U.S. patent application number 15/370568 was filed with the patent office on 2017-03-23 for antiviral compounds.
The applicant listed for this patent is Hoffmann-La Roche Inc.. Invention is credited to Steven Joseph Berthel, Zhi Chen, Feng Chi, Elbert Chin, David Shawn Erickson, Stephen Deems Gabriel, Buelent Kocer, Eric Mertz, Jean-Marc Plancher, Robert J. Weikert.
Application Number | 20170081312 15/370568 |
Document ID | / |
Family ID | 50190453 |
Filed Date | 2017-03-23 |
United States Patent
Application |
20170081312 |
Kind Code |
A1 |
Berthel; Steven Joseph ; et
al. |
March 23, 2017 |
ANTIVIRAL COMPOUNDS
Abstract
The present invention discloses compounds of Formula (I):
wherein the variables in Formula (I) are defined as described
herein. Also disclosed are pharmaceutical compositions containing
such compounds and methods for using the compounds of Formula (I)
in the prevention or treatment of HCV infection. ##STR00001##
Inventors: |
Berthel; Steven Joseph;
(Mendham Township, NJ) ; Chen; Zhi; (Livingston,
NJ) ; Chi; Feng; (Basking Ridge, NJ) ; Chin;
Elbert; (San Mateo, CA) ; Erickson; David Shawn;
(Leonia, NJ) ; Gabriel; Stephen Deems;
(Morristown, NJ) ; Kocer; Buelent; (Maulburg,
DE) ; Mertz; Eric; (Fair Lawn, NJ) ; Plancher;
Jean-Marc; (Hagenthal-le-Bas, FR) ; Weikert; Robert
J.; (Basel, CH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Hoffmann-La Roche Inc. |
Nutley |
NJ |
US |
|
|
Family ID: |
50190453 |
Appl. No.: |
15/370568 |
Filed: |
December 6, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14766905 |
Aug 10, 2015 |
9540345 |
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PCT/EP2014/054015 |
Mar 3, 2014 |
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15370568 |
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61772943 |
Mar 5, 2013 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 471/04 20130101;
A61K 31/506 20130101; A61K 45/06 20130101; A61K 31/496 20130101;
C07D 401/12 20130101; A61P 31/12 20180101; A61K 31/4196 20130101;
A61P 1/16 20180101; A61K 31/4439 20130101; A61K 38/21 20130101;
C07D 409/12 20130101; C07D 249/14 20130101; A61K 31/501 20130101;
C07D 403/12 20130101; A61P 31/14 20180101; A61K 31/437 20130101;
C07D 487/10 20130101 |
International
Class: |
C07D 401/12 20060101
C07D401/12; A61K 38/21 20060101 A61K038/21; A61K 31/4439 20060101
A61K031/4439; C07D 403/12 20060101 C07D403/12; A61K 31/506 20060101
A61K031/506; C07D 409/12 20060101 C07D409/12; A61K 31/4196 20060101
A61K031/4196; A61K 31/501 20060101 A61K031/501; C07D 471/04
20060101 C07D471/04; A61K 31/437 20060101 A61K031/437; A61K 31/496
20060101 A61K031/496; A61K 45/06 20060101 A61K045/06; C07D 487/10
20060101 C07D487/10 |
Claims
1-20. (canceled)
21. A compound of formula I: ##STR00189## wherein: A is unsaturated
or partially unsaturated monocyclic or bicyclic heteroaryl or
monocyclic or spirocyclic heterocycloalkyl; each R.sup.1 is
independently halo, halo lower alkyl, or lower alkyl sulfonyl; m is
0, 1, or 2; each R.sup.2 is independently halo, lower alkoxy, oxo,
amino, lower alkyl, C(.dbd.O)OR.sup.2', S(.dbd.O)R.sup.2',
S(.dbd.O).sub.2NHR.sup.2', hydroxyl lower alkyl,
C(.dbd.O)NHR.sup.2', or C(.dbd.O)R.sup.2'; n is 0, 1, or 2; and
R.sup.2' is lower alkyl, halo lower alkyl, or adamantyl, or a
pharmaceutically acceptable salt thereof.
22. The compound of claim 21, wherein m is 1.
23. The compound of claim 22, wherein m is 2.
24. The compound of claim 23, wherein one R.sup.1 is halo and the
other is halo lower alkyl.
25. The compound of claim 23, wherein both R.sup.1 are halo.
26. The compound of claim 21, wherein n is 0.
27. The compound of claim 21, wherein n is 1.
28. The compound of claim 21, wherein n is 2.
29. The compound of claim 21, wherein A is pyridinyl, pyrimidinyl,
pyridazinyl, triazolyl, tetrazolyl, piperazinyl,
2,6-diaza-spiro[3.3]heptanyl, azetidinyl, thiophenyl, or
pyrazolyl.
30. The compound of claim 29, wherein R.sup.2 is halo, lower
alkoxy, oxo, amino, lower alkyl, C(.dbd.O)OR.sup.2',
S(.dbd.O).sub.2R.sup.2', S(.dbd.O).sub.2NHR.sup.2', hydroxyl lower
alkyl, C(.dbd.O)NHR.sup.2', or C(.dbd.O)R.sup.2'.
31. The compound of claim 30, wherein R.sup.2' is lower alkyl or
halo lower alkyl.
32. A method for preventing a Hepatitis C Virus (HCV) infection,
comprising the step of administering to a patient in need thereof a
therapeutically effective amount of a compound of claim 1.
33. The method of claim 33, further comprising administering to a
patient in need thereof a therapeutically effective amount of an
immune system suppressant.
34. A method for treating a Hepatitis C Virus (HCV) infection,
comprising the step of administering to a patient in need thereof a
therapeutically effective amount of a compound of claim 1.
35. The method of claim 32 or 34, further comprising administering
a combination of antiviral agents that inhibits replication of
HCV.
36. The method of claim 32, further comprising administering an
immune system modulator or an antiviral agent that inhibits
replication of HCV, or a combination thereof.
37. The method of claim 36, wherein the immune system modulator is
an interferon or a chemically derivatized interferon.
38. The method of claim 36, wherein the antiviral agent is selected
from the group consisting of a HCV protease inhibitor, a HCV
polymerase inhibitor, a HCV helicase inhibitor, a HCV NS5A
inhibitor, or any combination thereof.
39. A pharmaceutical composition, comprising a therapeutically
effective amount of a compound of claim 21 and a pharmaceutically
acceptable excipient.
40. The method of claim 34, further comprising administering an
immune system modulator or an antiviral agent that inhibits
replication of HCV, or a combination thereof.
41. The method of claim 40, wherein the immune system modulator is
an interferon or a chemically derivatized interferon.
42. The method of claim 40, wherein the antiviral agent is selected
from the group consisting of a HCV protease inhibitor, a HCV
polymerase inhibitor, a HCV helicase inhibitor, a HCV NS5A
inhibitor, or any combination thereof.
Description
[0001] The present invention provides compounds of Formula I useful
as inhibitors of hepatitis C virus (HCV), as inhibitors of HCV
infection, and for the prevention and treatment of hepatitis C
infection.
[0002] Hepatitis C virus (HCV) infection is a major health problem
that affects 170 million people worldwide and 3-4 million people in
the United States (Armstrong, G. L., et al., Ann. Intern. Med.
2006, 144:705-714; Lauer, G. M., et al., N. Eng. J. Med. 2001,
345:41-52). HCV infection leads to chronic liver disease, such as
cirrhosis and hepatocellular carcinoma in a substantial number of
infected individuals. Chronic HCV infection associated liver
cirrhosis and hepatocellular carcinoma are also the leading cause
of liver transplantation in the United States. Current treatments
for HCV infection include immunotherapy with pegylated
interferon-.alpha. in combination with the nucleoside-analog
ribavirin. Pegylated interferon-.alpha. in combination with
ribavirin and one of the two recently approved HCV NS3 protease
inhibitors Incivek or Victrelis is the current standard of care for
the treatment of genotype 1 HCV infected patients, the most
difficult to treat patient population. However, current HCV
treatments are compromised by suboptimal sustained virological
response rates and associated with severe side effects, as well as
resistance to the protease inhibitors. Therefore there is a clear
need for improved antiviral drugs with better efficacy, safety, and
resistance profiles.
[0003] The infection of human hepatocytes by HCV, also known as HCV
entry, is mediated by the functional interactions of
virally-encoded envelope glycoproteins E1 and E2 and host cell
co-receptors, followed by a receptor-mediated endocytosis
processes. This HCV entry step is a putative target for therapeutic
intervention. Several virally-encoded enzymes are also putative
targets for therapeutic intervention, including a metalloprotease
(NS2-3), a serine protease (NS3, amino acid residues 1-180), a
helicase (NS3, full length), an NS3 protease cofactor (NS4A), a
membrane protein (NS4B), a zinc metalloprotein (NS5A) and an
RNA-dependent RNA polymerase (NS5B).
[0004] Systems have been developed to study the biology of HCV
entry into host cells. Pseudotyping systems where the E1 and E2
glycoproteins are used to functionally replace the glycoproteins of
retroviruses have been developed (Bartoech, B., Dubuisson, J. and
Cosset, F.-L. J. Exp. Med. 2003, 197:633-642; Hsu, M. et al. Proc.
Natl. Acad. Sci. USA. 2003, 100:7271-7276). These systems yield HCV
pseudoparticles that bind to and enter host cells in a manner which
is believed to be analogous to the natural virus, thus making them
a convenient tool to study the viral entry steps as well as to
identify inhibitors blocking this process.
[0005] There is a clear and long-felt need to develop effective
therapeutics for treatment of HCV infection. Specifically, there is
a need to develop compounds that selectively inhibit HCV viral
entry and replication and that are useful for treating HCV-infected
patients and protecting liver transplant patients from HCV
re-infection. This application discloses novel compounds that are
effective in prevention of HCV infection. Additionally, the
disclosed compounds provide advantages for pharmaceutical uses, for
example, with respect to their mechanism of action, binding,
prevention of infection, inhibition efficacy, and target
selectivity.
SUMMARY OF THE INVENTION
[0006] The application provides compound of formula I
##STR00002##
A is unsaturated or partially unsaturated monocyclic or bicyclic
heteroaryl or monocyclic or spirocyclic heterocycloalkyl; each
R.sup.1 is independently halo, halo lower alkyl, or lower alkyl
sulfonyl; m is 0, 1, or 2; each R.sup.2 is independently halo,
lower alkoxy, oxo, amino, lower alkyl, C(.dbd.O)OR.sup.2',
S(.dbd.O).sub.2R.sup.2', S(.dbd.O).sub.2NHR.sup.2', hydroxyl lower
alkyl, C(.dbd.O)NHR.sup.2', or C(.dbd.O)R.sup.2'; n is 0, 1, or 2;
and R.sup.2' is lower alkyl, halo lower alkyl, or adamantly; or a
pharmaceutically acceptable salt thereof.
[0007] The application provides a method for preventing a Hepatitis
C Virus (HCV) infection comprising administering to a patient in
need thereof a therapeutically effective amount of a compound of
Formula I.
[0008] The application provides a method for treating a Hepatitis C
Virus (HCV) infection comprising administering to a patient in need
thereof a therapeutically effective amount of a compound of Formula
I.
[0009] The application provides a composition comprising a compound
of Formula I and a pharmaceutically acceptable excipient.
DETAILED DESCRIPTION OF THE INVENTION
Definition
[0010] The phrase "a" or "an" entity as used herein refers to one
or more of that entity; for example, a compound refers to one or
more compounds or at least one compound. As such, the terms "a" (or
"an"), "one or more", and "at least one" can be used
interchangeably herein.
[0011] As used in this specification, whether in a transitional
phrase or in the body of the claim, the terms "comprise(s)" and
"comprising" are to be interpreted as having an open-ended meaning.
That is, the terms are to be interpreted synonymously with the
phrases "having at least" or "including at least". When used in the
context of a process, the term "comprising" means that the process
includes at least the recited steps, but may include additional
steps. When used in the context of a compound or composition, the
term "comprising" means that the compound or composition includes
at least the recited features or components, but may also include
additional features or components.
[0012] As used herein, unless specifically indicated otherwise, the
word "or" is used in the "inclusive" sense of "and/or" and not the
"exclusive" sense of "either/or".
[0013] The term "independently" is used herein to indicate that a
variable is applied in any one instance without regard to the
presence or absence of a variable having that same or a different
definition within the same compound. Thus, in a compound in which
R'' appears twice and is defined as "independently carbon or
nitrogen", both R''s can be carbon, both R''s can be nitrogen, or
one R'' can be carbon and the other nitrogen.
[0014] When any variable occurs more than one time in any moiety or
formula depicting and describing compounds employed or claimed in
the present invention, its definition on each occurrence is
independent of its definition at every other occurrence. Also,
combinations of substituents and/or variables are permissible only
if such compounds result in stable compounds.
[0015] The symbols "*" at the end of a bond or "" drawn through a
bond each refer to the point of attachment of a functional group or
other chemical moiety to the rest of the molecule of which it is a
part. Thus, for example:
##STR00003##
[0016] A bond drawn into ring system (as opposed to connected at a
distinct vertex) indicates that the bond may be attached to any of
the suitable ring atoms.
[0017] The term "optional" or "optionally" as used herein means
that a subsequently described event or circumstance may, but need
not, occur, and that the description includes instances where the
event or circumstance occurs and instances in which it does not.
For example, "optionally substituted" means that the optionally
substituted moiety may incorporate a hydrogen atom or a
substituent.
[0018] If a substituent is designated to be "absent", the
substituent is not present.
[0019] The term "about" is used herein to mean approximately, in
the region of, roughly, or around. When the term "about" is used in
conjunction with a numerical range, it modifies that range by
extending the boundaries above and below the numerical values set
forth. In general, the term "about" is used herein to modify a
numerical value above and below the stated value by a variance of
20%.
[0020] Certain compounds may exhibit tautomerism. Tautomeric
compounds can exist as two or more interconvertable species.
Prototropic tautomers result from the migration of a covalently
bonded hydrogen atom between two atoms. Tautomers generally exist
in equilibrium and attempts to isolate an individual tautomers
usually produce a mixture whose chemical and physical properties
are consistent with a mixture of compounds. The position of the
equilibrium is dependent on chemical features within the molecule.
For example, in many aliphatic aldehydes and ketones, such as
acetaldehyde, the keto form predominates while; in phenols, the
enol form predominates. Common prototropic tautomers include
keto/enol (--C(.dbd.O)--CH--.revreaction.--C(--OH).dbd.CH--),
amide/imidic acid (--C(.dbd.O)--NH--.revreaction.--C(--OH).dbd.N--)
and amidine (--C(.dbd.NR)--NH--.revreaction.--C(--NHR).dbd.N--)
tautomers. The latter two are particularly common in heteroaryl and
heterocyclic rings and the present invention encompasses all
tautomeric forms of the compounds.
[0021] Technical and scientific terms used herein have the meaning
commonly understood by one of skill in the art to which the present
invention pertains, unless otherwise defined. Reference is made
herein to various methodologies and materials known to those of
skill in the art. Standard reference works setting forth the
general principles of pharmacology include Goodman and Gilman's The
Pharmacological Basis of Therapeutics, 10.sup.th Ed., McGraw Hill
Companies Inc., New York (2001). Any suitable materials and/or
methods known to those of skill can be utilized in carrying out the
present invention. However, preferred materials and methods are
described. Materials, reagents and the like to which reference are
made in the following description and examples are obtainable from
commercial sources, unless otherwise noted.
[0022] The definitions described herein may be appended to form
chemically-relevant combinations, such as "heteroalkylaryl,"
"haloalkylheteroaryl," "arylalkylheterocyclyl," "alkylcarbonyl,"
"alkoxyalkyl," and the like. When the term "alkyl" is used as a
suffix following another term, as in "phenylalkyl," or
"hydroxyalkyl," this is intended to refer to an alkyl group, as
defined above, being substituted with one to two substituents
selected from the other specifically-named group. Thus, for
example, "phenylalkyl" refers to an alkyl group having one to two
phenyl substituents, and thus includes benzyl, phenylethyl, and
biphenyl. An "alkylaminoalkyl" is an alkyl group having one to two
alkylamino substituents. "Hydroxyalkyl" includes 2-hydroxyethyl,
2-hydroxypropyl, 1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl,
2,3-dihydroxybutyl, 2-(hydroxymethyl), 3-hydroxypropyl, and so
forth. Accordingly, as used herein, the term "hydroxyalkyl" is used
to define a subset of heteroalkyl groups defined below. The term
-(ar)alkyl refers to either an unsubstituted alkyl or an aralkyl
group. The term (hetero)aryl or (het)aryl refers to either an aryl
or a heteroaryl group.
[0023] The term "carbonyl" or "acyl" as used herein denotes a group
of formula --C(.dbd.O)R wherein R is hydrogen or lower alkyl as
defined herein.
[0024] The term "ester" as used herein denotes a group of formula
--C(.dbd.O)OR wherein R is lower alkyl as defined herein.
[0025] The term "alkyl" as used herein denotes an unbranched or
branched chain, saturated, monovalent hydrocarbon residue
containing 1 to 10 carbon atoms. The term "lower alkyl" denotes a
straight or branched chain hydrocarbon residue containing 1 to 6
carbon atoms. "C.sub.1-10 alkyl" as used herein refers to an alkyl
composed of 1 to 10 carbons. Examples of alkyl groups include, but
are not limited to, lower alkyl groups include methyl, ethyl,
propyl, i-propyl, n-butyl, i-butyl, t-butyl or pentyl, isopentyl,
neopentyl, hexyl, heptyl, and octyl.
[0026] When the term "alkyl" is used as a suffix following another
term, as in "phenylalkyl," or "hydroxyalkyl," this is intended to
refer to an alkyl group, as defined above, being substituted with
one to two substituents selected from the other specifically-named
group. Thus, for example, "phenylalkyl" denotes the radical
R'R''--, wherein R' is a phenyl radical, and R'' is an alkylene
radical as defined herein with the understanding that the
attachment point of the phenylalkyl moiety will be on the alkylene
radical. Examples of arylalkyl radicals include, but are not
limited to, benzyl, phenylethyl, 3-phenylpropyl. The terms
"arylalkyl" or "aralkyl" are interpreted similarly except R' is an
aryl radical. The terms "(het)arylalkyl" or "(het)aralkyl" are
interpreted similarly except R' is optionally an aryl or a
heteroaryl radical.
[0027] The terms "haloalkyl" or "halo lower alkyl" or "lower
haloalkyl" refers to a straight or branched chain hydrocarbon
residue containing 1 to 6 carbon atoms wherein one or more carbon
atoms are substituted with one or more halogen atoms.
[0028] The term "alkylene" or "alkylenyl" as used herein denotes a
divalent saturated linear hydrocarbon radical of 1 to 10 carbon
atoms (e.g., (CH.sub.2).sub.n) or a branched saturated divalent
hydrocarbon radical of 2 to 10 carbon atoms (e.g., --CHMe- or
--CH.sub.2CH(i-Pr)CH.sub.2--), unless otherwise indicated. Except
in the case of methylene, the open valences of an alkylene group
are not attached to the same atom. Examples of alkylene radicals
include, but are not limited to, methylene, ethylene, propylene,
2-methyl-propylene, 1,1-dimethyl-ethylene, butylene,
2-ethylbutylene.
[0029] The term "alkoxy" as used herein means an --O-alkyl group,
wherein alkyl is as defined above such as methoxy, ethoxy,
n-propyloxy, i-propyloxy, n-butyloxy, i-butyloxy, t-butyloxy,
pentyloxy, hexyloxy, including their isomers. "Lower alkoxy" as
used herein denotes an alkoxy group with a "lower alkyl" group as
previously defined. "C.sub.1-10 alkoxy" as used herein refers to
an-O-alkyl wherein alkyl is C.sub.1-10.
[0030] The terms "haloalkoxy" or "halo lower alkoxy" or "lower
haloalkoxy" refers to a lower alkoxy group, wherein one or more
carbon atoms are substituted with one or more halogen atoms.
[0031] The term "hydroxyalkyl" as used herein denotes an alkyl
radical as herein defined wherein one to three hydrogen atoms on
different carbon atoms is/are replaced by hydroxyl groups.
[0032] The term "sulfinyl" as used herein denotes a --SO--
group.
[0033] The term "sulfonyl" as used herein denotes a --SO.sub.2--
group.
[0034] The terms "alkylsulfonyl" and "arylsulfonyl" as used herein
refers to a group of formula --S(.dbd.O).sub.2R wherein R is alkyl
or aryl respectively and alkyl and aryl are as defined herein. The
term "heteroalkylsulfonyl" as used herein refers herein denotes a
group of formula --S(.dbd.O).sub.2R wherein R is "heteroalkyl" as
defined herein.
[0035] The term "lower alkyl sulfonylamido" as used herein refers
to a group of formula --S(.dbd.O).sub.2NR.sub.2 wherein each R is
independently hydrogen or C.sub.1-3 alkyl, and lower alkyl is as
defined herein.
[0036] The term "trifluoromethyl sulfonyl" as used herein refers to
a group of formula --S(.dbd.O).sub.2CF.sub.3.
[0037] The term "trifluoromethyl sulfinyl" as used herein refers to
a group of formula --S(.dbd.O)CF.sub.3.
[0038] The term "trifluoromethyl sulfanyl" as used herein refers to
a group of formula --SCF.sub.3.
[0039] The term "nitro" as used herein refers to a group of formula
--N.sup.+(.dbd.O)O.sup.-.
[0040] The term "carboxyl" as used herein refers to a group of
formula --C(.dbd.O)R.sub.2 wherein each R is independently hydrogen
or C.sub.1-3 alkyl, and lower alkyl is as defined herein.
[0041] The term "cycloalkyl" denotes a monovalent saturated
monocyclic or bicyclic hydrocarbon group of 3 to 10 ring carbon
atoms. In particular embodiments cycloalkyl denotes a monovalent
saturated monocyclic hydrocarbon group of 3 to 8 ring carbon atoms.
Bicyclic means consisting of two saturated carbocycles having one
or more carbon atoms in common. Particular cycloalkyl groups are
monocyclic. Examples for monocyclic cycloalkyl are cyclopropyl,
cyclobutanyl, cyclopentyl, cyclohexyl or cycloheptyl. Examples for
bicyclic cycloalkyl are bicyclo[2.2.1]heptanyl, or
bicyclo[2.2.2]octanyl.
[0042] The term "amino" as used herein denotes a group of the
formula --NR'R'' wherein R' and R'' are independently hydrogen,
alkyl, alkoxy, cycloalkyl, heterocycloalkyl, aryl or heteroaryl.
Alternatively, R' and R'', together with the nitrogen to which they
are attached, can form a heterocycloalkyl. The term "primary amino"
denotes a group wherein both R' and R'' are hydrogen. The term
"secondary amino" denotes a group wherein R' is hydrogen and R'' is
not. The term "tertiary amino" denotes a group wherein both R' and
R'' are not hydrogen. Particular secondary and tertiary amines are
methylamine, ethylamine, propylamine, isopropylamine, phenylamine,
benzylamine dimethylamine, diethylamine, dipropylamine and
diisopropylamine.
[0043] The term "amido" as used herein denotes a group of the
formula --C(.dbd.O)NR'R'' or --NR'C(.dbd.O)R'' wherein R' and R''
are independently hydrogen, alkyl, alkoxy, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl.
[0044] The term "heteroaryl" denotes a monovalent aromatic
heterocyclic mono- or bicyclic ring system of 5 to 12 ring atoms,
comprising 1, 2, 3 or 4 heteroatoms selected from N, O and S, the
remaining ring atoms being carbon. Examples of heteroaryl moieties
include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl,
thiazolyl, triazolyl oxadiazolyl, thiadiazolyl, tetrazolyl,
pyridinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl,
triazinyl, azepinyl, diazepinyl, isoxazolyl, benzofuranyl,
isothiazolyl, benzothienyl, indolyl, isoindolyl, isobenzofuranyl,
benzimidazolyl, benzoxazolyl, benzoisoxazolyl, benzothiazolyl,
benzoisothiazolyl, benzooxadiazolyl, benzothiadiazolyl,
benzotriazolyl, purinyl, quinolinyl, isoquinolinyl, quinazolinyl,
or quinoxalinyl.
[0045] The term "heterocycloalkyl" denotes a monovalent saturated
or partly unsaturated mono- or bicyclic ring system of 3 to 9 ring
atoms, comprising 1, 2, or 3 ring heteroatoms selected from N, O
and S, the remaining ring atoms being carbon. In particular
embodiments, heterocycloalkyl is a monovalent saturated monocyclic
ring system of 4 to 7 ring atoms, comprising 1, 2, or 3 ring
heteroatoms selected from N, O and S, the remaining ring atoms
being carbon. Examples for monocyclic saturated heterocycloalkyl
are aziridinyl, oxiranyl, azetidinyl, oxetanyl, pyrrolidinyl,
tetrahydrofuranyl, tetrahydro-thienyl, pyrazolidinyl,
imidazolidinyl, oxazolidinyl, isoxazolidinyl thiazolidinyl,
piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl,
morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholin-4-yl,
azepanyl, diazepanyl, homopiperazinyl, or oxazepanyl. Examples for
bicyclic saturated heterocycloalkyl are 8-aza-bicyclo[3.2.1]octyl,
quinuclidinyl, 8-oxa-3-aza-bicyclo[3.2.1]octyl,
9-aza-bicyclo[3.3.1]nonyl, 3-oxa-9-aza-bicyclo[3.3.1]nonyl, or
3-thia-9-aza-bicyclo[3.3.1]nonyl. Examples for partly unsaturated
heterocycloalkyl are dihydrofuryl, imidazolinyl, dihydro-oxazolyl,
tetrahydro-pyridinyl, or dihydropyranyl.
Inhibitors of HCV Entry
[0046] The application provides a compound of formula I
##STR00004##
A is unsaturated or partially unsaturated monocyclic or bicyclic
heteroaryl or monocyclic or spirocyclic heterocycloalkyl; each
R.sup.1 is independently halo, halo lower alkyl, or lower alkyl
sulfonyl; m is 0, 1, or 2; each R.sup.2 is independently halo,
lower alkoxy, oxo, amino, lower alkyl, C(.dbd.O)OR.sup.2',
S(.dbd.O).sub.2R.sup.2', S(.dbd.O).sub.2NHR.sup.2', hydroxyl lower
alkyl, C(.dbd.O)NHR.sup.2', or C(.dbd.O)R.sup.2'; n is 0, 1, or 2;
and R.sup.2' is lower alkyl, halo lower alkyl, or adamantly; or a
pharmaceutically acceptable salt thereof.
[0047] The application provides a compound of formula I, wherein m
is 1.
[0048] The application provides a compound of formula I, wherein m
is 2.
[0049] The application provides a compound of formula I, wherein
one R.sup.1 is halo and the other is halo lower alkyl.
[0050] The application provides a compound of formula I, wherein m
is 2 and one R.sup.1 is halo and the other is halo lower alkyl.
[0051] The application provides a compound of formula I, wherein
both R.sup.1 are halo.
[0052] The application provides a compound of formula I, wherein m
is 2 and both R.sup.1 are halo.
[0053] The application provides any of the above compounds of
formula I, wherein n is 0.
[0054] The application alternatively provides any of the above
compounds of formula I, wherein n is 1.
[0055] The application alternatively provides any of the above
compounds of formula I, wherein n is 2.
[0056] The application provides any of the above compounds of
formula I, wherein A is pyridinyl, pyrimidinyl, pyridazinyl,
triazolyl, tetrazolyl, piperazinyl, 2,6-diaza-spiro[3.3]heptanyl,
azetidinyl, thiophenyl, or pyrazolyl.
[0057] The application provides any of the above compounds of
formula I, wherein R.sup.2 is halo, lower alkoxy, oxo, amino, lower
alkyl, C(.dbd.O)OR.sup.2', S(.dbd.O).sub.2R.sup.2',
S(.dbd.O).sub.2NHR.sup.2', hydroxyl lower alkyl,
C(.dbd.O)NHR.sup.2', or C(.dbd.O)R.sup.2'.
[0058] The application provides any of the above compounds of
formula I, wherein R.sup.2' is lower alkyl or halo lower alkyl.
[0059] The application provides a compound selected from the group
consisting of: [0060]
N.sup.3-[3,5-Dichloro-4-(6-methoxy-pyridin-3-yl)-phenyl]-1H-[1,2,4]triazo-
le-3,5-diamine; [0061]
N.sup.3-[3,5-Dichloro-4-(5-methanesulfonyl-pyridin-3-yl)-phenyl]-1H-[1,2,-
4]triazole-3,5-diamine; [0062]
5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-1H-pyridi-
n-2-one; [0063]
N.sup.3-[4-(6-Amino-pyridin-3-yl)-3,5-dichloro-phenyl]-1H-[1,2,4]triazole-
-3,5-diamine; [0064]
N.sup.3-[4-(2-Amino-pyrimidin-5-yl)-3,5-dichloro-phenyl]-1H-[1,2,4]triazo-
le-3,5-diamine; [0065]
N3-[3,5-Dichloro-4-(2-methoxy-pyridin-4-yl)-phenyl]-1H-[1,2,4]triazole-3,-
5-diamine; [0066]
4-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-1H-pyridi-
n-2-one; [0067]
N-{5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-pyridi-
n-2-yl}-methanesulfonamide; [0068]
N.sup.5-[3-Fluoro-4-(6-fluoro-pyridin-3-yl)-5-trifluoromethyl-phenyl]-1H--
[1,2,4]triazole-3,5-diamine; [0069]
N.sup.5-(3-Fluoro-4-pyridin-3-yl-5-trifluoromethyl-phenyl)-1H-[1,2,4]tria-
zole-3,5-diamine; [0070]
N.sup.3-[3,5-Dichloro-4-(6-methanesulfonyl-pyridin-3-yl)-phenyl]-1H-[1,2,-
4]triazole-3,5-diamine; [0071]
6-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-2,6-diaza-
-spiro[3.3]heptane-2-carboxylic acid tert-butyl ester; [0072]
N.sup.3-(3-Chloro-4-pyridazin-3-yl-phenyl)-1H-[1,2,4]triazole-3,5-diamine-
; [0073]
N.sup.3-[3-Chloro-4-(1-methyl-1H-tetrazol-5-yl)-phenyl]-1H-[1,2,4-
]triazole-3,5-diamine; [0074]
N.sup.3-[3-Chloro-4-(2-methyl-2H-[1,2,4]triazol-3-yl)-phenyl]-1H-[1,2,4]t-
riazole-3,5-diamine; [0075]
N.sup.3-(3,5-Dichloro-4-pyrazol-1-yl-phenyl)-1H-[1,2,4]triazole-3,5-diami-
ne; [0076]
4-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-
-3,6-dihydro-2H-pyridine-1-carboxylic acid tert-butyl ester; [0077]
N.sup.3-[3,5-Dichloro-4-(1,2,3,6-tetrahydro-pyridin-4-yl)-phenyl]-1H-[1,2-
,4]triazole-3,5-diamine; [0078]
4-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-6-trifluoromethyl-phe-
nyl]-3,6-dihydro-2H-pyridine-1-carboxylic acid tert-butyl ester;
[0079]
N.sup.3-[3-Chloro-4-(1,2,3,6-tetrahydro-pyridin-4-yl)-5-trifluoromethyl-p-
henyl]-1H-[1,2,4]triazole-3,5-diamine; [0080]
N.sup.5-(3,5-Dichloro-4-[1,2,4]triazolo[4,3-a]pyridin-3-yl-phenyl)-1H-[1,-
2,4]triazole-3,5-diamine; [0081]
N-{3-Chloro-4-[6-(propane-2-sulfonyl)-pyridin-3-yl]-5-trifluoromethyl-phe-
nyl}-4H-[1,2,4]triazole-3,5-diamine; [0082]
5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-6-trifluoromethyl-phe-
nyl]-pyridine-2-sulfonic acid tert-butylamide; [0083]
5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-6-trifluoromethyl-phe-
nyl]-pyridine-2-sulfonic acid
(2,2,2-trifluoro-1,1-dimethyl-ethyl)-amide; [0084]
5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-6-trifluoromet-
hyl-phenyl]-pyridine-2-sulfonic acid adamantan-1-ylamide; [0085]
N.sup.3-[2-Chloro-4'-(4-methyl-piperazin-1-yl)-6-trifluoromethyl-biphenyl-
-4-yl]-1H-[1,2,4]triazole-3,5-diamine; [0086]
N.sup.3-(3-Chloro-4-pyridazin-3-yl-phenyl)-1H-[1,2,4]triazole-3,5-diamine-
; [0087]
1-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-phenyl]-1H-pyridin-2-o-
ne; [0088]
1-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-phenyl]-1H--
pyridin-2-one; [0089]
1-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-1H-pyridi-
n-2-one; [0090]
5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-6-trifluoromethyl-phe-
nyl]-pyridine-2-carboxylic acid methylamide; [0091]
N.sup.3-(3,5-Dichloro-4-pyridazin-4-yl-phenyl)-1H-[1,2,4]triazole-3,5-dia-
mine; [0092]
1-{3-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-thioph-
en-2-yl}-ethanone; [0093]
N3-(3,5-Dichloro-4-pyridin-4-yl-phenyl)-1H-[1,2,4]triazole-3,5-diamine;
[0094]
N3-[3,5-Dichloro-4-(5-chloro-thiophen-2-yl)-phenyl]-1H-[1,2,4]tria-
zole-3,5-diamine; [0095]
N3-(3,5-Dichloro-4-pyridin-3-yl-phenyl)-1H-[1,2,4]triazole-3,5-diamine;
[0096]
N3-[3,5-Dichloro-4-(1H-pyrazol-3-yl)-phenyl]-1H-[1,2,4]triazole-3,-
5-diamine; [0097]
N3-(3,5-Dichloro-4-pyrimidin-5-yl-phenyl)-1H-[1,2,4]triazole-3,5-diamine;
[0098]
N3-[3,5-Dichloro-4-(2-methoxy-pyrimidin-5-yl)-phenyl]-1H-[1,2,4]tr-
iazole-3,5-diamine; [0099]
N3-[3,5-Dichloro-4-(6-trifluoromethyl-pyridin-3-yl)-phenyl]-1H-[1,2,4]tri-
azole-3,5-diamine; [0100]
N3-[3,5-Dichloro-4-(1-methyl-3-trifluoromethyl-1H-pyrazol-4-yl)-phenyl]-1-
H-[1,2,4]triazole-3,5-diamine; [0101]
N3-[3,5-Dichloro-4-(5-chloro-pyridin-3-yl)-phenyl]-1H-[1,2,4]triazole-3,5-
-diamine; and [0102]
N3-[3,5-Dichloro-4-(6-methoxy-pyridin-2-yl)-phenyl]-1H-[1,2,4]triazole-3,-
5-diamine.
[0103] The application provides a method for preventing a Hepatitis
C Virus (HCV) infection comprising administering to a patient in
need thereof a therapeutically effective amount of a compound of
Formula I.
[0104] The application provides the above method, further
comprising administering to a patient in need thereof a
therapeutically effective amount of an immune system
suppressant.
[0105] The application provides a method for treating a Hepatitis C
Virus (HCV) infection comprising administering to a patient in need
thereof a therapeutically effective amount of a compound of Formula
I.
[0106] The application provides any of the above methods, further
comprising administering a combination of antiviral agents that
inhibits replication of HCV.
[0107] The application provides any of the above methods, further
comprising administering an immune system modulator or an antiviral
agent that inhibits replication of HCV, or a combination
thereof.
[0108] The application provides the above method, wherein the
immune system modulator is an interferon or a chemically
derivatized interferon.
[0109] The application provides any of the above methods, further
comprising administering an immune system modulator or an antiviral
agent that inhibits replication of HCV, or a combination thereof,
wherein the antiviral agent is selected from the group consisting
of a HCV protease inhibitor, a HCV polymerase inhibitor, a HCV
helicase inhibitor, a HCV NS5A inhibitor, or any combination
thereof.
[0110] The application provides a composition comprising a compound
of Formula I and a pharmaceutically acceptable excipient.
[0111] The application provides the use of the compound of Formula
I in the preparation of a medicament for the prevention of HCV.
[0112] The application provides the use of the compound of Formula
I in the preparation of a medicament for the treatment of HCV.
[0113] The application provides any compound, composition, method
or use as described herein.
Compounds
[0114] Examples of representative compounds encompassed by the
present invention and within the scope of the invention are
provided in the following Table. These examples and preparations
which follow are provided to enable those skilled in the art to
more clearly understand and to practice the present invention. They
should not be considered as limiting the scope of the invention,
but merely as being illustrative and representative thereof.
[0115] In general, the nomenclature used in this application is
based on AUTONOM.TM. v.4.0, a Beilstein Institute computerized
system for the generation of IUPAC systematic nomenclature. If
there is a discrepancy between a depicted structure and a name
given that structure, the depicted structure is to be accorded more
weight. In addition, if the stereochemistry of a structure or a
portion of a structure is not indicated with, for example, bold or
dashed lines, the structure or portion of the structure is to be
interpreted as encompassing all stereoisomers of it.
[0116] TABLE I depicts examples of compounds according to generic
Formula I:
TABLE-US-00001 TABLE I # Nomenclature Structure 1
N.sup.3-[3,5-Dichloro-4-(6-methoxy- pyridin-3-yl)-phenyl]-1H-
[1,2,4]triazole-3,5-diamine ##STR00005## 2
N.sup.3-[3,5-Dichloro-4-(5- methanesulfonyl-pyridin-3-yl)-phenyl]-
1H-[1,2,4]triazole-3,5-diamine ##STR00006## 3
5-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2,6-dichloro-phenyl]-1H- pyridin-2-one ##STR00007## 4
N.sup.3-[4-(6-Amino-pyridin-3-yl)-3,5-
dichloro-phenyl]-1H-[1,2,4]triazole- 3,5-diamine ##STR00008## 5
N.sup.3-[4-(2-Amino-pyrimidin-5-yl)-3,5-
dichloro-phenyl]-1H-[1,2,4]triazole- 3,5-diamine ##STR00009## 6
N3-[3,5-Dichloro-4-(2-methoxy- pyridin-4-yl)-phenyl]-1H-
[1,2,4]triazole-3,5-diamine ##STR00010## 7
4-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2,6-dichloro-phenyl]-1H- pyridin-2-one ##STR00011## 8
N-{5-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2,6-dichloro-phenyl]-pyridin- 2-yl}-methanesulfonamide
##STR00012## 9 N.sup.5-[3-Fluoro-4-(6-fluoro-pyridin-3-
yl)-5-trifluoromethyl-phenyl]-1H- [1,2,4]triazole-3,5-diamine
##STR00013## 10 N.sup.5-(3-Fluoro-4-pyridin-3-yl-5
trifluoromethyl-phenyl)-1H- [1,2,4]triazole-3,5-diamine
##STR00014## 11 N.sup.3-[3,5-Dichloro-4-(6-
methanesulfonyl-pyridin-3-yl)-phenyl]-
1H-[1,2,4]triazole-3,5-diamine ##STR00015## 12
6-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2,6-dichloro-phenyl]-2,6-
diaza-spiro[3.3]heptane-2-carboxylic acid tert-butyl ester
##STR00016## 13 N.sup.3-(3-Chloro-4-pyridazin-3-yl-
phenyl)-1H-[1,2,4]triazole-3,5-diamine ##STR00017## 14
N.sup.3-[3-Chloro-4-(1-methyl-1H-tetrazol-
5-yl)-phenyl]-1H-[1,2,4]triazole-3,5- diamine ##STR00018## 15
N.sup.3-[3-Chloro-4-(2-methyl-2H- [1,2,4]triazol-3-yl)-phenyl]-1H-
[1,2,4]triazole-3,5-diamine ##STR00019## 16
N.sup.3-(3,5-Dichloro-4-pyrazol-1-yl-
phenyl)-1H-[1,2,4]triazole-3,5-diamine ##STR00020## 17
4-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2,6-dichloro-phenyl]-3,6- dihydro-2H-pyridine-1-carboxylic
acid tert-butyl ester ##STR00021## 18
N.sup.3-[3,5-Dichloro-4-(1,2,3,6-
tetrahydro-pyridin-4-yl)-phenyl]-1H- [1,2,4]triazole-3,5-diamine
##STR00022## 19 4-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2-chloro-6-trifluoromethyl-
phenyl]-3,6-dihydro-2H-pyridine-1- carboxylic acid tert-butyl ester
##STR00023## 20 N.sup.3-[3-Chloro-4-(1,2,3,6-tetrahydro-
pyridin-4-yl)-5-trifluoromethyl-
phenyl]-1H-[1,2,4]triazole-3,5-diamine ##STR00024## 21
N.sup.5-(3,5-Dichloro-4-[1,2,4]triazolo[4,3-
a]pyridin-3-yl-phenyl)-1H- [1,2,4]triazole-3,5-diamine ##STR00025##
22 N-{3-Chloro-4-[6-(propane-2- sulfonyl)-pyridin-3-yl]-5-
trifluoromethyl-phenyl}-4H- [1,2,4]triazole-3,5-diamine
##STR00026## 23 5-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2-chloro-6-trifluoromethyl- phenyl]-pyridine-2-sulfonic
acid tert- butylamide ##STR00027## 24
5-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2-chloro-6-trifluoromethyl- phenyl]-pyridine-2-sulfonic
acid (2,2,2-trifluoro-1,1-dimethyl- ethyl)-amide ##STR00028## 25
5-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2-chloro-6-trifluoromethyl- phenyl]-pyridine-2-su1fonic
acid adamantan-1-ylamide ##STR00029## 26
N.sup.3-[2-Chloro-4'-(4-methyl-piperazin-
1-yl)-6-trifluoromethyl-biphenyl-4-yl]-
1H-[1,2,4]triazole-3,5-diamine ##STR00030## 27
N.sup.3-(3-Chloro-4-pyridazin-3-yl-
phenyl)-1H-[1,2,4]triazole-3,5-diamine ##STR00031## 28
1-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-phenyl]-1H-pyridin-2-one ##STR00032## 29
1-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2-chloro-phenyl]-1H-pyridin- 2-one ##STR00033## 30
1-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2,6-dichloro-phenyl]-1H- pyridin-2-one ##STR00034## 31
N*3*-[3,5-Dichloro-4-(1,4,5,6-tetra
hydro-pyrimidin-2-yl)-phenyl]-1H- [1,2,4]triazole-3,5-diamine
##STR00035## 32 5-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2-chloro-6-trifluoromethyl- phenyl]-pyridine-2-carboxylic
acid methylamide ##STR00036## 33
N.sup.3(3,5-Dichloro-4-pyridazin-4-yl-
phenyl)-1H-[1,2,4]triazole-3,5-diamine ##STR00037## 34
1-{3-[4-(5-Amino-1H-[1,2,4]triazol-3-
ylamino)-2,6-dichloro-phenyl]- thiophen-2-yl}-ethanone ##STR00038##
35 N3-(3,5-Dichloro-4-pyridin-4-yl-
phenyl)-1H-[1,2,4]triazole-3,5-diamine ##STR00039## 36
N3-[3,5-Dichloro-4-(5-chloro- thiophen-2-yl)-phenyl]-1H-
[1,2,4]triazole-3,5-diamine ##STR00040## 37
N3-(3,5-Dichloro-4-pyridin-3-yl
phenyl)-1H-[1,2,4]triazole-3,5-diamine ##STR00041## 38
N3-[3,5-Dichloro-4-(1H-pyrazol-3-yl)-
phenyl]-1H-[1,2,4]triazole-3,5-diamine ##STR00042## 39
N3-(3,5-Dichloro-4-pyrimidin-5-yl-
phenyl)-1H-[1,2,4]triazole-3,5-diamine ##STR00043## 40
N3-[3,5-Dichloro-4-(2-methoxy- pyrimidin-5-yl)-phenyl]-1H-
[1,2,4]triazole-3,5-diamine ##STR00044## 41 N3-[3,5-Dichloro-4-(6-
trifluoromethyl-pyridin-3-yl)-phenyl]-
1H-[1,2,4]triazole-3,5-diamine ##STR00045## 42
N3-[3,5-Dichloro-4-(1-methyl-3- trifluoromethyl-1H-pyrazol-4-yl)-
phenyl]-1H-[1,2,4]triazole-3,5-diamine ##STR00046## 43
N3-[3,5-Dichloro-4-(5-chloro-pyridin-
3-yl)-phenyl]-1H-[1,2,4]triazole-3,5- diamine ##STR00047## 44
N3-[3,5-Dichloro-4-(6-methoxy- pyridin-2-yl)-phenyl]-1H-
[1,2,4]triazole-3,5-diamine ##STR00048##
[0117] The following schemes depict general methods for obtaining
compounds of Formula I.
##STR00049##
##STR00050##
Dosage and Administration:
[0118] The compounds of the present invention may be formulated in
a wide variety of oral administration dosage forms and carriers.
Oral administration can be in the form of tablets, coated tablets,
dragees, hard and soft gelatin capsules, solutions, emulsions,
syrups, or suspensions. Compounds of the present invention are
efficacious when administered by other routes of administration
including continuous (intravenous drip) topical parenteral,
intramuscular, intravenous, subcutaneous, transdermal (which may
include a penetration enhancement agent), buccal, nasal, inhalation
and suppository administration, among other routes of
administration. The preferred manner of administration is generally
oral using a convenient daily dosing regimen which can be adjusted
according to the degree of affliction and the patient's response to
the active ingredient.
[0119] A compound or compounds of the present invention, as well as
their pharmaceutically useable salts, together with one or more
conventional excipients, carriers, or diluents, may be placed into
the form of pharmaceutical compositions and unit dosages. The
pharmaceutical compositions and unit dosage forms may be comprised
of conventional ingredients in conventional proportions, with or
without additional active compounds or principles, and the unit
dosage forms may contain any suitable effective amount of the
active ingredient commensurate with the intended daily dosage range
to be employed. The pharmaceutical compositions may be employed as
solids, such as tablets or filled capsules, semisolids, powders,
sustained release formulations, or liquids such as solutions,
suspensions, emulsions, elixirs, or filled capsules for oral use;
or in the form of suppositories for rectal or vaginal
administration; or in the form of sterile injectable solutions for
parenteral use. A typical preparation will contain from about 5% to
about 95% active compound or compounds (w/w). The term
"preparation" or "dosage form" is intended to include both solid
and liquid formulations of the active compound and one skilled in
the art will appreciate that an active ingredient can exist in
different preparations depending on the target organ or tissue and
on the desired dose and pharmacokinetic parameters.
[0120] The term "excipient" as used herein refers to a compound
that is useful in preparing a pharmaceutical composition, generally
safe, non-toxic and neither biologically nor otherwise undesirable,
and includes excipients that are acceptable for veterinary use as
well as human pharmaceutical use. The compounds of this invention
can be administered alone but will generally be administered in
admixture with one or more suitable pharmaceutical excipients,
diluents or carriers selected with regard to the intended route of
administration and standard pharmaceutical practice.
[0121] "Pharmaceutically acceptable" means that which is useful in
preparing a pharmaceutical composition that is generally safe,
non-toxic, and neither biologically nor otherwise undesirable and
includes that which is acceptable for veterinary as well as human
pharmaceutical use.
[0122] A "pharmaceutically acceptable salt" form of an active
ingredient may also initially confer a desirable pharmacokinetic
property on the active ingredient which were absent in the non-salt
form, and may even positively affect the pharmacodynamics of the
active ingredient with respect to its therapeutic activity in the
body. The phrase "pharmaceutically acceptable salt" of a compound
means a salt that is pharmaceutically acceptable and that possesses
the desired pharmacological activity of the parent compound. Such
salts include: (1) acid addition salts, formed with inorganic acids
such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric
acid, phosphoric acid, and the like; or formed with organic acids
such as acetic acid, propionic acid, hexanoic acid,
cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic
acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric
acid, tartaric acid, citric acid, benzoic acid,
3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,
methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic
acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid,
4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,
4-toluenesulfonic acid, camphorsulfonic acid,
4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic
acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary
butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic
acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic
acid, and the like; or (2) salts formed when an acidic proton
present in the parent compound either is replaced by a metal ion,
e.g., an alkali metal ion, an alkaline earth ion, or an aluminum
ion; or coordinates with an organic base such as ethanolamine,
diethanolamine, triethanolamine, tromethamine, N-methylglucamine,
and the like.
[0123] Solid form preparations include powders, tablets, pills,
capsules, cachets, suppositories, and dispersible granules. A solid
carrier may be one or more substances which may also act as
diluents, flavoring agents, solubilizers, lubricants, suspending
agents, binders, preservatives, tablet disintegrating agents, or an
encapsulating material. In powders, the carrier generally is a
finely divided solid which is a mixture with the finely divided
active component. In tablets, the active component generally is
mixed with the carrier having the necessary binding capacity in
suitable proportions and compacted in the shape and size desired.
Suitable carriers include but are not limited to magnesium
carbonate, magnesium stearate, talc, sugar, lactose, pectin,
dextrin, starch, gelatin, tragacanth, methylcellulose, sodium
carboxymethylcellulose, a low melting wax, cocoa butter, and the
like. Solid form preparations may contain, in addition to the
active component, colorants, flavors, stabilizers, buffers,
artificial and natural sweeteners, dispersants, thickeners,
solubilizing agents, and the like.
[0124] Liquid formulations also are suitable for oral
administration include liquid formulation including emulsions,
syrups, elixirs, aqueous solutions, aqueous suspensions. These
include solid form preparations which are intended to be converted
to liquid form preparations shortly before use. Emulsions may be
prepared in solutions, for example, in aqueous propylene glycol
solutions or may contain emulsifying agents such as lecithin,
sorbitan monooleate, or acacia. Aqueous solutions can be prepared
by dissolving the active component in water and adding suitable
colorants, flavors, stabilizing, and thickening agents. Aqueous
suspensions can be prepared by dispersing the finely divided active
component in water with viscous material, such as natural or
synthetic gums, resins, methylcellulose, sodium
carboxymethylcellulose, and other well-known suspending agents.
[0125] The compounds of the present invention may be formulated for
parenteral administration (e.g., by injection, for example bolus
injection or continuous infusion) and may be presented in unit dose
form in ampoules, pre-filled syringes, small volume infusion or in
multi-dose containers with an added preservative. The compositions
may take such forms as suspensions, solutions, or emulsions in oily
or aqueous vehicles, for example solutions in aqueous polyethylene
glycol. Examples of oily or nonaqueous carriers, diluents, solvents
or vehicles include propylene glycol, polyethylene glycol,
vegetable oils (e.g., olive oil), and injectable organic esters
(e.g., ethyl oleate), and may contain formulatory agents such as
preserving, wetting, emulsifying or suspending, stabilizing and/or
dispersing agents. Alternatively, the active ingredient may be in
powder form, obtained by aseptic isolation of sterile solid or by
lyophilisation from solution for constitution before use with a
suitable vehicle, e.g., sterile, pyrogen-free water.
[0126] The compounds of the present invention may be formulated for
topical administration to the epidermis as ointments, creams or
lotions, or as a transdermal patch. Ointments and creams may, for
example, be formulated with an aqueous or oily base with the
addition of suitable thickening and/or gelling agents. Lotions may
be formulated with an aqueous or oily base and will in general also
containing one or more emulsifying agents, stabilizing agents,
dispersing agents, suspending agents, thickening agents, or
coloring agents. Formulations suitable for topical administration
in the mouth include lozenges comprising active agents in a
flavored base, usually sucrose and acacia or tragacanth; pastilles
comprising the active ingredient in an inert base such as gelatin
and glycerin or sucrose and acacia; and mouthwashes comprising the
active ingredient in a suitable liquid carrier.
[0127] The compounds of the present invention may be formulated for
administration as suppositories. A low melting wax, such as a
mixture of fatty acid glycerides or cocoa butter is first melted
and the active component is dispersed homogeneously, for example,
by stirring. The molten homogeneous mixture is then poured into
convenient sized molds, allowed to cool, and to solidify.
[0128] The compounds of the present invention may be formulated for
vaginal administration. Pessaries, tampons, creams, gels, pastes,
foams or sprays containing in addition to the active ingredient
such carriers as are known in the art to be appropriate.
[0129] The compounds of the present invention may be formulated for
nasal administration. The solutions or suspensions are applied
directly to the nasal cavity by conventional means, for example,
with a dropper, pipette or spray. The formulations may be provided
in a single or multidose form. In the latter case of a dropper or
pipette, this may be achieved by the patient administering an
appropriate, predetermined volume of the solution or suspension. In
the case of a spray, this may be achieved for example by means of a
metering atomizing spray pump.
[0130] The compounds of the present invention may be formulated for
aerosol administration, particularly to the respiratory tract and
including intranasal administration. The compound will generally
have a small particle size for example of the order of five (5)
microns or less. Such a particle size may be obtained by means
known in the art, for example by micronization. The active
ingredient is provided in a pressurized pack with a suitable
propellant such as a chlorofluorocarbon (CF), for example,
dichlorodifluoromethane, trichlorofluoromethane, or
dichlorotetrafluoroethane, or carbon dioxide or other suitable gas.
The aerosol may conveniently also contain a surfactant such as
lecithin. The dose of drug may be controlled by a metered valve.
Alternatively the active ingredients may be provided in a form of a
dry powder, for example a powder mix of the compound in a suitable
powder base such as lactose, starch, starch derivatives such as
hydroxypropylmethyl cellulose and polyvinylpyrrolidine (PVP). The
powder carrier will form a gel in the nasal cavity. The powder
composition may be presented in unit dose form for example in
capsules or cartridges of e.g., gelatin or blister packs from which
the powder may be administered by means of an inhaler.
[0131] When desired, formulations can be prepared with enteric
coatings adapted for sustained or controlled release administration
of the active ingredient. For example, the compounds of the present
invention can be formulated in transdermal or subcutaneous drug
delivery devices. These delivery systems are advantageous when
sustained release of the compound is necessary and when patient
compliance with a treatment regimen is crucial. Compounds in
transdermal delivery systems are frequently attached to a
skin-adhesive solid support. The compound of interest can also be
combined with a penetration enhancer, e.g., Azone
(1-dodecylaza-cycloheptan-2-one). Sustained release delivery
systems are inserted subcutaneously into to the subdermal layer by
surgery or injection. The subdermal implants encapsulate the
compound in a lipid soluble membrane, e.g., silicone rubber, or a
biodegradable polymer, e.g., polylactic acid.
[0132] Suitable formulations along with pharmaceutical carriers,
diluents and excipients are described in Remington: The Science and
Practice of Pharmacy 1995, edited by E. W. Martin, Mack Publishing
Company, 19th edition, Easton, Pa. A skilled formulation scientist
may modify the formulations within the teachings of the
specification to provide numerous formulations for a particular
route of administration without rendering the compositions of the
present invention unstable or compromising their therapeutic
activity.
[0133] The modification of the present compounds to render them
more soluble in water or other vehicle, for example, may be easily
accomplished by minor modifications (salt formulation,
esterification, etc.), which are well within the ordinary skill in
the art. It is also well within the ordinary skill of the art to
modify the route of administration and dosage regimen of a
particular compound in order to manage the pharmacokinetics of the
present compounds for maximum beneficial effect in patients.
[0134] The term "therapeutically effective amount" as used herein
means an amount required to reduce symptoms of the disease in an
individual. The dose will be adjusted to the individual
requirements in each particular case. That dosage can vary within
wide limits depending upon numerous factors such as the severity of
the disease to be treated, the age and general health condition of
the patient, other medicaments with which the patient is being
treated, the route and form of administration and the preferences
and experience of the medical practitioner involved. For oral
administration, a daily dosage of between about 0.01 and about 1000
mg/kg body weight per day should be appropriate in monotherapy
and/or in combination therapy. A preferred daily dosage is between
about 0.1 and about 500 mg/kg body weight, more preferred 0.1 and
about 100 mg/kg body weight and most preferred 1.0 and about 10
mg/kg body weight per day. Thus, for administration to a 70 kg
person, the dosage range would be about 7 mg to 0.7 g per day. The
daily dosage can be administered as a single dosage or in divided
dosages, typically between 1 and 5 dosages per day. Generally,
treatment is initiated with smaller dosages which are less than the
optimum dose of the compound. Thereafter, the dosage is increased
by small increments until the optimum effect for the individual
patient is reached. One of ordinary skill in treating diseases
described herein will be able, without undue experimentation and in
reliance on personal knowledge, experience and the disclosures of
this application, to ascertain a therapeutically effective amount
of the compounds of the present invention for a given disease and
patient.
[0135] The pharmaceutical preparations are preferably in unit
dosage forms. In such form, the preparation is subdivided into unit
doses containing appropriate quantities of the active component.
The unit dosage form can be a packaged preparation, the package
containing discrete quantities of preparation, such as packeted
tablets, capsules, and powders in vials or ampoules. Also, the unit
dosage form can be a capsule, tablet, cachet, or lozenge itself, or
it can be the appropriate number of any of these in packaged
form.
Indications and Method of Treatment
Indication
[0136] The application provides a method for preventing a Hepatitis
C Virus (HCV) infection comprising administering to a patient in
need thereof a therapeutically effective amount of a compound of
Formula I.
[0137] The application provides the above method, further
comprising administering to a patient in need thereof a
therapeutically effective amount of an immune system
suppressant.
[0138] The application provides a method for treating a Hepatitis C
Virus (HCV) infection comprising administering to a patient in need
thereof a therapeutically effective amount of a compound of Formula
I.
[0139] The application provides any of the above methods, further
comprising administering an immune system modulator or an antiviral
agent that inhibits replication of HCV, or a combination
thereof.
[0140] The application provides the above method, wherein the
immune system modulator is an interferon or a chemically
derivatized interferon.
[0141] The application provides any of the above methods, further
comprising administering an immune system modulator or an antiviral
agent that inhibits replication of HCV, or a combination thereof,
wherein the antiviral agent is selected from the group consisting
of a HCV protease inhibitor, a HCV polymerase inhibitor, a HCV
helicase inhibitor, a HCV NS5A inhibitor, or any combination
thereof.
Combination Therapy
[0142] The compounds of the invention and their isomeric forms and
pharmaceutically acceptable salts thereof are useful in treating
and preventing HCV infection alone or when used in combination with
other compounds targeting viral or cellular elements or functions
involved in the HCV lifecycle. Classes of compounds useful in the
invention include, without limitation, all classes of HCV
antivirals.
[0143] For combination therapies, mechanistic classes of agents
that can be useful when combined with the compounds of the
invention include, for example, nucleoside and non-nucleoside
inhibitors of the HCV polymerase, protease inhibitors, helicase
inhibitors, NS4B inhibitors, NS5A inhibitors and medicinal agents
that functionally inhibit the internal ribosomal entry site (IRES)
and other medicaments that inhibit HCV cell attachment or virus
entry, HCV RNA translation, HCV RNA transcription, replication or
HCV maturation, assembly or virus release. Specific compounds in
these classes and useful in the invention include, but are not
limited to, macrocyclic, heterocyclic and linear HCV protease
inhibitors such as telaprevir (VX-950), boceprevir (SCH-503034),
narlaprevir (SCH-9005 18), ITMN-191 (R-7227), TMC-435350 (a.k.a.
TMC-435), MK-7009, BI-201335, BI-2061 (ciluprevir), BMS-650032,
ACH-1625, ACH-1095 (HCV NS4A protease co-factor inhibitor), VX-500,
VX-8 13, PHX-1766, PHX2054, IDX-136, IDX-3 16, ABT-450 EP-0 13420
(and congeners) and VBY-376; the Nucleosidic HCV polymerase
(replicase) inhibitors useful in the invention include, but are not
limited to, R7128, PSI-785 1, IDX-184, IDX-102, R1479, UNX-08 189,
PSI-6130, PSI-938 and PSI-879 and various other nucleoside and
nucleotide analogs and HCV inhibitors including (but not limited
to) those derived as 2'-C-methyl modified nucleos(t)ides, 4'-aza
modified nucleos(t)ides, and 7'-deaza modified nucleos(t)ides.
Non-nucleosidic HCV polymerase (replicase) inhibitors useful in the
invention, include, but are not limited to, HCV-796, HCV-371,
VCH-759, VCH-916, VCH-222, ANA-598, MK-3281, ABT-333, ABT-072,
PF-00868554, BI-207127, GS-9190, A-837093, JKT-109, GL-59728 and
GL-60667.
[0144] In addition, compounds of the invention can be used in
combination with cyclophyllin and immunophyllin antagonists (e.g.,
without limitation, DEBIO compounds, NM-811 as well as cyclosporine
and its derivatives), kinase inhibitors, inhibitors of heat shock
proteins (e.g., HSP90 and HSP70), other immunomodulatory agents
that can include, without limitation, interferons (-alpha, -beta,
-omega, -gamma, -lambda or synthetic) such as Intron A, Roferon-A,
Canferon-A300, Advaferon, Infergen, Humoferon, Sumiferon MP,
Alfaferone, IFN-.beta., Feron and the like; polyethylene glycol
derivatized (pegylated) interferon compounds, such as PEG
interferon-.alpha.-2a (Pegasys), PEG interferon-.alpha.-2b
(PEGIntron), pegylated IFN-.alpha.-con1 and the like; long acting
formulations and derivatizations of interferon compounds such as
the albumin-fused interferon, Albuferon, Locteron, and the like;
interferons with various types of controlled delivery systems
(e.g., ITCA-638, omega-interferon delivered by the DUROS
subcutaneous delivery system); compounds that stimulate the
synthesis of interferon in cells, such as resiquimod and the like;
interleukins; compounds that enhance the development of type 1
helper T cell response, such as SCV-07 and the like; TOLL-like
receptor agonists such as CpG-10101 (actilon), isotorabine, ANA773
and the like; thymosin .alpha.-1; ANA-245 and ANA-246; histamine
dihydrochloride; propagermanium; tetrachlorodecaoxide; ampligen;
IMP-321; KRN-7000; antibodies, such as civacir, XTL-6865 and the
like and prophylactic and therapeutic vaccines such as InnoVac C,
HCV E1E2/MF59 and the like. In addition, any of the above-described
methods involving administering an NS5A inhibitor, a Type I
interferon receptor agonist (e.g., an IFN-.alpha.) and a Type II
interferon receptor agonist (e.g., an IFN-.gamma.) can be augmented
by administration of an effective amount of a TNF-.alpha.
antagonist. Exemplary, non-limiting TNF-.alpha. antagonists that
are suitable for use in such combination therapies include ENBREL,
REMICADE, and HUMIRA.
[0145] In addition, compounds of the invention can be used in
combination with antiprotozoans and other antivirals thought to be
effective in the treatment of HCV infection such as, without
limitation, the prodrug nitazoxanide. Nitazoxanide can be used as
an agent in combination with the compounds disclosed in this
invention as well as in combination with other agents useful in
treating HCV infection such as peginterferon .alpha.-2a and
ribavirin.
[0146] Compounds of the invention can also be used with alternative
forms of interferons and pegylated interferons, ribavirin or its
analogs (e.g., tarabavarin, levoviron), microRNA, small interfering
RNA compounds (e.g., SIRPLEX-140-N and the like), nucleotide or
nucleoside analogs, immunoglobulins, hepatoprotectants,
anti-inflammatory agents and other inhibitors of NS5A. Inhibitors
of other targets in the HCV lifecycle include NS3 helicase
inhibitors; NS4A co-factor inhibitors; antisense oligonucleotide
inhibitors, such as ISIS-14803, AVI-4065 and the like;
vector-encoded short hairpin RNA (shRNA); HCV specific ribozymes
such as heptazyme, RPI, 13919 and the like; entry inhibitors such
as HepeX-C, HuMax-HepC and the like; alpha glucosidase inhibitors
such as celgosivir, UT-231B and the like; KPE-02003002 and BIVN 401
and IMPDH inhibitors. Other illustrative HCV inhibitor compounds
include those disclosed in the following publications: U.S. Pat.
Nos. 5,807,876; 6,498,178; 6,344,465; and 6,054,472; PCT Patent
Application Publication Nos. WO97/40028; WO98/4038 1; WO00/56331,
WO02/04425; WO03/007945; WO03/010141; WO03/000254; WO01/32153;
WO00/06529; WO00/18231; WO00/10573; WO00/13708; WO01/85172;
WO03/037893; WO03/037894; WO03/037895; WO02/100851; WO02/100846;
WO99/01582; WO00/09543; WO02/18369; WO98/17679, WO00/056331;
WO98/22496; WO99/07734; WO05/073216, WO05/073195 and
WO08/021927.
[0147] Additionally, combinations of, for example, ribavirin and
interferon, may be administered as multiple combination therapy
with at least one of the compounds of the invention. The present
invention is not limited to the aforementioned classes or compounds
and contemplates known and new compounds and combinations of
biologically active agents. It is intended that combination
therapies of the present invention include any chemically
compatible combination of a compound of this inventive group with
other compounds of the inventive group or other compounds outside
of the inventive group, as long as the combination does not
eliminate the anti-viral activity of the compound of this inventive
group or the anti-viral activity of the pharmaceutical composition
itself.
[0148] Combination therapy can be sequential, that is treatment
with one agent first and then a second agent (for example, where
each treatment comprises a different compound of the invention or
where one treatment comprises a compound of the invention and the
other comprises one or more biologically active agents) or it can
be treatment with both agents at the same time (concurrently).
Sequential therapy can include a reasonable time after the
completion of the first therapy before beginning the second
therapy. Treatment with both agents at the same time can be in the
same daily dose or in separate doses. Combination therapy need not
be limited to two agents and may include three or more agents. The
dosages for both concurrent and sequential combination therapy will
depend on absorption, distribution, metabolism and excretion rates
of the components of the combination therapy as well as other
factors known to one of skill in the art. Dosage values will also
vary with the severity of the condition to be alleviated. It is to
be further understood that for any particular subject, specific
dosage regimens and schedules may be adjusted over time according
to the individual's need and the judgment of the one skilled in the
art administering or supervising the administration of the
combination therapy.
[0149] The application provides a method for preventing a Hepatitis
C Virus (HCV) infection comprising administering to a patient in
need thereof a therapeutically effective amount of a compound of
Formula I.
[0150] The application provides the above method, further
comprising administering to a patient in need thereof a
therapeutically effective amount of an immune system
suppressant.
[0151] The application provides a method for treating a Hepatitis C
Virus (HCV) infection comprising administering to a patient in need
thereof a therapeutically effective amount of a compound of Formula
I.
[0152] The application provides any of the above methods, further
comprising administering an immune system modulator or an antiviral
agent that inhibits replication of HCV, or a combination
thereof.
[0153] The application provides the above method, wherein the
immune system modulator is an interferon or a chemically
derivatized interferon.
[0154] The application provides any of the above methods, further
comprising administering an immune system modulator or an antiviral
agent that inhibits replication of HCV, or a combination thereof,
wherein the antiviral agent is selected from the group consisting
of a HCV protease inhibitor, a HCV polymerase inhibitor, a HCV
helicase inhibitor, a HCV NS5A inhibitor, or any combination
thereof.
Examples
[0155] Commonly used abbreviations include: acetyl (Ac),
azo-bis-isobutyrylnitrile (AIBN), atmospheres (Atm),
9-borabicyclo[3.3.1]nonane (9-BBN or BBN),
2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP),
tert-butoxycarbonyl (Boc), di-tert-butyl pyrocarbonate or boc
anhydride (BOC.sub.2O), benzyl (Bn), butyl (Bu), Chemical Abstracts
Registration Number (CASRN), benzyloxycarbonyl (CBZ or Z), carbonyl
diimidazole (CDI), 1,4-diazabicyclo[2.2.2]octane (DABCO),
diethylaminosulfur trifluoride (DAST), dibenzylideneacetone (dba),
1,5-diazabicyclo[4.3.0]non-5-ene (DBN),
1,8-diazabicyclo[5.4.0]undec-7-ene (DBU),
N,N'-dicyclohexylcarbodiimide (DCC), 1,2-dichloethane (DCE),
dichloromethane (DCM), 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone
(DDQ), diethyl azodicarboxylate (DEAD),
di-iso-propylazodicarboxylate (DIAD), di-iso-butylaluminumhydride
(DIBAL or DIBAL-H), di-iso-propylethylamine (DIPEA), N,N-dimethyl
acetamide (DMA), 4-N,N-dimethylaminopyridine (DMAP),
N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO),
1,1'-bis-(diphenylphosphino)ethane (dppe),
1,1'-bis-(diphenylphosphino)ferrocene (dppf),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI),
2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), ethyl (Et),
ethyl acetate (EtOAc), ethanol (EtOH),
2-ethoxy-2H-quinoline-1-carboxylic acid ethyl ester (EEDQ), diethyl
ether (EtzO), ethyl isopropyl ether (EtOiPr),
O-(7-azabenzotriazole-1-yl)-N, N,N'N'-tetramethyluronium
hexafluorophosphate acetic acid (HATU), acetic acid (HOAc),
1-N-hydroxybenzotriazole (HOBt), high pressure liquid
chromatography (HPLC), iso-propanol (IPA), isopropylmagnesium
chloride (iPrMgCl), hexamethyl disilazane (HMDS), liquid
chromatography mass spectrometry (LCMS), lithium hexamethyl
disilazane (LiHMDS), meta-chloroperoxybenzoic acid (m-CPBA),
methanol (MeOH), melting point (mp), MeSO.sub.2-- (mesyl or Ms),
methyl (Me), acetonitrile (MeCN), m-chloroperbenzoic acid (MCPBA),
mass spectrum (ms), methyl t-butyl ether (MTBE), methyl
tetrahydrofuran (MeTHF), N-bromosuccinimide (NBS), n-Butyllithium
(nBuLi), N-carboxyanhydride (NCA), N-chlorosuccinimide (NCS),
N-methylmorpholine (NMM), N-methylpyrrolidone (NMP), pyridinium
chlorochromate (PCC), Dichloro-((bis-diphenylphosphino)ferrocenyl)
palladium(II) (Pd(dppf)Cl.sub.2), palladium(II) acetate
(Pd(OAc).sub.2), tris(dibenzylideneacetone)dipalladium(0)
(Pd.sub.2(dba).sub.3), pyridinium dichromate (PDC), phenyl (Ph),
propyl (Pr), iso-propyl (i-Pr), pounds per square inch (psi),
pyridine (pyr),
1,2,3,4,5-Pentaphenyl-1'-(di-tert-butylphosphino)ferrocene
(Q-Phos), room temperature (ambient temperature, rt or RT),
sec-Butyllithium (sBuLi), tert-butyldimethylsilyl or t-BuMe.sub.2Si
(TBDMS), tetra-n-butylammonium fluoride (TBAF), triethylamine (TEA
or Et.sub.3N), 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO),
triflate or CF.sub.3SO.sub.2-- (Tf), trifluoroacetic acid (TFA),
1,1'-bis-2,2,6,6-tetramethylheptane-2,6-dione (TMHD),
O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium tetrafluoroborate
(TBTU), thin layer chromatography (TLC), tetrahydrofuran (THF),
trimethylsilyl or Me.sub.3Si (TMS), p-toluenesulfonic acid
monohydrate (TsOH or pTsOH), 4-Me-CH.sub.4SO.sub.2-- or tosyl (Ts),
and N-urethane-N-carboxyanhydride (UNCA). Conventional nomenclature
including the prefixes normal (n), iso (i-), secondary (sec-),
tertiary (tert-) and neo have their customary meaning when used
with an alkyl moiety. (J. Rigaudy and D. P. Klesney, Nomenclature
in Organic Chemistry, IUPAC 1979 Pergamon Press, Oxford).
General Conditions
[0156] Compounds of the invention can be made by a variety of
methods depicted in the illustrative synthetic reactions described
below in the Examples section.
[0157] The starting materials and reagents used in preparing these
compounds generally are either available from commercial suppliers,
such as Aldrich Chemical Co., or are prepared by methods known to
those skilled in the art following procedures set forth in
references such as Fieser and Fieser's Reagents for Organic
Synthesis; Wiley & Sons: New York, 1991, Volumes 1-15; Rodd's
Chemistry of Carbon Compounds, Elsevier Science Publishers, 1989,
Volumes 1-5 and Supplementals; and Organic Reactions, Wiley &
Sons: New York, 1991, Volumes 1-40. It should be appreciated that
the synthetic reaction schemes shown in the Examples section are
merely illustrative of some methods by which the compounds of the
invention can be synthesized, and various modifications to these
synthetic reaction schemes can be made and will be suggested to one
skilled in the art having referred to the disclosure contained in
this application.
[0158] The starting materials and the intermediates of the
synthetic reaction schemes can be isolated and purified if desired
using conventional techniques, including but not limited to,
filtration, distillation, crystallization, chromatography, and the
like. Such materials can be characterized using conventional means,
including physical constants and spectral data.
[0159] Unless specified to the contrary, the reactions described
herein are typically conducted under an inert atmosphere at
atmospheric pressure at a reaction temperature range of from about
-78.degree. C. to about 150.degree. C., often from about 0.degree.
C. to about 125.degree. C., and more often and conveniently at
about room (or ambient) temperature, e.g., about 20.degree. C.
[0160] Various substituents on the compounds of the invention can
be present in the starting compounds, added to any one of the
intermediates or added after formation of the final products by
known methods of substitution or conversion reactions. If the
substituents themselves are reactive, then the substituents can
themselves be protected according to the techniques known in the
art. A variety of protecting groups are known in the art, and can
be employed. Examples of many of the possible groups can be found
in "Protective Groups in Organic Synthesis" by Green et al., John
Wiley and Sons, 1999. For example, nitro groups can be added by
nitration and the nitro group can be converted to other groups,
such as amino by reduction, and halogen by diazotization of the
amino group and replacement of the diazo group with halogen. Acyl
groups can be added by Friedel-Crafts acylation. The acyl groups
can then be transformed to the corresponding alkyl groups by
various methods, including the Wolff-Kishner reduction and
Clemmenson reduction. Amino groups can be alkylated to form mono-
and di-alkylamino groups; and mercapto and hydroxy groups can be
alkylated to form corresponding ethers. Primary alcohols can be
oxidized by oxidizing agents known in the art to form carboxylic
acids or aldehydes, and secondary alcohols can be oxidized to form
ketones. Thus, substitution or alteration reactions can be employed
to provide a variety of substituents throughout the molecule of the
starting material, intermediates, or the final product, including
isolated products.
Preparative Examples
Intermediate 1
Procedure 1
N*3*-(4-Bromo-3-chloro-5-trifluoromethyl-phenyl)-1H-[1,2,4]triazole-3,5-di-
amine (Intermediate 1)
##STR00051##
[0161]
2-bromo-1-chloro-5-isothiocyanato-3-(trifluoromethyl)benzene
##STR00052##
[0163] To a suspension of
4-bromo-3-chloro-5-(trifluoromethyl)aniline (15 g, 54.7 mmol, Eq:
1.00) in dichloromethane (13.2 g, 10.0 ml, 155 mmol, Eq: 25.3) at
0, was added 1,1'-thiocarbonyldiimidazole (11.7 g, 65.6 mmol, Eq:
1.2) The reaction was gradually warmed to room temperature and
stirred overnight. The reaction was concentrated and
chromatographed (220 g Redisep, 5 to 15% dichloromethane/hexane) to
give 13.84 g (80%) pale yellow oil.
(Z)-methyl
N-4-bromo-3-chloro-5-(trifluoromethyl)phenyl-N'-cyanocarbamimid-
othioate
##STR00053##
[0165] To a solution of
2-bromo-1-chloro-5-isothiocyanato-3-(trifluoromethyl)benzene (13.84
g, 43.7 mmol, Eq: 1.00) in dimethoxyethane (100 mL) was added to
sodium hydrogen cyanamide (3.36 g, 52.5 mmol, Eq: 1.2) and methanol
(10 mL). After 30 minutes, methyl iodide (15.9 g, 7 ml, 112 mmol,
Eq: 2.56) was added to the magenta-colored soln and the reaction
was stirred overnight at room temperature. The reaction mixture was
concentrated to dryness and dissolved in .about.50 mL acetonitrile.
Added 100 mL water to give a white precipitate. Filtered white
solid, rinsed with water and air-dried o/n to give 16.0 g (99%) of
white solid.
N*3*-(4-Bromo-3-chloro-5-trifluoromethyl-phenyl)-1H-[1,2,4]triazole-3,5-di-
amine (Intermediate 1)
##STR00054##
[0167] In a 500 mL round-bottomed flask, (Z)-methyl
N-4-bromo-3-chloro-5-(trifluoromethyl)phenyl-N'-cyanocarbamimidothioate
(1.45 g, 3.89 mmol, Eq: 1.00) was combined with ethanol (15 ml) to
give a white suspension. Hydrazine (1.25 g, 1.22 ml, 38.9 mmol, Eq:
10) was added and the reaction mixture was heated to 70.degree. C.
and stirred for 3 h. The reaction was cooled and water (.about.40
mL) was added to the reaction with shaking. The resulting
suspension was filtered, washed with water and vacuum oven dried at
45C over weekend. Obtained a white solid as desired product (1.12
g, 81% yield). Another sample was collected from mother liquor as
an pink solid (148 mg, .about.90 pure, 9.6% yield)
[0168] MS m/z 356 [M+H]
Intermediate 2
Procedure 1
N*3*-(4-Bromo-3,5-dichloro-phenyl)-1H-[1,2,4]triazole-3,5-diamine
(Intermediate 2)
##STR00055##
[0169] (Z)-methyl
N-4-bromo-3,5-dichlorophenyl-N'-cyanocarbamimidothioate
##STR00056##
[0171] A solution of sodium methoxide (2.6 ml, 1.3 mmol, Eq: 1.23)
was added to cyanamide (50 mg, 1.19 mmol, Eq: 1.12) and stirred at
room temperature for 15 minutes.
2-bromo-1,3-dichloro-5-isothiocyanatobenzene (300 mg, 1.06 mmol,
Eq: 1.00) was added to the reaction mixture and stirred for 1 hr.
Iodomethane (331 ms, 146 .mu.l, 2.33 mmol, Eq: 2.2) was added and
the pale yellow solution was stirred overnight at room temperature.
The resulting suspension was filtered and air dried to give 154 mg
(43%) of desired product as a light brown solid.
N*3*-(4-Bromo-3,5-dichloro-phenyl)-1H-[1,2,4]triazole-3,5-diamine
(Intermediate 2)
##STR00057##
[0173] A solution of (Z)-methyl
N-4-bromo-3,5-dichlorophenyl-N'-cyanocarbamimidothioate (154 mg,
454 .mu.mol, Eq: 1.00) and hydrazine (153 mg, 150 .mu.l, 4.78 mmol,
Eq: 10.5) in ethanol (5 mL) was heated at 65.degree. C. After 3 hr,
LCMS ok, no sm. Cooled to rt and stirred solution over weekend. The
reaction mixture was concentrated and chromatographed (11 g
Supelco, 0 to 10% MeOH/CH2Cl2) to give 80 mg (55%) of desired
product as an off-white solid.
[0174] .sup.1H NMR (300 MHz, DMSO) .quadrature.: 11.35 (s, 1H),
9.33 (s, 1H), 7.75 (s, 2H), 6.05 (s, 2H) ppm
Intermediate 3
Procedure 1
N.sup.5-(4-bromo-3-fluoro-5-trifluoromethylphenyl)-1H-[1,2,4]-triazole-3,5-
-diamine (Intermediate 3)
##STR00058##
[0175]
2-bromo-1-fluoro-5-isothiocyanato-3-trifluoromethylbenzene
##STR00059##
[0177] 4-bromo-3-fluoro-5-trifluoromethylaniline (4.22 g, 16.4
mmol, Eq: 1.00) and calcium carbonate (3.44 g, 1.17 ml, 34.3 mmol,
Eq: 2.1) were suspended in 50% aqueous dichlormethane (20 ml)
mixture. The thick suspension was stirred vigorously at 0.degree.
C. Thiophosgene (2.07 g, 138 ml, 18.0 mmol, Eq: 1.1) was added
slowly dropwise to the mixture. After the addition the mixture was
stirred at 0.degree. C. for 1.5 hr then stirred overnight at room
temperature. The solids were filtered and the filtrate was
extracted with dichloromethane. The combined organic phases were
washed with water, brine, dried over sodium sulfate and
concentrated in vacuo to afford 4.71 g (96%) of the desired
material as a light brown solid.
[0178] .sup.1H NMR (300 MHz, DMSO-d.sub.6) ppm 7.84 (s, 1H) 7.96
(dd, J=9.06, 2.27 Hz, 1H)
(4-Bromo-3-fluoro-5-trifluoromethyl-phenylamino)-(methyl-.quadrature..sup.-
4sulfanylidene)-methyl-cyanamide
##STR00060##
[0180] 2-bromo-1-fluoro-5-isothiocyanato-3-trifluoromethylbenzene
(4.71 g, 15.7 mmol, Eq: 1.00) was dissolved in anhydrous methanol
(30 ml). Sodium hydrogencyanamide (1.00 g, 15.7 mmol, Eq: 1) was
added and the reaction was stirred for 1 hr at ambient temperature.
Methyl iodide (4.46 g, 1.96 ml, 31.4 mmol, Eq: 2) was added
dropwise and the reaction was stirred overnight at ambient
temperature. The light brown suspension was filtered to afford 1.91
g (34%) of the desired product as a pink solid.
[0181] MS +m/z: 357.7. (M+1)
[0182] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .quadrature. ppm 2.78
(s, 3H) 7.87 (s, 1H) 7.97 (dd, J=1.00 Hz, 1H) 10.38 (br. s, 1H)
Prepared of
N.sup.5-(4-bromo-3-fluoro-5-trifluoromethylphenyl)-1H-[1,2,4]-triazole-3,-
5-diamine (Intermediate 3)
##STR00061##
[0184] Hydrazine (1.71 g, 53.4 mmol, Eq: 10) was added to a stirred
suspension of
(4-Bromo-3-fluoro-5-trifluoromethyl-phenylamino)-(methyl-.quadrature..sup-
.4sulfanylidene)-methyl-cyanamide (1.9 g, 5.34 mmol, Eq: 1.00) in
ethanol (30 ml). The mixture was heated to 70.degree. C. for 1 hr.
The reaction mixture was concentrated to a reduced volume (.about.5
ml) and water (.about.10 ml) was added dropwise while stirring. The
suspension was stirred for 30 min. The precipitate was filtered and
washed with water (.about.50 ml), then dried under high vacuum at
70.degree. C. for two hours to filtered to afford 1.73 g (95%) of
the desired product as a light pink solid.
[0185] MS +m/z: 339.9. (M+1)
[0186] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .quadrature. ppm 6.03
(s, 2H) 7.81 (s, 1H) 7.86 (d, J=12.13 Hz, 1H) 9.52 (s, 1H) 11.40
(s, 1H)
N*3*-[3,5-Dichloro-4-(6-methoxy-pyridin-3-yl)-phenyl]-1H-[1,2,4]triazole-3-
,5-diamine (Compound 1)
##STR00062##
[0187] 3,5-dichloro-4-(6-methoxypyridin-3-yl)aniline
##STR00063##
[0189] A microwave vial containing 4-bromo-3,5-dichloroaniline (250
mg, 1.04 mmol, Eq: 1.00), 6-methoxypyridin-3-ylboronic acid (206
mg, 1.35 mmol, Eq: 1.3), sodium carbonate (275 mg, 2.59 mmol, Eq:
2.5) and bis(triphenylphosphine)palladium (II) chloride (42.0 mg,
59.8 .mu.mol, Eq: 0.0577) was degassed with Argon for 15 min.
Dimethoxyethane (4 mL) and water (1 mL) was added and the reaction
was heated for 30 min with the microwave at 115 deg. The reaction
mixture was concentrated, diluted with ethyl acetate, and washed
with brine. Dried org extract with sodium sulfate and
chromatographed (40 g Redisep, 100% hexane to 10% ethyl
acetate/hexane) to give 206 mg (74%) of desired product as a
colorless oil.
5-(2,6-dichloro-4-isothiocyanatophenyl)-2-methoxypyridine
##STR00064##
[0191] To a suspension of calcium carbonate (192 mg, 1.92 mmol, Eq:
2.51) and thiophosgene (105 mg, 70 .mu.l, 913 mol, Eq: 1.19) in
dichloromethane (10.0 ml)/water (10.0 ml) at 0 deg, was added
3,5-dichloro-4-(6-methoxypyridin-3-yl)aniline (206 mg, 765 .mu.mol,
Eq: 1.00) The reaction was gradually warmed to room temperature and
stirred overnight. Added 24 mL 1N HCl slowly. Separated organic
layer and dried over sodium sulfate to give 186 mg (78%) of desired
product as a pale yellow oil.
N-((3,5-dichloro-4-(6-methoxypyridin-3-yl)phenylamino)(methylthio)methyl)c-
yanamide
##STR00065##
[0193] To a solution of
5-(2,6-dichloro-4-isothiocyanatophenyl)-2-methoxypyridine (186 mg,
598 .mu.mol, Eq: 1.00) in MeOH (5 mL) was added to sodium hydrogen
cyanamide (48 mg, 750 .mu.mol, Eq: 1.25). After 30 minutes, methyl
iodide (170 mg, 75 .mu.l, 1.2 mmol, Eq: 2.01) was added and the
reaction was stirred overnight at room temperature. The reaction
mixture was concentrated and chromatographed (24 g Redisep, 10 to
40% ethyl acetate/hexane) to give 131 mg (60%) of desired product
as a white solid.
N*3*-[3,5-Dichloro-4-(6-methoxy-pyridin-3-yl)-phenyl]-1H-[1,2,4]triazole-3-
,5-diamine (Compound 1)
##STR00066##
[0195] To a solution of
N-((3,5-dichloro-4-(6-methoxypyridin-3-yl)phenylamino)(methylthio)methyl)-
cyanamide (131 mg, 355 .mu.mol, Eq: 1.00) in ethanol (5 mL) was
added hydrazine (123 mg, 120 .mu.l, 3.82 mmol, Eq: 10.8). The
reaction mixture was heated at 60 deg o/n. The resulting suspension
was filtered to give 80 mg of desired product as a white solid. The
filtrate precipitated over time and was filtered to give an
additional 49 mg of white solid, for a total of 129 mg (100%).
[0196] MS m/z 351 [M+H]
N3*-[3,5-Dichloro-4-(5-methanesulfonyl-pyridin-3-yl)-phenyl]-1H-[1,2,4]tri-
azole-3,5-diamine (Compound 2)
##STR00067##
[0198] A microwave vial containing
N3-(4-bromo-3,5-dichlorophenyl)-1H-1,2,4-triazole-3,5-diamine
Intermediate 2 (100 mg, 310 .mu.mol, Eq: 1.00),
5-(methylsulfonyl)pyridin-3-ylboronic acid (98 mg, 488 .mu.mol, Eq:
1.57), sodium carbonate (85 mg, 802 .mu.mol, Eq: 2.59) and
Pd(Ph.sub.3P).sub.4 (38 mg, 32.9 .mu.mol, Eq: 0.106) was degassed
for 15 minutes with Argon. Dioxane (2 mL) and water (0.5 mL) was
added, and the suspension was degassed for 5 minutes with
sonication, and the reaction was heated at 125.degree. for 1 hr
with microwave. The reaction mixture was diluted with ethyl
acetate, washed with brine, dried over sodium sulfate, and
chromatographed (24 g Supelco, 0 to 10% MeOH/DCM) to give 20 mg of
desired product as a yellow solid.
[0199] MS m/z 399 [M+H]
5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-1H-pyridin-
-2-one (Compound 3)
##STR00068##
[0201] A solution of
N3-(3,5-dichloro-4-(6-methoxypyridin-3-yl)phenyl)-1H-1,2,4-triazole-3,5-d-
iamine Compound 1 (95 mg, 271 .mu.mol, Eq: 1.00) and HBr (231 mg,
155 .mu.l, 1.37 mmol, Eq: 5.06) in AcOH (5 mL) was heated at 100
deg for 2 days in a sealed tube. The reaction mixture was carefully
poured into ice NaHCO.sub.3, extracted with ethyl acetate, and
dried with sodium sulfate to give 42 mg (46%) of desired product as
an off-white solid.
[0202] MS m/z 337 [M+H]
N*3*-[4-(6-Amino-pyridin-3-yl)-3,5-dichloro-phenyl]-1H-[1,2,4]triazole-3,5-
-diamine (Compound 4)
##STR00069##
[0204] A microwave vial containing
N3-(4-bromo-3,5-dichlorophenyl)-1H-1,2,4-triazole-3,5-diamine
Intermediate 2 (100 mg, 310 .mu.mol, Eq: 1.00),
5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (104
mg, 473 .mu.mol, Eq: 1.53), sodium carbonate (92 mg, 868 .mu.mol,
Eq: 2.8) and Pd(Ph.sub.3P).sub.4 (58 mg, 50.2 .mu.mol, Eq: 0.162)
was degassed for 15 minutes with Argon. Dioxane (2 mL) and water
(0.5 mL) was added, and the suspension was degassed for 5 minutes
with sonication, and the reaction was heated at 125.degree. for 1
hr with microwave. The reaction was concentrated, diluted with
ethyl acetate, washed with brine, dried with sodium sulfate, and
purified by preparative plate chromatography (10% MeOH/DCM) to give
26 mg (25%) of desired product as a light yellow solid.
[0205] MS m/z 336 [M+H]
N*3*-[4-(2-Amino-pyridin-5-yl)-3,5-dichloro-phenyl]-1H-[1,2,4]triazole-3,5-
-diamine (Compound 5)
##STR00070##
[0207] A microwave vial containing
N3-(4-bromo-3,5-dichlorophenyl)-1H-1,2,4-triazole-3,5-diamine
Intermediate 2 (100 mg, 310 .mu.mol, Eq: 1.00),
5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine
(108 mg, 489 .mu.mol, Eq: 1.58), sodium carbonate (88 mg, 830
.mu.mol, Eq: 2.68) and Pd(Ph.sub.3P).sub.4 (49 ag, 42.4 .mu.mol,
Eq: 0.137) was degassed for 15 minutes with Argon. Dioxane (2 mL)
and water (0.5 mL) was added, and the suspension was degassed for 5
minutes with sonication, and the reaction was heated at 125.degree.
for 1.5 hr with microwave. The reaction mixture was concentrated,
diluted with ethyl acetate, washed with brine, dried with sodium
sulfate, and chromatographed (11 g Supelco, 0 to 10% MeOH/DCM) to
give 9.5 mg (9%) of desired product as a light yellow solid.
[0208] MS m/z 337 [M+H]
N3-(3,5-dichloro-4-(2-methoxypyridin-4-yl)phenyl)-1H-1,2,4-triazole-3,5-di-
amine (Compound 6)
##STR00071##
[0210] To a solution of
N-((3,5-dichloro-4-(2-methoxypyridin-4-yl)phenylamino)(methylthio)methyl)-
cyanamide (192 mg, 520 .mu.mol, Eq: 1.00) in ethanol (7 mL) was
added hydrazine (184 mg, 180 .mu.l, 5.74 mmol, Eq: 11.0). The
reaction mixture was heated at 60 deg o/n. The reaction was
concentrated and chromatographed (23 g Supelco, 0 to 10% MeOH/DCM)
to give 126 mg (69%) of desired product as a white solid.
##STR00072##
3,5-dichloro-4-(2-methoxypyridin-4-yl)aniline
##STR00073##
[0212] A microwave vial containing 4-bromo-3,5-dichloroaniline (350
mg, 1.45 mmol, Eq: 1.00), 2-methoxypyridin-4-ylboronic acid (289
mg, 1.89 mmol, Eq: 1.3), sodium carbonate (394 mg, 3.72 mmol, Eq:
2.56) and bis(triphenylphosphine)palladium (II) chloride (65 mg,
92.6 .mu.mol, Eq: 0.0637) was degassed with Argon for 15 min. DME
(8 mL) and water (2 mL) was added and the reaction was heated for
30 min with the microwave at 115 deg. The reaction mixture was
concentrated, diluted with ethyl acetate, washed with brine, dried
with sodium sulfate and chromatographed (40 g Redisep, 100% to 10%
ethyl acetate/hexane) to give 254 mg (65%) of desired product as a
colorless oil.
4-(2,6-dichloro-4-isothiocyanatophenyl)-2-methoxypyridine
##STR00074##
[0214] To a suspension of calcium carbonate (264 mg, 2.64 mmol, Eq:
2.79) and thiophosgene (135 mg, 90 .mu.l, 1.17 mmol, Eq: 1.24) in
dichloromethane (10.0 mL)/water (10.0 mL) at 0 deg, was added
3,5-dichloro-4-(2-methoxypyridin-4-yl)aniline (254 mg, 944 .mu.mol,
Eq: 1.00) The reaction was gradually warmed to room temperature and
stirred overnight. Added 2.5 mL 1N HCl slowly. Separated organic
layer and dried over sodium sulfate to give 240 mg (82%) of desired
product as a light yellow oil.
N-((3,5-dichloro-4-(2-methoxypyridin-4-yl)phenylamino)(methyltho)methyl)cy-
anamide
##STR00075##
[0216] To a solution of
4-(2,6-dichloro-4-isothiocyanatophenyl)-2-methoxypyridine (240 mg,
771 .mu.mol, Eq: 1.00) in MeOH (6 mL) was added to sodium hydrogen
cyanamide (61.9 mg, 967 .mu.mol, Eq: 1.25). After 30 minutes,
methyl iodide (227 mg, 100 .mu.l, 1.6 mmol, Eq: 2.07) was added and
the reaction was stirred overnight at room temperature. The
resulting suspension was filtered to give 94 mg of desired product
as a white solid. The filtrate was concentrated and chromatographed
(24 g Redisep, 10 to 40% ethyl acetate/hexane) to give an
additional 98 mg of desired product as an off-white solid, for a
total of 192 mg (68%) of product.
N3-(3,5-dichloro-4-(2-methoxypyridin-4-yl)phenyl)-1H-1,2,4-triazole-3,5-di-
amine (Compound 6)
##STR00076##
[0218] To a solution of
N-((3,5-dichloro-4-(2-methoxypyridin-4-yl)phenylamino)(methylthio)methyl)-
cyanamide (192 mg, 520 .mu.mol, Eq: 1.00) in ethanol (7 mL) was
added hydrazine (184 mg, 180 .mu.l, 5.74 mmol, Eq: 11.0). The
reaction mixture was heated at 60 deg o/n. The reaction was
concentrated and chromatographed (23 g Supelco, 0 to 10% MeOH/DCM)
to give 126 mg (69%) of desired product as a white solid.
[0219] MS m/z 352 [M+H]
4-[4-(5-Amino-1H-[1,2,4]triazole-3-ylamino)-2,6-dichloro-phenyl]-1H-pyridi-
n-2-one (Compound 7)
##STR00077##
[0221] 5/9 10 am A solution of
N3-(3,5-dichloro-4-(2-methoxypyridin-4-yl)phenyl)-1H-1,2,4-triazole-3,5-d-
iamine Compound 6 (126 mg, 359 .mu.mol, Eq: 1.00) and HBr (298 mg,
200 .mu.l, 1.77 mmol, Eq: 4.93) in AcOH (5 mL) was heated at 100
deg for 2 d in a sealed tube. The reaction mixture was carefully
poured into ice NaHCO.sub.3, extracted with ethyl acetate, dried
with sodium sulfate to give 64 mg (53%) of desired product as an
off-white solid.
[0222] MS m/z 337 [M+H]
N-{5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-pyridin-
-2-yl}-methanesulfonamide (Compound 8)
##STR00078##
[0223]
N-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)meth-
anesulfonamide
##STR00079##
[0225] To a solution of
5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (500
mg, 2.27 mmol, Eq: 1.00) and pyridine (538 mg, 550 .mu.l, 6.8 mmol,
Eq: 2.99) in DCM (10 mL) at 0 deg, was added Ms-Cl (323 mg, 220
.mu.l, 2.82 mmol, Eq: 1.24). The reaction mixture was gradually
warmed to room temperature and stirred overnight. The reaction
mixture was diluted with DCM, washed with 1N HCl, and dried over
sodium sulfate to give 118 mg (17%) of white solid, containing
desired product and impurities.
N-{5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-pyridin-
-2-yl}-methanesulfonamide
##STR00080##
[0227] A microwave vial containing
N3-(4-bromo-3,5-dichlorophenyl)-1H-1,2,4-triazole-3,5-diamine
Compound 7 (100 mg, 310 .mu.mol, Eq: 1.00),
N-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)methanesul-
fonamide (118 mg, 396 .mu.mol, Eq: 1.28), sodium carbonate (82.0
mg, 774 .mu.mol, Eq: 2.5) and Pd(Ph.sub.3P).sub.4 (42 mg, 36.3
.mu.mol, Eq: 0.117) was degassed for 15 minutes with Argon. Dioxane
(2 mL) and water (0.5 mL) was added, and the suspension was
degassed for 5 minutes with sonication, and the reaction was heated
at 125.degree. for 1.5 hr with microwave.
[0228] The reaction mixture was concentrated, diluted with ethyl
acetate, washed with brine, dried with sodium sulfate, and
chromatographed (24 g Redisep, 0 to 10% MeOH/DCM) to give a brown
solid containing product and impurities. The solid was triturated
with MeOH/DCM to give 30 mg (24%) of desired product as a light
brown solid.
[0229] MS m/z 414 [M+H]
N.sup.5-(3-Fluoro-4-(6-fluoropyridin-3-yl)-5-trifluoromethylphenyl)-1H-[1,-
2,4]-triazole-3,5-diamine (Compound 9)
##STR00081##
[0231] To a 15 mL microwave vial was added
N.sup.5-(4-bromo-3-fluoro-5-trifluoromethylphenyl)-1H-[1,2,4]-triazole-3,-
5-diamine Intermediate 3 (50 mg, 147 .mu.mol, Eq: 1.00),
6-fluoropyridin-3-ylboronic acid (31.1 mg, 221 .mu.mol, Eq: 1.5)
and sodium hydroxide (735 .mu.l, 735 .mu.mol, Eq: 5) in DME (1.5
ml). Pd(Ph.sub.3P).sub.4 (8.49 mg, 7.35 .mu.mol, Eq: 0.05) was
added, the mixture was purged with argon, the vial was capped and
heated in the microwave at 120.degree. C. for 30 minutes. The
reaction mixture was diluted with dichloromethane, filtered through
celite and concentrated in vacuo. The residue was taken up in MeOH,
filtered through a 4 micron filter and the crude material was
purified by preparative HPLC (20% ACN: 0.3% TFA in water to 100%
ACN) to afford 11 mg (20%) of the desired product as a white
solid.
[0232] MS +m/z: 357.0. (M+1)
N.sup.5-(3-Fluoro-4-(pyridin-3-yl)-5-trifluoromethylphenyl)-1H-[1,2,4]-tri-
azole-3,5-diamine (Compound 10)
##STR00082##
[0234] Prepared by a similar procedure to Compound 8, except
substituted pyridine-3-boronic acid for 6-fluoropyridin-3-ylboronic
acid to afford 5 mg (7%) of the desired material as a white
solid.
[0235] MS +m/z: 339.0. (M+1)
N.sup.5-(3,5-Dichloro-4-(6-methanesulfonyl-pyridin-3-yl)-phenyl)-1H-[1,2,4-
]-triazole-3,5-diamine (Compound 11)
##STR00083##
[0237] To a 15 mL microwave vial was added
N3-(4-bromo-3,5-dichlorophenyl)-1H-1,2,4-triazole-3,5-diamine
Intermediate 2 (201 mg, 622 .mu.mol, Eq: 1.00),
6-(methylsulfonyl)pyridin-3-ylboronic acid (125 mg, 622 .mu.mol,
Eq: 1.00) and Cs.sub.2CO.sub.3 (608 mg, 1.87 mmol, Eq: 3) in
n-butanol (3.00 ml) and water (600 .mu.l). PdCl.sub.2(DPPF) (50.8
mg, 62.2 .mu.mol, Eq: 0.1) was added, the mixture was purged with
argon, the vial was capped and heated in the microwave at
135.degree. C. for 30 minutes. The mixture was diluted with
dichloromethane, added Na.sub.2SO.sub.4 and filtered through
celite. The filtrate was taken up in methanol, filtered and
concentrated. The crude material was purified by preparative HPLC
(0.1% TFA in water/0.1% TFA in AcCN). 95% to 10% over 25 minutes to
afford 13 mg (5%) of the desired material as a white solid.
[0238] MS +m/z: 398.9/400.8. (M+1)
6-[4-(5-Amino-2H-[1,2,4]triazol-3-ylamino)-2,6-dichlorophenyl]-2,6-diaza-s-
piro[3.3]heptane-2-carboxylic acid tert-butyl ester (Compound
12)
##STR00084##
[0239]
6-(2,6-Dichloro-4-nitrophenyl]-2,6-diaza-spiro[3.3]heptane-2-carbox-
ylic acid tert-butyl ester
##STR00085##
[0241] In a 50 ml round-bottomed flask,
1,3-dichloro-2-fluoro-5-nitrobenzene (467 mg, 2.22 mmol, Eq: 1.00),
tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (529 mg, 2.67
mmol, Eq: 1.2) and Cs.sub.2CO.sub.3 (1.81 g, 5.56 mmol, Eq: 2.5)
were combined with DMF (10 ml) to give a yellow suspension. The
mixture was heated to 90.degree. C. and stirred under argon for 18
hours. TLC indicated total conversion. Cooled and the reaction
mixture was diluted with H.sub.2O and EtOAc. The aqueous layer was
washed with EtOAc (2.times.40 ml). The organic layers were
combined, washed with H.sub.2O (2.times.25 mL), brine (1.times.25
mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo to
afford the desired product in quantitative yield as a bright yellow
powder.
[0242] MS -m/z: 386.9/389.0. (M-1)
6-(4-Amino-2,6-dichlorophenyl]-2,6-diaza-spiro[3.3]heptane-2-carboxylic
acid tert-butyl ester
##STR00086##
[0244] In a 50 mL round-bottomed flask, tert-butyl
6-(2,6-dichloro-4-nitrophenyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate
(700 mg, 1.8 mmol, Eq: 1.00) and ammonium formate (955 mg, 15.1
mmol, Eq: 8.4) were combined with methanol (100 ml) and water (15
ml) to give a yellow suspension. Zinc (495 mg, 7.57 mmol, Eq: 4.2)
was added and the suspension was stirred at 25.degree. C. for 18
hours. Additional ammonium formate (1.02 g, 16.2 mmol, Eq: 9) and
zinc (472 mg, 7.21 mmol, Eq: 4) were added with methanol (50 ml)
and water (10 ml) and the yellow suspension was stirred at
25.degree. C. for 5 hours. The reaction mixture was filtered and
the filter cake was washed with methanol. The filtrate was
concentrated in vacuo. The crude material was triturated with hot
EtOAc, filtered and stripped. The dark yellow filtrate was stripped
in vacuo and the crude material was purified by flash
chromatography (silica gel, 80 g, 0% to 30% EtOAc in heptane). The
dark yellow oil was dried under vacuum at 25C overnight to afford
399 mg (62%) the desired product as a yellow crystalline solid.
[0245] MS +m/z: 358/360. (M+1)
6-(2,6-Dichloro-4-isothiocyanato-phenyl]-2,6-diaza-spiro[3.3]heptane-2-car-
boxylic acid tert-butyl ester
##STR00087##
[0247] Tert-butyl
6-(4-amino-2,6-dichlorophenyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate
(137 mg, 382 .mu.mol, Eq: 1.00) and calcium carbonate (80.4 ms, 803
.mu.mol, Eq: 2.1) were suspended in a 50% aqueous dichloromethane
(3 ml). Thiophosgene (48.4 mg, 32.2 .mu.L, 421 .mu.mol, Eq: 1.1)
was added dropwise to the mixture at 25.degree. C. After the
addition the mixture was stirred at 25.degree. C. for 22 hours. The
reaction was filtered and the filter cake was washed with
dichloromethane. The filtrate was separated and the aqueous layer
extracted with dichloromethane. The organic layers were dried over
Na.sub.2SO.sub.4 and concentrated in vacuo to give a yellow
crystalline solid. The crude material was used without any further
purification.
tert-butyl
6-(2,6-dichloro-4-(cyanamido(methylthio)methyleneamino)phenyl)--
2,6-diazaspiro[3.3]heptane-2-carboxylate
##STR00088##
[0249] In a 250 mL round-bottomed flask, tert-butyl
6-(2,6-dichloro-4-isothiocyanatophenyl)-2,6-diazaspiro[3.3]heptane-2-carb-
oxylate (153 mg, 382 .mu.mol, Eq: 1.00) was combined with methanol
(2 ml) to give an yellow suspension. Sodium cyanamide (26.9 mg, 420
.mu.mol, Eq: 1.1) was added and the reaction was stirred at
25.degree. C. for 1.5 hour under argon. Methyl iodide (65.1 mg,
28.7 .mu.l, 459 .mu.mol, Eq: 1.2) was added and the reaction
mixture was stirred at 25.degree. C. for 17 hours under argon. The
reaction mixture was concentrated in vacuo and used without any
further purification.
[0250] MS +m/z: 457/459. (M+1)
6-[4-(5-Amino-2H-[1,2,4]triazol-3-ylamino)-2,6-dichlorophenyl]-2,6-diaza-s-
piro[3.3]heptane-2-carboxylic acid tert-butyl eater (Compound
12)
##STR00089##
[0252] In a 50 mL round-bottomed flask, tert-butyl
6-(2,6-dichloro-4-(cyanamido(methylthio)methyleneamino)
phenyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (174 mg, 381
.mu.mol, Eq: 1.00) was combined with ethanol (3 ml) to give a
yellow solution. Hydrazine monohydrate (191 mg, 185 .mu.l, 3.81
mmol, Eq: 10) was added and the reaction mixture was heated to
70.degree. C. and stirred for 1.5 hour. The crude reaction mixture
was concentrated in vacuo and the residue was diluted with water
and stirred overnight. The solid was filtered triturated with MeOH.
The suspension was filtered and the filtrate was concentrated. The
crude material was purified by preparative HPLC (0.1% TFA in
water/0.1% TFA in AcCN). 95% to 10% over 25 mins. The clean
fractions were pooled, basified with 3 drops of 1M NaOH and
stripped in vacuo to give a white powder. The powder was filtered
and washed with water to remove salts. The clean product was dried
under vacuum at 45-C overnight to afford 13.5 mg (8%) of the
desired product as a white solid.
[0253] MS +m/z: 440.0/441.9. (M+1)
{3-Aminomethyl-1-[4-(5-amino-2H-[1,2,4]triazol-3-ylamino)-2,6-dichlorophen-
yl]-azetidin-3-yl}-methanol (Compound 13)
##STR00090##
[0255] In a 100 mL round-bottomed flask, tert-butyl
6-(4-(5-amino-1H-[1,2,4]-triazol-3-ylamino)-2,6-dichlorophenyl)-2,6-diaza-
spiro[3.3]heptane-2-carboxylate Compound 11 (21 mg, 47.7 .mu.mol,
Eq: 1.00) was combined with dichloromethane (2 ml) methanol (0.5
ml) and TFA (1.5 ml) to give a pink solution. The reaction was
stirred at 25.degree. C. for 3 days. The reaction was concentrated
in vacuo and the residue was purified by preparative HPLC (0.1% TFA
in water/0.1% TFA in AcCN). 95% to 10% over 16 mins. The colorless
glass was dried under vacuum at 45.degree. C. to afford 2.3 mg
(14%) of the desired product as a white solid.
[0256] MS -m/z: 356/358 (M-1)
[0257]
N3-(3-chloro-4-(pyridazin-3-yl)phenyl)-1H-1,2,4-triazole-3,5-diamin-
e (Compound 14)
##STR00091##
3-Chloro-4-pyridazin-3-yl-phenylamine
##STR00092##
[0259] In a 250 mL round-bottomed flask,
3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline
(950 mg, 3.75 mmol, Eq: 1.00), 3-chloropyridazine (437 mg, 3.82
mmol, Eq: 1.02) and tetrakis(triphenylphosphine)palladium(0) (437
mg, 378 .mu.mol, Eq: 0.101) were combined with toluene (38.0 mL) to
give a brown solution. A 2.0 M aqueous solution of sodium carbonate
(7.6 ml, 15.2 mmol, Eq: 4.06) and ethanol (7.6 mL) were added. The
reaction mixture was stirred at 115.degree. C. for 5 hours.
[0260] The reaction mixture was cooled to room temperature, and it
was stirred at room temperature overnight. The reaction mixture was
partitioned between ethyl acetate (10 mL) and water (10 mL). The
organic phase was dried (Na.sub.2SO.sub.4), filtered, then
concentrated over silica gel. The silica-supported crude product
was loaded onto a 115 gram silica gel column (Analogix). Flash
chromatography (70% ethyl acetate in hexanes) gave
3-chloro-4-pyridazin-3-yl-phenylamine (700 mg, 90%) containing
triphenylphosphine oxide as a minor impurity.
3-(2-Chloro-4-isothiocyanato-phenyl)-pyridazine
##STR00093##
[0262] In a 1 L round-bottomed flask,
3-chloro-4-(pyridazin-3-yl)aniline (0.7 g, 3.4 mmol, Eq: 1.00) and
1,1'-thiocarbonyldiimidazole (700 mg, 3.93 mmol, Eq: 1.15) were
combined with methylene chloride (20 mL) to give a brown solution.
This mixture was stirred at room temperature over 22 hours. The
reaction mixture was concentrated over silical gel. The
silica-supported crude product was loaded onto a 115 g Analogix
column. Flash chromatography (50% ethyl acetate-hexanes) provided
3-(2-chloro-4-isothiocyanato-phenyl)-pyridazine (400 mg, 47%) as a
white crystalline solid.
(Z)-Methyl
N-3-chloro-4-(pyridazin-3-yl)phenyl-N'-cyanocarbamimidothioate
##STR00094##
[0264] In a 50 mL pear-shaped flask, cyanamide (204 mg, 4.84 mmol,
Eq: 3.0) and a 0.5 M solution of sodium methoxide in methanol (4.84
mL, 2.42 mmol, Eq: 1.5) were combined with methanol (6.5 mL) to
give a colorless solution. This mixture was stirred at room
temperature for 15 minutes. After this time, the mixture was added
dropwise to a suspension of
3-(2-chloro-4-isothiocyanatophenyl)pyridazine (400 mg, 1.61 mmol,
Eq: 1.00) and 6.5 mL methanol. Upon addition of the
cyanamide-sodium methoxide mixture, the suspension quickly became a
slightly yellow solution. This mixture was stirred at room
temperature for 1 hour. After this time, iodomethane (344 mg, 151
.mu.l, 2.42 mmol, Eq: 1.5) was added. The reaction mixture was
stirred at room temperature overnight. In the morning, a yellow
precipitate was present. The reaction mixture was filtered, and the
collected solids were washed with a minimal amount of methanol.
After further drying on the buchner funnel, this product was
collected. A second crop was obtained when the mother liquor
yielded a yellow precipitate. The two products were combined to
give (Z)-methyl
N-3-chloro-4-(pyridazin-3-yl)phenyl-N'-cyanocarbamimidothioate (229
mg, 46%) as a yellow powder.
N3-(3-chloro-4-(pyridazin-3-yl)phenyl)-1H-1,2,4-triazole-3,5-diamine
(Compound 14)
##STR00095##
[0266] In a 50 mL round-bottomed flask, (Z)-methyl
N-3-chloro-4-(pyridazin-3-yl)phenyl-N'-cyanocarbamimidothioate (229
mg, 754 .mu.mol, Eq: 1.00) and hydrazine (245 mg, 240 .mu.l, 7.64
mmol, Eq: 10.1) were combined with ethanol (7.5 ml) to give a
yellow suspension. After stirring for about ten minutes, nearly all
of the starting material dissolved to give a yellow solution. The
reaction mixture was heated at 85.degree. C. After only 15 minutes
heating, the reaction mixture was again a yellow suspension. The
reaction was heated at 85.degree. C. for a total of 5 hours. The
reaction mixture was cooled to room temperature, and the
precipitate was collected using vacuum filtration to provide
N3-(3-chloro-4-(pyridazin-3-yl)phenyl)-1H-1,2,4-triazole-3,5-diamine
(153 mg, 71%) as a yellow powder. MS cald. for
C.sub.12H.sub.10ClN.sub.7 [(M+H)]: 288, obsd. 288.0.
N3-(3-Chloro-4-(1-methyl-1H-tetraol-5-yl)phenyl)-1H-1,2,4-triazole-3,5-dia-
mine (Compound 15)
##STR00096##
[0267] 2-Chloro-N-methyl-4-nitro-benzamide
##STR00097##
[0269] In a 500 mL round-bottomed flask, 2-chloro-4-nitrobenzoic
acid (5.0 g, 24.8 mmol, Eq: 1.00) and thionyl chloride (40.8 g, 25
ml, 343 mmol, Eq: 13.8) were combined to give a white suspension.
This mixture was refluxed neat for 4 hours. After this time, the
reaction mixture was concentrated on the rotary evaporator. The
crude product was combined with 45 mL methylene chloride and cooled
to 0.degree. C. using an ice-water bath. Methylamine hydrochloride
(2.01 g, 29.8 mmol) was added followed by N,N-diisopropylethylamine
(8.66 mL, 49.6 mmol). The reaction mixture was stirred and warmed
to room temperature as the ice bath gradually melted. The reaction
mixture was stirred over night for 16 hours. After this time, the
reaction mixture was washed with water then brine. The organic
phase was dried over MgSO.sub.4, filtered, and concentrated to
afford 2-chloro-N-methyl-4-nitro-benzamide (258 mg, 63%) as a
yellow solid. The crude product was used in the next step without
further purification.
5-(2-Chloro-4-nitro-phenyl)-1-methyl-1H-tetrazole
##STR00098##
[0271] In a 500 mL round-bottomed flask,
2-chloro-N-methyl-4-nitro-benzamide (6.31 g, 29.4 mmol) and thionyl
chloride (5.25 g, 3.22 ml, 44.1 mmol, Eq: 1.5) were combined with
toluene (200 mL). The resultant mixture was refluxed overnight. In
the morning, the mixture was cooled to room temperature, and the
mixture was stirred at room temperature over the weekend. The
reaction mixture was concentrated on the rotary evaporator. The
crude residue was then combined with 20 mL additional toluene, and
concentrated again. A mixture of the crude product from above and
40 mL acetonitrile was added to a 0.degree. C. mixture of
azidotrimethylsilane (5.44 ml, 41.2 mmol, Eq: 1.4) in acetonitrile
(80 mL). The reaction mixture was stirred at 0.degree. C. for 2
hours, and then warmed to room temperature. The reaction mixture
was stirred at room temperature overnight. The reaction mixture was
concentrated, and the residue was partitioned between methylene
chloride and water. The organic phase was washed with brine, dried
over MgSO.sub.4, filtered, and concentrated over silica gel. The
silica gel supported crude product was loaded onto a 220 gram
analogix column. Flash chromatography (45% ethyl acetate-hexanes)
provided 5-(2-chloro-4-nitro-phenyl)-1-methyl-1H-tetrazole (4.14 g,
59%) as a yellow, crystalline solid.
3-Chloro-4-(1-methyl-1H-tetraol-5-yl)-phenylamine
##STR00099##
[0273] In a 100 mL round-bottomed flask,
5-(2-chloro-4-nitrophenyl)-1-methyl-1H-tetrazole (0.6 g, 2.5 mmol,
Eq: 1.00), iron (700 mg, 12.5 mmol, Eq: 5.01) and ammonium chloride
(1.34 g, 25.0 mmol, Eq: 10.00) were combined with methanol (6.7 ml)
to give a grey suspension. Water (3.3 mL) was added. The reaction
mixture was heated at 75.degree. C. for 1 hour. The reaction
mixture was cooled to room temperature, and then concentrated. The
crude product was partitioned between saturated aqueous sodium
bicarbonate and ethyl acetate. The organic phase was dried over
Na.sub.2SO.sub.4, filtered, and concentrated to afford
3-chloro-4-(1-methyl-1H-tetrazol-5-yl)-phenylamine (490 mg, 93%) as
a colorless oil. The crude product was taken onto the next step
without additional purification.
5-(2-chloro-4-isothiocyanatophenyl)-1-methyl-1H-tetrazole
##STR00100##
[0275] In a 500 mL round-bottomed flask,
3-chloro-4-(1-methyl-1H-tetrazol-5-yl)aniline (490 mg, 2.34 mmol,
Eq: 1.00) and 1,1'-thiocarbonyldiimidazole (480 mg, 2.69 mmol, Eq:
1.15) were combined with methylene chloride (20 ml) to give a light
brown suspension. The reaction mixture was stirred at room
temperature over the weekend under a balloon of argon. The reaction
mixture was concentrated over silica gel, and the silica gel
supported crude product was loaded onto a 115 gram Analogix column.
Flash chromatography (40% ethyl acetate-hexanes) afforded
5-(2-chloro-4-isothiocyanatophenyl)-1-methyl-1H-tetrazole (334 mg,
57%) as a colorless film.
(Z)-methyl
N-3-chloro-4-(1-methyl-1H-tetrazol-5-yl)phenyl-N'-cyanocarbamim-
idothioate
##STR00101##
[0277] In a 250 mL pear-shaped flask, a 0.5 M solution of sodium
methoxide in methanol (4.0 mL, 2.00 mmol, Eq: 1.51) and cyanamide
(167 mg, 3.98 mmol, Eq: 3.0) were combined with methanol (13 mL) to
give a colorless solution. This mixture was stirred at room
temperature for 15 minutes. This mixture was then added dropwise to
a mixture of
5-(2-chloro-4-isothiocyanatophenyl)-1-methyl-1H-tetrazole (334 mg,
1.33 mmol, Eq: 1.00) and 13 mL methanol. The resulting mixture was
stirred at room temperature for one hour. After this time, methyl
iodide (284 mg, 125 .mu.l, 2.00 mmol, Eq: 1.51) was added. The
reaction mixture was stirred at room temperature overnight. In the
morning, a white precipitate had formed. This product was collected
using vacuum filtration, furnishing (Z)-methyl
N-3-chloro-4-(1-methyl-1H-tetrazol-5-yl)phenyl-N'-cyanocarbamimidothioate
(258 mg, 63%) as a white powder.
N3-(3-Chloro-4-(1-methyl-1H-tetrazol-5-yl)phenyl)-1H-1,2,4-triazole-3,5-di-
amine (Compound 15)
##STR00102##
[0279] In a 100 mL round-bottomed flask, (Z)-methyl
N-3-chloro-4-(1-methyl-1H-tetrazol-5-yl)phenyl-N'-cyanocarbamimidothioate
(258 mg, 838 .mu.mol, Eq: 1.00) and hydrazine (265 .mu.L, 8.43
mmol, Eq: 10.1) were combined with ethanol (8.5 mL). The reaction
mixture was refluxed for three hours. After this time, the reaction
mixture was cooled to room temperature, then stirred at room
temperature overnight. In the morning, a white precipitate came out
of solution, giving a thick white suspension. The precipitate was
collected via vacuum filtration, giving
N3-(3-chloro-4-(1-methyl-1H-tetrazol-5-yl)phenyl)-1H-1,2,4-triazol-
e-3,5-diamine. MS cald. for C.sub.10H.sub.10ClN.sub.9 [(M+H)]: 292,
obsd. 292.0.
N3-(3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)phenyl)-1H-1,2,4-triazole-3-
,5 diamine (Compound 16)
##STR00103##
[0280]
2-Chloro-N-[1-dimethylamino-meth-(Z)-ylidene]-4-nitro-benzamide
##STR00104##
[0282] In a 500 mL round-bottom flask, 2-chloro-4-nitrobenzamide
(3.5 g, 17.4 mmol, Eq: 1.00) and N,N-dimethylformamide dimethyl
acetal (23 mL, 173 mmol, Eq: 9.92) were combined to give an
off-white suspension. This mixture was heated at 130.degree. C.
overnight. In the morning, the solvent was evaporated off, giving
2-chloro-N-[1-dimethylamino-meth-(Z)-ylidene]-4-nitro-benzamide as
a yellow solid. This crude product was used in subsequent steps
without further purification.
5-(2-chloro-4-nitrophenyl)-1-methyl-1H-1,2,4-triazole
##STR00105##
[0284] In a 500 mL round-bottomed flask,
(Z)-2-chloro-N-((dimethylamino)methylene)-4-nitrobenzamide (2.0 g,
7.82 mmol, Eq: 1.00) and methylhydrazine (5.35 mL, 102 mmol, Eq:
13) were combined with glacial acetic acid (90 mL) to give an
off-white suspension. The reaction mixture was heated at 90.degree.
C. for 1.5 hours. After this time, the reaction mixture was cooled
to about 50.degree. C., and subsequently concentrated on the rotary
evaporator. The crude product was partitioned between ethyl acetate
and saturated aqueous NaHCO.sub.3. The organic phase was dried over
MgSO.sub.4, filtered, and concentrated. The crude product was
purified using flash chromatography (120 gram Analogix column,
15-35% ethyl acetate-hexanes) to provide
5-(2-chloro-4-nitrophenyl)-1-methyl-1H-1,2,4-triazole (481 ag, 26%)
as a yellow solid.
3-Chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)aniline
##STR00106##
[0286] In a 250 mL round-bottomed flask,
5-(2-chloro-4-nitrophenyl)-1-methyl-1H-1,2,4-triazole (481 mg, 2.02
mmol, Eq: 1.00), iron (113 mg, 2.02 mmol, Eq: 1.00) and ammonium
chloride (108 mg, 2.02 mmol; Eq: 1.00) were combined with methanol
(5.4 mL) to give a brown suspension. Water (2.7 mL) was added. The
reaction mixture was heated at 85.degree. C. After only 15 minutes,
the reaction mixture had already changed to a rust-colored
suspension. The reaction mixture was stirred at 85.degree. C. for
three hours. After this time, TLC indicated that some starting
material still remained. The reaction mixture was cooled to room
temperature. Additional iron (108 mg iron, 1.93 mmol) and ammonium
chloride (113 mg, 2.11 mmol) were added. The reaction mixture was
heated for another 1 hour, and then cooled to room temperature. The
reaction mixture was stirred at room temperature overnight. After
this time, TLC indicated that the reaction was complete. The
mixture was concentrated, and the residue was partitioned between
ethyl acetate and saturated aqueous sodium bicarbonate. The organic
phase was dried over Na.sub.2SO.sub.4, filtered, and concentrated
to afford 3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)aniline as a
yellow oil.
5-(2-Chloro-4-isothiocyanatophenyl)-1-methyl-1H-1,2,4-triazole
##STR00107##
[0288] In a 1 L round-bottom flask,
3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)aniline (421 mg, 2.02
mmol, Eq: 1.00) and 1,1'-thiocarbonyldiimidazole (414 mg, 2.32
mmol, Eq: 1.15) were combined with methylene chloride (20.2 mL) to
give a brown suspension. This reaction mixture was stirred at room
temperature over the weekend. The reaction mixture was concentrated
over silica gel. The silica-gel supported crude product was loaded
onto a 120 gram silica gel column. Flash chromatography (35% ethyl
acetate-hexanes) gave
5-(2-chloro-4-isothiocyanatophenyl)-1-methyl-1H-1,2,4-triazole (280
mg, 55%).
(Z)-Methyl
N-3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)phenyl-N'-cyanocar-
bamimidothioate
##STR00108##
[0290] In a 10 mL pear-shaped flask, a 0.5 M solution of sodium
methoxide in methanol (840 L, 420 .mu.mol, Eq: 1.5) and cyanamide
(35.0 mg, 833 .mu.mol, Eq: 2.98) were combined to give a colorless
solution. This mixture was stirred at room temperature for 15
minutes. After this time, the mixture was added to a solution of 11
mL methanol and
5-(2-chloro-4-isothiocyanatophenyl)-1-methyl-1H-1,2,4-triazole (70
mg, 279 .mu.mol, Eq: 1.00). The reaction mixture was stirred at
room temperature for 1 hour. Methyl iodide (104 .mu.L, 1.68 mmol)
was added, and the reaction mixture was stirred at room temperature
overnight. In the morning, the reaction mixture was a clear,
colorless solution. The solvent was evaporated without heating so
that there was 1 mL of methanol remaining. This mixture was diluted
with methylene chloride (10 mL). Upon addition of methylene
chloride, the reaction mixture became cloudy. The diluted reaction
mixture was concentrated over silica gel with minimal heating. The
silica gel supported crude product was loaded onto a 25 gram silica
gel column. Flash chromatography (80%-100% ethyl acetate-hexanes)
provided (Z)-methyl
N-3-chloro-4-(1-methyl-1H-1,2,4-triazole-5-yl)phenyl-N'-cyanocarbamimidot-
hioate (28 mg, 33%) as a white solid.
N3-(3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)phenyl)-1H-1,2,4-triazole-3-
,5-diamine (Compound 16)
##STR00109##
[0292] In a 10 mL round-bottom flask, (Z)-methyl
N-3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)phenyl-N'-cyano-carbamimidot-
hioate (28 mg, 91.3 i.mu.mol, Eq: 1.00) and hydrazine (30.6 mg, 30
.mu.L, 955 .mu.mol, Eq: 10.5) were combined with ethanol (1 mL) to
give a colorless solution. The reaction mixture was heated at
75.degree. C. for hours. The reaction mixture was concentrated to
afford
N3-(3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)phenyl)-1H-1,2,4-triazole--
3,5-diamine (23 mg, 87%) as a white powder. MS cald. for
C.sub.11H.sub.11ClN.sub.8 [(M+H)]: 291, obsd. 291.0.
N3-(3,5-dichloro-4-(1H-pyrazol-1-yl)phenyl)-1H-1,2,4-triazole-3,5-diamine
(Compound 17)
##STR00110##
[0293] 1-(2,6-Dichloro-4-nitro-phenyl)-1H-pyrazole
##STR00111##
[0295] In a 250 mL round-bottom flask,
1,3-dichloro-2-fluoro-5-nitrobenzene (1.5 g, 7.14 mmol, Eq: 1.00),
1H-pyrazole (486 mg, 7.14 mmol, Eq: 1.00) and potassium carbonate
(1.5 g, 10.9 mmol, Eq: 1.52) were combined with DMF (30 mL) to give
a yellow suspension. This mixture was heated at 115.degree. C. for
three hours. The reaction mixture was cooled to room temperature.
The cooled mixture was poured into .about.100 mL of ice water. A
cream-colored solid precipitated out. The precipitate was collected
via vacuum filtration and air dried to provide
1-(2,6-dichloro-4-nitro-phenyl)-1H-pyrazole (1.73 g, 94%) as an
off-white solid.
3,5-Dichloro-4-pyrazol-1-yl-phenylamine
##STR00112##
[0297] In a 250 mL round-bottom flask,
1-(2,6-dichloro-4-nitrophenyl)-1H-pyrazole (1.73 g, 6.7 mmol, Eq:
1.00), iron (1.80 g, 32.2 mmol, Eq: 4.81) and ammonium chloride
(3.6 g, 67.3 mmol, Eq: 10.0) were combined with methanol (18 mL) to
give a grey suspension. Water (9 mL) was added. The reaction
mixture was heated at 85.degree. C. for 6 hours. The reaction
mixture was cooled to room temperature, and then partitioned
between ethyl acetate and saturated aqueous ammonium chloride. The
organic phase was dried (Na.sub.2SO.sub.4), filtered, and finally
concentrated to afford 3,5-dichloro-4-pyrazol-1-yl-phenylamine
(1.29 g, 84%) as a beige solid.
1-(2,6-Dichloro-4-isothiocyanato-phenyl)-1H-pyrazole
##STR00113##
[0299] In a 1 L round-bottomed flask,
3,5-dichloro-4-(1H-pyrazol-1-yl)aniline (1.5 g, 6.58 mmol, Eq:
1.00) and 1,1'-Thiodicarbonylimidazole (1.30 g, 7.29 mmol, Eq:
1.11) were combined with methylene chloride (50 mL) to give a light
yellow suspension. The reaction mixture was stirred at room
temperature for 24 hours. The reaction mixture was concentrated
over silica gel, and the silica gel supported crude product was
loaded onto a 120 gram silica gel column. Flash chromatography (15%
ethyl acetate-hexanes) afforded
1-(2,6-dichloro-4-isothiocyanato-phenyl)-1H-pyrazole (886 mg, 50%)
as a light yellow oil.
(Z)-Methyl
N-3-chloro-4-(1-methyl-1H-pyrazol-1-yl)-phenyl-N'-cyanocarbamim-
idothioate
##STR00114##
[0301] In a 100 mL round-bottomed flask, cyanamide (75 mg, 1.78
mmol, Eq: 3.01) and a 0.5 M solution of sodium methoxide in
methanol (1.77 mL, 886 .mu.mol, Eq: 1.5) were combined to give a
colorless solution. This mixture was stirred at room temperature
for 15 minutes. After this time, the cyanamide mixture was added
rapidly dropwise to a solution of
1-(2,6-dichloro-4-isothiocyanatophenyl)-1H-pyrazole (160 mg, 592
.mu.mol, Eq: 1.00) in methanol (10 mL). This mixture was stirred at
room temperature for 1 hour. Finally, iodomethane (126 mg, 55.6
.mu.L, 888 .mu.mol, Eq: 1.5) was added. The reaction mixture was
diluted with 10 mL of methylene chloride and concentrated over
celite. The celite-supported crude product was loaded onto a 40
gram silica gel column. Flash chromatography (15%-75% ethyl
acetate-hexanes) provided (Z)-methyl
N-3-chloro-4-(1-methyl-1H-pyrazol-1-yl)-phenyl-N'-cyanocarbamimidothioate
(62 mg, 32%) as a white solid.
N3-(3,5-dichloro-4-(1H-pyrazol-1-yl)phen
##STR00115##
[0303] In a 25 mL round-bottomed flask,
(Z)-meth:N'-cyanocarbamimidothioate (62 mg, 190 .mu.mmol, Eq: 10.1)
were combined with ethanwas refluxed for 16 hours. The reaction mi
ethanol-methylene chloride solution. This mixture was filtered and
The product was further dried down in the vacuum oven at
.about.75.degree. C. to give
N3-(3,5-dichloro-4-(1H-pyrazol-1-yl)phenyl)-1H-1,2,4-triazole-3,5-
-diamine (58 mg, 99%) as a light brown powder.
[0304] MS cald. for C.sub.11H.sub.9Cl.sub.2N.sub.7 [(M+H)]: 311,
obsd. 311.8.
tert-Butyl
4-(4-(5-amino-1H-1,2,4-triazol-3-ylamino)-2,6-dichlorophenyl)-5-
,6-dihydropyridine-1(2H)-carboxylate (Compound 18)
##STR00116##
[0305]
4-(4-Amino-2,6-dichloro-phenyl)-3,6-dihydro-2H-pyridine-1-carboxyli-
c acid tertbutyl ester
##STR00117##
[0307] In a sealed tube,
1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-ylboronic acid
(450 mg, 1.98 mmol, Eq: 0.955),
[1,1-bis(diphenylphosphino)ferrocene]dichlorpalladium(II) (150 mg,
205 .mu.mol, Eq: 0.0988) and 4-bromo-3,5-dichloroaniline (0.5 g,
2.08 mmol, Eq: 1.00) were combined with DMF (7 mL) to give a dark
red suspension. Potassium carbonate (860 mg, 6.22 mmol, Eq: 3.00)
was added. The reaction mixture was heated at 115.degree. C.
overnight. The reaction mixture was combined with water, then the
dark suspension was extracted with ethyl acetate. The organic phase
was dried (MgSO.sub.4), filtered, and concentrated to a black oil.
This product was dissolved in CH.sub.2Cl.sub.2 and the mixture was
concentrated over silica gel. The silica gel supported crude
product was loaded onto an 80 gram silica gel column. Flash
chromatography (5%-20% ethyl acetate-hexanes) provided
4-(4-amino-2,6-dichloro-phenyl)-3,6-dihydro-2H-pyridine-1-carboxylic
acid tert-butyl ester (0.267 g, 38%) as a light yellow oil.
4-(2,6-Dichloro-4-isothiocyanato-phenyl)-3,6-dihydro-2H-pyridine-carboxyli-
c acid tert-butyl ester
##STR00118##
[0309] In a 100 mL round-bottomed flask, tert-butyl
4-(4-amino-2,6-dichlorophenyl)-5,6-dihydropyridine-1(2H)-carboxylate
(0.229 g, 667 .mu.mol, Eq: 1.00) and 1,1'-thiocarbonyldiimidazole
(140 mg, 786 .mu.mol, Eq: 1.18) were combined with methylene
chloride (4 mL) to give a brown suspension. The reaction mixture
was stirred at room temperature over the weekend. The reaction was
concentrated over silica gel. The silica gel supported crude
product was loaded onto a 80 gram SiliCycle column. Flash
chromatography afforded tert-butyl
4-(4-(5-amino-1H-1,2,4-triazol-3-ylamino)-2,6-dichlorophenyl)-5,6-dihydro-
pyridine-1(2H)-carboxylate (216 mg, 84%) as a yellow oil.
(Z)-tert-Butyl
4-(2,6-dichloro-4-(cyanamido(methylthio)methylene-amino)phenyl)-5,6-dihyd-
ropyridine-1(2H)-carboxylate
##STR00119##
[0311] In a 100 mL round-bottomed flask, cyanamide (70 mg, 1.67
mmol, Eq: 2.98) and a 0.5 M solution of sodium methoxide in
methanol (1.7 mL, 850 .mu.mol, Eq: 1.52) were combined to give a
colorless solution. This mixture was stirred at room temperature
over 15 minutes. After this time, a solution of tert-butyl
4-(2,6-dichloro-4-isothiocyanatophenyl)-5,6-dihydropyridine-1(2H)-carboxy-
late (0.215 g, 558 .mu.mol, Eq: 1.00) in methanol (5.6 mL) was
added. The reaction mixture was stirred at room temperature for 1
hour. Finally, methyl iodide (118 mg, 52.0 .mu.L, 832 .mu.mol, Eq:
1.49) was added and the reaction mixture was stirred at room
temperature overnight. The reaction mixture was concentrated
directly over celite. The celite-supported crude product was loaded
onto a 40 gram SiliCycle column. Flash chromatography (30%-65%
ethyl acetate-hexanes) afforded (Z)-tert-butyl
4-(2,6-dichloro-4-(cyanamido(methylthio)methylene-amino)phenyl)-5,6-dihyd-
ropyridine-1(2H)-carboxylate (33 mg, 13%) as a colorless oil.
tert-Butyl
4-(4-(5-amino-1H-1,2,4-triazol-3-ylamino)-2,6-dichlorophenyl)-5-
,6-dihydropyridine-1(2H)-carboxylate (Compound 18)
##STR00120##
[0313] In a 50 mL pear-shaped flask, (Z)-tert-butyl
4-(2,6-dichloro-4-(cyanamido(methylthio)methylene-amino)phenyl)-5,6-dihyd-
ropyridine-1 (2H)-carboxylate (33 mg, 74.8 .mu.mol, Eq: 1.00) and
hydrazine (24.5 mg, 24 .mu.L, 765 .mu.mol, Eq: 10.2) were combined
with ethanol (1 mL) to give a colorless solution. The reaction
mixture was heated at 75.degree. C. for 2.5 hours. After this time,
LCMS showed the reaction to be complete. The reaction mixture was
concentrated to afford an oil. This product was re-dissolved in
CDCl.sub.3 and CD.sub.3OD. Solvents were evaporated again to afford
tert-butyl
4-(4-(5-amino-1H-1,2,4-triazol-3-ylamino)-2,6-dichlorophenyl)-5,6-dihydro-
pyridine-1(2H)-carboxylate (22 mg, 69%) as an off-white oily
solid.
[0314] MS [(M+H).sup.+] obsd. 425.0.
N3-(3,5-dichloro-4-(1,2,3,6-tetrahydropyridin-4-yl)phenyl)-1H-1,2,4-triazo-
le-3,5-diamine (Compound 19)
##STR00121##
[0316] In a 5 mL vial, tert-butyl
4-(4-(5-amino-1H-1,2,4-triazol-3-ylamino)-2,6-dichlorophenyl)-5,6-dihydro-
pyridine-1(2H)-carboxylate Compound 17 (16 mg, 37.6 mol, Eq: 1.00)
and 4.0 M HCl in dioxane (250 .mu.L, 1.00 mmol, Eq: 26.6) were
combined with methylene chloride (250 .mu.L) to give a light brown
suspension. The reaction mixture was stirred at room temperature
over 3 hours. The mixture was then partioned between ethyl acetate
and saturated aqueous NaHCO.sub.3.
[0317] The organic phase was dried (Na.sub.2SO.sub.4), filtered and
concentrated to afford
N3-(3,5-dichloro-4-(1,2,3,6-tetrahydropyridin-4-yl)phenyl)-1H-1,2,4-triaz-
ole-3,5-diamine as a colorless oil. MS [(M-H).sup.-] obsd.
323.1.
tert-Butyl
4-(4-(5-amino-1H-1,2,4-triazol-3-ylamino)-2-chloro-6-(trifluoro-
methyl)phenyl)-5,6-dihydropyridine-1(2H)-carboxylate (Compound
20)
##STR00122##
[0318] tert-Butyl
4-(4-(4-amino-2-chloro-6-(trifluoromethyl)phenyl)-5,6-dihydropyridine-1(2-
H)-carboxylate
##STR00123##
[0320] In a 250 mL round-bottomed flask,
4-bromo-3-chloro-5-(trifluoromethyl)aniline (2.55 g, 9.29 mmol, Eq:
1.00), tert-butyl
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-
-carboxylate (2.76 g, 8.92 mmol, Eq: 0.96) and potassium carbonate
(3.85 g, 27.9 mmol, Eq: 3.0) were combined with DMF (30 mL).
1,1'-Bis(diphenylphosphino)ferrocene-palladium(I) dichloromethane
complex (1.14 g, 1.39 mmol, Eq: 0.15) was added. The reaction
mixture was stirred overnight at 125.degree. C. The reaction
mixture was cooled to room temperature and poured into water. The
resulting suspension was extracted with ethyl acetate. The organic
phase was dried over MgSO.sub.4, filtered, and concentrated over
silica gel. The silica gel supported crude product was split into
two lots. Each lot was purified separately on a 120 gram SiliCycle
column using flash chromatography (15%-40% ethyl acetate-hexanes)
to afford tert-butyl
4-(4-amino-2-chloro-6-(trifluoromethyl)phenyl)-5,6-dihydropyridine-1(2H)--
carboxylate (2.23 g, 64%) as a light yellow oil.
tert-Butyl
4-(2-chloro-4-isothiocynato-6-(trifluoromethyl)phenyl)-5,6-dihy-
dropyridine-1(2H)-carboxylate
##STR00124##
[0322] In a 1 L round-bottomed flask, tert-butyl
4-(4-amino-2-chloro-6-(trifluoromethyl)phenyl)-5,6-dihydropyridine-1(2H)--
carboxylate (2.23 g, 5.92 mmol, Eq: 1.00) and
di(1H-imidazol-1-yl)methanethione (1.2 g, 6.73 mmol, Eq: 1.14) were
combined with methylene chloride (60 mL) to give a light yellow
solution. The reaction mixture was stirred overnight at room
temperature. The reaction mixture was concentrated over silica gel.
The silica gel supported crude product was loaded onto a 120 gram
silica gel column. Flash chromatography (10%-20% ethyl
acetate-hexanes) provided tert-butyl
4-(2-chloro-4-isothiocyanato-6-(trifluoromethyl)phenyl)-5,6-dihydropyridi-
ne-1(2H)-carboxylate (1.67 g, 67%) as a colorless oil.
(Z)-tert-Butyl
4-(2-chloro-4-(cyanamido(methylthio)-methyleneamino)-6-(trifluoromethyl)p-
henyl)-5,6-dihydropyridine-1(2H)-carboxylate
##STR00125##
[0324] In a 250 mL round-bottomed flask, a 0.5 M solution of sodium
methoxide in methanol (7.16 mL, 3.58 mmol, Eq: 1.5) and cyanamide
(301 mg, 7.16 mmol, Eq: 3.0) were combined to give a colorless
solution. The mixture was stirred at room temperature for 20
minutes. A solution of tert-butyl
4-(2-chloro-4-isothiocyanato-6-(trifluoromethyl)phenyl)-5,6-dihydropyridi-
ne-1(2H)-carboxylate (1.0 g, 2.39 mmol) in 18 mL of methanol was
added dropwise via a syringe. The reaction mixture was stirred at
room temperature over 1.5 hours. After this time, methyl iodide
(224 .mu.L, 3.58 mmol, Eq: 1.5) was added. The reaction mixture was
stirred at room temperature overnight. The reaction mixture was
concentrated over celite.
[0325] The celite supported crude product was loaded onto a 80 gram
silica gel column. Flash chromatography (15%-50% ethyl
acetate-hexanes gave (Z)-tert-butyl
4-(2-chloro-4-(cyanamido-(methylthio)methyleneamino)-6-(trifluoremethyl)p-
henyl)-5,6-dihydropyridine-1(2H)-carboxylate (0.776 g, 68%) as a
light yellow oil.
tert-Butyl
4-(4-(5-amino-1H-1,2,4-triazol-3-ylamino)-2,6-dichlorophenyl)-5-
,6-dihydropyridine-1(2H)-carboxylate (Compound 20)
##STR00126##
[0327] In a 250 mL round-bottomed flask, (Z)-tert-butyl
4-(2-chloro-4-(cyanamido(methylthio)-methyleneamino)-6-(trifluoromethyl)p-
henyl)-5,6-dihydropyridine-1(2H)carboxylate (0.776 g, 1.63 mmol,
Eq: 1.00) and hydrazine (524 mg, 513 .mu.L, 16.3 mmol, Eq: 10) were
combined with ethanol (16 mL) to give a colorless solution. The
reaction mixture was refluxed for 4 hours, then cooled to room
temperature and stirred for another 3 hours. After this time, LC-MS
analysis showed a small amount of starting material and a possible
intermediate. Heating was continued for another 14 hours. After
this time the reaction was cooled to room temperature and
concentrate to a pink oil. This crude product was precipitated from
10 mL of 30% ethanol-methylene chloride. This first crop of
precipitate was isolated by filtration. The mother liquor from
above eventually solidified over the weekend. These solids were
triturated with cold ethanol, and then the slurry was filtered.
Collected solids were washed with cold ethanol and then dried using
vacuum filtration to provide a second crop. The first and second
crops were combined to afford tert-butyl
4-(4-(5-amino-1H-1,2,4-triazol-3-ylamino)-2-chloro-6-(trifluoromethyl)phe-
nyl)-5,6-dihydropyridine-1(2H)-carboxylate (183 mg, 24%) as an
off-white solid. MS cald. for
C.sub.19H.sub.22ClF.sub.3N.sub.6O.sub.2 [(M+H).sup.+]: 459, obsd.
459.0.
N3-[3-Chloro-4-(1,2,3,6-tetrahydro-pyridin-4-yl)-5-trifluoroethyl-phenyl]--
1H-[1,2,4]triazole-3,5-diamine hydrochloride (Compound 21)
##STR00127##
[0329] In a 20 mL pear-shaped flask, tert-butyl
4-(4-(5-amino-1H-1,2,4-triazol-3-ylamino)-2-chloro-6-(trifluoromethyl)phe-
nyl)-5,6-dihydropyridine-1(2H)-carboxylate Compound 19 (20 mg, 43.6
.mu.mol, Eq: 1.00) and a 4 M solution of hydrochloric acid in
dioxane (500 .mu.L, 2.00 mmol, Eq: 45.9) were combined with dioxane
(500 .mu.L) to give a colorless solution. After only a few minutes,
oily solids precipitated at the bottom of the reaction flask. The
reaction mixture was stirred at room temperature for 5 hours. After
this time, the reaction mixture was allowed to settle without
stirring. The liquids were decanted, giving only the oily solid in
the flask. This product was dissolved in methanol and then
transferred to a vial and concentrated to afford
N3-[3-chloro-4-(1,2,3,6-tetrahydro-pyridin-4-yl)-5-trifluoromethyl-
-phenyl]-1H-[1,2,4]triazole-3,5-diamine hydrochloride. MS cald. for
C.sub.14H.sub.14ClF.sub.3N.sub.6 [(M+H).sup.+]: 359, obsd.
359.0.
N*3*-(3,5-Dichloro-4-[1,2,4]triazolo[4,3-a]pyridin-3-yl-phenyl)-1H-[1,2,4]-
triazole-3,5-diamine (Compound 22)
##STR00128##
[0330]
4-([1,2,4]triazolo[4,3a]pyridin-3-yl)-3,5-dichloroaniline
##STR00129##
[0332] A solution of 2,6-dichloro-4-nitro-benzaldehyde (105 mg,
0.48 mmol) and 2-hydrazinylpyridine (57 mg, 0.52 mmol) in
CH.sub.3OH was heated to 50.degree. C. for 3 hours after which all
volatiles were removed under reduced pressure. The resulting solid
was suspended in THF (5 mL) and chloramine-T (164 mg, 0.67 mmol)
was added to yield a deep burgundy colored reaction mixture which
was heated to 50.degree. C. for one hour. All volatiles were again
removed and the desired product,
3-(2,6-dichloro-4-nitrophenyl)-[1,2,4]triazolo[4,3-a]pyridine (120
mg, 81%) was isolated as an orange solid by silica gel
chromatography (EtOAc).
[0333] A solution of
3-(2,6-dichloro-4-nitrophenyl)-[1,2,4]triazolo[4,3-a]pyridine (105
mg, 0.34 mmol) in a 1:1 mixture of THF:satd NH.sub.4Cl (4 mL) was
treated with zinc dust (67 mg; 1.0 mmol) and the reaction mixture
was stirred for 2 hours. Filtration followed by removal of
volatiles under reduced pressure gave the crude reaction product.
4-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3,5-dichloroaniline (26 mg,
27%) was isolated as a yellow oil by silica gel chromatography
(EtOAc).
N*3*-(3,5-Dichloro-4-[1,2,4]triazolo[4,3-a]pyridin-3-yl-phenyl)-1H-[1,2,4]-
triazole-3,5-diamine (Compound 22)
##STR00130##
[0335] 4-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3,5-dichloroaniline
(58 mg, 0.28 mmol) and thiocarbonyldiimidazole (58 mg, 0.21 mmol)
were stirred overnight in a solution of CH.sub.2C.sub.2(2 mL) after
which the product,
3-(2,6-dichloro-4-isothiocyanatophenyl)-[1,2,4]triazolo[4,3-a]pyridine,
was isolated directly by column chromatography (66% EtOAc in
hexane) as a white solid (37 mg, 55%).
[0336] Sodium hydrogencyanamide (8.4 mg, 0.13 mmol) was added to a
solution of
3-(2,6-dichloro-4-isothiocyanatophenyl)-[1,2,4]triazolo[4,3-a]pyridine
(35 mg, 0.11 mmol) in 1:1 CH.sub.3OH:CH.sub.3CN (2 mL). The
reaction mixture was stirred for 3 hours after which no starting
material was detected by TIC. Iodomethane (13.6 .mu.L; 0.22 mmol)
was then added and the reaction mixture was stirred overnight. All
volatiles were removed under reduced pressure to yield a waxy solid
from which (Z)-methyl
N-4-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3,5-dichlorophenyl-N'-cyanocarba-
mimidothioate (29 mg; 71%) was purified by silica gel
chromatography (AcOEt)
[0337] (Z)-Methyl
N-4-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-3,5-dichlorophenyl-N'-cyanocarba-
mimidothioate (26 mg, 0.069 mmol) and hydrazine (10.8 L, 0.34 mmol)
were heated in ethanol for 3 hours after which all volatiles were
removed under reduced pressure. From the crude product,
N*3*-(3,5-dichloro-4-[1,2,4]triazolo[4,3-a]pyridin-3-yl-phenyl)-1H-[1,2,4-
]triazole-3,5-diamine was isolated as a white solid by column
chromatography (21 mg, 84%). MH+=361.0
N*3*-{3-Chloro-4-[6-(propane-2-sulfonyl)-pyridin-3-yl]-5-trifluoromethyl-p-
henyl}-1H-[1,2,4]triazole-3,5-diamine (Compound 23)
##STR00131##
[0338] 5-Bromo-2-isopropylsulfanyl-pyridine
##STR00132##
[0340] To a solution of 5-bromo-2-chloropyridine (1.03 g, 5.35
mmol, Eq: 1) in dimethylformamide (20 ml) was added sodium
2-propanethiolate (3 g, 30.6 mmol, Eq: 5.71) and the resulting
mixture stirred at room temperature for 1H. The resulting mixture
was poured into water (20 ml) and extracted with methylene chloride
(3.times.50 ml). The combined organic phases were washed with brine
(1.times.100 ml) and dried over magnesium sulfate. The mixture was
filtered and evaporated to give crude
5-bromo-2-isopropylsulfanyl-pyridine as a yellow oil.
5-Bromo-2-(propane-2-sulfonyl)-pyridine
##STR00133##
[0342] To a solution of crude 5-bromo-2-isopropylsulfanyl-pyridine
(1.24 g, 5.34 mmol, Eq: 1) in dichloromethane (10 ml) was added
3-chloroperoxybenzoic acid (4.61 g, 13.4 mmol, Eq: 2.5) in 5
portions over 15 minutes. The resulting mixture was stirred at room
temperature for 30 minutes before being quenched with a 1N sodium
hydroxide solution (25 ml). The resulting mixture was extracted
with dichloromethane (3.times.50 ml) and the combined organic
phases washed with water (100 ml) and brine (100 ml) and dried over
magnesium sulfate. The mixture was filtered and evaporated to give
a white solid which was purified by flash chromatography (Analogix
Intelliflash 310, Analogix SF15-12 g column, 0-20% ethyl
acetatehexane). Like fractions were combined and evaporated to give
5-bromo-2-(propane-2-sulfonyl)-pyridine as a white solid (995 mg,
71%).
2-(Propane-2-sulfonyl)-5-(4,4,55-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyr-
idine
##STR00134##
[0344] To a solution of 5-bromo-2-(propane-2-sulfonyl)-pyridine
(995 mg, 3.77 mmol, Eq: 1),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (2.96
g, 11.6 mmol, Eq: 3.09), and potassium acetate (1.69 g, 17.3 mmol,
Eq: 4.58) in dioxane (15 mL) was added
1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex (295 mg, 403 .mu.mol, Eq: 0.107). The
resulting mixture was heated to 85.degree. C. overnight under an
argon atmosphere. The resulting mixture was cooled to room
temperature, diluted with ethyl acetate (50 ml) and evaporated. The
residue was taken up in ethyl acetate (100 ml), washed with brine
(100 ml) and dried over sodium sulfate. The mixture was filtered
and to the filtrate was added silica gel (.about.3 g). The mixture
was evaporated and purified by flash chromatography (Analogix
Intelliflash, VersaPak Spherical Silica column 20-45 .mu.M, 23 g,
10-50% ethyl acetate/hexane). Like fractions were combined and
evaporated to give crude
2-(propane-2-sulfonyl)-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-p-
yridine as a pale yellow solid (1.1221 g, 54% pure by HNMR,
52%).
N*3*-{3-Chloro-4-[6-(propane-2-sulfonyl)-pyridin-3-yl]-5-trifluoromethyl-p-
henyl}-1H-[1,2,4]triazole-3,5-diamine (Compound 23)
##STR00135##
[0346] A mixture of
N3-(4-bromo-3-chloro-5-(trifluoromethyl)phenyl)-1H-1,2,4-triazole-3,5-dia-
mine Intermediate 1 (407 mg, 1.14 mmol, Eq: 1.08),
2-(isopropylsulfonyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyri-
dine (607 mg, 1.05 mmol, Eq: 1.00) a 3M potassium carbonate
solution (702 .mu.l, 2.11 mmol, Eq: 2), dimethoxyethane (2 ml)
p-dioxane (2 ml) and tetrakis(triphenylphosphine)palladium(0) (277
mg, 240 .mu.mol, Eq: 0.228) was placed in a microwave reaction tube
and degassed with argon under sonication for 15 minutes. The tube
was sealed and the resulting mixture was heated in a microwave
reactor to 128.degree. C. and held at that temperature for 3 h. The
reaction mixture was cooled and diluted with ethyl acetate (2 mL)
and water (1 mL) and the phases separated. The aqueous phase was
extracted with ethyl acetate (3.times.1 mL) and the combined
organic phases were washed with brine and dried over sodium
sulfate. The resulting mixture was filtered and evaporated to give
a brown residue which was chromatographed on silica (Analogix
Intelliflash 310, RediSep silica 24 g, 4-10%
methanol/dichloromethane gradient). Like fractions were combined
and evaporated and the residue purified by SFC chromatography
(THAR/SFC/Waters Multigram II system eluted with 25%
methanol/carbon dioxide at 70 ml/mn on a CYANO column 3.times.25 cm
with detection at 220 nM and 35 degrees and 100 bar backpressure)
to give
N*3*-{3-chloro-4-[6-(propane-2-sulfonyl)-pyridin-3-yl]-5-trifluoromethyl--
phenyl}-1H-[1,2,4]triazole-3,5-diamine as a light yellow solid (125
mg, 26%). MS +m/z: 460.8 (M+H).sup.+
5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-6-trifluoromethyl-phen-
yl]-pyridine-2-sulfonic acid tert-butylamide (Compound 24)
##STR00136##
[0347] 2-(benzylthio)-5-bromopyridine
##STR00137##
[0349] To a solution of benzyl mercaptan (5.29 g, 5.00 ml, 42.6
mmol, Eq: 1.1), in dry tetrahydrofuran (75 ml) was added sodium
hydride (1.41 g, 55.7 mmol, Eq: 1.44) in 3 portions over 30
minutes. The resulting white slurry was allowed to stir at room
temperature for 30 minutes before 25-dibromopyridine (9.17 g, 38.7
mmol, Eq: 1) was added in one portion. The resulting mixture was
allowed to stir at room temperature for 3 hours. The resulting
mixture was quenched with water (75 ml) and extracted with ether
(3.times.100 mL). The ether layers were combined, washed with a
saturated sodium bicarbonate solution (250 ml) and dried over
magnesium sulfate. The mixture was filtered and evaporated to give
2-(benzylthio)-5-bromopyridine as a yellow oil (10.68 g).
5-Bromo-pyridine-2-sulfonic acid tert-butylamide
##STR00138##
[0351] A mixture of crude 2-(benzylthio)-5-bromopyridine (2.94 g,
10.5 mmol, Eq: 1.00), water (30.0 ml), and carbon tetrachloride
(125 ml) was cooled to 0.degree. C. in an ice bath and vigorously
stirred while chlorine gas was bubbled slowly through the mixture.
After 3 minutes the reaction mixture was saturated with chlorine
and turned bright yellow green. Bubbling was continued for 10
minutes followed by 5 additional minutes of stirring under a
chlorine blanket before the mixture was sparged of chlorine by
bubbling argon into the solution for 10 minutes. The mixture was
diluted with dichloromethane (100 ml) and the phases separated. The
organic phase was washed with water (100 ml), a saturated sodium
bicarbonate solution (100 ml) and brine. The organic phase was
treated with tert-butylamine (3.84 g, 5.56 ml, 52.5 mmol, Eq: 5),
and stirred at room temperature overnight. The resulting mixture
was washed with water (2.times.100 ml), a saturated sodium
bicarbonate solution (2.times.100 ml) and brine (1.times.200 ml)
and dried over magnesium sulfate. The mixture was filtered and
evaporated and the residue purified by flash chromatography
(Analogix Intelliflash 310, SF15-12 g column, 0-20% ethyl
acetate/hexane gradient). Like fractions were combined and
evaporated to give to 5-bromo-pyridine-2-sulfonic acid
tert-butylamide as a pale yellow solid (565 mg, 18%).
5-(4,4,5,5-Tetramethyl-[1,3,2]dioxaorolan-2-yl)-pyridine-2-sulfonic
acid tert-butylamide
##STR00139##
[0353] To a solution of 5-bromo-pyridine-2-sulfonic acid
tert-butylamide (565 mg, 1.93 mmol, Eq: 1.00),
4,4,4',4',5,5,5,5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (1.51 g,
5.95 mmol, Eq: 3.09), and potassium acetate (866 mg, 8.83 mmol, Eq:
4.58) in dioxane (20 ml) was added
1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex (151 mg, 206 .mu.mol, Eq: 0.107) and the
resulting mixture heated to 85.degree. C. overnight under an argon
atmosphere. The resulting mixture was cooled to room temperature,
diluted with ethyl acetate (25 ml) and evaporated. The residue was
taken up in ethyl acetate (50 ml), washed with brine (50 ml) and
dried over sodium sulfate. The mixture was filtered and to the
filtrate was added silica gel (.about.1.5 g). The mixture was
evaporated and purified by flash chromatography (Analogix
Intelliflash, VersaPak Spherical Silica column 20-45 uM, 11 g
column, 10-50% ethyl acetate/hexane). Like fractions were combined
and evaporated to give crude
5-(4,4,5,5-tetramethyl-[1,3,2]dioxaorolan-2-yl)-pyridine-2-sulfonic
acid tert-butylamide as an off-white solid (404 mg, 37% pure,
23%).
5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-6-trifluoromethyl-phen-
yl]-pyridine-2-sulfonic acid tert-butylamide (Compound 24)
##STR00140##
[0355] A mixture of
N3-(4-bromo-3-chloro-5-(trifluoromethyl)phenyl)-1H-1,2,4-triazole-3,5-dia-
mine Intermediate 1 (253 mg, 709 .mu.mol, Eq: 1),
5-(4,4,5,5-tetramethyl-[1,3,2]dioxaorolan-2-yl)-pyridine-2-sulfonic
acid tert-butylamide (635 mg, 709 .mu.mol, Eq: 1.00), a 3M
potassium carbonate solution (473 .mu.l, 1.42 mmol, Eq: 2),
dimethoxyethane (1.5 ml) p-dioxane (1 ml) and
tetrakis(triphenylphosphine)palladium(0) (152 mg, 131 .mu.mol, Eq:
0.185) was placed in a microwave reaction tube and degassed with
argon under sonication for 15 minutes. The tube was sealed and the
resulting mixture was heated in a microwave reactor to 128.degree.
C. and held at that temperature for 3 h. The reaction mixture was
cooled and diluted with ethyl acetate (3 mL) and water (8 mL) and
the phases separated. The aqueous phase was extracted with ethyl
acetate (3.times.15 mL) and the combined organic phases were washed
with brine and dried over sodium sulfate. The resulting mixture was
filtered and evaporated to give a brown residue which was
chromatographed on silica (Analogix Intelliflash 310, RediSep
silica 24 g, 4-10% methanol/dichloromethane gradient). Like
fractions were combined and evaporated and the residue purified by
SFC chromatography (THAR/SFC/Waters Multigram II system eluted with
25% methanol/carbon dioxide at 70 ml/mn on a CYANO column
3.times.25 cm with detection at 220 nM and 35 degrees and 100 bar
backpressure) to give
5-(4-(5-amino-1H-1,2,4-triazol-3-ylamino)-2-chloro-6-(trifluoromethyl)phe-
nyl)-N-tert-butylpyridine-2-sulfonamide as a light yellow solid (45
mg, 13%). MS +m/z: 489.9 (M+H).sup.+
5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-6-trifluoromethyl-phen-
yl]-pyridine-2-sulfonic acid
(2,2,2-trifluoro-1,1-dimethyl-ethyl)-amide (Compound 25)
##STR00141##
[0356] 5-Bromo-pyridine-2-sulfonyl chloride
##STR00142##
[0358] A mixture of crude 2-(benzylthio)-5-bromopyridine (prepared
in Compound 2, 7.74 g, 27.6 mmol, Eq: 1.00), water (80.0 ml), and
carbon tetrachloride (300 ml) was cooled to 0.degree. C. in an ice
bath and vigorously stirred while chlorine gas was bubbled slowly
through the mixture. After 3 minutes the reaction mixture was
saturated with chlorine and turned bright yellow green. Bubbling
was continued for 10 minutes followed by 5 additional minutes of
stirring under a chlorine blanket before the mixture was sparged of
chlorine by bubbling argon into the solution for 10 minutes. The
mixture was diluted with dichloromethane (100 ml) and the phases
separated. The organic phase was washed with water (100 ml), a
saturated sodium bicarbonate solution (100 ml) and brine and then
dried over magnesium sulfate. The mixture was filtered and
evaporated to give 5-bromopyridine-2-sulfonyl chloride as a
semi-crystalline solid (4.34 g, 61%).
5-Bromo-pyridine-2-sulfonic acid
(2,2,2-trifluoro-1,1-dimethyl-ethyl)-amide
##STR00143##
[0360] To a solution of 5-bromopyridine-2-sulfonyl chloride (2.17
g, 6.34 mmol, Eq: 1) in dry pyridine (10 ml) was added
1,1,1-trifluoro-2-methylpropan-2-amine (806 mg, 6.34 mmol, Eq: 1)
and the resulting mixture stirred at room temperature overnight.
The mixture was evaporated and the residue absorbed onto silica and
purified by flash chromatography (Analogix Intelliflash 310,
Redisep Rf 40 g column, 0-30% ethyl acetate/hexane gradient). Like
fractions were combined and evaporated to give
5-bromo-pyridine-2-sulfonic acid
(2,2,2-trifluoro-1,1-dimethyl-ethyl)-amide as a light yellow solid
(660 ag, 30%).
5-(4,4,5,5-Tetraethyl-[1,3,2]dioxaborolan-2-yl)-pyridine-2-sulfonic
acid (2,2,2-trifluoro-1,1-dimethyl-ethyl)-amide
##STR00144##
[0362] To a solution of 5-bromo-pyridine-2-sulfonic acid
(2,2,2-trifluoro-1,1-dimethyl-ethyl)-amide (660 mg, 1.9 mmol, Eq:
1), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane)
(1.49 g, 5.87 mmol, Eq: 3.09), and potassium acetate (855 mg, 8.71
mmol, Eq: 4.58) in dioxane (20 ml) was added
1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex (149 mg, 203 .mu.mol, Eq: 0.107) and the
resulting mixture heated to 85.degree. C. overnight under an argon
atmosphere. The resulting mixture was cooled to room temperature,
diluted with ethyl acetate (25 ml) and evaporated. The residue was
taken up in ethyl acetate (50 ml), washed with brine (50 ml) and
dried over sodium sulfate. The mixture was filtered and to the
filtrate was added silica gel (.about.1.5 g). The mixture was
evaporated and purified by flash chromatography (Analogix
Intelliflash RediSep Rf 24 g column, 10-50% ethyl acetate/hexane).
Like fractions were combined and evaporated to give crude
5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine-2-sulfonic
acid (2,2-trifluoro-1,1-dimethyl-ethyl)-amide as an off-white solid
(637 mg, 70% pure by HNMR, 60%).
5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-6-trifluoromethyl-phen-
yl]-pyridine-2-sulfonic acid
(2,2,2-trifluoro-1,1-dimethyl-ethyl)-amide (Compound 25)
##STR00145##
[0364] A mixture of
N3-(4-bromo-3-chloro-5-(trifluoromethyl)phenyl)-1H-1,2,4-triazole-3,5-dia-
mine Intermediate 1 (190 mg, 533 .mu.mol, Eq: 1.00),
5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine-2-sulfonic
acid (2,2,2-trifluoro-1,1-dimethyl-ethyl)-amide (300 mg, 533
.mu.mol, Eq: 1.00) a 3M potassium carbonate solution (355 .mu.l,
1.07 mmol, Eq: 2), dimethoxyethane (1 ml) p-dioxane (1 ml) and
tetrakis(triphenylphosphine)palladium(0) (114 mg, 98.5 .mu.mol, Eq:
0.185) was placed in a microwave reaction tube and degassed with
argon under sonication for 15 minutes. The tube was sealed and the
resulting mixture was heated in a microwave reactor to 128.degree.
C. and held at that temperature for 3 h. The reaction mixture was
cooled and diluted with ethyl acetate (2 mL) and water (1 mL) and
the phases separated. The aqueous phase was extracted with ethyl
acetate (3.times.1 mL) and the combined organic phases were washed
with brine and dried over sodium sulfate. The resulting mixture was
filtered and evaporated to give a brown residue which was
chromatographed on silica (Analogix Intelliflash 310, RediSep
silica 24 g, 4-10% methanol/dichloromethane gradient). Like
fractions were combined and evaporated and the residue purified by
SFC chromatography (THAR/SFC/Waters Multigram II system eluted with
20% methanol/carbon dioxide at 70 ml/mn on a SILICA column
3.times.25 cm with detection at 220 nM and 35 degrees and 100 bar
backpressure) to give
5-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-6-trifluoromethyl-phe-
nyl]-pyridine-2-sulfonic acid
(2,2,2-trifluoro-1,1-dimethyl-ethyl)-amide as an off white solid
(22.4 mg, 8%). MS +m/z: 453.9 (M+H).sup.+
4-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-6-trifluoromethyl-phen-
yl]-pyridine-2-sulfonic acid adamantan-1-ylamide (Compound 26)
##STR00146##
[0365] 5-Bromo-pyridine-2-sulfonic acid adamantan-1-ylamide
##STR00147##
[0367] To a solution of 5-bromopyridine-2-sulfonyl chloride
(prepared in example 3, 2.17 g, 6.34 mmol, Eq: 1) in pyridine (10
ml) was added 1-adamantamine (960 mg, 6.34 mmol, Eq: 1) and the
resulting mixture stirred at room temperature overnight. The
mixture was evaporated and the residue absorbed onto silica and
purified by flash chromatography (Analogix Intelliflash 310,
RedisepRf 40 g column, 0-30% ethyl acetate/hexane). Like fractions
were combined and evaporated to give 5-bromo-pyridine-2-sulfonic
acid adamantan-1-ylamide (900 mg, 38%).
5-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-sulfonic
acid adamantan-1-ylamide
##STR00148##
[0369] To a solution of 5-bromo-pyridine-2-sulfonic acid
adamantan-1-ylamide (900 mg, 2.42 mmol, Eq: 1),
bis(pinacolato)diboron (1.9 g, 7.49 mmol, Eq: 3.09), and potassium
acetate (1.09 g, 11.1 mmol, Eq: 4.58) in dioxane was added
1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex (190 mg, 259 .mu.mol, Eq: 0.107). The
resulting mixture was heated to 85.degree. C. overnight under an
argon atmosphere. The resulting mixture was cooled to room
temperature, diluted with ethyl acetate (25 ml) and evaporated. The
residue was taken up in ethyl acetate (50 ml), washed with brine
(50 ml) and dried over sodium sulfate. The mixture was filtered and
to the filtrate was added silica gel (.about.1.5 g). The mixture
was evaporated and purified by flash chromatography (Analogix
Intelliflash, RediSep Rf 24 g column, 10-50% ethyl acetate/hexane).
Like fractions were combined and evaporated to give crude
5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine-2-sulfoni-
c acid adamantan-1-ylamide as a pale yellow solid (759 mg, 71% pure
by HNMR, 53%).
4-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chlor-6-trifluoromethyl-pheny-
l]-pyridine-2-sulfonic acid adamantan-1-ylamide (Compound 26)
##STR00149##
[0371] A mixture of
N3-(4-bromo-3-chloro-5-(trifluoromethyl)phenyl)-1H-1,2,4-triazole-3,5-dia-
mine (256 mg, 717 .mu.mol, Eq: 1),
5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine-2-sulfonic
acid adamantan-1-ylamide (300 mg, 717 .mu.mol, Eq: 1) a 3M
potassium carbonate solution (478 .mu.l, 1.43 mmol, Eq: 2),
dimethoxyethane (1 ml) p-dioxane (1 ml) and
tetrakis(triphenylphosphine)palladium(0) (114 mg, 98.5 .mu.mol, Eq:
0.185) was placed in a microwave reaction tube and degassed with
argon under sonication for 15 minutes. The tube was sealed and the
resulting mixture was heated in a microwave reactor to 128.degree.
C. and held at that temperature for 3 h. The reaction mixture was
cooled and diluted with ethyl acetate (2 mL) and water (1 mL) and
the phases separated. The aqueous phase was extracted with ethyl
acetate (3.times.1 mL) and the combined organic phases were washed
with brine and dried over sodium sulfate. The resulting mixture was
filtered and evaporated to give a brown residue which was
chromatographed on silica (Analogix Intelliflash 310, RediSep
silica 24 g, 4-10% methanol/dichloromethane gradient). Like
fractions were combined and evaporated and the residue purified by
SFC chromatography (THAR/SFC/Waters Multigram II system eluted with
20% methanol/carbon dioxide at 70 ml/mn on a SILICA column
3.times.25 cm with detection at 220 nM and 35 degrees and 100 bar
backpressure) to give
4-[4-(5-amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-6-trifluoromethyl-phe-
nyl]-pyridine-2-sulfonic acid adamantan-1-ylamide as a light yellow
solid (44.4 mg, 11%). MS +m/z: 568 (M).sup.+
N3-(2-chloro-4'-(4-methylpiperazin-1-yl)-6-(trifluoromethyl)biphenyl-4-yl)-
-1H-1,2,4-triazole-3,5-diamine (Compound 27)
##STR00150##
[0373] A mixture of
N3-(4-bromo-3-chloro-5-(trifluoromethyl)phenyl)-1H-1,2,4-triazole-3,5-dia-
mine Intermediate 1 (590 mg, 1.65 mmol, Eq: 1),
1-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperaz-
ine (500 mg, 1.65 mmol, Eq: 1) a 3M potassium carbonate solution
(1.1 ml, 3.31 mmol, Eq: 2), dimethoxyethane (1.67 ml) p-dioxane
(1.67 ml) and tetrakis(triphenylphosphine)palladium(0) (354 mg, 306
.mu.mol, Eq: 0.185) was placed in a microwave reaction tube and
degassed with argon under sonication for 15 minutes. The tube was
sealed and the resulting mixture was heated in a microwave reactor
to 128.degree. C. and held at that temperature for 3 h. The
reaction mixture was cooled, the tube opened and
1-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperaz-
ine (200 mg, 1.65 mmol, Eq: 1) was added. The tube was resealed and
heated in a microwave reactor to 128.degree. C. for 2 h. The
reaction mixture was cooled and diluted with ethyl acetate and the
organic phase washed with brine and dried over magnesium sulfate.
The mixture was filtered and evaporated to give a brown residue
which was chromatographed on silica (Analogix Intelliflash 310,
RediSep silica 24 g, 4-10% methanol/dichloromethane gradient). Like
fractions were combined and evaporated and the residue purified by
HPLC (Gilson, Supelcosil ABZ+Plus column, 25 cm.times.21.2 mm 12
.mu.M, 25-100% acetonitrile/water gradient 0.3% formic acid). Like
fractions were combined and lyophilized to give
N3-(2-chloro-4'-(4-methylpiperazin-1-yl)-6-(trifluoromethyl)biphenyl-4-yl-
)-1H-1,2,4-triazole-3,5-diamine as a white lyophilized solid (20.23
mg, 44.8 .mu.mol, 2.71%). MS +m/z: 543.0 (M+H)
1-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-phenyl]-1H-pyridin-2-one
(Compound 28)
##STR00151##
[0374] 1-(4-Nitro-phenyl)-1H-pyridin-2-one
##STR00152##
[0376] In a 50 mL round-bottomed flask, potassium tert-butoxide
(885 mg, 7.89 mmol, Eq: 1.50) and pyridin-2(1H)-one (500 mg, 5.26
mmol, Eq: 1.00) were combined with DMF (10.0 mL) to give a light
brown solution at 0.degree. C. under nitrogen.
1-fluoro-4-nitrobenzene (742 mg, 5.26 mmol, Eq: 1.00) was added.
The reaction was stirred overnight at room temperature. The
reaction mixture was diluted with CH.sub.2Cl.sub.2 (50 mL), washed
with H.sub.2O (25 mL) and brine (25 mL). The organic layer was
dried over anhydrous MgSO.sub.4, filtered and volatiles were
removed under reduced pressure to yield an oil from which the
compound was isolated by column chromatography
(Hexanes/EtOAc=70/30) to give an off-white solid 1 g (88%). MH+
217.1
1-(4-Amino-phenyl)-1H-pyridin-2-one
##STR00153##
[0378] In a 100 mL round bottle, 1-(4-nitrophenyl)pyridin-2(1H)-one
(1.0 g, 4.63 mmol, Eq: 1.00) and zinc (1.51 g, 23.1 mmol, Eq: 5.00)
were combined with a solution of saturated NH.sub.4Cl aqueous
solution/THF (1:1) (50 ml), the mixture was stirred at for
overnight. Filter out the solid, extracted with CH.sub.2Cl.sub.2
(50 mL.times.2), the organic layer was dried over anhydrous
Na.sub.2SO.sub.4; the solution was concentrated under vacuum to
afford the crude product 740 mg (86%). MH+ 188.3
1-(4-Isothiocyanato-phenyl)-1H-pyridin-2-one
##STR00154##
[0380] In a 100 mL round-bottomed flask,
di(1H-imidazol-1-yl)methanethione (718 mg, 4.03 mmol, Eq: 1.5) was
combined with CH.sub.2Cl.sub.2 (30 mL) to give a colorless
solution. 1-(4-Aminophenyl)pyridin-2(1H)-one (500 mg, 2.69 mmol,
Eq: 1.00) in CH.sub.2Cl.sub.2 (20 mL) was added dropwise at
0.degree. C. The reaction was allowed to warm to room temperature,
and allowed to stir overnight. Concentrate the solution, the
compound was isolated by column chromatography
(Hexanes/EtOAc=80/20) to give the product 580 mg (95%).
MH+228.9
Methylsulfanyl-[4-(2-oxo-2H-pyridin-1-yl)-phenylamino]-methyl-cyanamide
##STR00155##
[0382] In a 100 mL round-bottomed flask,
1-(4-isothiocyanatophenyl)pyridin-2(1H)-one (580 mg, 2.54 mmol, Eq:
1.00) in MeOH (20 mL), sodium hydrogencyanamide (184 mg, 2.87 mmol,
Eq: 1.13) was added. The suspension turned to clear after a few
minutes, the reaction was allowed to stir at room temperature for 1
hour, iodomethane (721 mg, 5.08 mmol, Eq: 2) was added, the
reaction mixture was allowed to stir at room temperature overnight.
Concentrate the solution, the compound was isolated by column
chromatography (CH.sub.2Cl.sub.2/MeOH=95/5) to give an off-white
solid 120 mg (17%). MH+ 284.9
1-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-phenyl]-1H-pyridin-2-one
(Compound 28)
##STR00156##
[0384] In a 100 mL round-bottomed flask,
methylsulfanyl-[4-(2-oxo-2H-pyridin-1-yl)-phenylamino]-methyl-cyanamide
(120 mg, 419 .mu.mol, Eq: 1.00) in EtOH (30 mL), hydrazine (134 mg,
4.19 mmol, Eq: 10.00) was added. The reaction was heated to
65.degree. C. for 3 hours. The reaction mixture was concentrated,
added H.sub.2O (20 mL) to the residue, filtered out the solid and
washed the solid with H.sub.2O (30 mL) and CH.sub.2Cl.sub.2 (10
mL), air-dried the solid overnight to give an off-white solid 36 mg
(32%). MH+ 268.9
1-[4-(5-Amino-1H-[1,2,4]triol-3-ylamino)-2-chloro-phenyl]-1H-pyridin-2-one
(Compound 29)
##STR00157##
[0385] 1-(2-Chloro-4-nitro-phenyl)-1H-pyridin-2-one
##STR00158##
[0387] In a 50 mL round-bottomed flask, potassium tert-butoxide
(7.08 g, 63.1 mmol, Eq: 1.50) and pyridin-2(1H)-one (4 g, 42.1
mmol, Eq: 1.00) were combined with DMF (50.0 mL) to give a light
brown solution at 0.degree. C. under nitrogen. N2.
2-chloro-1-fluoro-4-nitrobenzene (7.38 g, 42.1 mmol, Eq: 1.00) was
added. The reaction was stirred overnight at room temperature. The
reaction mixture was diluted with CH.sub.2Cl.sub.2 (50 mL), washed
with H.sub.2O (25 mL) and brine (25 mL). The organic layer was
dried over anhydrous MgSO.sub.4, filtered and volatiles were
removed under reduced pressure to yield an oil from which the
compound was isolated by column chromatography
(Hexanes/EtOAc=70/30) to give an off-white solid 4.07 g (39%). MH+
250.9
1-(4-Amino-2-chloro-phenyl)-1H-pyridin-2-one
##STR00159##
[0389] In a 100 mL round bottle,
1-(2-chloro-4-nitrophenyl)pyridin-2(1H)-one (4.07 g, 16.2 mmol, Eq:
1.00) and zinc (5.31 g, 81.2 mmol, Eq: 5.00) were combined with a
solution of saturated NH.sub.4Cl aqueous solution/THF (1:1) (50
mL), the mixture was stirred at for overnight. Filter out the
solid, extracted with CH.sub.2Cl.sub.2 (50 mL.times.2), the organic
layer was dried over anhydrous Na.sub.2SO.sub.4; the solution was
concentrated under vacuum to afford the crude product 3.3 g (92%).
MH+ 221.0
1-(2-Chloro-4-isothiocyanato-phenyl)-1H-pyridin-2-one
##STR00160##
[0391] In a 100 mL round-bottomed flask,
di(1H-imidazol-1-yl)methanethione (2.42 g, 13.6 mmol, Eq: 1.5) was
combined with CH.sub.2Cl.sub.2 (30 mL) to give a colorless
solution. 1-(4-amino-2-chlorophenyl)pyridin-2(1H)-one (2 g, 9.06
mmol, Eq: 1.00) in CH.sub.2Cl.sub.2 (20 mL) was added dropwise at
0.degree. C. The reaction was allowed to warm to room temperature,
and allowed to stir overnight. Concentrate the solution, the
compound was isolated by column chromatography
(Hexanes/EtOAc=50/50) to give the product 1.1 g (46%). MH+
262.9
[3-Chloro-4-(2-oxo-2H-pyridin-1-yl)-phenylamino]-methylsulfanyl-methyl-cya-
namide
##STR00161##
[0393] In a 100 mL round-bottomed flask,
1-(2-chloro-4-isothiocyanatophenyl)pyridin-2(1H)-one (1.1 g, 4.19
mmol, Eq: 1.00) in MeOH (20 mL), sodium hydrogencyanamide (303 mg,
4.73 mmol, Eq: 1.13) was added. The suspension turned to clear
after a few minutes, the reaction was allowed to stir at room
temperature for 1 hour, iodomethane (1.19 g, 8.37 mmol, Eq: 2) was
added, the reaction mixture was allowed to stir at room temperature
overnight. Concentrate the solution, the compound was isolated by
column chromatography (CH.sub.2Cl.sub.2/MeOH=95/5) to give an
off-white solid 590 mg (44%). MH+ 320.9
1-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2-chloro-phenyl]-1H-pyridin-2-o-
ne (Compound 29)
##STR00162##
[0395] In a 100 mL round-bottomed flask,
[3-chloro-4-(2-oxo-2H-pyridin-1-yl)-phenylamino]-methylsulfanyl-methyl-cy-
anamide (590 mg, 1.84 mmol, Eq: 1.00) in EtOH (30 mL), hydrazine
(589 mg, 18.4 mmol, Eq: 10.00) was added. The reaction was heated
to 65.degree. C. for 3 hours. The reaction mixture was
concentrated, added H.sub.2O (20 mL) to the residue, filtered out
the solid and washed the solid with H.sub.2O (30 mL) and
CH.sub.2Cl.sub.2 (10 mL), air-dried the solid overnight to give an
off-white solid 494 mg (89%). MH+ 303.0
1-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-1H-pyridin-
-2-one (Compound 30)
##STR00163##
[0396] 1-(2,6-Dichloro-4-nitro-phenyl)-1H-pyridin-2-one
##STR00164##
[0398] In a 50 mL round-bottomed flask, potassium tert-butoxide
(3.54 g, 31.5 mmol, Eq: 1.50) and pyridin-2(1H)-one (2 g, 21.0
mmol, Eq: 1.00) were combined with DMF (25.0 ml) to give a light
brown suspension at 0.degree. C. under nitrogen.
1,3-dichloro-2-fluoro-5-nitrobenzene (4.42 g, 21.0 mmol, Eq: 1.00)
was added. The reaction was stirred overnight at room temperature.
The reaction mixture was diluted with CH.sub.2C.sub.2 (50 mL),
washed with H.sub.2O (25 mL) and brine (25 mL). The organic layer
was dried over anhydrous MgSO.sub.4, filtered and volatiles were
removed under reduced pressure to yield an oil from which the
compound was isolated by column chromatography
(Hexanes/EtOAc=70/30) to give an off-white solid 2.8 g (47%). MH+
284.9
1-(4-Amino-2,6-dichloro-phenyl)-1H-pyridin-2-one
##STR00165##
[0400] In a 100 mL round bottle,
1-(2,6-dichloro-4-nitrophenyl)pyridin-2(1H)-one (2.8 g, 9.82 mmol,
Eq: 1.00) and zinc (3.21 g, 49.1 mmol, Eq: 5.00) were combined with
a solution of saturated NH.sub.4Cl aqueous solution/THF (1:1) (50
mL), the mixture was stirred at for overnight. Filter out the
solid, extracted with CH.sub.2Cl.sub.2 (50 mL.times.2), the organic
layer was dried over anhydrous Na.sub.2SO.sub.4; the solution was
concentrated under vacuum to afford the crude product 1.3 g (50%).
MH+ 255.9
1-(2,6-Dichloro-4-isothiocyanato-phenyl)-1H-pyridin-2-one
##STR00166##
[0402] In a 100 mL round-bottomed flask,
di(1H-imidazol-1-yl)methanethione (1.26 g, 7.06 mmol, Eq: 1.5) was
combined with CH.sub.2Cl.sub.2 (30 mL) to give a colorless
solution. 1-(4-amino-2,6-dichlorophenyl)-pyridin-2(1H)-one (1.2 g,
4.7 mmol, Eq: 1.00) in CH.sub.2Cl.sub.2 (20 mL) was added dropwise
at 0.degree. C. The reaction was allowed to warm to room
temperature, and allowed to stir overnight. Concentrate the
solution, the compound was isolated by column chromatography
(Hexanes/EtOAc=50/50) to give the product 520 mg (37%). MH+
296.8
[3,5-Dichloro-4-(2-oxo-2H-pyridin-1-yl)-phenylamino]-methylsulfanyl-methyl-
-cyanamide
##STR00167##
[0404] In a 100 mL round-bottomed flask,
1-(2,6-dichloro-4-isothiocyanatophenyl)pyridin-2(1H)-one (520 mg,
1.75 mmol, Eq: 1.00) in MeOH (20 mL), sodium hydrogencyanamide (127
mg, 1.98 mmol, Eq: 1.13) was added. The suspension turned to clear
after a few minutes, the reaction was allowed to stir at room
temperature for 1 hour, iodomethane (497 mg, 3.5 mmol, Eq: 2) was
added, the reaction mixture was allowed to stir at room temperature
overnight. Concentrate the solution, the compound was isolated by
column chromatography (CH.sub.2Cl.sub.2/MeOH=95/5) to give an
off-white solid 200 mg (32%). MH+ 352.9
1-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-1H-pyridin-
-2-one (Compound 30)
##STR00168##
[0406] In a 100 mL round-bottomed flask,
[3,5-dichloro-4-(2-oxo-2H-pyridin-1-yl)-phenylamino]-methylsulfanyl-methy-
l-cyanamide (200 mg, 563 .mu.mol, Eq: 1.00) in EtOH (30 mL),
hydrazine (180 mg, 5.63 mmol, Eq: 10.00) was added. The reaction
was heated to 65.degree. C. for 3 hours. The reaction mixture was
concentrated, added H.sub.2O (20 mL) to the residue, filtered out
the solid and washed the solid with H.sub.2O (30 mL) and
CH.sub.2Cl.sub.2 (10 mL), air-dried the solid overnight to give an
off-white solid 36 mg (19%). MH+ 336.9
N3-(3,5-dichloro-4-(1,4,5,6-tetrahydropyrimidin-2-yl)phenyl-1H-1,2,4-triaz-
ole-3-diamine. trifluoroacetic salt (Compound 31)
##STR00169##
[0407] 2,6-dichloro-4-nitrobenzonitrile
##STR00170##
[0409] A solution of copper(i) cyanide (2.6 g, 29.0 mmol, Eq: 2) in
DMSO (5 mL) was heated at 60.degree. for 1 hr. Tert-butyl nitrite
(5.98 g, 6.9 ml, 58.0 mmol, Eq: 4.00) and a solution
2,6-dichloro-4-nitroaniline (3 g, 14.5 mmol, Eq: 1.00) in DMSO (5
mL) was added and the reaction was stirred for 3 hr. The reaction
mixture was poured into ice water and extracted with ethyl acetate
3.times.. The organic extract was washed with brine and dried over
sodium sulfate. Chromatography (200 g Analogix, 100% hex to 5%
EtOAc/hex) gave 515 mg (16%) of desired product as a light brown
solid.
4-amino-2,6-dichlorobenzonitrile
##STR00171##
[0411] A solution of 2,6-dichloro-4-nitrobenzonitrile (2.46 g, 11.3
mmol, Eq: 1.00), iron (3.17 g, 56.7 mmol, Eq: 5) and ammonium
chloride (6.06 g, 113 mmol, Eq: 10) in methanol (30 mL)/water (10
mL) was heated at reflux o/n. TLC shows incomplete reaction.
Continued heating at 100 deg for 6 hr, then 60 deg overnight. The
reaction mixture was filtered over Celite. The filtrate was
suspended in ethyl acetate to give insoluble solid. The slurry was
concentrated to dryness, suspended in water, filtered, and rinsed
with water. The solid was transferred to a round bottom flask,
suspended in benzene, and concentrated. An additional portion of
benzene was added and concentrated once more to give 1.37 g (65%)
of desired product as a light brown solid.
2,6-dichloro-4-isothiocyanatobenzonitrile
##STR00172##
[0413] A suspension of 4-amino-2,6-dichlorobenzonitrile (500 mg,
2.67 mmol, Eq: 1.00), thiophosgene (1.35 g, 900 .mu.l, 11.7 mmol,
Eq: 439), triethylamine (875 mg, 1.2 ml, 8.64 mmol, Eq: 3.23) in
benzene (30 ml) was heated at reflux overnight. The brown reaction
mixture was concentrated and chromatographed (80 g Analogix, 0 to
5% ethyl acetate/hexane) to give 436 mg (71%) of desired product as
a light brown solid.
(Z)-methyl
N'-cyano-N-(3,5-dichloro-4-cyanophenyl)carbamimidothioate
##STR00173##
[0415] Sodium methoxide (0.5M in methanol) (5.7 ml, 2.85 mmol, Eq:
1.22) was added to cyanamide (108 mg, 2.58 mmol, Eq: 1.1). After 15
minutes, the solution was added to a solution of
2,6-dichloro-4-isothiocyanatobenzonitrile (537 mg, 2.34 mmol, Eq:
1.00) in methanol (5 mL). After 1 hr, methyl iodide (704 mg, 310
.mu.l, 4.96 mmol, Eq: 2.11) was added and the reaction was stirred
overnight at room temperature. The resulting suspension was
filtered and dried to give 43 mg of desired product as a gray
solid. The filtrate was concentrated and chromatographed (40 g
Analogix, 50% EtOAc/hex to 75% EtOAc/hex) to give 191 mg of desired
product as a yellow solid. The solids were combined to give 234 mg
(35%) of desired product.
4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-dichlorobenzonitrile
##STR00174##
[0417] A solution of (Z)-methyl
N'-cyano-N-(3,5-dichloro-4-cyanophenyl)carbamimidothioate (234 mg,
821 .mu.mol, Eq: 1.00) and hydrazine (263 mg, 258 .mu.l, 8.21 mmol,
Eq: 10) in ethanol (10 mL) was heated at 65.degree. C. o/n. The
reaction mixture was concentrated and chromatographed (23 Supelco,
100% DCM to 5% to 10% MeOH/DCM to give 156 mg (71%) of desired
product as an off-white solid.
[0418] .sup.1H NMR (300 MHz, DMSO) .delta.: 11.55 (s, 1H), 9.99 (s,
1H), 7.74 (s, 2H), 6.14 (s, 2H) ppm
N3-(3,5-dichloro-4-(1,4,5,6-tetrahydropyrimidin-2-yl)phenyl)-1H-1,2,4-tria-
zole-3,5-diamine. trifluoroacetic salt (Compound 31)
##STR00175##
[0420] A mixture of
4-(5-amino-1H-1,2,4-triazol-3-ylamino)-2,6-dichlorobenzonitrile (85
ag, 316 .mu.mol, Eq: 1.00), propane-1,3-diamine (70.2 mg, 948
.mu.mol, Eq: 3) and 4-methylbenzenesulfonic acid, monohydrate (60.1
mg, 316 .mu.mol, Eq: 1.00) was stirred at 210.degree. C. for 5
h.
[0421] The reaction mixture is cooled down and the resulting solid
was dried in vacuo then purified by reverse phase HLPC to afford 46
mg (33%) of the desired product as a white solid.
[0422] MS -m/z: 438.0 (M-H).sup.-
5-(4-(5-amino-1H-1,2,4-triazol-3-ylamino)-2-chloro-6-(trifluoromethyl)phen-
yl)-N-methylpicolinamide (Compound 32
##STR00176##
[0424] A mixture of
N3-(4-bromo-3-chloro-5-(trifluoromethyl)phenyl)-1H-1,2,4-triazole-3,5-dia-
mine Intermediate 1 (200 mg, 561 .mu.mol, Eq: 1.00),
N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamide
[945863-21-8] (198 mg, 757 .mu.mol, Eq: 1.35) and
(triphenylphosphine)palladium (0) (64.8 mg, 56.1 .mu.mol, Eq: 0.1)
was degassed (vacuum/nitrogen cycles) then degassed dry dioxane
(1.85 ml) (nitrogen bubbling with sonication) and a degassed
(nitrogen bubbling with sonication) 2M solution of sodium carbonate
in water (561 .mu.l, 1.12 mmol, Eq: 2) were added. The reaction
mixture was sealed and stirred at 105.degree. C. for 18 h. The
reaction mixture was adsorbed unto silica (Ig), concentrated and
purified on silica gel (silica 24 g, dichloromethane/methanol 97:3
to 70:30). One fraction was isolated and dried in vacuo to afford
45 mg (20%) of the desired product as a yellow semi-solid
solid.
[0425] MS +m/z: 411.9 (M+H).sup.+
N3-(3,5-dichloro-4-(pyridazin-4-yl)phenyl)-1H-1,2,4-triazole-3,5-diamine
(Compound 33)
##STR00177##
[0427] To a degassed (cycle vacuum/nitrogen) mixture of
N3-(4-bromo-3,5-dichlorophenyl)-1H-1,2,4-triazole-3,5-diamine
Intermediate 2 (80 mg, 248 .mu.mol, Eq: 1.00) and
Pd(PPh.sub.3).sub.2Cl.sub.2 (17.4 mg, 24.8 .mu.mol, Eq: 0.1) was
added degassed (nitrogen bubbling with sonication) dry
dimethylformamide (3 ml) followed by 4-(tributylstannyl)pyridazine
(229 mg, 195 .mu.l, 619 .mu.mol, Eq: 2.5). The reaction mixture was
further degassed (vacuum/nitrogen cycles).
[0428] The reaction mixture was stirred at 100.degree. C. over 3
days then adsorbed unto silica (0.7 g) and purified on silicagel
(column 12 g, dichloromethane/methanol 100:0 to 60:40). 37 mg of a
brown solid was obtained. The compound was triturated in methanol
(1 mL) then washed with methanol (2.times.1 mL). The solid was
dried in vacuo to afford 30 mg (38%) of the desired product as a
brown-red solid.
[0429] NMR (300 MHz, DMSO d.sup.6): 11.38 (1H, a); 9.45 (1H, s);
9.34 (1H, dd, J=1, 6 Hz); 9.22 (1H, broad s); 7.78 (21, s); 7.74
(1H, dd, J=3, 6 Hz); 6.06 (2H, s).
1-{3-[4-(5-Amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-thiophe-
n-2-yl}-ethanone (Compound 34)
##STR00178##
[0431] To a solution of
N-3-(4-bromo-3,5-dichlorophenyl)-1H-1,2,4-triazole-3,5-diamine
Intermediate 2 (80.7 mg, 0.25 mmol, Eq: 1.00) in dioxane (1.25 mil)
was added tetrakis(triphenylphosphine)palladium(0) (23.1 mg, 20
.mu.mol, Eq: 0.08), potassium carbonate (167 .mu.l, 3M, Eq: 2), and
2-acetyl-3-thienylboronic acid (85 mg, 0.5 mmol, Eq: 2). The
mixture was degassed twice under Nitrogen, then was heated at
100.degree. C. for 16 hours. After being cooled to room
temperature, the solvent was removed in vacuo. To the crude product
was added a few drops of acetic acid, followed by addition of
mixture of MeOH/CH3CN/H2O(45%/45%/10%)(2 ml). The resulting
suspension was centrifuged, and the solution was separated and
purified by reverse phase HPLC (0.1% HOAc in acetonitrile and
water). Product fractions were collected and lyophilized to afford
1-{3-[4-(5-amino-1H-[1,2,4]triazol-3-ylamino)-2,6-dichloro-phenyl]-thioph-
en-2-yl}-ethanone (28.5 mg, 31%) as a white solid. MS m/z: 369
(M+H)+.
[0432] The following compounds 35-44 were all prepared in an
analogous manner to example 34:
N-3-(3,5-Dichloro-4-(pyridin-4-yl)phenyl)-1H-1,2,4-triazole-3,5-diamine
(Compound 35)
##STR00179##
[0434] Starting from Pyridin-4-ylboronic acid, yield=13%, MS m/z
322 (M+H)
N-3-[3,5-Dichloro-4-(5-chloro-thiophen-2-yl)-phenyl]-1H-[1,2,4]triazole-3,-
5-diamine (Compound 36)
##STR00180##
[0436] Starting from 5-Chlorothiophene-2-boronic acid, yield=2%. MS
m/z 362 (M+H)
N-3-(3,5-Dichloro-4-pyridin-3-yl-phenyl)-1H-[1,2,4]triazole-3,5-diamine
(Compound 37)
##STR00181##
[0438] Starting from Pyridine-3-boronic acid, yield=15%, MS m/z 322
(M+H)
N-3-[3,5-Dichloro-4-(1H-pyrazol-3-yl)-phenyl]-1H-[1,2,4]triazole-3,5-diami-
ne (Compound 38)
##STR00182##
[0440] Starting from Pyrazole-3-boronic acid, yield=1%, MS m/z 311
(M+H)
N-3-(3,5-Dichloro-4-pyrimidin-5-yl-phenyl)-1H-[1,2,4]triazole-3,5-diamine
(Compound 39)
##STR00183##
[0442] Starting from Pyrimidine-5-boronic acid, yield=4%, MS m/z
323 (M+H)
N-3-[3,5-Dichloro-4-(2-methoxy-pyrimidin-5-yl)-phenyl]-1H-[1,2,4]triazole--
3,5-diamine (Compound 40)
##STR00184##
[0444] Starting from 2-Methoxypyrimidine-5-boronic acid, yield=26%,
MS m/z 353 (M+H)
N-3-[3,5-Dichloro-4-(6-trifluoromethyl-pyridin-3-yl)-phenyl]-1H-[1,2,4]tri-
azole-3,5-diamine (Compound 41)
##STR00185##
[0446] Starting from 2-Trifluormethyl-5-pyridineboric acid,
yield=61%, MS m/z 390 (M+H)
N-3-[3,5-Dichloro-4-(1-methyl-3-trifluoromethyl-1H-pyrazol-4-yl)-phenyl]-1-
H-[1,2,4]triazole-3,5-diamine (Compound 42)
##STR00186##
[0447] Starting from 1-Methyl-3-trifluoromethylpyrazole-4-boronic
acid, yield=3%, MS m/z 393 (M+H)
N-3-[3,5-Dichloro-4-(5-chloro-pyridin-3-yl)-phenyl]-1H-[1,2,4]triazole-3,5-
-diamine (Compound 43)
##STR00187##
[0449] Starting from 5-Chloropyridin-3-boronic acid, yield=31%, MS
m/z 357 (M+H)
N-3-[3,5-Dichloro-4-(6-methoxy-pyridin-2-yl)-phenyl]-1H-[1,2,4]triazole-3,-
5-diamine (Compound 44)
##STR00188##
[0451] Starting from 6-Methoxypyridine-2-boronic acid, yield=1%, MS
m/z 352 (M+H)
Biological Examples
[0452] Determination of compounds HCV GT1b and GT1a entry
inhibitory activity using the pseudotyped HCV particle (HCVpp)
reporter assay.
[0453] Mammalian expression plasmids for the generation of
pseudotyped virus particles.
[0454] Plasmids expressing HCV E1 and E2 envelope proteins of GT1a
H77 strain (Proc Natl Acad Sci USA 1997 94:8738-43) or GT1b Con1
strain (Science 1999 285:110-3) were constructed by cloning the
nucleic acids encoding the last 60 amino acids of HCV core protein
and all of the HCV E1 and E2 proteins into pcDNA3.1(+) vector.
Plasmid pVSV-G expressing the glycoprotein G of the vesicular
stomatitis virus (VSV G) is from Clontech (cat #631530). The HIV
packaging construct expressing the firefly luciferase reporter gene
was modified based on the envelope defective
pNL.4.3.Luc-R.sup.-.E.sup.- vector (Virology 1995 206:935-44) by
further deleting part of the HIV envelope protein.
[0455] Generation of pseudotyped virus particles in transiently
transfected HEK-293T cells.
[0456] Pseudotyped HCV GT1a and GT1b particles (HCVpp) and the
pseudotyped VSV G particles (VSVpp) were generated from transiently
transfected HEK-293T cells (ATCC cat# CRL-573). For generating
HCVpp, the HEK-293T cells were transfected with equal amounts of
plasmids expressing the HCV envelope proteins and the HIV packaging
genome by using polyethylenimine (Polysciences cat#23966) as
transfection reagent. For generating VSVpp, the HEK-293T cells were
transfected with equal amounts of plasmids expressing VSV G and the
HIV packaging genome by using polyethylenimine. 24 hours after the
transfection, the cell culture medium containing the transfection
mixture was replaced with fresh Dulbecco's Modified Eagle Medium
(DMEM-Glutamax.TM.-I; Invitrogen cat #10569-010) supplemented with
10% Fetal Bovine Serum (Invitrogen cat #10082-147) and 2 mM
L-glutamine (Invitrogen cat #25030-081). The supernatant was
collected 48 hours after the transfection and filtered through a
sterile 0.45 .mu.m filter. Aliquots of the supernatant was frozen
and stored at -80.degree. C. until use.
[0457] Huh7-high CD81 cells with high CD81 expression level were
enriched by flow cytometry sorting using FITC-labeled CD81 antibody
JS-81 (BD Biosciences ca 561956) to allow more efficient HCV entry.
The Huh7-high CD81 cells were cultured in Dulbecco's Modified Eagle
Medium (DMEM-Glutamax.TM.; Invitrogen cat #10569-010). The medium
was supplemented with 10% Fetal Bovine Serum (Invitrogen cat
#10082-147) and 1% penicillin/streptomycin (Invitrogen cat
#15070-063). Cells were maintained at 37.degree. C. in a humidified
5% CO.sub.2 atmosphere.
[0458] Determination of compound HCVpp entry inhibitory activity in
Huh7-high CD81 cells.
[0459] Huh7-high CD81 cells were plated at a cell density of 8000
cells per well in 96 well plates (Perkin Elmer, cat #6005660).
Cells were plated in 100 .mu.l of Dulbecco's Modified Eagle Medium
(DMEM-Glutamax.TM.-I, Invitrogen Cat #10569-010) supplemented with
10% Fetal Bovine Serum (Invitrogen Cat #10082-147) and 1%
penicillin/streptomycin (Invitrogen cat #15070-063). Cells were
allowed to equilibrate for 24 hours at 37.degree. C. and 5% CO2 at
which time compounds and pseudotyped viruses were added. On the day
of the assay, HCVpp aliquots were thawed in 37.degree. C. water
bath and kept at 4.degree. C. until use. Compounds (or medium as a
control) were diluted in 3 fold dilution series in
DMEM-Glutamax.TM.-I with 2% DMSO and 2% penicillin/streptomycin.
The 100 .mu.l plating medium in each culture well was removed
followed by the addition of 50 .mu.l compound dilutions and 50
.mu.l thawed HCVpp. Firefly luciferase reporter signal was read 72
hours after the addition of compounds and HCVpp using the
Steady-Glo luciferase Assay System (Promega, cat # E2520) following
the manufacturer's instruction. EC50 values were defined as the
compound concentration at which a 50% reduction in the levels of
firefly luciferase reporter was observed as compared to control
samples in the absence of compound and was determined by non-linear
fitting of compound dose-response data.
[0460] Determination of compound selectivity in Huh7-high CD81
cells.
[0461] Huh7 hCD81 cell assay plates and compound dilutions were set
up in the same format as in the HCVpp assay. 24 hours after cell
plating, thawed VSVpp was diluted by 800 fold in
DMEM-Glutamax.TM.-I supplemented with 10% fetal bovine serum. After
removal of the cell plating medium from the culture wells, 50 .mu.l
compound dilutions and 50 .mu.l diluted VSVpp were added to the
wells. Firefly luciferase reporter signal was read 72 hours after
the addition of compounds and VSVpp using the Steady-Glo luciferase
Assay System (Promega, cat # E2520). EC50 values were defined as
the compound concentration at which a 50% reduction in the levels
of firefly luciferase reporter was observed as compared to control
samples in the absence of compound and was determined by non-linear
fitting of compound dose-response data. The EC50 was approximated
if maximum percentage inhibition was less than 90% and more than
70%.
[0462] Representative assay data can be found in Table II
below:
TABLE-US-00002 TABLE II Compound # HCVpp GT-1b (EC.sub.50, .mu.M)
VSVpp (EC.sub.50, .mu.M) 1 0.767 3.7 2 0.386 10.0 3 0.364 10.0 4
0.173 1.4 5 0.1 0.4 6 0.33 10.0 7 0.809 10.0 8 0.386 10.0 9 0.998
31.1 10 1.238 34.2 11 0.271 2.2 12 0.216 10.0 13 10 10.0 14 6.52
15.0 15 6.724 16 34.772 17 4.144 34.6 18 0.088 10.0 19 0.505 10.0
20 0.073 13.6 21 0.568 10.0 22 2.038 10.0 23 0.179 7.5 24 0.008 4.1
25 0.005 10.0 26 0.005 3.4 27 0.301 10.0 28 100 100.0 29 13.718 8.1
30 5.747 5.6 31 5.454 4.7 32 0.169 21.8 33 0.78 10.6 34 0.144 1.8
35 0.338 2.5 36 0.558 11.9 37 0.413 8.1 38 3.387 35.8 39 0.536 5.1
40 0.52 3.3 41 0.703 32.6 42 0.0527 2.4 43 0.771 5.3 44 1.156
7.8
[0463] The foregoing invention has been described in some detail by
way of illustration and example, for purposes of clarity and
understanding. It will be obvious to one of skill in the art that
changes and modifications may be practiced within the scope of the
appended claims.
[0464] Therefore, it is to be understood that the above description
is intended to be illustrative and not restrictive. The scope of
the invention should, therefore, be determined not with reference
to the above description, but should instead be determined with
reference to the following appended claims, along with the full
scope of equivalents to which such claims are entitled.
[0465] All patents, patent applications and publications cited in
this application are hereby incorporated by reference in their
entirety for all purposes to the same extent as if each individual
patent, patent application or publication were so individually
denoted.
* * * * *