceptable carriers such as sterile isotonic aqueous or nonaqueous
solutions, dispersions, suspensions or emulsions, or sterile
powders which may be reconstituted into sterile injectable
solutions or dispersions just prior to use, which may contain
antioxidants, buffers, bacteriostats, solutes which render the
formulation isotonic with the blood of the intended recipient or
suspending or thickening agents.
[0146] Examples of suitable aqueous and nonaqueous carriers that
may be employed in the pharmaceutical compositions of the invention
include water, ethanol, glycol ethers, polyols (such as glycerol,
propylene glycol, polyethylene glycol, and the like), and suitable
mixtures thereof, vegetable oils, such as olive oil, and injectable
organic esters, such as ethyl oleate. Proper fluidity can be
maintained, for example, by the use of coating materials, such as
lecithin, by the maintenance of the required particle size in the
case of dispersions, and by the use of surfactants.
[0147] These compositions may also contain adjuvants such as
preservatives, wetting agents, emulsifying agents and dispersing
agents. Prevention of the action of microorganisms may be ensured
by the inclusion of various antibacterial and antifungal agents,
for example, paraben, chlorobutanol, phenol sorbic acid, and the
like. It may also be desirable to include isotonic agents, such as
sugars, sodium chloride, and the like into the compositions. In
addition, prolonged absorption of the injectable pharmaceutical
form may be brought about by the inclusion of agents that delay
absorption such as aluminum monostearate and gelatin.
[0148] In some cases, in order to prolong the effect of a drug, it
is desirable to slow the absorption of the drug from subcutaneous
or intramuscular injection. This may be accomplished by the use of
a liquid suspension of crystalline or amorphous material having
poor water solubility. The rate of absorption of the drug then
depends upon its rate of dissolution which, in turn, may depend
upon crystal size and crystalline form. Alternatively, delayed
absorption of a parenterally-administered drug form is accomplished
by dissolving or suspending the drug in an oil vehicle.
[0149] Injectable depot forms are made by forming microencapsule
matrices of the subject compounds in biodegradable polymers such as
polylactide-polyglycolide. Depending on the ratio of drug to
polymer, and the nature of the particular polymer employed, the
rate of drug release can be controlled. Examples of other
biodegradable polymers include poly(orthoesters) and
poly(anhydrides). Depot injectable formulations are also prepared
by entrapping the drug in liposomes or microemulsions that are
compatible with body tissue.
[0150] The preparations of the present invention may be given
orally, parenterally, topically, or rectally. They are of course
given by forms suitable for each administration route. For example,
they are administered in tablets or capsule form, by injection,
inhalation, eye lotion, ointment, suppository, etc., administration
by injection, infusion or inhalation; topical by lotion or
ointment; and rectal by suppositories.
[0151] The phrases "parenteral administration" and "administered
parenterally" as used herein means modes of administration other
than enteral and topical administration, usually by injection, and
includes, without limitation, intravenous, intramuscular,
intraarterial, intrathecal, intracapsular, intraorbital,
intracardiac, intradermal, intraperitoneal, transtracheal,
subcutaneous, subcuticular, intraarticular, subcapsular,
subarachnoid, intraspinal and intrasternal injection and infusion.
Intravenous infusion is sometimes a preferred method of delivery
for compounds of the invention. Infusion may be used to deliver a
single daily dose or multiple doses. In some embodiments, a
compound of the invention is administered by infusion over an
interval between 15 minutes and 4 hours, typically between 0.5 and
3 hours. Such infusion may be used once per day, twice per day or
up to three times per day.
[0152] The phrases "systemic administration," "administered
systemically," "peripheral administration" and "administered
peripherally" as used herein mean the administration of a compound,
drug or other material other than directly into the central nervous
system, such that it enters the patient's system and, thus, is
subject to metabolism and other like processes, for example,
subcutaneous administration.
[0153] These compounds may be administered to humans and other
animals for therapy by any suitable route of administration,
including orally, nasally, as by, for example, a spray, rectally,
intravaginally, parenterally, intracisternally and topically, as by
powders, ointments or drops, including buccally and
sublingually.
[0154] Regardless of the route of administration selected, the
compounds of the present invention, which may be used in a suitable
hydrated form, and/or the pharmaceutical compositions of the
present invention, are formulated into pharmaceutically acceptable
dosage forms by conventional methods known to those of skill in the
art.
[0155] Actual dosage levels of the active ingredients in the
pharmaceutical compositions of this invention may be varied so as
to obtain an amount of the active ingredient which is effective to
achieve the desired therapeutic response for a particular patient,
composition, and mode of administration, without being toxic to the
patient.
[0156] The selected dosage level will depend upon a variety of
factors including the activity of the particular compound of the
present invention employed, or the ester, salt or amide thereof,
the route of administration, the time of administration, the rate
of excretion of the particular compound being employed, the
duration of the treatment, other drugs, compounds and/or materials
used in combination with the particular compound employed, the age,
sex, weight, condition, general health and prior medical history of
the patient being treated, and like factors well known in the
medical arts.
[0157] A physician or veterinarian having ordinary skill in the art
can readily determine and prescribe the effective amount of the
pharmaceutical composition required. For example, the physician or
veterinarian could start doses of the compounds of the invention
employed in the pharmaceutical composition at levels lower than
that required in order to achieve the desired therapeutic effect
and gradually increase the dosage until the desired effect is
achieved.
[0158] In general, a suitable daily dose of a compound of the
invention will be that amount of the compound that is the lowest
dose effective to produce a therapeutic effect. Such an effective
dose will generally depend upon the factors described above.
Generally, intravenous and subcutaneous doses of the compounds of
this invention for a patient, when used for the indicated effects,
will range from about 0.0001 to about 100 mg per kilogram of body
weight per day, more preferably from about 0.01 to about 50 mg per
kg per day, and still more preferably from about 0.1 to about 20 mg
per kg per day. An effective amount is that amount which prevents
or treats a viral infection, such as HBV.
[0159] If desired, the effective daily dose of the active compound
may be administered as a single dose per day, or as two, three,
four, five, six or more sub-doses administered separately at
appropriate intervals throughout the day, optionally, in unit
dosage forms. Compounds delivered orally or by inhalation, are
commonly administered in one to four doses per day. Compounds
delivered by injection are typically administered once per day, or
once every other day. Compounds delivered by infusion are typically
administered in one to three doses per day. When multiple doses are
administered within a day, the doses may be administered at
intervals of about 4 hours, about 6 hours, about 8 hours or about
12 hours.
[0160] While it is possible for a compound of the present invention
to be administered alone, it is preferable to administer the
compound as a pharmaceutical composition such as those described
herein. Thus methods of using the compounds of the invention
include administering the compound as a pharmaceutical composition,
wherein at least one compound of the invention is admixed with a
pharmaceutically acceptable carrier prior to administration.
Use of Compounds of the Invention in Combination with
Immunomodulators
[0161] The compounds and compositions described herein can be used
or administered in combination with one or more therapeutic agents
that act as immunomodulators, e.g., an activator of a costimulatory
molecule, or an inhibitor of an immune-inhibitory molecule, or a
vaccine. The Programmed Death 1 (PD-1) protein is an inhibitory
member of the extended CD28/CTLA4 family of T cell regulators
(Okazaki et al. (2002) Curr Opin Immunol 14: 391779-82; Bennett et
al. (2003) J. Immunol. 170:711-8). PD-1 is expressed on activated B
cells, T cells, and monocytes. PD-1 is an immune-inhibitory protein
that negatively regulates TCR signals (Ishida, Y. et al. (1992)
EMBO J. 11:3887-3895; Blank, C. et al. (Epub 2006 Dec. 29) Immunol.
Immunother. 56(5):739-745), and is up-regulated in chronic
infections. The interaction between PD-1 and PD-L1 can act as an
immune checkpoint, which can lead to, e.g., a decrease in
infiltrating lymphocytes, a decrease in T-cell receptor mediated
proliferation, and/or immune evasion by cancerous or infected cells
(Dong et al. (2003) J. Mol. Med. 81:281-7; Blank et al. (2005)
Cancer Immunol. Immunother. 54:307-314; Konishi et al. (2004) Clin.
Cancer Res. 10:5094-100). Immune suppression can be reversed by
inhibiting the local interaction of PD-1 with PD-L1 or PD-L2; the
effect is additive when the interaction of PD-1 with PD-L2 is
blocked as well (Iwai et al. (2002) Proc. Nat'l. Acad. Sci. USA
99:12293-7; Brown et al. (2003) J. Immunol. 170:1257-66).
Immunomodulation can be achieved by binding to either the
immune-inhibitory protein (e.g., PD-1) or to binding proteins that
modulate the inhibitory protein (e.g., PD-L1, PD-L2).
[0162] In one embodiment, the combination therapies of the
invention include an immunomodulator that is an inhibitor or
antagonist of an inhibitory molecule of an immune checkpoint
molecule. In another embodiment the immunomodulator binds to a
protein that naturally inhibits the immuno-inhibitory checkpoint
molecule. When used in combination with antiviral compounds, these
immunomodulators can enhance the antiviral response, and thus
enhance efficacy relative to treatment with the antiviral compound
alone.
[0163] The term "immune checkpoints" refers to a group of molecules
on the cell surface of CD4 and CD8 T cells. These molecules can
effectively serve as "brakes" to down-modulate or inhibit an
adaptive immune response. Immune checkpoint molecules include, but
are not limited to, Programmed Death 1 (PD-1), Cytotoxic
T-Lymphocyte Antigen 4 (CTLA-4), B7H1, B7H4, OX-40, CD137, CD40,
and LAG3, which directly inhibit immune cells. Immunotherapeutic
agents which can act as immune checkpoint inhibitors useful in the
methods of the present invention, include, but are not limited to,
inhibitors of PD-L1, PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT,
LAIR1, CD160, 2B4 and/or TGFR beta. Inhibition of an inhibitory
molecule can be performed by inhibition at the DNA, RNA or protein
level. In some embodiments, an inhibitory nucleic acid (e.g., a
dsRNA, siRNA or shRNA), can be used to inhibit expression of an
inhibitory molecule. In other embodiments, the inhibitor of an
inhibitory signal is a polypeptide, e.g., a soluble ligand, or an
antibody or antigen-binding fragment thereof, that binds to the
inhibitory molecule.
[0164] By "in combination with," it is not intended to imply that
the therapy or the therapeutic agents must be administered at the
same time and/or formulated for delivery together, although these
methods of delivery are within the scope described herein. The
immunomodulator can be administered concurrently with, prior to, or
subsequent to, one or more compounds of the invention, and
optionally one or more additional therapies or therapeutic agents.
The therapeutic agents in the combination can be administered in
any order. In general, each agent will be administered at a dose
and/or on a time schedule determined for that agent. It will
further be appreciated that the therapeutic agents utilized in this
combination may be administered together in a single composition or
administered separately in different compositions. In general, it
is expected that each of the therapeutic agents utilized in
combination be utilized at levels that do not exceed the levels at
which they are utilized individually. In some embodiments, the
levels utilized in combination will be lower than those utilized
individually.
[0165] In certain embodiments, the antiviral compounds described
herein are administered in combination with one or more
immunomodulators that are inhibitors of PD-1, PD-L1 and/or PD-L2.
Each such inhibitor may be an antibody, an antigen binding fragment
thereof, an immunoadhesin, a fusion protein, or an oligopeptide.
Examples of such immunomodulators are known in the art.
[0166] In some embodiments, the immunomodulator is an anti-PD-1
antibody chosen from MDX-1106, Merck 3475 or CT-011.
[0167] In some embodiments, the immunomodulator is an immunoadhesin
(e.g., an immunoadhesin comprising an extracellular or PD-1 binding
portion of PD-LI or PD-L2 fused to a constant region (e.g., an Fc
region of an immunoglobulin sequence).
[0168] In some embodiments, the immunomodulator is a PD-1 inhibitor
such as AMP-224.
[0169] In some embodiments, the immunomodulator is a PD-LI
inhibitor such as anti-PD-LI antibody.
[0170] In some embodiments, the immunomodulator is an anti-PD-LI
binding antagonist chosen from YW243.55.S70, MPDL3280A, MEDI-4736,
MSB-0010718C, or MDX-1105. MDX-1105, also known as BMS-936559, is
an anti-PD-LI antibody described in WO2007/005874. Antibody
YW243.55.S70 is an anti-PD-LI described in WO 2010/077634.
[0171] In some embodiments, the immunomodulator is nivolumab (CAS
Registry Number: 946414-94-4). Alternative names for nivolumab
include MDX-1106, MDX-1106-04, ONO-4538, or BMS-936558. Nivolumab
is a fully human IgG4 monoclonal antibody which specifically blocks
PD-1. Nivolumab (clone 5C4) and other human monoclonal antibodies
that specifically bind to PD-1 are disclosed in U.S. Pat. No.
8,008,449, EP2161336 and WO2006/121168.
[0172] In some embodiments, the immunomodulator is an anti-PD-1
antibody Pembrolizumab. Pembrolizumab (also referred to as
Lambrolizumab, MK-3475, MK03475, SCH-900475 or KEYTRUDA.RTM.;
Merck) is a humanized IgG4 monoclonal antibody that binds to PD-1.
Pembrolizumab and other humanized anti-PD-1 antibodies are
disclosed in Hamid, O. et al. (2013) New England Journal of
Medicine 369 (2): 134-44, U.S. Pat. No. 8,354,509, WO2009/114335,
and WO2013/079174.
[0173] In some embodiments, the immunomodulator is Pidilizumab
(CT-011; Cure Tech), a humanized IgG1k monoclonal antibody that
binds to PD1. Pidilizumab and other humanized anti-PD-1 monoclonal
antibodies are disclosed in WO2009/101611.
[0174] Other anti-PD1 antibodies useful as immunomodulators for use
in the methods disclosed herein include AMP 514 (Amplimmune), and
anti-PD1 antibodies disclosed in U.S. Pat. No. 8,609,089, US
2010028330, and/or US 20120114649. In some embodiments, the
anti-PD-L1 antibody is MSB0010718C. MSB0010718C (also referred to
as A09-246-2; Merck Serono) is a monoclonal antibody that binds to
PD-L1.
[0175] In some embodiments, the immunomodulator is MDPL3280A
(Genentech/Roche), a human Fc optimized IgG1 monoclonal antibody
that binds to PD-L1. MDPL3280A and other human monoclonal
antibodies to PD-L1 are disclosed in U.S. Pat. No. 7,943,743 and
U.S Publication No.: 20120039906. Other anti-PD-L1 binding agents
useful as immunomodulators for methods of the invention include
YW243.55.S70 (see WO2010/077634), MDX-1105 (also referred to as
BMS-936559), and anti-PD-L1 binding agents disclosed in
WO2007/005874.
[0176] In some embodiments, the immunomodulator is AMP-224
(B7-DCIg; Amplimmune; e.g., disclosed in WO2010/027827 and
WO2011/066342), is a PD-L2 Fc fusion soluble receptor that blocks
the interaction between PD1 and B7-H1.
[0177] In some embodiments, the immunomodulator is an anti-LAG-3
antibody such as BMS-986016. BMS-986016 (also referred to as
BMS986016) is a monoclonal antibody that binds to LAG-3. BMS-986016
and other humanized anti-LAG-3 antibodies are disclosed in US
2011/0150892, WO2010/019570, and WO2014/008218
[0178] In certain embodiments, the combination therapies disclosed
herein include a modulator of a costimulatory molecule or an
inhibitory molecule, e.g., a co-inhibitory ligand or receptor.
[0179] In one embodiment, the costimulatory modulator, e.g.,
agonist, of a costimulatory molecule is chosen from an agonist
(e.g., an agonistic antibody or antigen-binding fragment thereof,
or soluble fusion) of OX40, CD2, CD27, CDS, ICAM-1, LFA-1
(CD11a/CD18), ICOS (CD278), 4-1BB (CD137), GITR, CD30, CD40, BAFFR,
HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3 or CD83
ligand.
[0180] In another embodiment, the combination therapies disclosed
herein include an immunomodulator that is a costimulatory molecule,
e.g., an agonist associated with a positive signal that includes a
costimulatory domain of CD28, CD27, ICOS and/or GITR.
[0181] Exemplary GITR agonists include, e.g., GITR fusion proteins
and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies),
such as, a GITR fusion protein described in U.S. Pat. No.
6,111,090, European Patent No.: 090505B1, U.S. Pat. No. 8,586,023,
PCT Publication Nos.: WO 2010/003118 and 2011/090754, or an
anti-GITR antibody described, e.g., in U.S. Pat. No. 7,025,962,
European Patent No.: 1947183B1, U.S. Pat. Nos. 7,812,135,
8,388,967, 8,591,886, European Patent No.: EP 1866339, PCT
Publication No.: WO 2011/028683, PCT Publication No.: WO
2013/039954, PCT Publication No.: WO2005/007190, PCT Publication
No.: WO 2007/133822, PCT Publication No.: WO2005/055808, PCT
Publication No.: WO 99/40196, PCT Publication No.: WO 2001/03720,
PCT Publication No.: WO99/20758, PCT Publication No.:
WO2006/083289, PCT Publication No.: WO 2005/115451, U.S. Pat. No.
7,618,632, and PCT Publication No.: WO 2011/051726.
[0182] In one embodiment, the immunomodulator used is a soluble
ligand (e.g., a CTLA-4-Ig), or an antibody or antibody fragment
that binds to PD-L1, PD-L2 or CTLA4. For example, the anti-PD-1
antibody molecule can be administered in combination with an
anti-CTLA-4 antibody, e.g., ipilimumab, for example. Exemplary
anti-CTLA4 antibodies include Tremelimumab (IgG2 monoclonal
antibody available from Pfizer, formerly known as ticilimumab,
CP-675,206); and Ipilimumab (CTLA-4 antibody, also known as
MDX-010, CAS No. 477202-00-9).
[0183] In one embodiment, an anti-PD-1 antibody molecule is
administered after treatment with a compound of the invention as
described herein.
[0184] In another embodiment, an anti-PD-1 or PD-L1 antibody
molecule is administered in combination with an anti-LAG-3 antibody
or an antigen-binding fragment thereof. In another embodiment, the
anti-PD-1 or PD-L1 antibody molecule is administered in combination
with an anti-TIM-3 antibody or antigen-binding fragment thereof. In
yet other embodiments, the anti-PD-1 or PD-L1 antibody molecule is
administered in combination with an anti-LAG-3 antibody and an
anti-TIM-3 antibody, or antigen-binding fragments thereof. The
combination of antibodies recited herein can be administered
separately, e.g., as separate antibodies, or linked, e.g., as a
bispecific or trispecific antibody molecule. In one embodiment, a
bispecific antibody that includes an anti-PD-1 or PD-L1 antibody
molecule and an anti-TIM-3 or anti-LAG-3 antibody, or
antigen-binding fragment thereof, is administered. In certain
embodiments, the combination of antibodies recited herein is used
to treat a cancer, e.g., a cancer as described herein (e.g., a
solid tumor). The efficacy of the aforesaid combinations can be
tested in animal models known in the art. For example, the animal
models to test the synergistic effect of anti-PD-1 and anti-LAG-3
are described, e.g., in Woo et al. (2012) Cancer Res.
72(4):917-27).
[0185] Exemplary immunomodulators that can be used in the
combination therapies include, but are not limited to, e.g.,
afutuzumab (available from Roche.RTM.); pegfilgrastim
(Neulasta.RTM.); lenalidomide (CC-5013, Revlimid.RTM.); thalidomide
(Thalomid.RTM.), actimid (CC4047); and cytokines, e.g., IL-21 or
IRX-2 (mixture of human cytokines including interleukin 1,
interleukin 2, and interferon .gamma., CAS 951209-71-5, available
from IRX Therapeutics).
[0186] Exemplary doses of such immunomodulators that can be used in
combination with the antiviral compounds of the invention include a
dose of anti-PD-1 antibody molecule of about 1 to 10 mg/kg, e.g., 3
mg/kg, and a dose of an anti-CTLA-4 antibody, e.g., ipilimumab, of
about 3 mg/kg.
[0187] Examples of embodiments of the methods of using the
antiviral compounds of the invention in combination with an
immunomodulator include these, which may be used along with a
compound of Formula I or any subgenus or species thereof that is
disclosed herein:
[0188] i. A method to treat a viral infection in a subject,
comprising administering to the subject a compound of Formula (I)
as described herein, and an immunomodulator.
[0189] ii. The method of embodiment i, wherein the immunomodulator
is an activator of a costimulatory molecule or an inhibitor of an
immune checkpoint molecule.
[0190] iii. The method of either of embodiments i and ii, wherein
the activator of the costimulatory molecule is an agonist of one or
more of OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS
(CD278), 4-1BB (CD137), GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT,
NKG2C, SLAMF7, NKp80, CD160, B7-H3 and CD83 ligand.
[0191] iv. The method of any of embodiments i-iii above, wherein
the inhibitor of the immune checkpoint molecule is chosen from
PD-1, PD-L1, PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1,
CD160, 2B4 and TGFR beta.
[0192] v. The method of any of any of embodiments i-iii, wherein
the inhibitor of the immune checkpoint molecule is chosen from an
inhibitor of PD-1, PD-L1, LAG-3, TIM-3 or CTLA4, or any combination
thereof.
[0193] vi. The method of any of embodiments i-v, wherein the
inhibitor of the immune checkpoint molecule is a soluble ligand or
an antibody or antigen-binding fragment thereof, that binds to the
immune checkpoint molecule.
[0194] vii. The method of any of embodiments i-vi, wherein the
antibody or antigen-binding fragment thereof is from an IgG1 or
IgG4 (e.g., human IgG1 or IgG4).
[0195] viii. The method of any of embodiments i-vii, wherein the
antibody or antigen-binding fragment thereof is altered, e.g.,
mutated, to increase or decrease one or more of: Fc receptor
binding, antibody glycosylation, the number of cysteine residues,
effector cell function, or complement function.
[0196] ix. The method of any of embodiments i-viii, wherein the
antibody molecule is a bispecific or multispecific antibody
molecule that has a first binding specificity to PD-1 or PD-L1 and
a second binding specificity to TIM-3, LAG-3, or PD-L2.
[0197] x. The method of any of embodiments i-ix, wherein the
immunomodulator is an anti-PD-1 antibody chosen from Nivolumab,
Pembrolizumab or Pidilizumab.
[0198] xi. The method of any of embodiments i-x, wherein the
immunomodulator is an anti-PD-L1 antibody chosen from YW243.55.S70,
MPDL3280A, MEDI-4736, MSB-0010718C, or MDX-1105.
[0199] xii. The method of any of embodiments i-x, wherein the
immunomodulator is an anti-LAG-3 antibody molecule.
[0200] xiii. The method of embodiment xii, wherein the anti-LAG-3
antibody molecule is BMS-986016.
[0201] xiv. The method of any of embodiments i-x, wherein the
immunomodulator is an anti-PD-1 antibody molecule administered by
injection (e.g., subcutaneously or intravenously) at a dose of
about 1 to 30 mg/kg, e.g., about 5 to 25 mg/kg, about 10 to 20
mg/kg, about 1 to 5 mg/kg, or about 3 mg/kg., e.g., once a week to
once every 2, 3, or 4 weeks.
[0202] xv. The method of embodiment xiv, wherein the anti-PD-1
antibody molecule is administered at a dose from about 10 to 20
mg/kg every other week.
[0203] xvi. The method of embodiment xv, wherein the anti-PD-1
antibody molecule, e.g., nivolumab, is administered intravenously
at a dose from about 1 mg/kg to 3 mg/kg, e.g., about 1 mg/kg, 2
mg/kg or 3 mg/kg, every two weeks.
[0204] xvii. The method of embodiment xv, wherein the anti-PD-1
antibody molecule, e.g., nivolumab, is administered intravenously
at a dose of about 2 mg/kg at 3-week intervals.
[0205] The compounds of the invention share certain structural
features with compounds reported to have the same utility as the
compounds of the invention. For example, Example 132 in
WO2015/113990 has this structure:
##STR00017##
and similar biological activity to the compounds of the invention.
As illustrated herein, certain compounds of the invention have
improved solubility and safety profiles when compared to the
reference example in common screens that are used to predict
suitability for development.
[0206] The compounds as described herein may be synthesized by the
general synthetic routes below, specific examples of which are
described in more detail in the Examples.
General Synthetic Procedures
[0207] All starting materials, building blocks, reagents, acids,
bases, dehydrating agents, solvents, and catalysts utilized to
synthesize the compounds of the invention are either commercially
available or can be produced by organic synthesis methods known to
one of ordinary skill in the art (Houben-Weyl 4th Ed. 1952, Methods
of Organic Synthesis, Thieme, Volume 21). General methods for
synthesis of compounds of the invention are illustrated by the
Examples below, and by methods disclosed in published PCT
applications WO2015/113990 and WO2015/173164.
LIST OF ABBREVIATIONS
[0208] Ac acetyl [0209] ACN Acetonitrile [0210] AcOEt/EtOAc Ethyl
acetate [0211] AcOH acetic acid [0212] aq aqueous [0213] Bn benzyl
[0214] Bu butyl (nBu=n-butyl, tBu=tert-butyl) [0215] CDI
Carbonyldiimidazole [0216] DBU 1,8-Diazabicyclo[5.4.0]-undec-7-ene
[0217] Boc2O di-tert-butyl dicarbonate [0218] DCE
1,2-Dichloroethane [0219] DCM Dichloromethane [0220] DIAD
Diisopropyl azodicarboxylate [0221] DiBAI-H Diisobutylaluminum
Hydride [0222] DIPEA N-Ethyldiisopropylamine [0223] DMA N,
N-dimethylacetamide [0224] DMAP Dimethylaminopyridine [0225] DMF
N,N'-Dimethylformamide [0226] DMSO Dimethylsulfoxide [0227] EDC
1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide [0228] EI
Electrospray ionisation [0229] Et.sub.2O Diethylether [0230]
Et.sub.3N Triethylamine [0231] Ether Diethylether [0232] EtOAc
Ethyl acetate [0233] EtOH Ethanol [0234] FA Formic acid [0235] FC
Flash Chromatography [0236] h hour(s) [0237] HCl Hydrochloric acid
[0238] HOBt 1-Hydroxybenzotriazole [0239] HPLC High Performance
Liquid Chromatography [0240] H.sub.2O Water [0241] IPA isopropanol
[0242] L liter(s) [0243] LC-MS Liquid Chromatography Mass
Spectrometry [0244] LiHMDS Lithium bis(trimethylsilyl)amide [0245]
Me methyl [0246] MeI Iodomethane [0247] MeOH Methanol [0248] mg
milligram [0249] min minute(s) [0250] mL milliliter [0251] MS Mass
Spectrometry [0252] Pd/C palladium on charcoal [0253] PG protecting
group [0254] Ph phenyl [0255] Ph.sub.3P triphenyl phosphine [0256]
Prep Preparative [0257] Rf ratio of fronts [0258] RP reverse phase
[0259] Rt Retention time [0260] rt Room temperature [0261] SFC
Supercritical Fluid Chromatography [0262] SiO.sub.2 Silica gel
[0263] T3P.RTM. Propylphosphonic acid anhydride [0264] TBAF
Tetrabutylammonium fluoride [0265] TBDMS t-Butyldimethylsilyl
[0266] TEA Triethylamine [0267] TFA Trifluoroacetic acid [0268] THF
Tetrahydrofuran [0269] TLC Thin Layer Chromatography [0270] TsCl
toluene sulfonyl chloride
[0271] Within the scope of this text, a readily removable group
that is not a constituent of the particular desired end product of
the compounds of the present invention is designated a "protecting
group," unless the context indicates otherwise. The protection of
functional groups by such protecting groups, the protecting groups
themselves, and their cleavage reactions are described for example
in standard reference works, such as e.g., Science of Synthesis:
Houben-Weyl Methods of Molecular Transformation. Georg Thieme
Verlag, Stuttgart, Germany. 2005. 41627 pp. (URL:
http://www.science-of-synthesis.com (Electronic Version, 48
Volumes)); J. F. W. McOmie, "Protective Groups in Organic
Chemistry", Plenum Press, London and New York 1973, in T. W. Greene
and P. G. M. Wuts, "Protective Groups in Organic Synthesis", Third
edition, Wiley, New York 1999, in "The Peptides"; Volume 3
(editors: E. Gross and J. Meienhofer), Academic Press, London and
New York 1981, in "Methoden der organischen Chemie" (Methods of
Organic Chemistry), Houben Weyl, 4th edition, Volume 15/I, Georg
Thieme Verlag, Stuttgart 1974, in H.-D. Jakubke and H. Jeschkeit,
"Aminosauren, Peptide, Proteine" (Amino acids, Peptides, Proteins),
Verlag Chemie, Weinheim, Deerfield Beach, and Basel 1982, and in
Jochen Lehmann, "Chemie der Kohlenhydrate: Monosaccharide und
Derivate" (Chemistry of Carbohydrates: Monosaccharides and
Derivatives), Georg Thieme Verlag, Stuttgart 1974. A characteristic
of protecting groups is that they can be removed readily (i.e.,
without the occurrence of undesired secondary reactions) for
example by solvolysis, reduction, photolysis or alternatively under
physiological conditions (e.g., by enzymatic cleavage).
[0272] Salts of compounds of the present invention having at least
one salt-forming group may be prepared in a manner known per se.
For example, salts of compounds of the present invention having
acid groups may be formed, for example, by treating the compounds
with metal compounds, such as alkali metal salts of suitable
organic carboxylic acids, e.g., the sodium salt of 2-ethyl hexanoic
acid, with organic alkali metal or alkaline earth metal compounds,
such as the corresponding hydroxides, carbonates or hydrogen
carbonates, such as sodium or potassium hydroxide, carbonate or
hydrogen carbonate, with corresponding calcium compounds or with
ammonia or a suitable organic amine, stoichiometric amounts or only
a small excess of the salt-forming agent preferably being used.
Acid addition salts of compounds of the present invention are
obtained in customary manner, e.g., by treating the compounds with
an acid or a suitable anion exchange reagent. Internal salts of
compounds of the present invention containing acid and basic
salt-forming groups, e.g., a free carboxy group and a free amino
group, may be formed, e.g., by the neutralization of salts, such as
acid addition salts, to the isoelectric point, e.g., with weak
bases, or by treatment with ion exchangers.
[0273] Salts can be converted in customary manner into the free
compounds; metal and ammonium salts can be converted, for example,
by treatment with suitable acids, and acid addition salts, for
example, by treatment with a suitable basic agent.
[0274] Mixtures of isomers obtainable according to the invention
can be separated in a manner known per se into the individual
isomers; diastereoisomers can be separated, for example, by
partitioning between polyphasic solvent mixtures, recrystallization
and/or chromatographic separation, for example over silica gel or
by, e.g., medium pressure liquid chromatography over a reversed
phase column, and racemates can be separated, for example, by the
formation of salts with optically pure salt-forming reagents and
separation of the mixture of diastereoisomers so obtainable, for
example by means of fractional crystallization, or by
chromatography over optically active column materials.
[0275] Intermediates and final products can be worked up and/or
purified according to standard methods, e.g., using chromatographic
methods, distribution methods, (re-) crystallization, and the
like.
EXAMPLES
[0276] The invention is illustrated by the following examples,
which should not be construed as limiting. The assays used
throughout the Examples are well established in the art:
demonstration of efficacy in these assays is generally regarded as
predictive of efficacy in subjects.
General Conditions:
[0277] Mass spectra were run on LC-MS systems using electrospray
ionization. These were WATERS Acquity Single Quard Detector.
[M+H].sup.+ refers to mono-isotopic molecular weights. NMR spectra
were run on open access Varian 400 or Varian 500 NMR spectrometers.
Spectra were measured at 298K and were referenced using the solvent
peak. Chemical shifts for .sup.1H NMR are reported in parts per
million (ppm). Mass spectra were run on LC-MS systems with one of
the following conditions:
[0278] 1. Waters Acquity UPLC-H class system equipped with SQD
detector.
Column: ACQUITY UPLC HSS C18 (50*2.1) mm, 1.8 u.
[0279] Column temperature: Ambient.
Mobile Phase: A) 0.1% FA+5 mM Ammonium Acetate in Water.
[0280] B) 0.1% FA in Acetonitrile. Gradient: 5-5% solvent B in 0.40
min, 5-35% solvent B in 0.80 min, 35-55% solvent B in 1.2 min,
[0281] 55-100% solvent B in 2.5 min. Flow rate: 0.55 mL/min.
Compounds were detected by a Waters Photodiode Array Detector.
[0282] 2. Waters LCMS system equipped with ZQ 2000 detector.
Column: X-BRIDGE C18 (50*4.6) mm, 3.5 u.
[0283] Column temperature: Ambient.
Mobile Phase: A) 0.1% NH.sub.3 in Water.
[0284] B) 0.1% NH.sub.3 in Acetonitrile. Gradient: 5-95% solvent B
in 5.00 min. Flow rate: 1.0 mL/min. Compounds were detected by a
Waters Photodiode Array Detector.
[0285] 3. Waters ACQUITY UPLC system and equipped with a ZQ 2000 MS
system.
Column: Kinetex by Phenomenex, 2.6 um, 2.1.times.50 mm
[0286] Column temperature: 50.degree. C. Gradient: 2-88% (or
00-45%, or 65-95%) solvent B over a 1.29 min period Flow rate: 1.2
mL/min. Compounds were detected by a Waters Photodiode Array
Detector.
Example 1: Synthesis of Racemic
10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,3,3a,7,12b-hexahy-
drocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylic acid
[rac-1]
##STR00018##
[0287] Step 1:
5-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclopentanone
[1.1a]
##STR00019##
[0289] A mixture of Pd(OAc).sub.2 (8.16 mg, 0.036 mmol), sodium
tert-butoxide (0.454 g, 4.72 mmol),
dicyclohexyl(2'-methyl-[1,1'-biphenyl]-2-yl)phosphane (32 mg),
2,2-dimethylcyclopentanone (0.547 ml, 4.36 mmol) and
4-bromo-1-methoxy-2-(3-methoxypropoxy)benzene (1 g, 3.63 mmol) in
toluene (4.0 mL) was heated in a sealed vial at 50.degree. C. for
18 hours. The mixture was diluted with EtOAc and filtered. The
filtrate was concentrated and the remaining oil was purified by
silica gel column chromatography, EtOAc/heptane 5 to 50%, to give
product (500 mg, 45% yield). LC-MS (m/z): 307.2 [M+H].sup.+.
.sup.1H NMR (400 MHz, CDCl.sub.3): 7.72-7.28 (m, 1H), 6.94-6.61 (m,
2H), 4.10 (m, 2H), 3.91-3.77 (m, 4H), 3.65-3.49 (m, 2H), 3.41-3.27
(m, 3H), 2.36 (d, J=4.2 Hz, 1H), 2.19-1.88 (m, 4H), 1.88-1.71 (m,
1H), 1.21-1.11 (m, 3H), 1.07 (s, 3H)
Step 2:
5-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclopentanam-
ine [1.1b]
##STR00020##
[0291] To the mixture of
5-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclopentanone
(320 mg, 1.0 mmol) in MeOH (3 mL) was added acetic acid ammonia
salt (1.6 g, 20.9 mmol) and sodium cyanoborohydride (656 mg, 10.4
mmol). The mixture was stirred at 80.degree. C. for 8 hours and
then was concentrated under reduced pressure. The remaining
material was diluted with EtOAc, washed with water and brine, dried
over Na.sub.2SO.sub.4 and concentrated. The crude material was used
in the next step with no further purification. LCMS (m/z): 308.0
[M+H].sup.+.
Step 3: N-(5-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2
dimethylcyclopentyl)formamide [1.1c]
##STR00021##
[0293] To the mixture of
5-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclopentanamine
(320 mg, 1.041 mmol) in dioxane (3 ml) was added formic acid (0.160
mL, 4.16 mmol). The mixture was stirred at 100.degree. C. for 6
hours. The mixture was concentrated to afford the crude product
which was used in the next step with no further purification. LCMS
(m/z): 336.2 [M+H].sup.+.
Step 4:
7-methoxy-8-(3-methoxypropoxy)-3,3-dimethyl-2,3,3a,9b-tetrahydro-1-
H-cyclopenta[c]isoquinoline [1.1d-I] and [1.1d-II]
##STR00022##
[0295] To a mixture of
N-(5-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclopentyl)forma-
mide (349 mg, 1.04 mmol) in acetonitrile (1.8 ml) was added
POCl.sub.3 (140 .mu.l, 1.50 mmol). The mixture was stirred at
85.degree. C. for 2 hours and then concentrated. The residue was
dissolved in EtOAc and basified by adding ammonium hydroxide
solution. The phases were separated and the organic layer was
washed with brine, dried over Na.sub.2SO.sub.4 and concentrated.
The remaining material was purified by silica gel chromatography,
acetone/heptane 5 to 50% to give product rac-1.1d-I and
rac-1.1d-II.
[0296] Trans isomer rac-1.1d-II: (70 mg, 21% yield). LCMS (m/z):
318.3 [M+H].sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3): 8.22 (s, 1H),
6.93-6.83 (m, 1H), 6.70 (s, 1H), 4.16 (t, J=6.4 Hz, 2H), 3.94-3.83
(m, 3H), 3.57 (q, J=4.7 Hz, 2H), 3.35 (d, J=1.5 Hz, 4H), 2.84 (s,
2H), 2.21-1.98 (m, 4H), 1.84-1.70 (m, 3H), 1.68-1.53 (m, 3H), 1.22
(s, 4H), 1.06 (s, 4H)
[0297] Cis isomer rac-1.1d-I: (54 mg, 16% yield). LCMS (m/z): 318.3
[M+H].sup.+. 1H NMR (400 MHz, CDCl.sub.3): 8.17 (s, 1H), 6.75 (s,
1H), 6.67 (s, 1H), 4.14 (t, J=6.2 Hz, 3H), 3.92-3.83 (m, 3H), 3.77
(d, J=10.3 Hz, 1H), 3.57 (t, J=5.4 Hz, 3H), 3.35 (d, J=1.6 Hz, 3H),
3.25 (q, J=9.4 Hz, 1H), 2.42-2.21 (m, 2H), 2.19-2.01 (m, 3H), 1.64
(dd, J=20.6, 7.8 Hz, 4H), 1.45 (t, J=8.0 Hz, 2H), 1.24 (d, J=6.1
Hz, 4H), 0.89 (s, 4H).
[0298] The relative configuration of rac-1.1d-I and rac-1.1d-II
were confirmed by nuclear Overhauser effect (nOe) experiments.
Step 5: Ethyl
10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,3,3a,7,8,8a,12b-o-
ctahydrocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylate
[rac-1.1e]
##STR00023##
[0300] To a mixture of
7-methoxy-8-(3-methoxypropoxy)-3,3-dimethyl-2,3,3a,9b-tetrahydro-1H-cyclo-
penta[c]isoquinoline (cis isomer, rac-1.1d-I) (51 mg, 0.161 mmol)
in EtOH (0.6 ml) was added (Z)-ethyl
2-(ethoxymethylene)-3-oxobutanoate (90 mg, 0.482 mmol). The mixture
was stirred at 110.degree. C. for 16 hours. After cooling, the
mixture was concentrated and the crude material was used in the
next step with no further purification. LCMS (m/z): 458.0
[M+H].sup.+.
Step 6: Ethyl
10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,3,3a,7,12b-hexahy-
drocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylate
[rac-1.1f]
##STR00024##
[0302] To a mixture of ethyl
10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,3,3a,7,8,8a,
12b-octahydrocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylate
(73.7 mg, 0.161 mmol) in DME (0.3 ml) was added p-chloranil (39.6
mg, 0.161 mmol). The mixture was stirred at 110.degree. C. for 2
hours. After cooling to rt, the mixture was filtered and the solid
was washed with cold DME. After drying, the desired product (28 mg,
38% yield) was a light yellow solid. LCMS (m/z): 456.0
[M+H].sup.+.
Step 7: racemic
10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,3,3a,7,12b-hexahy-
drocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylic acid
[rac-1]
##STR00025##
[0304] To a mixture of ethyl
10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,3,3a,7,12b-hexahy-
drocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylate (28 mg,
0.061 mmol) in THF (0.4 ml), MeOH (0.4 ml) and water (0.4 ml) was
added LiOH (4.42 mg, 0.184 mmol). After stirring at rt for 2 hours,
the mixture was concentrated and then acidified by adding 3.0 N HCl
aq solution. To the mixture was added EtOAc. The organic layer was
washed with water and brine, dried, and concentrated. The crude
residue was purified by reverse phase HPLC to give product (5 mg,
19% yield). LCMS (m/z): 428.2 [M+H].sup.+. 1H NMR (400 MHz,
CD.sub.3CN): 8.41 (s, 1H), 7.33 (d, J=22.5 Hz, 2H), 6.97 (s, 1H),
4.38 (d, J=8.5 Hz, 1H), 4.17 (dtd, J=13.1, 9.6, 4.8 Hz, 2H), 3.91
(s, 3H), 3.80 (t, J=8.9 Hz, 1H), 3.53 (t, J=6.2 Hz, 3H), 3.32 (s,
3H), 2.32 (q, J=7.7 Hz, 4H), 2.13-2.00 (m, 3H), 1.65 (dt, J=13.2,
6.8 Hz, 2H), 1.45 (dt, J=12.9, 7.9 Hz, 1H), 1.22 (s, 3H), 0.48 (s,
3H).
Chiral Separation:
(3aS,12bR)-10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,3,3a,7-
,12b-hexahydrocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylic
acid [1.1] and
(3aR,12bS)-10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo--
1,2,3,3a,7,12b-hexahydrocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxyli-
c acid [1.2]
##STR00026##
[0306] Compound rac-1 (40 mg, 0.090 mmol) was separated by chiral
HPLC (Column: AD 21.times.250 mm, Heptane/IPA=30/70, flow rate 20
ml/min) to afford the two enantiomers 1.1 and 1.2.
[0307] Compound 1.1: tR 9.55 min; 8 mg 20% yield. LCMS (m/z): 428.2
[M+H].sup.+. 1 H NMR (400 MHz, CD.sub.3CN): 8.41 (s, 1H), 7.33 (d,
J=22.5 Hz, 2H), 6.97 (s, 1H), 4.38 (d, J=8.5 Hz, 1H), 4.17 (dtd,
J=13.1, 9.6, 4.8 Hz, 2H), 3.91 (s, 3H), 3.80 (t, J=8.9 Hz, 1H),
3.53 (t, J=6.2 Hz, 3H), 3.32 (s, 3H), 2.32 (q, J=7.7 Hz, 4H),
2.13-2.00 (m, 3H), 1.65 (dt, J=13.2, 6.8 Hz, 2H), 1.45 (dt, J=12.9,
7.9 Hz, 1H), 1.22 (s, 3H), 0.48 (s, 3H).
[0308] Compound 1:2: tR 18.90 min, 8 mg, 20% yield. LCMS (m/z):
428.2 [M+H].sup.+. 1 H NMR (400 MHz, CD.sub.3CN): 8.41 (s, 1H),
7.33 (d, J=22.5 Hz, 2H), 6.97 (s, 1H), 4.38 (d, J=8.5 Hz, 1H), 4.17
(dtd, J=13.1, 9.6, 4.8 Hz, 2H), 3.91 (s, 3H), 3.80 (t, J=8.9 Hz,
1H), 3.53 (t, J=6.2 Hz, 3H), 3.32 (s, 3H), 2.32 (q, J=7.7 Hz, 4H),
2.13-2.00 (m, 3H), 1.65 (dt, J=13.2, 6.8 Hz, 2H), 1.45 (dt, J=12.9,
7.9 Hz, 1H), 1.22 (s, 3H), 0.48 (s, 3H).
Example 2: Synthesis of Racemic
10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,3,3a,7,12b-hexahy-
drocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylic acid
[rac-2]
##STR00027##
[0310] Using the trans-fused isomer from Step 4 of Example 1, the
title compound was prepared by the same method used to make Example
1. LCMS (m/z): 428.2 [M+H].sup.+. 1 H NMR (400 MHz, CD.sub.3CN):
.delta. 8.72 (s, 1H), 7.34 (s, 1H), 7.23 (s, 1H), 6.84 (s, 1H),
4.17 (q, J=6.4 Hz, 2H), 3.90 (s, 3H), 3.74 (d, J=13.5 Hz, 1H),
3.60-3.42 (m, 4H), 3.32 (s, 4H), 2.31 (dq, J=17.6, 7.9, 6.3 Hz,
2H), 2.10-2.00 (m, 3H), 1.52 (s, 4H), 1.29 (s, 4H)
Example 3.1: Synthesis of
(3aS,12bR)-8-fluoro-10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1-
,2,3,3a,7,12b-hexahydrocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylic
acid
Step 1: ethyl
(Z)-2-(ethoxymethylene)-4,4-difluoro-3-((trimethylsilyl)oxy)but-3-enoate
[3.1a]
##STR00028##
[0312] Under an argon atmosphere, a mixture of Mg (3.69 g, 152
mmol)) and TMSCl (19.43 mL, 152 mmol) was treated with ultrasound
irradiation for 15 min. To the mixture was added DMF (30 mL)),
ethyl (Z)-2-(ethoxymethylene)-4,4,4-trifluoro-3-oxobutanoate (4.56
g, 19 mmol) was added dropwise at 50.degree. C. under an argon
atmosphere. The reaction mixture was stirred for additional 3 min.
at 50.degree. C. After removal of excess TMSCl in vacuo, the crude
mixture was filtered and the filtrate (containing 3.1a and DMF) was
used in the next step without further purification.
Step 2: ethyl
8-fluoro-10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,3,3a,7,1-
2b-hexahydrocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylate
[3.1b-1] and [3.1b-2]
##STR00029##
[0314] To a suspension of ZnI.sub.2 (920 mg, 2.88 mmol)) and 1.1d-I
(915 mg, 2.88 mmol) in dry MeCN (10 mL), a solution of crude 3.1a
(5091 mg, 17.30 mmol) in dry DMF (30 mL) was added dropwise at
50.degree. C., and the reaction mixture was stirred overnight. The
reaction mixture was poured into 10% HCl and extracted with DCM.
The organic layer was washed with brine, and dried over MgSO.sub.4.
After filtration, the organic layer was concentrated and the crude
oil purified by silica gel chromatography (0-10% MeOH/EtOAc) to
afford rac-3.1 b (1.2 g, 2.53 mmol, 88% yield)) as a pale yellow
solid. The material was then purified by chiral SFC (AD column,
flow rate 100 ml/min, CO.sub.2/EtOH=70/30, 256 bar) to provide to
provide products 3.1b-1 (tR 2.4 min) and 3.1b-2 (tR 4.4 min, 280
mg). LC-MS (m/z): 474.2 [M+H].sup.+.
Step 3:
(3aS,12bR)-8-fluoro-10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl--
7-oxo-1,2,3,3a,7,12b-hexahydrocyclopenta[c]pyrido[2,1-a]isoquinoline-6-car-
boxylic acid [3.1]
##STR00030##
[0316] To the solution of 3.1b-2 (330 mg, 0.697 mmol) in THF (1 mL)
was added NaOH (1.394 mL, 1.394 mmol). The reaction mixture was
stirred for 2 h, then the reaction was acidified with 1.5 ml 1N HCl
and extracted with dichloromethane. The organic layer was washed
with brine, and dried over MgSO.sub.4. After filtration and
concentration, the resultant solid was recrystallized from hot
EtOH/Water (5 ml; 5 ml) and the solids collected by vacuum
filtration. The material was further lyophilized from MeCN and
water to give product (230 mg, 0.511 mmol, 73.3%) as a tan solid.
LC-MS (m/z): 446.4 [M+H].sup.+. .sup.1H NMR (500 MHz, DMSO-d.sup.3)
.delta. 8.82 (s, 1H), 7.56 (s, 1H), 7.20 (s, 1H), 4.52 (d, J=5.5
Hz, 1H), 4.20-4.06 (m, 2H), 3.82 (s, 3H), 3.31 (dd, J=15.9, 4.8 Hz,
2H), 3.26 (s, 3H), 3.18 (d, J=15.9 Hz, 1H), 2.01 (p, J=6.3 Hz, 2H),
1.57-1.50 (m, 1H), 0.87 (d, J=6.6 Hz, 3H), 0.73 (d, J=6.7 Hz,
3H).
[0317] Compound 3.2 was synthesized from 3.1b-1 following step 3
procedure. LC-MS (m/z): 446.2 [M+H].sup.+.
Example 4.1: Synthesis of
(3aR,12bR)-10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-3,3a,7,12b-
-tetrahydro-2H-furo[3,2-c]pyrido[2,1-a]isoquinoline-6-carboxylic
acid
##STR00031##
[0318] Step 1:
4-(benzyloxy)-1-(4-methoxy-3-(3-methoxypropoxy)phenyl)-3,3-dimethylbutan--
2-one
##STR00032##
[0320] A 250 mL oven-dried round-bottomed flask was charged with
4-bromo-1-methoxy-2-(3-methoxypropoxy)benzene (6.7 g, 24.35 mmol),
XPhos (0.348 g, 0.731 mmol), Pd(OAc).sub.2 (0.082 g, 0.365 mmol)
and purged with nitrogen. Dioxane (Volume: 32.5 ml) was added and
the mixture stirred until homogeneous. LiHMDS (1.0 M in THF, 70.6
ml, 70.6 mmol) was added. 4-(Benzyloxy)-3,3-dimethylbutan-2-one
(10.05 g, 48.7 mmol) in 4 mL dioxane was added slowly. The flask
was fitted with a nitrogen-purged reflux condenser and then heated
to 71.degree. C. for 90 min. After cooling, the mixture was poured
into water and the pH brought to 3 with 4 N HCl. The aqueous layer
was extracted thrice with EtOAc, the combined organic layers were
dried over Na.sub.2SO.sub.4, and then concentrated onto 16 g
diatomaceous earth. The material was purified on a 120 g SiO2
Combiflash cartridge (0->50% EtOAc in heptanes). The major UV
active peak was concentrated to provide 4.1a (8.8 g, 21.97 mmol,
90% yield) as a yellow oil. LC-MS (m/z): 401.3 [M+H].sup.+. .sup.1H
NMR (400 MHz, CDCl.sub.3): 7.28-7.36 (m, 5H), 6.79 (d, J=8.2 Hz,
1H) 6.6-6.7 (m, 2H), 4.51 (s, 2H), 4.04 (t, J=6.5 Hz), 2H), 3.83
(s, 3H), 3.75 (s, 2H), 3.55 (t, J=6.2 Hz, 2H), 3.51 (s, 2H), 3.34
(s, 3H), 2.07 (quint, J=6.3 Hz, 2H), 1.20 (s, 6H).
Step 2:
4-(benzyloxy)-1-(4-methoxy-3-(3-methoxypropoxy)phenyl)-3,3-dimethy-
lbutan-2-amine [4.1b]
##STR00033##
[0322] A round-bottomed flask was charged with 4.1a (600. mg, 1.498
mmol) and methanol (1.5 mL). Ammonium acetate (1.732 g, 22.47 mmol)
was added and the mixture was stirred at rt for 30 mins then sodium
cyanoborohydride (471 mg, 7.49 mmol) was added. The mixture was
heated at 60.degree. C. until the reaction for 18 h. The reaction
was quenched by adding 6 N NaOH and stirring for 30 min. The
mixture was extracted twice with 2-Me-THF, the combined organic
layers dried over Na.sub.2SO.sub.4, and then filtered and
concentrated to provide crude 4.1 b (602 mg, 1.498 mmol, 100%
yield) that was used without further purification. LC-MS (m/z):
402.4 [M+H].sup.+.
Step 3:
N-(4-(benzyloxy)-1-(4-methoxy-3-(3-methoxypropoxy)phenyl)-3,3-dime-
thylbutan-2-yl)formamide [4.1c]
##STR00034##
[0324] 4.1 b (600 mg, 1.494 mmol) was taken up in DMF (2.3 ml) and
cooled to 0.degree. C. Triethylamine (1.04 mL, 7.5 mmol), was added
followed by EDC (573 mg, 2.99 mmol) and formic acid (229 .mu.l,
5.98 mmol). The mixture was stirred at rt for 2 hours. Water was
added and the mixture was extracted with EtOAc. The combined
organic layers were washed with 1.0 H aq HCl solution, brine, then
dried over Na.sub.2SO.sub.4 and concentrated. The crude material
was purified by silica gel (0->70% acetone in heptane) to give
4.1c (520 mg, 1.211 mmol, 81% yield) as a yellow oil. LC-MS (m/z):
430.3 [M+H].sup.+.
Step 4:
3-(1-(benzyloxy)-2-methylpropan-2-yl)-7-methoxy-6-(3-methoxypropox-
y)-3,4-dihydroisoquinoline [4.1d]
##STR00035##
[0326] A 200 mL round-bottomed flask was charged with 4.1c (4.71 g,
10.96 mmol) and purged with nitrogen. Acetonitrile (Volume: 43.8
ml) was added, and the flask cooled to 0-5.degree. C. POCl.sub.3
(1.532 ml, 16.44 mmol) was added dropwise. The flask was fitted
with a dry condenser and the mixture was heated to 70.degree. C.
for 1 hr. The mixture was cooled to rt and the volatiles were
removed on the rotary evaporator. The resultant oil was diluted
with EtOAc and water, then basified with sat. NH.sub.4OH until the
aqueous layer reached pH 11. The layers were separated and the
aqueous layer was extracted with EtOAc twice. The organic phases
were combined, dried over Na.sub.2SO.sub.4, filtered and
concentrated onto 6 g diatomaceous earth and purified on a 40 g
SiO2 Combiflash column (0->60% acetone/heptane) to provide 4.1d
(3.8 g, 84% yield) as an oil. LC-MS (m/z): 412.5 [M+H].sup.+.
Step 5: ethyl
6-(1-(benzyloxy)-2-methylpropan-2-yl)-10-methoxy-9-(3-methoxypropoxy)-2-o-
xo-6,7-dihydro-2H-pyrido[2,1-a]isoquinoline-3-carboxylate
[4.1e]
##STR00036##
[0328] To a solution of 4.1d (3.5 g, 8.50 mmol) in EtOH (Volume:
8.50 ml, Ratio: 1.000) was added (Z)-ethyl
2-(ethoxymethylene)-3-oxobutanoate (5.92 ml, 34.0 mmol) in a 20 mL
microwave vial. The mixture was then sealed and flushed with
nitrogen. The vial was heated at 110.degree. C. for 18 hours. The
reaction mixture was concentrated to dryness under vacuum. To the
residue was added DME (Volume: 8.50 ml, Ratio: 1.000) and
P-CHLORANIL (2.509 g, 10.21 mmol). The vial was sealed and heated
at 100.degree. C. for 1 hour. The solvent was removed and the
residue loaded onto 12 g diatomaceous earth and purified by silica
gel column chromatography (IPA/EtOAc 0->70%) to provide 4.1e
(3.5 g, 75%) as a clear oil. LC-MS (m/z): 550.5 [M+H].sup.+.
Step 6: ethyl
6-(1-hydroxy-2-methylpropan-2-yl)-10-methoxy-9-(3-methoxypropoxy)-2-oxo-6-
,7-dihydro-2H-pyrido[2,1-a]isoquinoline-3-carboxylate [4.1f]
##STR00037##
[0330] A mixture of 4.1e (600 mg, 1.092 mmol) and 10% Pd/C (349 mg,
0.327 mmol) in EtOH (Volume: 10 mL) was purged with H.sub.2 and
stirred for 4 h. After filtration, the filtrate was concentrated to
dryness to give the crude material which was purified by silica gel
chromatography (0-50% MeOH/EtOAc) to give 4.1f (353 mg, 0.768 mmol,
70.4% yield) as an oil. LC-MS (m/z): 460.3 [M+H].sup.+.
Step 7: ethyl
(3aR*,12bR*)-10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-3,3a,7,1-
2b-tetrahydro-2H-furo[3,2-c]pyrido[2,1-a]isoquinoline-6-carboxylate
[4.1g]
##STR00038##
[0332] To a solution of 4.1f (311 mg, 0.677 mmol) in acetonitrile
(9 mL) was added a solution of CuSO.sub.4 (108 mg, 0.677 mmol) and
K.sub.2S.sub.2O.sub.8 (366 mg, 1.354 mmol) in water (Volume: 1.9
mL). The resulting mixture was stirred at reflux for 1 hour. The
mixture was added 100 mL ethyl acetate, washed with water, brine
and dried over Na.sub.2SO.sub.4. The organic phase was concentrated
and the residue was used in the next step without further
purification. The crude residue was treated with a mixture of 2 mL
AcOH/0.1 mL H.sub.2SO.sub.4. After stirred for 20 mins at room
temperature, the mixture was added 100 ml DCM, washed with
saturated NaHCO.sub.3, dried over Na.sub.2SO.sub.4. The organic
phase was concentrated and the residue was purified by silica get
chromatography (0-70% IPA/EA) to give product 4.1g (180 mg,
60.4%).
Step 8:
(3aR,12bR)-10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-3,3-
a,7,12b-tetrahydro-2H-furo[3,2-c]pyrido[2,1-a]isoquinoline-6-carboxylic
acid [4.1]
##STR00039##
[0334] LiOH (3.28 ml, 6.56 mmol) in water was added to a solution
of 4.1g (1.2 g, 2.62 mmol) in THF (8.74 ml) and the mixture was
stirred for 30 min. The solution was then acidified by adding 4.0 N
HCl, diluted with water, and extracted thrice with DCM. The
combined organic layers were dried over Na.sub.2SO.sub.4, filtered
and concentrated. The material was purified by chiral
chromatography (AD column, SFC 5 mL/min, CO.sub.2/IPA=70/30), and
returned as 4.1 (1.36 min) and 4.2 (1.86 min). 4.1 (348 mg, 0.802
mmol, 30.6% yield) was a fluffy solid after lyophilization from
MeCN/H.sub.2O. LC-MS (m/z): 430.4 [M+H].sup.+. .sup.1H NMR (500
MHz, DMSO-d6): 8.60 (s, 1H), 7.66 (s, 1H), 7.56 (s, 1H), 7.06 (s,
1H), 5.54 (d, J=8.8 Hz, 1H), 4.96 (d, J=8.5 Hz, 1H), 4.11 (m, 2H),
3.91 (s, 3H), 3.70 (d, J=8.8 Hz, 1H), 3.48 (t, J=6.4 Hz, 2H), 3.37
(d, J=9.0 Hz, 1H), 3.25 (s, 3H), 1.99 (quint, J=6.6 Hz, 2H), 1.25
(s, 3H), 0.46 (s, 3H).
Example 5: Synthesis
10-chloro-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,3,3a,7,12b-hexahyd-
rocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylic acid [5.1]
and [5.2]
##STR00040##
[0335] Step 1: 4-bromo-1-chloro-2-(3-methoxypropoxy)benzene
[5a]
##STR00041##
[0337] To a mixture of 5-bromo-2-chlorophenol (15 g, 72.3 mmol),
and 1-bromo-3-methoxypropane (9.7 ml, 87 mmol) in DMF (40 ml) at rt
was added K.sub.2CO.sub.3 (20 g, 145 mmol), and the resultant
mixture was stirred at 50.degree. C. for 16 hours. The mixture was
then filtered, and the filtrate was concentrated. The residue was
purified by silica gel column chromatography, EtOAc/heptane 0 to
30% to give product (18.4 g, 91% yield). .sup.1H NMR (400 MHz,
Acetonitrile-d3): 7.38-7.20 (m, 2H), 7.10 (dd, J=8.4, 2.1 Hz, 1H),
4.13 (t, J=6.3 Hz, 2H), 3.54 (t, J=6.2 Hz, 2H), 3.31 (s, 3H),
2.09-1.98 (m, 2H).
Step 2:
5-(4-chloro-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclopentanone
[5b]
##STR00042##
[0339] A mixture of Pd(OAc).sub.2 (60 mg, 0.268 mmol), sodium
tert-butoxide (3.35 g, 34.9 mmol),
dicyclohexyl(2'-methyl-[1,1'-biphenyl]-2-yl)phosphane (230 mg),
2,2-dimethylcyclopentanone (4.04 ml, 32.2 mmol) and
4-bromo-1-methoxy-2-(3-methoxypropoxy)benzene (7.5 g, 26.8 mmol) in
toluene (30.0 mL) was heated in a sealed vial at 50.degree. C.
under nitrogen atmosphere for 6 hours. The mixture was diluted with
EtOAc and filtered. The filtrated solution was concentrated and the
remaining oil was purified by silica gel column chromatography,
EtOAc/heptane 5 to 50%, to give product (3 g, 36% yield). LC-MS
(m/z): 311.2 [M+H].sup.+.
Step 3:
5-(4-Chloro-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclopentanami-
ne [5c]
##STR00043##
[0341] To the mixture of
5-(4-chloro-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclopentanone
(3 g, 9.65 mmol) in MeOH (30 mL) was added acetic acid ammonia salt
(14.8 g, 192 mmol) and sodium cyanoborohydride (6.06 g, 97 mmol).
The mixture was stirred at 70.degree. C. for 16 hours and then was
concentrated under reduced pressure. The remaining material was
diluted with EtOAc, washed with water and brine, dried over
Na.sub.2SO.sub.4 and concentrated. The crude material was used in
the next step with no further purification. LCMS (m/z): 312.0
[M+H].sup.+.
Step 4: N-(5-(4-chloro-3-(3-methoxypropoxy)phenyl)-2,2
dimethylcyclopentyl)formamide [5d]
##STR00044##
[0343] To the mixture of
5-(4-chloro-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclopentanamine
(3 g, 9.65 mmol) in dioxane (25 ml) was added formic acid (1.48 ml,
38.6 mmol). The mixture was stirred at 100.degree. C. for 6 hours.
The mixture was concentrated to afford the crude product which was
used in the next step with no further purification. LCMS (m/z):
340.2 [M+H].sup.+.
Step 5:
7-chloro-8-(3-methoxypropoxy)-3,3-dimethyl-2,3,3a,9b-tetrahydro-1H-
-cyclopenta[c]isoquinoline [rac-5e-1 and rac-5e-2]
##STR00045##
[0345] To a mixture of
N-(5-(4-chloro-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclopentyl)formam-
ide (3.28 g, 9.65 mmol) in acetonitrile (17 ml) was added
POCl.sub.3 (1.35 ml, 14.5 mmol). The mixture was stirred at
85.degree. C. for 2 hours and then concentrated. The residue was
dissolved in EtOAc and basified by adding ammonium hydroxide
solution. The phases were separated and the organic layer was
washed with brine, dried over Na.sub.2SO.sub.4 and concentrated.
The remaining material was purified by silica gel chromatography,
acetone/heptane 5 to 50% to give product rac-5e-1 and rac-5e-2.
[0346] Less polar product: rac-5e-1 trans isomer (590 mg, 19%
yield). LCMS (m/z): 322.3 [M+H].sup.+.
[0347] More polar product: rac-5e-2 cis isomer (470 mg, 15% yield).
LCMS (m/z): 322.3 [M+H].sup.+. .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.15 (s, 1H), 7.25 (s, 2H), 6.69 (s, 1H), 4.14 (s, 3H),
3.78 (d, J=10.1 Hz, 1H), 3.60 (s, 3H), 3.43-3.32 (m, 4H), 3.26 (q,
J=9.5, 9.1 Hz, 2H), 2.47-2.19 (m, 3H), 2.16-2.02 (m, 4H), 1.66 (d,
J=10.1 Hz, 4H), 1.46 (t, J=8.6 Hz, 3H), 1.23 (s, 4H), 1.16-1.00 (m,
3H), 0.88 (s, 4H)
[0348] The relative configuration of rac-5e-1 and rac-5e-2 was
established by nuclear Overhauser effect (NOE) experiments.
Step 6: Ethyl
10-chloro-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,3,3a,7,8,8a,12b-oc-
tahydrocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylate
[rac-5f]
##STR00046##
[0350] To a mixture of
7-chloro-8-(3-methoxypropoxy)-3,3-dimethyl-2,3,3a,9b-tetrahydro-1H-cyclop-
enta[c]isoquinoline (cis isomer, rac-5e-2) (470 mg, 1.46 mmol) in
EtOH (5 ml) was added (Z)-ethyl 2-(ethoxymethylene)-3-oxobutanoate
(816 mg, 4.38 mmol). The mixture was stirred at 110.degree. C. for
16 hours. After cooling, the mixture was concentrated and the crude
material was used in the next step with no further purification.
LCMS (m/z): 462.0 [M+H].sup.+.
Step 7: Ethyl
(3aS,12bR)-10-chloro-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,3,3a,7,-
12b-hexahydrocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylate
[5g-1] and [5g-2]
##STR00047##
[0352] To a mixture of rac-5f ((673.7 mg, 1.461 mmol) in DME (3.5
ml) was added p-chloranil (359.6 mg, 1.461 mmol) and the mixture
was stirred at 110.degree. C. for 2 hours. After cooling to rt, the
mixture was concentrated and the residue was purified by silica gel
column chromatography, MeOH/DCM 0 to 6%, to give product (180 mg).
LCMS (m/z): 460.0 [M+H].sup.+. This product was separated by chiral
SFC (AD column, flow rate 100 ml/min, CO.sub.2/MeOH=75/25) to give
two enantiomers: 5g-1 (tR 3.55 min, 70 mg, 11% yield) and 5g-2 (tR
4.91 min, 70 mg, 11% yield).
Step 8:
(3aS,12bR)-10-chloro-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,-
3,3a,7,12b-hexahydrocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylic
acid [5.1]
##STR00048##
[0354] To a mixture of 5g-1 (70 mg, 0.152 mmol) in EtOH (1 ml) was
added NaOH (5 M, 0.152 ml, 0.761 mmol). After stirring at rt for 2
hours, the mixture was concentrated and then acidified by adding
3.0 N HCl aq solution. To the mixture was added EtOAc. The organic
layer was washed with water and brine, dried, and concentrated. The
crude residue was purified by silica gel column chromatography,
MeOH in DCM 0 to 5%, to give product (50 mg, 75% yield). LCMS
(m/z): 432.2 [M+H].sup.+. .sup.1H NMR (400 MHz, Acetonitrile-d3):
8.40 (s, 1H), 8.01 (s, 1H), 7.22 (s, 1H), 7.09 (s, 1H), 4.39 (d,
J=8.6 Hz, 1H), 4.24 (ddt, J=15.9, 9.6, 4.8 Hz, 2H), 4.08 (q, J=7.1
Hz, 4H), 3.83 (t, J=6.8 Hz, 1H), 3.57 (t, J=6.1 Hz, 2H), 3.32 (s,
3H), 2.48-2.24 (m, 3H), 2.15 (s, 2H), 2.07 (p, J=6.2 Hz, 3H), 1.66
(ddd, J=13.9, 8.1, 6.0 Hz, 1H), 1.56-1.40 (m, 1H), 1.28-1.16 (m,
8H), 0.47 (s, 3H).
Step 9:
(3aR,12bS)-10-chloro-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-1,2,-
3,3a,7,12b-hexahydrocyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylic
acid [5.2]
##STR00049##
[0356] To a mixture of 5g-2 (70 mg, 0.152 mmol) in EtOH (1 ml) was
added NaOH (5 M, 0.152 ml, 0.761 mmol). After stirring at rt for 2
hours, the mixture was concentrated and then acidified by adding
3.0 N HCl aq solution. To the mixture was added EtOAc. The organic
layer was washed with water and brine, dried, and concentrated. The
crude residue was purified by silica gel column chromatography,
MeOH in DCM 0 to 5%, to give product (59 mg, 79% yield). LCMS
(m/z): 432.2 [M+H].sup.+. .sup.1H NMR (400 MHz, Acetonitrile-d3):
8.40 (s, 1H), 8.01 (s, 1H), 7.22 (s, 1H), 7.09 (s, 1H), 4.39 (d,
J=8.6 Hz, 1H), 4.24 (ddt, J=15.9, 9.6, 4.8 Hz, 2H), 4.08 (q, J=7.1
Hz, 4H), 3.83 (t, J=6.8 Hz, 1H), 3.57 (t, J=6.1 Hz, 2H), 3.32 (s,
3H), 2.48-2.24 (m, 3H), 2.15 (s, 2H), 2.07 (p, J=6.2 Hz, 3H), 1.66
(ddd, J=13.9, 8.1, 6.0 Hz, 1H), 1.56-1.40 (m, 1H), 1.28-1.16 (m,
8H), 0.47 (s, 3H).
Example 6:
10-methoxy-11-(3-methoxypropoxy)-7-oxo-1,2,3,3a,7,12b-hexahydro-
cyclopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylic acid [6.1 and
6.2]
##STR00050##
[0357] Step 1.
2-(4-methoxy-3-(3-methoxypropoxy)phenyl)cyclopentan-1-one. [6a]
##STR00051##
[0359] To a solution of
4-bromo-1-methoxy-2-(3-methoxypropoxy)benzene (5 g, 18.24 mmol) in
1,4-Dioxane (100.0 mL) was added NaOAc (1.49 g 18.24 mmol),
cyclopentanone (4.59 g, 54.74 mmol), pyrrolidine (0.259 g, 3.64
mmol), P(O-Tol).sub.3 (0.222 g, 0.72 mmol.) and
1,1,3,3-Tetramethylbutylamine (0.471 g, 3.64 mmol) at room
temperature under nitrogen atmosphere. The reaction mixture was
purged with nitrogen. To the above reaction mixture, Pd(OAc).sub.2
(0.0817 g, 0.364 mmol) was added and the mixture was again purged
with nitrogen and then heated at 110.degree. C. for 15 hs. After
cooled at rt, the reaction mixture was filtered through celite bed,
the bed was further washed with ethyl acetate. The filtrate was
washed with water, brine, and dried over sodium sulfate, filtered,
and concentrated. The residue was purified by silica gel column
chromatography, (EtOAc/Hexane, 20-30%) to give product. LCMS (m/z):
279.35 [M+H].sup.+
Step 2.
1-(4-methoxy-3-(3-methoxypropoxy)phenyl)-3,3-dimethylbutan-2-amine-
.[6b]
##STR00052##
[0361] NH.sub.4OAc (8.73 g, 113 mmol) and NaBH.sub.3CN (0.949 g,
15.1 mmol) were added temperature to a solution of 6a (2.1 g, 7.55
mmol) in MeOH (15.1 mL) at rt and the resulting mixture was stirred
at room temperature for 18 hours. The reaction was then quenched by
adding 20% NaOH aqueous solution and stirred at rt for 20 minutes.
The reaction mixture was extracted with EtOAc. The organic layer
was washed with water, brine, dried over sodium sulfate and
concentrated to give product 6b (2.1 g crude). LCMS (m/z):281.2
[M+H].sup.+
Step 3.
N-(1-(4-methoxy-3-(3-methoxypropoxy)phenyl)-3,3-dimethylbutan-2-yl-
)formamide [6c]
##STR00053##
[0363] To a solution of 6b (2.0 g, 7.16 mmol) in DMF (11.9 mL) at
0.degree. C. was added EDC.HCl (2.73 g, 14.3 mmol), DIPEA (2.77 g,
21.5 mmol), followed by formic acid (1.31 g, 28.6 mmol). After
stirred at rt for 1 h, cold water was added to above reaction
mixture and the mixture was extracted with EtOAc. The organic layer
was washed with 10% NaHCO.sub.3 aq. solution, 10% HCl aq. solution,
water and brine. The separated organic layer was dried over sodium
sulfate, filtered and concentrated to give product. 6c (1.4 g). The
crude material was used in the next step with no further
purification. LCMS (m/z): 308.1 [M+H]+
Step 4.
7-methoxy-8-(3-methoxypropoxy)-2,3,3a,9b-tetrahydro-1H-cyclopenta[-
c]isoquinoline. [6d-1] and [6d-2]
##STR00054##
[0365] POCl.sub.3 (0.778 g, 5.07 mmol) was added to a solution of
6c (1.30 g, 4.23 mmol) in CH.sub.3CN (21.15 mL) at 0.degree. C. and
the reaction mixture was heated at 80.degree. C.) for 3 hours. The
reaction mixture was concentrated under vacuum and the residue was
dissolved in ethyl acetate. Ammonia solution (27% in water) was
added under stirring until the pH=11. The mixture was then
extracted with ethyl acetate. The combined ethyl acetate layers was
washed with water, brine, dried over sodium sulfate and
concentrated. The residue was purified by silica gel column
chromatography gave two isomers. 6d-1 (0.425 g, less polar product)
and 6d-2 (0.3 g, more polar isomer).
[0366] 6d-1: LCMS (m/z): 290.1 [M+H].sup.+. .sup.1H NMR (400 MHz,
CDCl.sub.3): 8.23 (d, J=3.1 Hz, 1H), 6.92 (s, 1H), 6.75 (d, J=8.8
Hz, 1H), 4.19 (t, J=6.5 Hz, 2H), 3.90 (s, 3H), 3.60 (t, J=6.0 Hz,
3H), 3.39 (d, J=9.1 Hz, 4H), 3.21-3.15 (m, 1H), 2.60-2.51 (m, 1H),
2.30-2.11 (m, 5H), 1.98-1.90 (m, 2H), 1.82 (dt, J=19.6, 11.4 Hz,
2H).
[0367] 6d-2: LCMS (m/z): 290.1 [M+H].sup.+. .sup.1H NMR (400 MHz,
CDCl.sub.3): 8.16 (s, 1H), 6.84 (s, 1H), 6.75 (s, 1H), 4.18 (t,
J=6.4 Hz, 2H), 3.90 (s, 3H), 3.60 (t, J=6.0 Hz, 2H), 3.38 (s, 3H),
2.91 (dd, J=17.4, 8.6 Hz, 1H), 2.34 (dd, J=14.8, 7.0 Hz, 1H),
2.20-2.10 (m, 3H), 2.08-1.95 (m, 3H), 1.67-1.55 (m, 3H), 1.60-1.44
(m, 2H), 0.97-0.82 (m, 2H).
Step 5. Ethyl
10-methoxy-11-(3-methoxypropoxy)-7-oxo-1,2,3,3a,7,12b-hexahydrocyclopenta-
[c]pyrido[2,1-a]isoquinoline-6-carboxylate [6e-1]
##STR00055##
[0369] A solution of 6d-1 (0.425 g, 1.45 mmol) and ethyl
(E)-2-(ethoxymethylene)-3-oxobutanoate (0.810 g, 4.35 mmol) in EtOH
(9 ml) was heated at 110.degree. C. for 18 hours. The mixture was
then concentrated under vacuum and the residue was dissolved in DME
(20.0 mL). p-chloranil (0.426 g, 1.73 mmol) was added and the
reaction mixture was heated to reflux for 2 hours. After volatile
solvent was removed under vacuum, diethyl ether was added. The
precipitate was collected by filtration to afford product. LCMS
(m/z): 428.3 [M+H].sup.+.
Step 6.
10-methoxy-11-(3-methoxypropoxy)-7-oxo-1,2,3,3a,7,12b-hexahydrocyc-
lopenta[c]pyrido[2,1-a]isoquinoline-6-carboxylic acid [6.1] and
[6.2]
##STR00056##
[0371] To a solution of 6e-1 (0.075 g, 0.175 mmol) in MeOH (6.0
mL), LiOH.2H.sub.2O (0.014 g, 0.35 mmol) was added followed by the
addition of water (2.0 mL) at rt. The reaction mixture was then
stirred at rt for 2 hours. After completion of reaction, the
reaction mixture was concentrated under vacuum. The residue was
dissolved into cold water and was acidified to pH 4-5 using dil.
HCl. The obtained solid was filtered, washed with cold water and
diethyl ether. The solid was dried well under vacuum to give off
white solid as the desired product 6.1 (0.029 g, 41% yield). LCMS
(m/z): [M+H].sup.+ 400.0. .sup.1H NMR (400 MHz, DMSO): 16.83 (s,
1H), 8.80 (s, 1H), 7.54 (d, J=3.3 Hz, 2H), 7.07 (s, 1H), 5.01-4.74
(m, 1H), 4.29-4.00 (m, 2H), 3.89 (s, 3H), 3.57 (s, 1H), 3.48 (t,
J=6.2 Hz, 2H), 3.34 (s, 3H), 2.37 (s, 1H), 2.23 (d, J=8.5 Hz, 1H),
2.08 (d, J=6.6 Hz, 1H), 2.01-1.90 (m, 2H), 1.64 (s, 1H), 1.49 (s,
2H).
[0372] Compound 6.2 was synthesized from compound 6d-2 following
the procedures Step 5-6 described for the synthesis of 6.1. LCMS
(m/z): [M+H].sup.+ 400.0. .sup.1H NMR (400 MHz, DMSO): 16.77 (s,
1H), 8.38 (s, 1H), 7.52 (d, J=20.7 Hz, 2H), 6.85 (s, 1H), 4.13 (t,
J=6.4 Hz, 2H), 3.89 (s, 3H), 3.48 (t, J=6.2 Hz, 2H), 3.26 (s, 3H),
3.09 (dd, J=19.7, 12.1 Hz, 1H), 2.38 (s, 1H), 2.10-1.92 (m, 5H),
1.70 (s, 1H).
[0373] Note: isomers 6.1 and 6.2 were isolated and tested
separately, but the stereochemistry of the isomers was not apparent
from their nmr data.
Example 7:
(3aR,12bR)-8-fluoro-10-methoxy-11-(3-methoxypropoxy)-3,3-di
methyl-7-oxo-3,3a,7,12b-tetrahydro-2H-furo[3,2-c]pyrido[2,1-a]isoquinolin-
e-6-carboxylic acid [7.1]
##STR00057##
[0374] Step 1:
4-hydroxy-1-(4-methoxy-3-(3-methoxypropoxy)phenyl)-3,3-dimethylbutan-2-on-
e [7.1a]
##STR00058##
[0376] A mixture of 4.1a (8.8 g, 21.97 mmol) and 10% Pd/C (2.1 g,
1.973 mmol) in MeOH (110 ml) was purged with vacuum and H.sub.2 was
stirred for 2 days. The atmosphere was purged with vacuum, and the
mixture was filtered through diatomaceous earth with MeOH washes,
and concentrated to provide a grey oil that was taken up in EtOAc
and passed through a small pad of SiO.sub.2, and the concentrated
to provide 7.1a (6.66 g, 21.46 mmol, 98% yield) as a yellow oil.
LC-MS (m/z): 311.3 [M+H].sup.+. .sup.1H NMR (500 MHz, CDCl.sub.3):
6.82 (d, J=8.24 Hz, 1H), 6.73 (s, 1H), 6.70 (d, J=8.10 Hz, 1H),
4.10 (t, J=6.41 Hz, 2H), 3.84 (s, 3H), 3.74 (s, 2H), 3.55-3.60 (m,
4H), 3.35 (s, 3H), 2.36 (br, 1H), 2.10 (quin, J=6.33 Hz, 2H), 1.23
(s, 6H).
Step 2:
2-(4-methoxy-3-(3-methoxypropoxy)phenyl)-4,4-dimethyldihydrofuran--
3(2H)-one [7.1b]
##STR00059##
[0378] A 2-neck, oven dried 250 mL round-bottomed flask was fitted
a pressure-equalizing addition funnel and with charged with 7.1a
(6.63 g, 21.36 mmol) and 50 mL DCM. The flask was cooled to
0.degree. C. The addition funnel was charged with Br.sub.2 (1.045
ml, 20.29 mmol) in 485 mL DCM, which was added dropwise over 25
min. Immediately after complete addition, the mixture was poured in
to 200 mL sat aq. Na.sub.2S2O.sub.3 and stirred for 20 min. The
mixture was extracted thrice with EtOAc. The combined organic
layers were dried over Na.sub.2SO.sub.4, filtered, and concentrate
onto 13 g diatomaceous earth. The crude material was purified on a
80 g RediSep SiO.sub.2 cartridge (0-60% EtOAc in heptanes) to
provide 7.1 b (3.89 g, 12.61 mmol, 59.1% yield) as a yellow oil
from the first major UV-active peak. LC-MS (m/z): 309.3
[M+H].sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3): 6.97 (s, 1H), 6.96
(d, J=7.96 Hz, 1H), 6.87 (br d, J=8.12 Hz, 1H), 4.76 (s, 1H),
4.08-4.17 (m, 3H), 3.96 (d, J=9.2 Hz, 1H), 3.85 (s, 3H), 3.57 (t,
J=6.13 Hz, 2H), 3.35 (s, 3H), 2.10 (quint, J=6.3 Hz, 2H), 1.20 (s,
3H), 1.16 (s, 3H).
Step 3:
N-(2-(4-methoxy-3-(3-methoxypropoxy)phenyl)-4,4-dimethyltetrahydro-
furan-3-yl)formamide [7.1c]
##STR00060##
[0380] A 250 mL round-bottomed flask was charged with 7.1b (3.7 g,
12.00 mmol) and MeOH (30.0 ml). NH.sub.4OAc (18.50 g, 240 mmol),
was added, followed by NaBH.sub.3CN (2.262 g, 36.0 mmol). The flask
was fitted with a condenser and refluxed overnight. At 20 h, the
mixture was cooled to rt and 30 mL 20 wt % NaOH (aq) was added.
After stirring for 1 h, diluted slightly with water, extracted
thrice with 2-MeTHF. The combined organic layers were dried over
Na.sub.2SO.sub.4, filtered and concentrated to provide a
heterogenous mixture. The mixture was taken up in DCM (30.0 ml).
NEt.sub.3 (10.03 ml, 71.9 mmol) was added, followed by formic acid
(1.840 ml, 48.0 mmol) and EDC (9.19 g, 48.0 mmol). At 1 h complete,
the mixture was poured into 20 mL water, 60 mL EtOAc and the layer
separated. The organic layer was washed twice with N NaHSO.sub.4,
once with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated onto diatomaceous earth. The crude material was
purified on a 40 g RediSep SiO.sub.2 cartridge (0->70% acetone
in heptane) to provide 7.1c (2.58 g, 7.65 mmol, 63.8% yield) as a
yellow oil. LC-MS (m/z): 338.3 [M+H].sup.+.
Step 4:
rac-(3aR,9bR)-7-methoxy-8-(3-methoxypropoxy)-3,3-dimethyl-2,3,3a,9-
b-tetrahydrofuro[3,2-c]isoquinoline [7.1d]
##STR00061##
[0382] A 200 mL round-bottomed flask was charged with 7.1c (2.6 g,
7.71 mmol) and purged with N.sub.2. MeCN (30.8 ml), followed by
POCl.sub.3 (1.077 ml, 11.56 mmol). The flask was fitted with a
condenser and then heated to 70.degree. C. At 30 min, the mixture
was cooled to rt and the volatiles were removed under reduced
pressure. The oil was diluted with EtOAc and water, and basisified
with sat. aq. NH.sub.4OH to pH 11. The layers were separated, and
the aqueous layer extracted twice with EtOAc. The combined organic
layers were dried over Na.sub.2SO.sub.4, filtered, and concentrate
onto 6 g diatomaceous earth. The crude material was purified on a
40 g RediSep SiO.sub.2 cartridge (0->70% acetone in heptane).
The most polar UV-active peak was isolated as 7.1d (900 mg, 2.82
mmol, 36.6% yield), which was shown to be the cis isomer by 2D NMR.
LC-MS (m/z): 320.3 [M+H]. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
8.27 (s, 1H), 6.98 (s, 1H), 6.86 (s, 1H), 4.97 (d, J=7.33 Hz, 1H),
4.14-4.23 (m, 2H), 3.87-3.94 (m, 4H), 3.48-3.60 (m, 3H), 3.39 (d,
J=7.80 Hz, 1H), 3.35 (s, 3H), 2.12 (quin, J=6.33 Hz, 2H), 1.37 (s,
3H), 1.14 (s, 3H).
Step 5: rac-ethyl
(3aR,12bR)-8-fluoro-10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-3-
,3a,7,12b-tetrahydro-2H-furo[3,2-c]pyrido[2,1-a]isoquinoline-6-carboxylate
[7.1e]
##STR00062##
[0384] To a suspension of ZnI.sub.2 (200 mg, 0.626 mmol) and 7.1d
(200 mg, 0.626 mmol) in dry MeCN (2 mL), was added a solution of
3.1a (553 mg, 1.879 mmol) in dry DMF (3 mL), dropwise, at
50.degree. C., and the reaction mixture was stirred overnight. The
reaction mixture was poured into 10% HCl and extracted with DCM.
The organic layer was washed with brine, and dried over MgSO.sub.4.
After filtration, the organic layer was concentrated onto
diatomaceous earth and purified on a 4 g RediSep SiO.sub.2
cartridge (0->60 IPA in EtOAc) to provide 7.1e (90 mg, 0.184
mmol, 30.6% yield) as a brown solid. LC-MS (m/z): 476.3
[M+H].sup.+.
Step 6:
(3aR,12bR)-8-fluoro-10-methoxy-1-(3-methoxypropoxy)-3,3-dimethyl-7-
-oxo-3,3a,7,12b-tetrahydro-2H-furo[3,2-c]pyrido[2,1-a]isoquinoline-6-carbo-
xylic acid [7.1]
##STR00063##
[0386] 7.1g (90 mg, 0.189 mmol) was suspended in THF (1.5 ml). Aq.
LiOH (500 .mu.l, 1.000 mmol) was added and the mixture was stirred
overnight. The pH was adjusted to 1 with 4 N HCl. The mixture was
extracted thrice with EtOAc. The combined organic layers were dried
over Na.sub.2SO.sub.4, filtered and concentrated to provide a light
brown solid. The material was purified by chiral SFC (AD column,
flow rate 100 ml/min, CO.sub.2/MeOH=80/20, 250 bar) to give two
enantiomers 7.1 (tR 4.56, 17.7 mg, 21%) and 7.2 (tR 2.89 min, 17.3
mg, 21%). Compound 7.1: LC-MS (m/z): 448.3 [M+H].sup.+. .sup.1H NMR
(500 MHz, CDCl.sub.3): 8.36 (s, 1H), 7.72 (s, 1H), 7.21 (s, 1H),
5.54 (d, J=7.6 Hz, 1H) 4.42 (d, J=6.4 Hz, 1H), 4.16-4.31 (m, 2H)
3.92 (s, 3H), 3.78 (d, J=8.5 Hz, 1H), 3.52-3.69 (m, 2H), 3.42 (br
d, J=9.22 Hz, 1H), 3.37 (s, 3H), 2.10-2.22 (m, 2H), 1.40 (s, 3H)
0.58 (s, 3H)
Example 8:
(3aS,12bR)-10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo--
3,3a,7,12b-tetrahydro-1H-furo[3,4-c]pyrido[2,1-a]isoquinoline-6-carboxylic
acid [8.1]
##STR00064##
[0387] Step 1:
3-hydroxy-1-(4-methoxy-3-(3-methoxypropoxy)phenyl)-3-methylbutan-2-one
[8.1a]
##STR00065##
[0389] A 200 mL round-bottomed flask was charged with
4-bromo-1-methoxy-2-(3-methoxypropoxy)benzene (13.0 g, 47.2 mmol),
NaOt-Bu (13.62 g, 142 mmol), xantphos (0.820 g, 1.417 mmol),
Pd.sub.2dba.sub.3 (0.649 g, 0.709 mmol) and THF (Volume: 140 mL).
To the mixture was added 3-hydroxy-3-methylbutan-2-one (9.65 g, 94
mmol). The flask was fitted with a reflux condenser and the mixture
was heated to 65.degree. C. in an aluminum chip bath for 3.5 h.
After cooling, the mixture was filtered through diatomaceous earth
with EtOAc and water washes. The pH of the aqueous layer was
adjusted to 2, the layers were separated, and the aqueous layer
extracted twice with EtOAc. The combined organic layers were dried
over Na.sub.2SO.sub.4, filter, and concentrated onto 6 g
diatomaceous earth. The material was purified by silica gel column
chromatography, EtOAc/heptane 0 to 70% to give product (5.7 g,
19.23 mmol, 40.7% yield). LC-MS (m/z): 297.3 [M+H].sup.+.
Step 2:
4-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2-dimethylfuran-3(2H)-o-
ne [8.1 b]
##STR00066##
[0391] 8.1a (1.43 g, 4.83 mmol) was dissolved in toluene (19.30
ml). Bredereck's reagent (1.993 ml, 8.69 mmol) was added, and the
flask fitted with reflux condenser and heated to 100.degree. C. for
1 h. After cooling to rt, 3 g of diatomaceous earth was added and
the volatiles were removed. The material was purified by silica gel
column chromatography, EtOAc/heptane 0 to 50% to provide product
8.1 b (1.26 g, 85% yield) as a yellow oil. LC-MS (m/z): 307.2
[M+H].sup.+. .sup.1H NMR (500 MHz, CHLOROFORM-d): 8.39 (s, 1H),
7.31 (d, J=1.89 Hz, 1H), 7.22 (dd, J=8.35, 2.05 Hz, 1H), 6.88 (d,
J=8.20 Hz, 1H), 4.16 (s, 2H), 3.87 (s, 3H), 3.58 (s, 2H), 3.36 (s,
3H), 2.13 (quin, J=6.38 Hz, 2H), 1.46 (s, 6H).
Step 3:
4-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2-dimethyldihydrofuran--
3(2H)-one [8.1c]
##STR00067##
[0393] 8.1b (1.26 g, 4.11 mmol) was taken up in EtOH (10.28 ml) and
THF (10.28 ml). The mixture was cooled to 0.degree. C. and
NaBH.sub.4 (0.202 g, 5.35 mmol) was added. After 2 h, the reaction
was quenched with saturated aq. NH.sub.4Cl and extracted with
EtOAc. The combined organic layers were dried over
Na.sub.2SO.sub.4, filtered and concentrated to provide an oil. The
oil was charged to a 100 mL round-bottomed flask and dissolved in
DCM (32 mL). The mixture was cooled to 0.degree. C. and DMP (4104
mg, 9.68 mmol) was added as a single portion. After 2 h, the
reaction mixture was filter through diatomaceous earth with DCM and
sat. aq. NaHCO.sub.3. The layers were separated. The aqueous layer
was extract twice with DCM. The combined organic layers were dried
over Na.sub.2SO.sub.4, filtered and concentrated to provide a
yellow oil which was purified by silica gel column chromatography,
EtOAc/heptane 0 to 70% to give product 8.1c (841 mg, 42.3% yield).
LC-MS (m/z): 309.2 [M+H].sup.+.
Step 4:
N-(4-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2-dimethyltetrahydro-
furan-3-yl)formamide [8.1 d]
##STR00068##
[0395] A 20 mL vial was charged with 8.1c (840 mg, 2.72 mmol) and
methanol (8 ml). Ammonium acetate (3.15 g, 40.9 mmol) was added and
the mixture stirred until homogeneous. Sodium cyanoborohydride
(0.342 g, 5.45 mmol) was added as a single portion. The yellow
solution was stirred overnight at 60.degree. C. At 20 h, The
mixture was cool to rt and quenched with 6 mL 5 M NaOH (20 wt %).
After 1 h, the mixture was extracted with EtOAc twice, dried over
Na.sub.2SO.sub.4, filtered and concentrated to provide a yellow
oil. The oil was taken up in formic acid (5.0 ml, 130 mmol) and
dioxane (7 ml) in an 8 mL vial. The vial was sealed and heated to
98.degree. C. 18 h. The volatiles were removed under reduced
pressure .degree. (50 C, 18 mbar) and the resultant oil azeotroped
twice with 30 mL toluene before a final concentration onto
diatomaceous earth with MeOH and DCM. The crude material was
purified by silica gel column chromatography, acetone/heptane
0->50% to give product 8.1d (230 mg, 25.1% yield) as a white
solid. LC-MS (m/z): 338.1 [M+H].sup.+.
Step 5:
7-methoxy-8-(3-methoxypropoxy)-3,3-dimethyl-1,3,3a,9b-tetrahydrofu-
ro[3,4-c]isoquinoline [8.1e]
##STR00069##
[0397] A 50 mL round-bottomed flask was charged with 8.1d (230 mg,
0.682 mmol) and purged with vacuum and back-filled with nitrogen.
MeCN (3.41 ml) was added, followed by POCl.sub.3 (0.095 ml, 1.022
mmol). The flask was fitted with a condenser and heated to
70.degree. C. At 2 h, cooled to rt and the volatiles were removed
under vacuum. The remaining oil was diluted with 40 mL EtOAc, and
10 mL water, and then basified with ammonium hydroxide solution
until pH 11. The layers were separated, and the aqueous layer
extracted EtOAc. The combined organic layers were dried over
Na.sub.2SO.sub.4, filtered and concentrated onto diatomaceous
earth. The crude material was purified by silica gel column
chromatography, acetone/heptane 0 to 50%, to give product 8.1e (202
mg, 93% yield). LC-MS (m/z): 320.2 [M+H].sup.+.
Step 6: ethyl
(3aS,12bR)-10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-3,3a,7,12b-
-tetrahydro-1H-furo[3,4-c]pyrido[2,1-a]isoquinoline-6-carboxylate
[8.1f]
##STR00070##
[0399] A 4 mL vial was charged with 8.1e (200 mg, 0.626 mmol) in
EtOH (1.5 ml). Added ethyl (E)-2-(ethoxymethylene)-3-oxobutanoate
(408 mg, 2.192 mmol). The vial was sealed, purged with N2, and
heated to 85.degree. C. overnight. An additional 400 mg ethyl
(E)-2-(ethoxymethylene)-3-oxobutanoate was added. After 5 h, the
volatiles were removed under vacuum. The oil was taken up in DME
(1.3 mL) and p-chloroanil (185 mg) was added. The mixture was
heated at 100.degree. C. for 30 min. After cooling at rt, the
solvent was removed on rotovap and 5 mL ether was added and the
solid filtered. The black solid was taken up in MeOH and loaded
onto diatomaceous earth, and then purified by silica gel column
chromatography, IPA/EtOAc 0 to 70 to give product. The
stereoisomers were separated by chiral HPLC (AD column, flow rate 1
mL/min, heptane/IPA=60/40). Peak 3 (tR 6.76 min) was isolated as
8.1f (13.5 mg, 0.030 mmol, 4.71% yield). LC-MS (m/z): 458.4
[M+H].sup.+.
Step 7:
(3aS,12bR)-10-methoxy-11-(3-methoxypropoxy)-3,3-dimethyl-7-oxo-3,3-
a,7,12b-tetrahydro-1H-furo[3,4-c]pyrido[2,1-a]isoquinoline-6-carboxylic
acid [8.1]
##STR00071##
[0401] To a solution of 8.1f (12.5 mg, 0.027 mmol) in THF (1 mL)
was added NaOH (0.022 mL, 0.109 mmol) and stirred overnight. The
solution was acidified by adding 4.0 N HCl aq solution and
extracted with EtOAc. The combined organic layers were dried over
Na.sub.2SO.sub.4 and concentrated to provide a yellow oil, which
was purified by HPLC (Kinetex column, 0.1% TFA in H.sub.2O/MeCN,
1.2 mL/min) to provide 8.1 (7.8 mg, 52%). .sup.1H NMR (500 MHz,
DMSO-d.sub.6): 8.62 (s, 1H), 7.64 (s, 1H), 7.57 (s, 1H), 7.11 (s,
1H), 5.04 (br d, J=8.75 Hz, 1H), 4.40 (br dd, J=9.81, 1.30 Hz, 1H),
4.17-4.26 (m, 3H), 4.11 (dt, J=9.75, 6.47 Hz, 2H), 3.94-4.00 (m,
1H), 3.91 (s, 3H), 3.50 (br t, J=6.27 Hz, 2H), 3.27 (s, 3H), 2.00
(dt, J=12.65, 6.21 Hz, 2H), 1.39 (s, 3H), 0.64 (s, 3H). LC-MS
(m/z): 430.1 [M+H].sup.+.
Example 9: Synthesis
11-methoxy-12-(3-methoxypropoxy)-4,4-dimethyl-8-oxo-2,3,4,4a,8,13b-hexahy-
dro-1H-pyrido[1,2-f]phenanthridine-7-carboxylic acid [9.1] and
[9.2]
##STR00072##
[0402] Step 1: 6-(4-methoxy-3-(3-methoxypropoxy)
phenyl)-2,2-dimethylcyclohexanone [9.1a]
##STR00073##
[0404] A mixture of Pd(OAc).sub.2 (14 mg, 0.064 mmol), sodium
tert-butoxide (0.795 g, 8.27 mmol),
dicyclohexyl(2'-methyl-[1,1'-biphenyl]-2-yl)phosphane (58 mg),
2,2-dimethylcyclohexanone (1.056 ml, 7.63 mmol) and
4-bromo-1-methoxy-2-(3-methoxypropoxy)benzene (1.75 g, 6.36 mmol)
in toluene (6.0 ml) was heated in a sealed vial under nitrogen
atmosphere at 50.degree. C. for 18 hours. The mixture was diluted
with EtOAc and washed with sat sodium bicarbonate. The organic
layer was separated, dried over Na.sub.2SO.sub.4, filtrated and
concentrated. The remaining oil was purified by silica gel column
chromatography, EtOAc/heptane 5 to 50%, to give product (1 g, 49.1%
yield). LC-MS (m/z): 321.2 [M+H].sup.+.
Step 2:
6-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclohexan-1--
amine [9.1 b]
##STR00074##
[0406] To the mixture of
5-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclohexanone
(500 mg, 1.56 mmol) in MeOH (5 ml) was added acetic acid ammonia
salt (2.4 g, 31.2 mmol) and sodium cyanoborohydride (981 mg, 15.4
mmol). The mixture was stirred at 70.degree. C. for 8 hours and
then was concentrated under reduced pressure. The remaining
material was diluted with EtOAc, washed with water and brine, dried
over Na.sub.2SO.sub.4 and concentrated. The crude material was used
in the next step without further purification. LCMS (m/z): 322.0
[M+H].sup.+.
Step 3: N-(5-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2
dimethylcyclohexyl)formamide [9.1c]
##STR00075##
[0408] To the mixture of
5-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclohexamine
(500 mg, 1.56 mmol) in dioxane (5 ml) was added formic acid (0.286
mL, 6.22 mmol). The mixture was stirred at 100.degree. C. for 6
hours. The mixture was concentrated to afford the crude product
which was used in the next step without further purification. LCMS
(m/z): 350.2 [M+H].sup.+.
Step 4:
8-methoxy-9-(3-methoxypropoxy)-4,4-dimethyl-1,2,3,4,4a,10b-hexahyd-
rophenanthridine [9.1d]
##STR00076##
[0410] To a mixture of
N-(5-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,2-dimethylcyclohexyl)formam-
ide (543 mg, 1.54 mmol) in acetonitrile (3 ml) was added
POCOl.sub.3 (217 .mu.l, 2.33 mmol). The mixture was stirred at
85.degree. C. for 2 hours and then concentrated. The residue was
dissolved in EtOAc and basified by ammonium hydroxide solution. The
phases were separated and the organic layer was washed with brine,
dried over Na.sub.2SO.sub.4 and concentrated. The remaining
material was purified by silica gel chromatography, acetone/heptane
5 to 50% to give the title product. (320 mg, 62% yield). LCMS
(m/z): 332.3 [M+H].sup.+.
Step 5: Ethyl
(11-methoxy-12-(3-methoxypropoxy)-4,4-dimethyl-8-oxo-2,3,4,4a,8,9,9a,13b--
octahydro-1H-pyrido[1,2-f]phenanthridine-7-carboxylate [9.1e]
##STR00077##
[0412] To a mixture of
8-methoxy-9-(3-methoxypropoxy)-4,4-dimethyl-1,2,3,4,4a,10b-hexahydrophena-
nthridine (140 mg, 0.422 mmol) in EtOH (1.6 ml) was added (Z)-ethyl
2-(ethoxymethylene)-3-oxobutanoate (236 mg, 1.267 mmol). The
mixture was stirred at 110.degree. C. for 16 hours. After cooling,
the mixture was concentrated and the crude material was used in the
next step without further purification. LCMS (m/z): 472.0
[M+H].sup.+.
Step 6: Ethyl
11-methoxy-12-(3-methoxypropoxy)-4,4-dimethyl-8-oxo-2,3,4,4a,8,13b-hexahy-
dro-1H-pyrido[1,2-f]phenanthridine-7-carboxylate [9.1f]
##STR00078##
[0414] To a mixture of 9.1e (199.7 mg, 0.422 mmol) in DME (1.0 ml)
was added p-chloranil (114.6 mg, 0.464 mmol). The mixture was
stirred at 110.degree. C. for 2 hours. After cooling to rt, the
mixture was filtered and the solid was washed with cold DME. After
drying, the desired product (100 mg, 50.5% yield over two steps)
was obtained as a light yellow solid. LCMS (m/z): 470.0
[M+H].sup.+.
Step 7:
11-methoxy-12-(3-methoxypropoxy)-4,4-dimethyl-8-oxo-2,3,4,4a,8,13b-
-hexahydro-1H-pyrido[1,2-f]phenanthridine-7-carboxylic acid [9.1]
and [9.2]
##STR00079##
[0416] To a mixture of 9.1f (50 mg, 0.106 mmol) in THF (0.6 ml),
MeOH (0.6 ml) and water (0.6 ml) was added LiOH (7.7 mg, 0.319
mmol). After stirring at rt for 2 hours, the mixture was
concentrated and then acidified by adding 3.0 N HCl aq solution. To
the resultant mixture was added EtOAc. The organic layer was washed
with water and brine, dried, and concentrated. The crude residue
was purified by reverse phase HPLC to give product (10 mg, 21%
yield). LCMS (m/z): 442.2 [M+H].sup.+. H NMR (400 MHz,
Acetonitrile-d3): 9.15 (s, 1H), 7.11 (s, 1H), 6.98 (s, 1H), 4.15
(dt, J=9.3, 4.7 Hz, 2H), 3.91 (s, 3H), 3.71 (d, J=12.4 Hz, 1H),
3.53 (t, J=6.2 Hz, 2H), 3.32 (s, 3H), 2.98 (td, J=12.2, 5.2 Hz,
1H), 2.74-2.54 (m, 1H), 2.12-2.00 (m, 2H), 1.96 (dt, J=4.5, 2.4 Hz,
5H), 1.46 (d, J=29.8 Hz, 9H).
[0417] The relative configuration of the product was established to
be as shown below by nuclear Overhauser effect (NOE) experiments,
but the absolute stereochemistry of each enantiomer has not been
confirmed.
##STR00080##
[0418] The racemic material was separated by chiral SFC (OD column,
flow rate 100 ml/min, CO.sub.2/MeOH=70/30) to give two enantiomers:
9.1 (tR 3.93 min) and 9.2 (tR 6.14 min). The following compounds
can be made by similar methods using starting materials that are
known in the art:
##STR00081## ##STR00082## ##STR00083## ##STR00084## ##STR00085##
##STR00086##
BIOLOGICAL EXAMPLES
HBV Cell Line
[0419] HepG2-Clone42, a Tet-inducible HBV-expressing cell line with
a stably integrated 1.3mer copy of the HBV ayw strain, was
generated based on the Tet-inducible HepAD38 cell line with slight
modifications. Ladner S K, et al., Antimicrobial Agents and
Chemotherapy. 41(8):1715-1720 (1997). HepG2-Clone42 cells were
cultured in DMEM/F-12+Glutamax.TM. (Life Technologies, Carlsbad,
Calif., USA), supplemented with 10% fetal bovine serum (Life
Technologies), G-418 (Corning, Manassas, Va., USA) at a final
concentration of 0.5 mg/mL, and 5 .mu.g/mL Doxycycline (Sigma, St.
Louis, Mo., USA) and maintained in 5% CO.sub.2 at 37.degree. C.
HBsAq Assay
[0420] HepG2-Clone42 cells were seeded in into black clear-bottom
96-well plates at a concentration of 6.0.times.10.sup.4 cells/well.
24 hours post-seeding, the cells were treated with 200 .mu.l/well
of media containing five-fold serial dilutions of compounds in
DMSO. DMSO alone was used as the no drug control. The final DMSO
concentration in all wells was 0.5%.
[0421] The HBsAg ELISA kit (Alpha Diagnostic International, San
Antonio, Tex., USE, Catalog #4110) was used to determine the level
(semi-quantitative) of secreted HBV sAg. The HBSAg ELISA assay was
performed following the manufacturer's protocol as described.
Step 1. Pipet 100 .mu.L each of compound or DMSO treated samples
into HBsAg ELISA plates. Seal plates and incubate at room temp for
60 minutes. Step 2. Aspirate samples and wash three times with Wash
Buffer. Dispense 100.mu. of antibody-HRP conjugate to each well.
Incubate at room temp for 30 minutes. Step 3. Aspirate samples and
wash three times with Wash Buffer. Add 100 .mu.L of TMB Substrate
to all wells and incubate 15 minutes at room temp. Step 4. Dispense
100 .mu.L of Stop Solution to each well. Measure absorbance of
ELISA plate at 450 nm.
Dose Response Curves
[0422] Dose-response curves were generated and the EC.sub.50 value
was defined as the compound concentration at which HBsAg secretion
was reduced 50% compared to the DMSO control.
[0423] EC.sub.50 values were determined as follows: [0424] 1.
Determine the percent of HBsAg secretion inhibition. Calculate the
percent inhibition on of HBsAg secretion inhibition using the
following equation:
[0424] 100.times.(X.sub.C-M.sub.B)/(M.sub.D-M.sub.B) [0425] where
X.sub.C is the absorbance signal from compound-treated well;
M.sub.B is average absorbance signal (background signal) for column
12 (no cells +HBsAg ELISA sample buffer) and M.sub.D is average
absorbance signal from DMSO-treated wells. Then calculate EC.sub.50
values by non-linear regression using a four parameter curve
logistic equation.
[0426] The curve fit model employed is XLFit Dose Response One Site
Model 204: y=(A+((B-A)/(1+(10{circumflex over ( )}((C-x)*D)))))
where A is the minimum y value, B is the maximum y value, C is the
log EC50 value, and D is the slope factor.
High Throughput Solubility Measurement
[0427] 1. Transfer 20 ul of 10 mM DMSO stock solution into a 96
deep well plate labeled as sample plate and 5 ul to another plate
labeled as compound standard plate. [0428] 2. Place the buffer
plate in a Multi-Tainer MT-4 container (FTS Systems). Freeze dry
overnight to remove DMSO. [0429] 3. Add 100 ul of Cl-free PBS (pH
6.8) to the dried compound in the buffer plate and 95 ul of DMSO to
the standard plate. [0430] 4. The buffer plate is be sonicated in a
water bath for 10 min. [0431] 5. The two plates are then placed
onto the VWR orbital shaker to equilibrate for 24 hours at room
temperature. [0432] 6. The buffer plate is centrifuged at 4000 rpm
for 30 min. [0433] 7. Transfer 10 ul aliquots of supernatant from
the buffer plate to a sample plate and dilute 5 fold. [0434] 8.
Inject both compound standard and sample into the UPLC/UV/CLND/MS
to generate multi detector qualitative and quantitive analytical
data. [0435] 9. Data was processed with Xcalibur. CLND equimolar
response was used for measuring compound concentration of DMSO
solution. UV270 nm or MS relative ratio was used for solubility
determination.
TABLE-US-00001 [0435] TABLE 1 HBsAg inhibition Compound Structure
HBsAg EC.sub.50 (nM) 1.1 ##STR00087## 0.1 1.2 ##STR00088## 13 rac-2
##STR00089## 460 3.1 ##STR00090## 0.3 3.2 ##STR00091## 740 4.1
##STR00092## 0.3 5.1 ##STR00093## 6 5.2 ##STR00094## 0.1 6.1
##STR00095## 9 6.2 ##STR00096## 5 7.1 ##STR00097## 0.8 7.2
##STR00098## 90 8.1 ##STR00099## 1 9.1 ##STR00100## 9 9.2
##STR00101## 303
TABLE-US-00002 TABLE 2 Pharmacokinetic data for selected compounds.
Compound 1.1 Mice.sup.a Rat.sup.b Dog.sup.c CL (mL/min kg) 24.6
15.3 6.4 Vss (L/kg) 3.6 1.3 1.7 T.sub.1/2term (h)* 1.6 2.4 5.9 AUC
(uM h) iv 1.5 2.5 3.1 AUC (uM h) PO 1.7 4.5 5.7 C.sub.max (uM) PO
0.57 1.72 1.9 T.sub.max p.o. (h) 0.5 1.33 0.22 Oral BA (% F) 59 97
93 .sup.aiv 1.0 mg/kg, PO 2.0 mg/kg. Formulation: solution in D5W
with PEG (20%) and solutol (5%) .sup.biv 1.0 mg/kg, PO 2.0 mg/kg.
Formulation: solution in D5W with PEG (20%) and solutol (5%)
.sup.civ 0.5 mg/kg, PO 1.0 mg/kg. Formulation: solution in D5W with
PEG (20%) and solutol (5%)
Compound 3.1
TABLE-US-00003 [0436] Mice.sup.a Rat.sup.b Dog.sup.c CL (mL/min kg)
6.9 2.1 1.3 Vss (L/kg) 1.0 0.6 0.6 T.sub.1/2term (h)* 3.8 9.2 7.2
AUC (uM h) iv 5.5 16.6 13.3 AUC (uM h) po 9.5 19.6 26.7 C.sub.max
(uM) PO 3.0 6.5 6.2 T.sub.max p.o. (h) 0.5 0.5 1.5 Oral BA (% F) 87
57 96 .sup.aiv 1.0 mg/kg, PO 2.0 mg/kg. Formulation: solution in
D5W with PEG (20%) and solutol (5%) .sup.biv 1.0 mg/kg, PO 2.0
mg/kg. Formulation: solution in D5W with PEG (20%) and solutol (5%)
.sup.civ 0.5 mg/kg, PO 1.0 mg/kg. Formulation: solution in D5W with
PEG (20%) and solutol (10%)
Compound 4.1
TABLE-US-00004 [0437] Mice.sup.a Rat.sup.b Dog.sup.c CL (mL/min kg)
12.7 4.9 2.7 Vss (L/kg) 1.5 0.75 1.7 T.sub.1/2term (h)* 1.8 1.2
12.3 AUC (uM h) iv 2.9 8.0 6.2 AUC (uM h) PO 7.2 10.2 12.8
C.sub.max (uM) PO 1.9 2.6 3.3 T.sub.max p.o. (h) 1.0 2.0 0.8 Oral
BA (% F) 100 65 94 .sup.aiv 1.0 mg/kg, PO 2.0 mg/kg. Formulation:
solution in D5W with PEG (20%) and solutol (5%) .sup.biv 1.0 mg/kg,
PO 2.0 mg/kg. Formulation: 75% PEG300 and 25% D5W .sup.civ 0.5
mg/kg, PO 1.0 mg/kg. Formulation: solution in D5W with 20% PEG and
10% solutol
Comparative Data
[0438] The following table provides data for solubility of
compounds of the invention in PBS (phosphate-buffered saline),
using a standard solubility screening method. It also provides data
for inhibition of a key cardiac sodium ion channel for selected
compounds: inhibition of Nav1.5 is often associated with
cardiotoxicity, thus compounds that exhibit little or no inhibition
of this sodium channel are less likely to cause adverse effects
than compounds that inhibit Nav1.5, and are predicted to be more
suitable for development as drugs. Thus the compound of Example 4.1
is predicted to be safer in this respect than the reference
compound.
[0439] Each of the compounds of the invention exhibited solubility
greater than 1 mM in PBS buffer. Compounds with higher solubility
possess lower risks in achieving the toxicological end points (in
animals) and the oral development pathway (for human) in terms of
the bioavailability criteria. A compound of similar structure (Ref.
Ex. #132 from WO2015/113990) lacking the fused ring of the claimed
compounds, was about 20-fold less soluble; thus the fused ring
provides compounds with improved physical properties for
formulation and/or development.
TABLE-US-00005 Example No. Solubility (mM) Nav1.5 EC.sub.50 (uM)
1.1 2.4 3.1 1.6 4.1 1.6 >500 ##STR00102## 0.086 46 (Ref. Example
#132)
* * * * *
References