U.S. patent application number 13/353864 was filed with the patent office on 2012-05-10 for compounds for the treatment of hepatitis c.
This patent application is currently assigned to Bristol-Myers Squibb Company. Invention is credited to John A. Bender, Piyasena Hewawasam, Yong Tu, Zhong Yang.
Application Number | 20120115847 13/353864 |
Document ID | / |
Family ID | 40263273 |
Filed Date | 2012-05-10 |
United States Patent
Application |
20120115847 |
Kind Code |
A1 |
Hewawasam; Piyasena ; et
al. |
May 10, 2012 |
Compounds for the Treatment of Hepatitis C
Abstract
The invention encompasses compounds of formula I as well as
compositions and methods of using the compounds. The compounds have
activity against hepatitis C virus (HCV) and are useful in treating
those infected with HCV. ##STR00001##
Inventors: |
Hewawasam; Piyasena;
(Middletown, CT) ; Tu; Yong; (Cheshire, CT)
; Bender; John A.; (Middletown, CT) ; Yang;
Zhong; (Southington, CT) |
Assignee: |
Bristol-Myers Squibb
Company
|
Family ID: |
40263273 |
Appl. No.: |
13/353864 |
Filed: |
January 19, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
12272974 |
Nov 18, 2008 |
8129367 |
|
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13353864 |
|
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60989524 |
Nov 21, 2007 |
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Current U.S.
Class: |
514/214.01 |
Current CPC
Class: |
A61P 31/00 20180101;
A61P 31/14 20180101; C07D 487/04 20130101; C07D 519/00
20130101 |
Class at
Publication: |
514/214.01 |
International
Class: |
A61K 31/55 20060101
A61K031/55; A61P 31/14 20060101 A61P031/14 |
Claims
1. A composition comprising a compound of formula I and a
pharmaceutically acceptable carrier ##STR00173## where R.sup.1 is
CONR.sup.5R.sup.6; R.sup.2 is ##STR00174## R.sup.3 is hydrogen,
halo, alkyl, alkenyl, hydroxy, benzyloxy, alkoxy, or haloalkoxy;
R.sup.4 is cycloalkyl; R.sup.5 is (alkyl)cycloalkylSO.sub.2;
R.sup.6 is hydrogen; and R.sup.8 is alkyl; or a pharmaceutically
acceptable salt thereof.
2. The composition of claim 1 where R.sup.4 is cyclohexyl; R.sup.5
is (methyl)cyclopropylSO.sub.2; and R.sup.8 is methyl; or a
pharmaceutically acceptable salt thereof.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a Continuation of U.S. Non-Provisional
Application Ser. No. 12/272,974 filed Nov. 18, 2008 which claims
the benefit of U.S. Provisional Application Ser. No. 60/989,524,
filed Nov. 21, 2007.
BACKGROUND OF THE INVENTION
[0002] Hepatitis C virus (HCV) is a major human pathogen, infecting
an estimated 170 million persons worldwide--roughly five times the
number infected by human immunodeficiency virus type 1. A
substantial fraction of these HCV infected individuals develop
serious progressive liver disease, including cirrhosis and
hepatocellular carcinoma (Lauer, G. M.; Walker, B. D. N. Engl. J.
Med. 2001, 345, 41-52).
[0003] HCV is a positive-stranded RNA virus. Based on a comparison
of the deduced amino acid sequence and the extensive similarity in
the 5'-untranslated region, HCV has been classified as a separate
genus in the Flaviviridae family. All members of the Flaviviridae
family have enveloped virions that contain a positive stranded RNA
genome encoding all known virus-specific proteins via translation
of a single, uninterrupted, open reading frame.
[0004] Considerable heterogeneity is found within the nucleotide
and encoded amino acid sequence throughout the HCV genome. At least
six major genotypes have been characterized, and more than 50
subtypes have been described. The major genotypes of HCV differ in
their distribution worldwide, and the clinical significance of the
genetic heterogeneity of HCV remains elusive despite numerous
studies of the possible effect of genotypes on pathogenesis and
therapy.
[0005] The single strand HCV RNA genome is approximately 9500
nucleotides in length and has a single open reading frame (ORF)
encoding a single large polyprotein of about 3000 amino acids. In
infected cells, this polyprotein is cleaved at multiple sites by
cellular and viral proteases to produce the structural and
non-structural (NS) proteins. In the case of HCV, the generation of
mature non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A, and
NS5B) is effected by two viral proteases. The first one is believed
to be a metalloprotease and cleaves at the NS2-NS3 junction; the
second one is a serine protease contained within the N-terminal
region of NS3 (also referred to as NS3 protease) and mediates all
the subsequent cleavages downstream of NS3, both in cis, at the
NS3-NS4A cleavage site, and in trans, for the remaining NS4A-NS4B,
NS4B-NS5A, NS5A-NS5B sites. The NS4A protein appears to serve
multiple functions, acting as a cofactor for the NS3 protease and
possibly assisting in the membrane localization of NS3 and other
viral replicase components. The complex formation of the NS3
protein with NS4A seems necessary to the processing events,
enhancing the proteolytic efficiency at all of the sites. The NS3
protein also exhibits nucleoside triphosphatase and RNA helicase
activities. NS5B (also referred to as HCV polymerase) is a
RNA-dependent RNA polymerase that is involved in the replication of
HCV. The HCV NS5B protein is described in "Structural Analysis of
the Hepatitis C Virus RNA Polymerase in Complex with
Ribonucleotides (Bressanelli; S. et al., Journal of Virology 2002,
3482-3492; and Defrancesco and Rice, Clinics in Liver Disease 2003,
7, 211-242.
[0006] Currently, the most effective HCV therapy employs a
combination of alpha-interferon and ribavirin, leading to sustained
efficacy in 40% of patients (Poynard, T. et al. Lancet 1998, 352,
1426-1432). Recent clinical results demonstrate that pegylated
alpha-interferon is superior to unmodified alpha-interferon as
monotherapy (Zeuzem, S. et al. N. Engl. J. Med. 2000, 343,
1666-1672). However, even with experimental therapeutic regimens
involving combinations of pegylated alpha-interferon and ribavirin,
a substantial fraction of patients do not have a sustained
reduction in viral load. Thus, there is a clear and important need
to develop effective therapeutics for treatment of HCV
infection.
DESCRIPTION OF THE INVENTION
[0007] One aspect of the invention is a compound of formula I
##STR00002##
where:
R.sup.1 is CONR.sup.5R.sup.6;
R.sup.2 is
##STR00003##
[0008] R.sup.3 is hydrogen, halo, alkyl, alkenyl, hydroxy,
benzyloxy, alkoxy, or haloalkoxy; R.sup.4 is cycloalkyl; R.sup.5 is
alkenylSO.sub.2, alkynylSO.sub.2, alkoxyalkylSO.sub.2,
(cycloalkyl)alkylSO.sub.2, (alkyl)cycloalkylSO.sub.2,
((cycloalkyl)alkyl)cycloalkylSO.sub.2, (benzyl)cycloalkylSO.sub.2,
(alkenyl)cycloalkylSO.sub.2, (alkynyl)cycloalkylSO.sub.2,
(trialkylsilyl)cycloalkylSO.sub.2,
(CO.sub.2R.sup.7)cycloalkylSO.sub.2, (PhCO)cycloalkylSO.sub.2,
(PhN(R.sup.7)CO)cycloalkylSO.sub.2, tetrahydrofuranylSO.sub.2,
tetrahydropyranylSO.sub.2, (tetrahydrofuranyl)alkylSO.sub.2,
(tetrahydropyranyl)alkylSO.sub.2, isoxazolidinylSO.sub.2,
Ar.sup.1SO.sub.2, (Ar.sup.1)alkylSO.sub.2, Ar.sup.2SO.sub.2, or
(CON(R.sup.7)(R.sup.7))alkyl; R.sup.6 is hydrogen or alkyl; R.sup.7
is hydrogen or alkyl; R.sup.8 is hydrogen, alkyl, cycloalkyl,
(cycloalkyl)alkyl, alkylcarbonyl, cycloalkylcarbonyl,
haloalkylcarbonyl, alkoxycarbonyl, alkylSO.sub.2,
cycloalkylSO.sub.2, haloalkylSO.sub.2, aminocarbonyl,
(alkylamino)carbonyl, (dialkylamino)carbonyl, benzyl,
benzyloxycarbonyl, or pyridinyl; Ar.sup.1 is phenyl, pyridinyl,
pyrimidinyl, quinolinyl, or isoquinolinyl, and is substituted with
0-3 substituents selected from halo, alkyl, alkoxy, and cyano; and
Ar.sup.2 is furanyl, thienyl, pyrrolyl, isoxazolyl, isothiazolyl,
pyrazolyl, oxazolyl, thiazolyl, imidazolyl, oxadiazolyl,
thiadiazolyl, triazolyl, or tetrazolyl, and is substituted with 0-3
substituents selected from halo, alkyl, alkoxy, and cyano; or a
pharmaceutically acceptable salt thereof. Another aspect of the
invention is a compound of formula I where R.sup.3 is hydrogen.
Another aspect of the invention is a compound of formula I where
R.sup.3 is methoxy. Another aspect of the invention is a compound
of formula I where R.sup.4 is cyclohexyl. Another aspect of the
invention is a compound of formula I where Ar.sup.1 is phenyl,
pyridinyl, quinolinyl, or isoquinolinyl, and is substituted with
0-3 substituents selected from halo, alkyl, alkoxy, and cyano.
Another aspect of the invention is a compound of formula I
according to the following stereochemistry.
##STR00004##
Another aspect of the invention is a compound of formula I
according to the following stereochemistry.
##STR00005##
[0009] Any scope of any variable, including R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, Ar.sup.1, and
Ar.sup.2, can be used independently with the scope of any other
instance of a variable.
[0010] Unless specified otherwise, these terms have the following
meanings. "Alkyl" means a straight or branched alkyl group composed
of 1 to 6 carbons. "Alkenyl" means a straight or branched alkyl
group composed of 2 to 6 carbons with at least one double bond.
"Alkynyl" means a straight or branched alkyl group composed of 2 to
6 carbons with at least one triple bond. "Cycloalkyl" means a
monocyclic ring system composed of 3 to 7 carbons. "Hydroxyalkyl,"
"alkoxy" and other terms with a substituted alkyl moiety include
straight and branched isomers composed of 1 to 6 carbon atoms for
the alkyl moiety. "Haloalkyl" and "haloalkoxy" include all
halogenated isomers from monohalo substituted alkyl to perhalo
substituted alkyl. "Aryl" includes carbocyclic and heterocyclic
aromatic substituents. Parenthetic and multiparenthetic terms are
intended to clarify bonding relationships to those skilled in the
art. For example, a term such as ((R)alkyl) means an alkyl
substituent further substituted with the substituent R.
[0011] The invention includes all pharmaceutically acceptable salt
forms of the compounds. Pharmaceutically acceptable salts are those
in which the counter ions do not contribute significantly to the
physiological activity or toxicity of the compounds and as such
function as pharmacological equivalents. These salts can be made
according to common organic techniques employing commercially
available reagents. Some anionic salt forms include acetate,
acistrate, besylate, bromide, chloride, citrate, fumarate,
glucouronate, hydrobromide, hydrochloride, hydroiodide, iodide,
lactate, maleate, mesylate, nitrate, pamoate, phosphate, succinate,
sulfate, tartrate, tosylate, and xinofoate. Some cationic salt
forms include ammonium, aluminum, benzathine, bismuth, calcium,
choline, diethylamine, diethanolamine, lithium, magnesium,
meglumine, 4-phenylcyclohexylamine, piperazine, potassium, sodium,
tromethamine, and zinc.
[0012] Some of the compounds of the invention possess asymmetric
carbon atoms (see, for example, the compound below). The invention
includes all stereoisomeric forms, including enantiomers and
diastereomers as well as mixtures of stereoisomers such as
racemates. Some stereoisomers can be made using methods known in
the art. Stereoisomeric mixtures of the compounds and related
intermediates can be separated into individual isomers according to
methods commonly known in the art. The use of wedges or hashes in
the depictions of molecular structures in the following schemes and
tables is intended only to indicate relative stereochemistry, and
should not be interpreted as implying absolute stereochemical
assignments.
##STR00006##
Synthetic Methods
[0013] The compounds may be made by methods known in the art
including those described below. Some reagents and intermediates
are known in the art. Other reagents and intermediates can be made
by methods known in the art using readily available materials. The
variables (e.g. numbered "R" substituents) used to describe the
synthesis of the compounds are intended only to illustrate how to
make and are not to be confused with variables used in the claims
or in other sections of the specification. Abbreviations used
within the schemes generally follow conventions used in the
art.
[0014] Methyl 2-bromo-3-cyclohexyl-1H-indole-6-carboxylate can be
hydrolyzed to 2-bromo-3-cyclohexyl-1H-indole-6-carboxylic acid (See
Scheme 1). This compound can be condensed with a variety of
sulfonyl ureas, using for example, 1,1'-carbonyldiimidazole in
combination with 1,8-diazabicyclo[5.4.0]undec-7-ene in anhydrous
THF. The resultant acyl sulfamides can be subjected to known
coupling reactions with a diversity of 2-formyl boronic acids or
esters, using for example, Suzuki coupling conditions, to provide
cyclic hemiaminal intermediates of the type depicted. These
compounds can be converted to indolobenzazepines derivatives by
treatment with methyl 2-(dimethoxyphosphoryl)acrylate under the
influence of cesium carbonate in DMF via consecutive Michael and
Horner Emmons reactions.
[0015] Related fused cyclopropyl ester derivatives can be generated
by methods known in the art, including treatment of the
indolobenzazepine esters with trimethyl sulfoxonium iodide under
strongly basic conditions in DMSO. The residual aliphatic ester
moiety in the resultant fused cyclopropanes can be hydrolyzed and
the product acids can be condensed with a variety of alkyl-fused
diamines. For example,
O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
tetrafluoroborate and diisopropyl ethyl amine in DMSO can give
alkyl fused diamine carboxamides.
##STR00007## ##STR00008##
[0016] N-protected diamines can also be coupled to the intermediate
indolobenzazepine acids and the resultant diamine carboxamides can
be deprotected using methods known in the art and derivatized using
a variety of synthetic protocols, some illustrative examples of
which are shown below (See Scheme 2).
##STR00009## ##STR00010##
[0017] An intermediate useful for the synthesis of some compounds
of the invention involves the preparation of the tert-butyl ester
indolobenzazepine shown in Scheme 3.
##STR00011## ##STR00012##
[0018] This methodology involves base catalyzed hydrolysis of the
indole methyl ester shown, followed by its reaction with either
thionyl chloride and potassium tertiary butoxide, or alkylation
with silver carbonate and tertiary butyl bromides. The resultant
compound can be transformed using chemistry analogous to that
outlined previously to provide the mixed ester indolobenzazepines
shown above.
[0019] These intermediates are useful in an alternative procedure
that can be employed for the preparation of acylsulfamide and
acylsulfonamide alkyl-fused diamines, as shown in Scheme 4.
Cyclopropanation of an intermediate t-butyl ester indolobenzazepine
and subsequent cleavage of the t-butyl ester group can generate the
acid which can be coupled to a diversity of sulfonamides and
sulfonylureas. Subsequent hydrolysis affords the related aliphatic
acid, which can be coupled with a diversity of alkyl-fused diamines
For example, O-(1H-benzotriazol-1-yl)-N,N, N',N'-tetramethyluronium
tetrafluoroborate and diisopropyl ethyl amine in DMSO can give the
alkyl fused diamine carboxamides.
##STR00013## ##STR00014##
[0020] Some examples exist as stereoisomeric mixtures. The
invention encompasses all stereoisomers of the compounds. Methods
of fractionating stereoisomeric mixtures are well known in the art,
and include but are not limited to; preparative chiral
supercritical fluid chromatography (SFC) and chiral high
performance liquid chromatography (HPLC). An example using this
approach is shown in scheme 5.
##STR00015## ##STR00016##
[0021] An additional method to achieve such separations involves
the preparation of mixtures of diastereomers which can be separated
using a variety of methods known in the art. One example of this
approach is shown below (Scheme 6).
##STR00017##
[0022] Some diastereomeric amides can be separated using reverse
phase HPLC. After hydroysis, the resultant optically active acids
can be coupled with fused diamine derivatives (Scheme 6). For
example, O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
tetrafluoroborate and diisopropyl ethyl amine in DMSO can be used
to give the alkyl fused carboxamides. Other standard acid amine
coupling methods can also be used to give optically active
carboxamides.
##STR00018##
Biological Methods
[0023] The compounds demonstrated activity against HCV NS5B as
determined in the following HCV RdRp assays.
[0024] HCV NS5B RdRp cloning, expression, and purification. The
cDNA encoding the NS5B protein of HCV, genotype 1b, was cloned into
the pET21a expression vector. The protein was expressed with an 18
amino acid C-terminal truncation to enhance the solubility. The E.
coli competent cell line BL21(DE3) was used for expression of the
protein. Cultures were grown at 37.degree. C. for .about.4 hours
until the cultures reached an optical density of 2.0 at 600 nm. The
cultures were cooled to 20.degree. C. and induced with 1 mM IPTG.
Fresh ampicillin was added to a final concentration of 50 .mu.g/ml
and the cells were grown overnight at 20.degree. C.
[0025] Cell pellets (3 L) were lysed for purification to yield
15-24 mgs of purified NS5B. The lysis buffer consisted of 20 mM
Tris-HCl, pH 7.4, 500 mM NaCl, 0.5% triton X-100, 1 mM DTT, 1 mM
EDTA, 20% glycerol, 0.5 mg/ml lysozyme, 10 mM MgCl.sub.2, 15 ug/ml
deoxyribonuclease I, and Complete TM protease inhibitor tablets
(Roche). After addition of the lysis buffer, frozen cell pellets
were resuspended using a tissue homogenizer. To reduce the
viscosity of the sample, aliquots of the lysate were sonicated on
ice using a microtip attached to a Branson sonicator. The sonicated
lysate was centrifuged at 100,000.times.g for 1 hr at 4.degree. C.
and filtered through a 0.2 .mu.m filter unit (Corning).
[0026] The protein was purified using two sequential chromatography
steps: Heparin sepharose CL-6B and polyU sepharose 4B (Pharmacia).
The chromatography buffers were identical to the lysis buffer but
contained no lysozyme, deoxyribonuclease I, MgCl2 or protease
inhibitor and the NaCl concentration of the buffer was adjusted
according to the requirements for charging the protein onto the
column. Each column was eluted with a NaCl gradient which varied in
length from 5-50 column volumes depending on the column type. After
the final chromatography step, the resulting purity of the enzyme
is >90% based on SDS-PAGE analysis. The enzyme was aliquoted and
stored at -80.degree. C.
[0027] Standard HCV NS5B RdRp enzyme assay. HCV RdRp genotype 1b
assays were run in a final volume of 60 .mu.l in 96 well plates
(Corning 3600). The assay buffer is composed of 20 mM Hepes, pH
7.5, 2.5 mM KCl, 2.5 mM MgCl.sub.2, 1 mM DTT, 1.6 U RNAse inhibitor
(Promega N2515), 0.01 mg/ml BSA (Sigma B6917), and 2% glycerol. All
compounds were serially diluted (3-fold) in DMSO and diluted
further in water such that the final concentration of DMSO in the
assay was 2%. HCV RdRp genotype 1b enzyme was used at a final
concentration of 28 nM. A polyA template was used at 6 nM, and a
biotinylated oligo-dT12 primer was used at 180 nM final
concentration. Template was obtained commercially (Amersham
27-4110). Biotinylated primer was prepared by Sigma Genosys. 3H-UTP
was used at 0.6 .mu.Ci (0.29 .mu.M total UTP). Reactions were
initiated by the addition of enzyme, incubated at 30.degree. C. for
60 min, and stopped by adding 25 .mu.l of 50 mM EDTA containing SPA
beads (4 .mu.g/.mu.l, Amersham RPNQ 0007). Plates were read on a
Packard Top Count NXT after >1 hr incubation at room
temperature.
[0028] Modified HCV NS5B RdRp enzyme assay. A modified enzyme assay
was performed essentially as described for the standard enzyme
assay except for the following: The biotinylated oligo dT12 primer
was precaptured on streptavidin-coated SPA beads by mixing primer
and beads in assay buffer and incubating at room temperature for
one hour. Unbound primer was removed after centrifugation. The
primer-bound beads were resuspended in 20 mM Hepes buffer, pH 7.5
and used in the assay at final concentrations of 20 nM primer and
0.67 .mu.g/.mu.l beads. Order of addition in the assay: enzyme
(1.75 nM) was added to diluted compound followed by the addition of
a mixture of template (0.36 nM), 3H-UTP (0.6 .mu.Ci, 0.29 .mu.M),
and primer-bound beads, to initiate the reaction; concentrations
given are final. Reactions were allowed to proceed for 4 hours at
30.degree. C.
[0029] IC.sub.50 values for compounds were determined using seven
different [I]. IC.sub.50 values were calculated from the inhibition
using the formula y=A+((B-A)/(1+((C/x) D))).
[0030] FRET Assay Preparation. The HCV FRET screening assay was
performed in 96-well cell culture plates. The FRET peptide
(Anaspec, Inc.) (Taliani et al., Anal. Biochem. 1996, 240, 60-67)
contains a fluorescence donor, EDANS, near one end of the peptide
and an acceptor, DABCYL, near the other end. The fluorescence of
the peptide is quenched by intermolecular resonance energy transfer
(RET) between the donor and the acceptor, but as the NS3 protease
cleaves the peptide the products are released from RET quenching
and the fluorescence of the donor becomes apparent. The assay
reagent was made as follows: 5.times. cell Luciferase cell culture
lysis reagent from Promega (#E153A) diluted to 1.times. with
dH.sub.2O, NaCl added to 150 mM final, the FRET peptide diluted to
20 .mu.M final from a 2 mM stock.
[0031] To prepare plates, HCV replicon cells, with or without a
Renilla luciferase reporter gene, were trypsinized and plated in a
96-well plate with titrated test compounds added in columns 3
through 12; columns 1 and 2 contained a control compound (HCV
control inhibitor), and the bottom row contained cells with DMSO
only. The plates were then placed in a CO.sub.2 incubator at
37.degree. C.
[0032] Assays. Subsequent to addition of the test compounds
described above (FRET Assay Preparation), at various times the
plate was removed and Alamar blue solution (Trek Diagnostics,
#00-100) was added to measure cellular toxicity. After reading in a
Cytoflour 4000 instrument (PE Biosystems), plates were rinsed with
PBS and then used for FRET assay by the addition of 30 ul of the
FRET peptide assay reagent described above (FRET Assay Preparation)
per well. The plate was then placed into the Cytoflour 4000
instrument which had been set to 340 excite/490 emission, automatic
mode for up to 20 cycles and the plate read in a kinetic mode.
Typically, the signal to noise using an endpoint analysis after the
reads was at least three-fold. Alternatively, after Alamar blue
reading, plates were rinsed with PBS, then used for luciferase
assay using the Promega Dual-Glo Luciferase Assay System or the
Promega EnduRen Live Cell Substrate assay.
[0033] Compound analysis was performed by quantification of the
relative HCV replicon inhibition and the relative cytotoxicity
values. To calculate cytoxicity values, the average Alamar Blue
fluorescence signals from the control wells were set as 100%
non-toxic. The individual signals in each of the compound test
wells were then divided by the average control signal and
multiplied by 100% to determine percent cytotoxicity. To calculate
the HCV replicon inhibition values, an average background value was
obtained from the two wells containing the highest amount of HCV
control inhibitor at the end of the assay period. These numbers
were similar to those obtained from naive Huh-7 cells. The
background numbers were then subtracted from the average signal
obtained from the control wells and this number was used as 100%
activity. The individual signals in each of the compound test wells
were then divided by the averaged control values after background
subtraction and multiplied by 100% to determine percent activity.
EC.sub.50 values were calculated as the concentration which caused
a 50% reduction in FRET or luciferase activity. The two numbers
generated for the compound plate, percent cytoxicity and percent
activity, were used to determine compounds of interest for further
analysis.
[0034] Representative data for compounds are reported in Table
1.
TABLE-US-00001 TABLE 1 Structure IC.sub.50 EC.sub.50 ##STR00019## B
B ##STR00020## B B ##STR00021## B B ##STR00022## B B ##STR00023## B
B ##STR00024## B B ##STR00025## B B ##STR00026## B B ##STR00027## B
B ##STR00028## B B ##STR00029## B B ##STR00030## B B ##STR00031## B
B ##STR00032## B B ##STR00033## B B ##STR00034## B B ##STR00035## B
B ##STR00036## B B ##STR00037## B B ##STR00038## B B ##STR00039## B
B ##STR00040## B E ##STR00041## B B ##STR00042## B B ##STR00043## B
B ##STR00044## B B ##STR00045## B B ##STR00046## B B ##STR00047## B
B ##STR00048## B B ##STR00049## B B ##STR00050## B B ##STR00051##
##STR00052## ##STR00053## B B ##STR00054## B B ##STR00055## B B
##STR00056## B B ##STR00057## B B ##STR00058## B B ##STR00059##
##STR00060##
A>0.5 .mu.M; B 0.0042 .mu.M-0.5 .mu.M; C<0.02 .mu.M but an
exact value was not determined; D>0.04 .mu.M but an exact value
was not determined; E<0.07 .mu.M but an exact value was not
determined; F>1.0 .mu.M but an exact value was not
determined
Pharmaceutical Compositions and Methods of Treatment
[0035] The compounds demonstrate activity against HCV NS5B and can
be useful in treating HCV and HCV infection. Therefore, another
aspect of the invention is a composition comprising a compound, or
a pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
[0036] Another aspect of the invention is a composition further
comprising a compound having anti-HCV activity.
[0037] Another aspect of the invention is a composition where the
compound having anti-HCV activity is an interferon. Another aspect
of the invention is where the interferon is selected from
interferon alpha 2B, pegylated interferon alpha, consensus
interferon, interferon alpha 2A, and lymphoblastoid interferon
tau.
[0038] Another aspect of the invention is a composition where the
compound having anti-HCV activity is a cyclosporin. Another aspect
of the invention is where the cyclosporin is cyclosporin A.
[0039] Another aspect of the invention is a composition where the
compound having anti-HCV activity is selected from the group
consisting of interleukin 2, interleukin 6, interleukin 12, a
compound that enhances the development of a type 1 helper T cell
response, interfering RNA, anti-sense RNA, Imiqimod, ribavirin, an
inosine 5'-monophospate dehydrogenase inhibitor, amantadine, and
rimantadine.
[0040] Another aspect of the invention is a composition where the
compound having anti-HCV activity is effective to inhibit the
function of a target selected from HCV metalloprotease, HCV serine
protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV
entry, HCV assembly, HCV egress, HCV NS5A protein, IMPDH, and a
nucleoside analog for the treatment of an HCV infection.
[0041] Another aspect of the invention is a composition comprising
a compound, or a pharmaceutically acceptable salt thereof, a
pharmaceutically acceptable carrier, an interferon and
ribavirin.
[0042] Another aspect of the invention is a method of inhibiting
the function of the HCV replicon comprising contacting the HCV
replicon with a compound or a pharmaceutically acceptable salt
thereof.
[0043] Another aspect of the invention is a method of inhibiting
the function of the HCV NS5B protein comprising contacting the HCV
NS5B protein with a compound or a pharmaceutically acceptable salt
thereof.
[0044] Another aspect of the invention is a method of treating an
HCV infection in a patient comprising administering to the patient
a therapeutically effective amount of a compound or a
pharmaceutically acceptable salt thereof. In another embodiment the
compound is effective to inhibit the function of the HCV replicon.
In another embodiment the compound is effective to inhibit the
function of the HCV NS5B protein.
[0045] Another aspect of the invention is a method of treating an
HCV infection in a patient comprising administering to the patient
a therapeutically effective amount of a compound, or a
pharmaceutically acceptable salt thereof, in conjunction with
(prior to, after, or concurrently) another compound having anti-HCV
activity.
[0046] Another aspect of the invention is the method where the
other compound having anti-HCV activity is an interferon.
[0047] Another aspect of the invention is the method where the
interferon is selected from interferon alpha 2B, pegylated
interferon alpha, consensus interferon, interferon alpha 2A, and
lymphoblastoid interferon tau.
[0048] Another aspect of the invention is the method where the
other compound having anti-HCV activity is a cyclosporin.
[0049] Another aspect of the invention is the method where the
cyclosporin is cyclosporin A.
[0050] Another aspect of the invention is the method where the
other compound having anti-HCV activity is selected from
interleukin 2, interleukin 6, interleukin 12, a compound that
enhances the development of a type 1 helper T cell response,
interfering RNA, anti-sense RNA, Imiqimod, ribavirin, an inosine
5'-monophospate dehydrogenase inhibitor, amantadine, and
rimantadine.
[0051] Another aspect of the invention is the method where the
other compound having anti-HCV activity is effective to inhibit the
function of a target selected from the group consisting of HCV
metalloprotease, HCV serine protease, HCV polymerase, HCV helicase,
HCV NS4B protein, HCV entry, HCV assembly, HCV egress, HCV NS5A
protein, IMPDH, and a nucleoside analog for the treatment of an HCV
infection.
[0052] Another aspect of the invention is the method where the
other compound having anti-HCV activity is effective to inhibit the
function of target in the HCV life cycle other than the HCV NS5B
protein.
[0053] "Therapeutically effective" means the amount of agent
required to provide a meaningful patient benefit as understood by
practitioners in the field of hepatitis and HCV infection.
[0054] "Patient" means a person infected with the HCV virus and
suitable for therapy as understood by practitioners in the field of
hepatitis and HCV infection.
[0055] "Treatment," "therapy," "regimen," "HCV infection," and
related terms are used as understood by practitioners in the field
of hepatitis and HCV infection.
[0056] The compounds of this invention are generally given as
pharmaceutical compositions comprised of a therapeutically
effective amount of a compound or its pharmaceutically acceptable
salt and a pharmaceutically acceptable carrier and may contain
conventional excipients. A therapeutically effective amount is that
which is needed to provide a meaningful patient benefit.
Pharmaceutically acceptable carriers are those conventionally known
carriers having acceptable safety profiles. Compositions encompass
all common solid and liquid forms including capsules, tablets,
losenges, and powders as well as liquid suspensions, syrups,
elixers, and solutions. Compositions are made using common
formulation techniques, and conventional excipients (such as
binding and wetting agents) and vehicles (such as water and
alcohols) are generally used for compositions.
[0057] Solid compositions are normally formulated in dosage units
and compositions providing from about 1 to 1000 mg of the active
ingredient per dose are preferred. Some examples of dosages are 1
mg, 10 mg, 100 mg, 250 mg, 500 mg, and 1000 mg. Generally, other
agents will be present in a unit range similar to agents of that
class used clinically. Typically, this is 0.25-1000 mg/unit.
[0058] Liquid compositions are usually in dosage unit ranges.
Generally, the liquid composition will be in a unit dosage range of
1-100 mg/mL. Some examples of dosages are 1 mg/mL, 10 mg/mL, 25
mg/mL, 50 mg/mL, and 100 mg/mL. Generally, other agents will be
present in a unit range similar to agents of that class used
clinically. Typically, this is 1-100 mg/mL.
[0059] The invention encompasses all conventional modes of
administration; oral and parenteral methods are preferred.
Generally, the dosing regimen will be similar to other agents used
clinically. Typically, the daily dose will be 1-100 mg/kg body
weight daily. Generally, more compound is required orally and less
parenterally. The specific dosing regime, however, will be
determined by a physician using sound medical judgement.
[0060] The invention also encompasses methods where the compound is
given in combination therapy. That is, the compound can be used in
conjunction with, but separately from, other agents useful in
treating hepatitis and HCV infection. In these combination methods,
the compound will generally be given in a daily dose of 1-100 mg/kg
body weight daily in conjunction with other agents. The other
agents generally will be given in the amounts used therapeutically.
The specific dosing regime, however, will be determined by a
physician using sound medical judgement.
[0061] Some examples of compounds suitable for compositions and
methods are listed in Table 2.
TABLE-US-00002 TABLE 2 Type of Brand Name Inhibitor or Target
Source Company Omega IFN IFN-.omega. Intarcia Therapeutics
BILN-2061 serine protease inhibitor Boehringer Ingelheim Pharma KG,
Ingelheim, Germany Summetrel antiviral Endo Pharmaceuticals
Holdings Inc., Chadds Ford, PA Roferon A IFN-.alpha.2a F.
Hoffmann-La Roche LTD, Basel, Switzerland Pegasys PEGylated
IFN-.alpha.2a F. Hoffmann-La Roche LTD, Basel, Switzerland Pegasys
and PEGylated IFN- F. Hoffmann-La Roche Ribavirin
.alpha.2a/ribavirin LTD, Basel, Switzerland CellCept HCV IgG F.
Hoffmann-La Roche immunosuppressant LTD, Basel, Switzerland
Wellferon lymphoblastoid IFN-.alpha.n1 GlaxoSmithKline plc,
Uxbridge, UK Albuferon-.alpha. albumin IFN-.alpha.2b Human Genome
Sciences Inc., Rockville, MD Levovirin ribavirin ICN
Pharmaceuticals, Costa Mesa, CA IDN-6556 caspase inhibitor Idun
Pharmaceuticals Inc., San Diego, CA IP-501 antifibrotic Indevus
Pharmaceuticals Inc., Lexington, MA Actimmune INF-.gamma. InterMune
Inc., Brisbane, CA Infergen A IFN alfacon-1 InterMune
Pharmaceuticals Inc., Brisbane, CA ISIS 14803 antisense ISIS
Pharmaceuticals Inc, Carlsbad, CA/Elan Phamaceuticals Inc., New
York, NY JTK-003 RdRp inhibitor Japan Tobacco Inc., Tokyo, Japan
Pegasys and PEGylated IFN-.alpha.2a/ Maxim Pharmaceuticals Ceplene
immune modulator Inc., San Diego, CA Ceplene immune modulator Maxim
Pharmaceuticals Inc., San Diego, CA Civacir HCV IgG Nabi
Biopharmaceuticals immunosuppressant Inc., Boca Raton, FL Intron A
IFN-.alpha.2b/.alpha.1-thymosin RegeneRx and Zadaxin
Biopharmiceuticals Inc., Bethesda, MD/ SciClone Pharmaceuticals
Inc, San Mateo, CA Levovirin IMPDH inhibitor Ribapharm Inc., Costa
Mesa, CA Viramidine Ribavirin Prodrug Ribapharm Inc., Costa Mesa,
CA Heptazyme ribozyme Ribozyme Pharmaceuticals Inc., Boulder, CO
Intron A IFN-.alpha.2b Schering-Plough Corporation, Kenilworth, NJ
PEG-Intron PEGylated IFN-.alpha.2b Schering-Plough Corporation,
Kenilworth, NJ Rebetron IFN-.alpha.2b/ribavirin Schering-Plough
Corporation, Kenilworth, NJ Ribavirin ribavirin Schering-Plough
Corporation, Kenilworth, NJ PEG-Intron/ PEGylated IFN-
Schering-Plough Ribavirin .alpha.2b/ribavirin Corporation,
Kenilworth, NJ Zadazim Immune modulator SciClone Pharmaceuticals
Inc., San Mateo, CA Rebif IFN-.beta.1a Serono, Geneva, Switzerland
IFN-.beta. and IFN-.beta. and EMZ701 Transition Therapeutics EMZ701
Inc., Ontario, Canada Batabulin .beta.-tubulin inhibitor Tularik
Inc., South San (T67) Francisco, CA Merimepodib IMPDH inhibitor
Vertex Pharmaceuticals (VX-497) Inc., Cambridge, MA Telaprevir NS3
serine protease Vertex Pharmaceuticals (VX-950, inhibitor Inc.,
Cambridge, MA/Eli LY-570310) Lilly and Co. Inc., Indianapolis, IN
Omniferon natural IFN-.alpha. Viragen Inc., Plantation, FL XTL-6865
monoclonal antibody XTL Biopharmaceuticals (XTL-002) Ltd., Rehovot,
Isreal HCV-796 NS5B Replicase Inhibitor Wyeth/Viropharma NM-283
NS5B Replicase Inhibitor Idenix/Novartis GL-59728 NS5B Replicase
Inhibitor Gene Labs/Novartis GL-60667 NS5B Replicase Inhibitor Gene
Labs/Novartis 2'C MeA NS5B Replicase Inhibitor Gilead PSI 6130 NS5B
Replicase Inhibitor Roche R1626 NS5B Replicase Inhibitor Roche SCH
503034 serine protease inhibitor Schering Plough NIM811 Cyclophilin
Inhibitor Novartis Suvus Methylene blue Bioenvision Multiferon Long
lasting IFN Viragen/Valentis Actilon TLR9 agonist Coley (CPG10101)
Interferon-.beta. Interferon-.beta.-1a Serono Zadaxin
Immunomodulator Sciclone Pyrazolo- HCV Inhibitors Arrow
Therapeutics Ltd. pyrimidine compounds and salts From WO-2005047288
26 May 2005 2'C Methyl NS5B Replicase Inhibitor Merck adenosine
GS-9132 HCV Inhibitor Achillion/Gilead (ACH-806)
DESCRIPTION OF SPECIFIC EMBODIMENTS
[0062] Unless otherwise specified, analytical LCMS data on the
following intermediates and examples were acquired using the
following columns and conditions. Stop time: Gradient time+1
minute; Starting conc: 0% B unless otherwise noted; Eluent A: 5%
CH.sub.3CN/95% H.sub.2O with 10 mM NH.sub.4OAc (for columns A, D
and E); 10% MeOH/90% H.sub.2O with 0.1% TFA (for columns B and C);
Eluent B: 95% CH.sub.3CN/5% H.sub.2O with 10 mM NH.sub.4OAc (for
columns A, D and E); 90% MeOH/10% H.sub.2O with 0.1% TFA (for
columns B and C); Column A: Phenomenex 10.mu. 4.6.times.50 mm C18;
Column B: Phenomenex C18 10.mu. 3.0.times.50 mm; Column C.
Phenomenex 4.6.times.50 mm C18 10.mu.; Column D: Phenomenex Lina
C18 5.mu. 3.0.times.50 mm; Column E. Phenomenex 5.mu. 4.6.times.50
mm C18.
[0063] As an artifact of the graphics software, some structures
have missing hydrogen atoms.
##STR00061##
[0064] 1H-Indole-6-carboxylic acid, 2-bromo-3-cyclohexyl-, methyl
ester. Freshly recrystallized pyridinium tribromide
(recrystallization from hot AcOH (5 mL per 1 g), rinsed with cold
AcOH and dried under high vacuum over KOH) was added in portions
(over 10 min.) to a stirring solution of methyl
3-cyclohexyl-1H-indole-6-carboxylate (60 g, 233 mmol) (prepared
using procedures describe in WO2004/065367) in CHCl.sub.3/THF (1:1,
1.25 L) at 2.degree. C. The reaction solution was stirred at
0-5.degree. C. for 2.5 h, and washed with sat. aq. NaHSO.sub.3 (1
L), 1 N HCl (1 L) and brine (1 L). The organic layer was dried
(MgSO.sub.4) and concentrated. The resulting red oil was diluted
with Et.sub.2O and concentrated. The resulting pink solid was
dissolved into Et.sub.2O (200 mL) treated with hexanes (300 mL) and
partially concentrated. The solids were collected by filtration and
rinsed with hexanes. The mother liquor was concentrated to dryness
and the procedure repeated. The solids were combined to yield
1H-indole-6-carboxylic acid, 2-bromo-3-cyclohexyl-, methyl ester
(64 g, 190 mmol, 82%) as a fluffy pink solid, which was used
without further purification. .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 8.47 (br s, 1H), 8.03 (d, J=1.4 Hz, 1H), 7.74 (dd, J=1.4,
8.8 Hz, 1H), 7.69 (d, J=8.8 Hz, 1H), 3.92 (s, 3H), 2.82 (tt, J=3.7,
11.7 Hz, 1H), 1.98-1.72 (m, 7H), 1.50-1.27 (m, 3H). 13CNMR (75 MHz,
CDCl.sub.3) .delta. 168.2, 135.6, 130.2, 123.1, 120.8, 120.3,
118.7, 112.8, 110.7, 52.1, 37.0, 32.2 (2), 27.0 (2), 26.1. LCMS:
m/e 334 (M-H).sup.-, ret time 3.34 min, column A, 4 minute
gradient.
##STR00062##
[0065] 1H-Indole-6-carboxylic acid, 2-bromo-3-cyclohexyl-. A
solution of methyl 2-bromo-3-cyclohexyl-1H-indole-6-carboxylate (20
g, 60 mmol) and LiOH (3.8 g, 160 mmol) in MeOH/THF/H.sub.2O (1:1:1,
300 mL) was heated at 90.degree. C. for 2 h. The reaction mixture
was cooled in an ice/H.sub.2O bath, neutralized with 1M HCl
(.about.160 mL) diluted with H.sub.2O (250 mL) and stirred for 1 h
at rt. The precipitates were collected by filtration rinse with
H.sub.2O and dried to yield 1H-indole-6-carboxylic acid,
2-bromo-3-cyclohexyl- (quant.) which was used without further
purification.
[0066] An alternative procedure that can by used to provide
1H-indole-6-carboxylic acid, 2-bromo-3-cyclohexyl- is described
below:
[0067] A solution of methyl
2-bromo-3-cyclohexyl-1H-indole-6-carboxylate (117 g, 349 mmol) and
LiOH.H.sub.2O (26.4 g, 629 mmol) in MeOH/THF/H.sub.2O (1:1:1, 1.8
L) was heated at reflux for 3 h. The reaction mixture was cooled in
an ice/H.sub.2O bath to .about.2.degree. C., neutralized with 1M
HCl (-650 mL) (added at such a rate that temperature did not exceed
5.degree. C.), diluted with H2O (1 L) and stirred while warming to
ambient temperature. The precipitates were collected by filtration
rinsed with H.sub.2O and dried to yield the mono THF solvate of
1H-indole-6-carboxylic acid, 2-bromo-3-cyclohexyl- (135.5 g, 345
mmol, 99%) as a yellow solid, which was used without further
purification. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 11.01 (br
s, 1H), 8.77 (s, 1H), 8.07 (d, J=1.5 Hz, 1H), 7.82 (dd, J=1.5, 8.8
Hz, 1H), 7.72 (d, J=8.8 Hz, 1H), 3.84-3.74 (m, 4H), 2.89 (m, 1H),
1.98-1.72 (m, 11H), 1.50-1.24 (m, 3H). 13CNMR (75 MHz, CDCl.sub.3)
.delta. 172.7, 135.5, 130.7, 122.3, 120.9 (2), 118.8, 113.3, 111.1,
67.9 (2), 37.0, 32.2 (2), 27.0 (2), 26.1, 25.5 (2). LCMS: m/e 320
(M-H).sup.-, ret time 2.21 min, column A, 4 minute gradient.
##STR00063##
[0068] 1H-Indole-6-carboxamide,
2-bromo-3-cyclohexyl-N-[(dimethylamino)sulfonyl]-.
1,1'-Carbonyldiimidazole (1.17 g, 7.2 mmol) was added to a stirred
solution of 2-bromo-3-cyclohexyl-1H-indole-6-carboxylic acid (2.03
g, 6.3 mmol) in THF (6 mL) at 22.degree. C. The evolution of
CO.sub.2 was instantaneous and when it slowed the solution was
heated at 50.degree. C. for 1 hr and then cooled to 22.degree. C.
N,N-Dimethylsulfamide (0.94 g, 7.56 mmol) was added followed by the
dropwise addition of a solution of DBU (1.34 g, 8.8 mmol) in THF (4
mL). Stirring was continued for 24 hr. The mixture was partitioned
between ethyl acetate and dilute HCl. The ethyl acetate layer was
washed with water followed by brine and dried over
Na.sub.2SO.sub.4. The extract was concentrated to dryness to leave
the title product as a pale yellow friable foam, (2.0 g, 74%,
>90% purity, estimated from NMR). .sup.1H NMR (300 MHz, DMSO-D6)
.delta. ppm 1.28-1.49 (m, 3H) 1.59-2.04 (m, 7H) 2.74-2.82 (m, 1H)
2.88 (s, 6H) 7.57 (dd, J=8.42, 1.46 Hz, 1H) 7.74 (d, J=8.78 Hz, 1H)
7.91 (s, 1H) 11.71 (s, 1H) 12.08 (s, 1H).
[0069] An alternative method for the preparation of
1H-indole-6-carboxamide,
2-bromo-3-cyclohexyl-N-[(dimethylamino)sulfonyl]-is described
below.
[0070] To a 1 L four necked round bottom flask equipped with a
mechanical stirrer, a temperature controller, a N2 inlet, and a
condenser, under N2, was added
2-bromo-3-cyclohexyl-1H-indole-6-carboxylic acid (102.0 g, 0.259
mol) and dry THF (300 mL). After stirring for 10 min, CDI (50.3 g,
0.31 mol) was added portion wise. The reaction mixture was then
heated to 50.degree. C. for 2 h. After cooling to 30.degree. C.,
N,N-dimethylaminosulfonamide (41.7 g, 0.336 mol) was added in one
portion followed by addition of DBU (54.1 mL, 0.362 mol) drop wise
over a period of 1 h. The reaction mixture was then stirred at rt
for 20 h. The solvent was removed in vacuo and the residue was
partitioned between EtOAc and 1 N HCl (1:1, 2 L). The organic layer
was separated and the aqueous layer was extracted with EtOAc (500
mL). The combined organic layers were washed with brine (1.5 L) and
dried over MgSO4. The solution was filtered and concentrated in
vacuo to give the crude product (111.0 g). The crude product was
suspended in EtOAc (400 mL) at 60.degree. C. To the suspension was
added heptane (2 L) slowly. The resulting suspension was stirred
and cooled to 0.degree. C. It was then filtered. The filter cake
was rinsed with small amount of heptane and house vacuum air dried
for 2 days. The product was collected as a white solid (92.0 g,
83%). .sup.1H NMR (MeOD, 300 MHz) .delta. 7.89 (s, H), 7.77 (d,
J=8.4 Hz, 1H), 7.55 (dd, J=8.4 and 1.8 Hz, 1H), 3.01 (s, 6H),
2.73-2.95 (m, 1H), 1.81-2.05 (m, 8H), 1.39-1.50 (m, 2H); m/z 429
(M+H)+.
##STR00064##
[0071] 1H-Indole-6-carboxamide,
3-cyclohexyl-N-[(dimethylamino)sulfonyl]-2-(2-formyl-4-methoxyphenyl)-.
A mixture of the
2-Bromo-3-cyclohexyl-N-[(dimethylamino)sulfonyl]-1H-indole-6-carboxamide
(4.28 g, 0.01 mol), 4-methoxy-2-formylphenyl boronic acid (2.7 g,
0.015 mol), 2-dicyclohexylphosphino-2',6'-dimethoxy-biphenyl (41
mg, 0.0001 mol), palladium acetate (11.2 mg), and finely ground
potassium carbonate (4.24 g, 0.02 mol) in toluene (30 mL) was
stirred under reflux and under nitrogen for 30 min, at which time
LC/MS analysis showed the reaction to be complete. The reaction
mixture was then diluted with ethyl acetate and water, and then
acidified with an excess of dilute HCl. The ethyl acetate layer was
then collected and washed with dilute HCl, water and brine. The
organic solution was then dried (magnesium sulfate), filtered and
concentrated to give a gum. The gum was diluted with hexanes (250
ml) and ethyl acetate (25 mL), and the mixture was stirred for 20
hr at 22.degree. C. during which time the product was transformed
into a bright yellow granular solid (4.8 g) which was used directly
without further purification.
[0072] An alternative procedure for the preparation of
1H-indole-6-carboxamide,
3-cyclohexyl-N-[(dimethylamino)sulfonyl]-2-(2-formyl-4-methoxyphenyl)-
is provided below:
[0073] To a slurried solution of
2-bromo-3-cyclohexyl-N-[(dimethylamino)sulfonyl]-indole-6-carboxamide
(54.0 g, 126 mmol), 4-methoxy-2-formylphenylboronic acid (29.5 g,
164 mmol) and LiCl (13.3 g, 315 mmol) in EtOH/toluene (1:1, 1 L)
was added a solution of Na.sub.2CO.sub.3 (40.1 g, 379 mmol) in
water (380 mL). The reaction mixture was stirred 10 min. and then
Pd(PPh3)4 (11.3 g, 10.0 mmol) was added. The reaction solution was
flushed with nitrogen and heated at 70.degree. C. (internal
monitoring) overnight and then cooled to rt. The reaction was
diluted with EtOAc (1 L) and EtOH (100 mL), washed carefully with
1N aqueous HCl (1 L) and brine (500 mL), dried (MgSO4), filtered
and concentrated. The residual solids were stirred with Et2O (600
mL) for 1 h and collected by filtration to yield
1H-indole-6-carboxamide,
3-cyclohexyl-N-[(dimethylamino)sulfonyl]-2-(2-formyl-4-methoxyphenyl)-
(52.8 g, 109 mmol, 87%) as a yellow powder which was used without
further purification. 1H NMR (300 MHz, d6-DMSO) .delta. 11.66 (s,
1H), 8.17 (s, 1H), 7.75 (d, J=8.4 Hz, 1H), 7.74 (d, J=8.4 Hz, 1H),
7.59 (dd, J=1.4, 8.4 Hz, 1H), 7.23-7.16 (m, 2H), 7.08 (dd, J=2.6,
8.4 Hz, 1H), 6.54 (d, J=8.8 Hz, 1H), 3.86 (s, 3H), 3.22-3.08 (m,
1H), 2.91 (s, 6H), 2.00-1.74 (m, 7H), 1.60-1.38 (m, 3H). 13CNMR (75
MHz, CDCl3) .delta. 165.7, 158.8, 147.2, 139.1, 134.3, 132.0,
123.4, 122.0, 119.2, 118.2, 114.8, 112.3, 110.4, 109.8, 79.6, 45.9,
37.2 (2), 34.7, 32.0 (2), 25.9 (2), 24.9. LCMS: m/e 482
(M-H).sup.-, ret time 2.56 min, column A, 4 minute gradient.
##STR00065##
[0074] 6H-Isoindolo[2,1-a]indole-3-carboxamide,
11-cyclohexyl-N-[(dimethylamino)sulfonyl]-6-ethoxy-8-methoxy-. To a
5 L four necked round bottom flask equipped with a temperature
controller, a condenser, a N2 inlet and a mechanical stirrer, was
charged toluene (900 mL), EtOH (900 mL),
2-bromo-3-cyclohexyl-N-(N,N-dimethylsulfamoyl)-1H-indole-6-carboxamide
(90 g, 0.21 mol), 2-formyl-4-methoxyphenylboronic acid (49.2 g,
0.273 mol) and LiCl (22.1 g, 0.525 mol). The resulting solution was
bubbled with N.sub.2 for 15 mins. A solution of Na.sub.2CO.sub.3
(66.8 g, 0.63 mol) in H.sub.2O (675 mL) was added and the reaction
mixture was bubbled with N.sub.2 for another (10 mins).
Pd(PPh.sub.3).sub.4 (7.0 g, 6.3 mmol) was added and the reaction
mixture was heated to 70.degree. C. for 20 h. After cooling to
35.degree. C., a solution of 1 N HCl (1.5 L) was added slowly. The
resulting mixture was transferred to a 6 L separatory funnel and
extracted with EtOAc (2.times.1.5 L). The combined organic extracts
were washed with brine (2 L), dried over MgSO4, filtered and
concentrated in vacuo to give a yellow solid, which was triturated
with 20% EtOAc in hexane (450 mL, 50.degree. C. to 0.degree. C.) to
give
3-cyclohexyl-N-(N,N-dimethylsulfamoyl)-2-(2-formyl-4-methoxyphenyl)-1H-in-
dole-6-carboxamide (65.9 g) as a yellow solid. HPLC purity,
98%.
[0075] The mother liquid from the trituration was concentrated in
vacuo. The residue was refluxed with EtOH (50 mL) for 3 h. The
solution was then cooled to 0.degree. C. The precipitates were
filtered and washed with cooled TBME (5.degree. C.) (20 mL). The
filter cake was house vacuum air dried to give a further quantity
of the title compound as a white solid (16.0 g). HPLC purity, 99%.
.sup.1H NMR (CDCl3, 300 MHz) .delta. 8.75 (s, 1H), 7.96 (s, 1H),
7.73 (d, J=8.4 Hz, 1H), 7.67 (d, J=8.4 Hz, 1H), 7.45 (dd, J=8.4 and
1.4 Hz, 1H), 7.09 (d, J=2.2 Hz, 1H), 6.98 (dd, J=8.4 and 2.2 Hz,
1H), 6.50 (s, 1H), 3.86 (s, 3H), 3.05 (s, 6H), 2.92-3.13 (m, 3H),
1.85-1.93 (m, 7H), 1.40-1.42 (m, 3H), 1.05 (t, J=7.1 Hz, 3H). m/z
512 (M+H)+.
##STR00066##
[0076] 1H-indole-6-carboxamide,
3-cyclohexyl-N-[(dimethylamino)sulfonyl]-2-(2-formyl-4-methoxyphenyl)-.
11-cyclohexyl-N-(N,N-dimethylsulfamoyl)-6-ethoxy-8-methoxy-6H-isoindolo[2-
,1-a]indole-3-carboxamide was dissolved in THF (75 mL). To the
solution was added a solution of 2 N HCl (300 mL). The mixture was
vigorously stirred under N2 at rt for 16 h. The resulting
suspension was filtered and washed with cooled TBME (2.times.30
mL). the filer cake was vacuum air dried overnight to give the
title compound as a yellow solid. HPLC purity, 99% .sup.1H NMR
(DMSO-d6, 300 MHz) .delta. 11.65 (s, 1H), 8.16 (s, 1H), 7.76 (d,
J=5.9 Hz, 1H), 7.73 (d, J=5.9 Hz, 1H), 7.58 (dd, J=8.5 and 1.5 Hz,
1H), 7.17-7.20 (m, 2H), 7.08 (dd, J=8.5 and 1.4 Hz, 1H), 6.55 (d,
J=8.6 Hz, 1H), 3.86 (s, 3H), 3.14-3.18 (m, 1H), 2.91 (s, 6H),
1.75-1.99 (m, 7H), 1.48-1.60 (m, 3H); m/z 484 (M+H)+.
##STR00067##
[0077] 7H-Indolo[2,1-a][2]benzazepine-6-carboxylic acid,
13-cyclohexyl-10-[[[(dimethylamino)sulfonyl]amino]carbonyl]-3-methoxy-,
methyl ester. A mixture of the
3-cyclohexyl-N-(N,N-dimethylsulfamoyl)-2-(2-formyl-4-methoxyphenyl)-1H-in-
dole-6-carboxamide (4.8 g, 0.01 mol), methyl
2-(dimethoxyphosphoryl)acrylate (9.7 g, 0.02 mol) and cesium
carbonate (7.1 g, 0.02 mol) in DMF (28 mL) was stirred for 20 hr at
an oil bath temperature of 55.degree. C. The mixture was poured
into ice-water and acidified with dilute HCl to precipitate the
crude product. The solid was collected, dried and flash
chromatographed on SiO.sub.2 (110 g) using an ethyl acetate and
methylene chloride (1:10) solution containing 2% acetic acid.
Homogeneous fractions were combined and evaporated to afford the
title compound as a pale yellow solid (3.9 g, 71% yield). MS: 552
(M=H+).
[0078] An alternate procedure for the preparation of
7H-indolo[2,1-a][2]benzazepine-6-carboxylic acid,
13-cyclohexyl-10-[[[(dimethylamino)sulfonyl]amino]carbonyl]-3-methoxy-,
methyl ester is provided below.
[0079] A solution of
11-cyclohexyl-N-[(dimethylamino)sulfonyl]-6-hydroxy-8-methoxy-6H-isoindol-
o[2,1-a]indole-3-carboxamide (cyclic hemiaminal) (63.0 g, 130
mmol), methyl 2-(dimethoxyphosphoryl)acrylate (60 g, 261 mmol),
cesium carbonate (106 g, 326 mmol) in DMF (400 mL) was heated at
60.degree. C. (bath temp) for 4.5 h. Additional methyl
2-(dimethoxyphosphoryl)acrylate (15 g, 65 mmol) and cesium
carbonate (21.2 g, 65 mmol) were added and the reaction was heated
at 60.degree. C. overnight then and cooled to rt. The stirring
reaction mixture was diluted with H.sub.2O (1 L), slowly
neutralized with 1N aqueous HCl (800 mL), stirred 3 h, and then the
precipitates were collected by filtration. The solids were
triturated with Et2O (800 mL) and dried to yield methyl
7H-indolo[2,1-a][2]benzazepine-6-carboxylic acid,
13-cyclohexyl-10-[[[(dimethylamino)sulfonyl]amino]carbonyl]-3-metho-
xy-, methyl ester (70.2 g, 127 mmol, 98%) as a yellow solid which
was used without further purification. 1H NMR (300 MHz, CDCl3)
.delta. 8.67 (s, 1H), 8.09 (s, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.80
(s, 1H), 7.50 (d, J=8.4 Hz, 1H), 7.42 (d, J=8.8 Hz, 1H), 7.08 (dd,
J=2.6, 8.8 Hz, 1H), 6.98 (d, J=2.6 Hz, 1H), 5.75-5.51 (m, 1H),
4.29-4.01 (m, 1H), 3.89 (s, 3H), 3.82 (s, 3H), 3.05 (s, 6H),
2.87-2.73 (m, 1H), 2.11-1.12 (m, 10H). LCMS: m/e 550 (M-H)-, ret
time 3.21 min, column A, 4 minute gradient.
##STR00068##
[0080] Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-1-
1-methoxy-, methyl ester, (+/-)-. DMSO (5 mL) was added to a
mixture of trimethylsulfoxonium iodide (199 mg, 0.906 mmol) and NaH
(38 mg in 60% oil dispersion, 0.953 mmol) in a round-bottomed
flask. The reaction mixture was stirred at rt for 0.5 hr.
7H-Indolo[2,1-a][2]benzazepine-6-carboxylic acid,
13-cyclohexyl-10-[[[(dimethylamino)sulfonyl]amino]carbonyl]-3-(methoxy)-,
methyl ester (125 mg, 0.227 mmol) was then added and the reaction
mixture was stirred at rt. for 3 hr., and then at 50.degree. C. for
a further 3 hr. The reaction was then quenched with water and
acidified with 1N HCl solution. The crude product then precipitated
as a light yellow solid which was collected by filtration and air
dried, (106 mg, 83% yield). 6 mg of this material was then purified
by Prep. HPLC to afford the title compound as a light yellow solid
(1.8 mg). MS m/z 566 (MH.sup.+), Retention time: 3.850 min. 1H NMR
(500 MHz, MeOD) .delta. ppm 0.28 (m, 0.36H) 1.19-2.20 (m, 11.64H)
2.70-3.02 (m, 2H) 3.03 (s, 2.16H) 3.05 (s, 3.84H) 3.49 (d, J=15.26
Hz, 0.64H) 3.54 (s, 1.92H) 3.83 (s, 1.08H) 3.91 (s, 3H) 4.08 (d,
J=15.26 Hz, 0.36H) 5.29 (d, J=15.26 Hz, 0.36H) 5.50 (d, J=14.95 Hz,
0.64H) 6.98-7.06 (m, 1H) 7.16 (d, J=2.44 Hz, 0.36H) 7.23 (d, J=2.44
Hz, 0.64H) 7.30 (d, J=8.55 Hz, 0.64H) 7.34 (d, J=8.55 Hz, 0.36H)
7.56 (dd, J=8.55, 1.53 Hz, 0.64H) 7.63 (dd, J=8.55, 1.53 Hz, 0.36H)
7.88 (d, J=8.55 Hz, 0.64H) 7.91 (d, J=8.55 Hz, 0.36H) 8.12 (s,
0.36H) 8.33 (d, J=1.53 Hz, 0.64H).
[0081] An alternative procedure for the preparation of the title
compounds is provided below.
[0082] To a flame dried, four necked, 1 L round bottom flask
equipped with a mechanical stirrer, N2 inlet and a thermometer,
under N2, was charged sodium hydride (95%) (3.09 g, 129.2 mmol) and
dry DMF (200 mL). With vigorous stirring, trimethylsulfoxonium
iodide (32.5 g, 147.3 mmol) portion wise during which time the
temperature rose to 30.degree. C. After stirring for 30 mins, a
solution of 7H-Indolo[2,1-a][2]benzazepine-6-carboxylic acid,
13-cyclohexyl-10-[[[(dimethylamino)sulfonyl]amino]carbonyl]-3-(methoxy)-,
methyl ester (33.8 g, 61.3 mmol) in dry DMF (70 mL) was added
quickly. The reaction mixture was stirred below 30.degree. C. for
30 mins and then poured into an ice cold solution of 1 N HCl (130
mL) in H2O (2 L) portion wise. After the resulting suspension was
mechanically stirred for 1 h, the precipitates were filtered and
the filter cake was washed with H2O (100 mL). The filter cake was
partitioned between EtOAc and 0.5 N HCl (1:1, 4 L). The organic
phase was separated, washed with H2O (1 L) and brine (1 L), dried
over MgSO.sub.4, filtered and concentrated in vacuo. The residue
was dissolved in EtOAc (150 mL), and the solution was filtered
through a silica gel pad (300 g in hexane) and rinsed with 50%
EtOAc in hexane (5 L). The filtrate was concentrated in vacuo to
give a slightly yellow solid which was triturated with 10% EtOAc in
TBME (220 mL) from 50.degree. C. to 0.degree. C. to give
cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-1-
1-methoxy-, methyl ester, (+/-)- as a white solid (26.1 g, 75%
yield). HPLC purity, 100%. .sup.1H NMR (DMSO-d.sub.6, 300 MHz)
.delta. 11.61 (s, 1H), 8.47 (s, 0.5H), 8.25 (s, 0.5H), 7.81-7.88
(m, 1H), 7.57-7.63 (m, 1H), 7.23-7.29 (m, 2H), 7.01-7.07 (m, 1H),
5.43 (d, J=15.0 Hz, 0.5H), 5.22 (d, J=15 Hz, 0.5H), 4.04 (dd,
J=15.4 and 6.6 Hz, 0.5H), 3.83 (s, 3H), 3.75 (s, 1H), 3.08-3.47 (m,
0.5H), 3.29 (s, 3H), 2.73-2.92 (m, 8H), 1.11-1.99 (m, 10.5H), 0.20
(m, 0.5H); m/z 566 (M+H).sup.+.
##STR00069##
[0083] Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-1-
1-methoxy-, methyl ester, (-)-. A sample of (+/-)
cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-1-
1-methoxy- methyl ester was dissolved in EtOH/CH.sub.3CN 1/1+0.5%
DEA at a concentration of 50 mg/ml. [The addition of DEA ensures
the compound remains in solution during the injection process].
This solution was then injected onto a Thar SFC-350 preparative SFC
under the conditions shown below.
[0084] Preparative conditions on Thar SFC-350: Column: Chiralcel
OJ-H 5.times.25 cm; mobile phase: 25% MeOH/CH3CN (1/1) in CO2;
pressure (bar): 100; flow rate (ml/min): 240; solution
concentration (mg/ml): 50; injection amount (ml): 4.5-5; Cycle time
(min/inj): 6.5-7; Temperature (.degree. C.): 45; throughput (g/hr):
.about.2; Detector wavelength (nm): 254.
[0085] From 371.4 g of racemic starting material, a total of 177.3
g of the desired second eluting (-) isomer was obtained, containing
.about.1 Meq of diethylamine This material was purified using the
following procedure. The mixture (24.7 g) dissolved in
dichloromethane (800 mL)) was washed sequentially with; 0.5 N HCl
(1.times.400 mL, 1.times.240 mL), H.sub.2O (2.times.240 mL), and
brine (2.times.240 mL). The organic layer was then dried (Anhy.
Na.sub.2SO.sub.4), filtered and evaporated to give 22.33 g of
(cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1,12b-dih-
ydro-11-methoxy-, methyl ester, (-)- as a yellow solid (92%
recovery). HPLC.sup.1>99% (Rt 2.38 min); LC/MS (ES.sup.+) 566.51
(M+H, 100); [.alpha.].sub.D.sup.25C-194.64.degree. (c 1.03, MeOH).
Anal. Calcd for C.sub.30H.sub.35N.sub.3O.sub.6S.0.33H.sub.2O: C,
63.04; H, 6.29; N, 7.35; S, 5.61; H.sub.2O, 1.04. Found: C, 63.07;
H, 6.01; N, 7.24; S, 5.58; H.sub.2O, 1.03. The NMR shows the
absence of Et.sub.2NH. The EE of this material was determined to be
>99% using the following analytical HPLC procedure.
[0086] Analytical conditions of ee determination on Thar analytical
SFC. Analytical Column. Chiralcel OJ (0.46.times.25 cm, 10 .mu.l);
BPR pressure: 100 bars; Temperature: 35.degree. C.; Flow rate: 3.0
ml/min; Mobile Phase: 15% MeOH/CH.sub.3CN (1/1) in CO.sub.2;
Detector Wavelength: 254 nm; Retention time (min): 4, 6.5.
##STR00070##
[0087] Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1-1,12b-dihydro-
-11-methoxy-, (-)-. To a solution of (-)
cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-1-
1-methoxy-, methyl ester (22.33 g, 39.5 mmol) in MeOH (300 mL) was
added 1 N NaOH (120 mL) slowly over 20 min., while maintaining the
reaction temperature <30.degree. C. The mixture was stirred at
rt under N.sub.2 for 18 h. The HPLC indicated the reaction was
complete. To the reaction solution was added 1 N HCl (130 mL).
After addition was complete, the pH of the reaction mixture was
about 2. The methanol in the reaction mixture was evaporated. Water
(300 mL) was added to the mixture which was then extracted with
CH.sub.2Cl.sub.2 (1.times.600 mL, 1.times.200 mL). The combined
extracts were washed with H.sub.2O (2.times.300 mL), brine
(2.times.300 mL), dried (Na.sub.2SO.sub.4) and evaporated to give
20.82 g (96% yield) of the title compound as a yellow solid. HPLC
conditions column. Phenomenoex Synergi Polar-RP 4 um 4.6.times.50
mm; UV: 220 nm; gradient time: 4 min; flow rate: 4 mL/min, 75-100%
B; solvent A: 10% MeOH/90% H.sub.2O with 0.2% H.sub.3PO.sub.4,
solvent B: 90% MeOH/10% H.sub.2O with 0.2% H.sub.3PO.sub.4. HPLC
>99% (Rt 1.80 min.) LC/MS (ES.sup.+) 552.25 (M+H, 100);
[.alpha.].sub.D.sup.25C-166.99.degree. (c 1.00, MeOH). GC analysis:
CH.sub.2Cl.sub.2 4.94%; Anal. Calcd for
C.sub.29H.sub.33N.sub.3O.sub.6S.0.16 H.sub.2O.0.35
CH.sub.2Cl.sub.2: C, 60.37; H, 5.87; N, 7.20; S, 5.49; H.sub.2O,
0.49; CH.sub.2Cl.sub.2, 5.02. Found: C, 59.95; H, 5.89; N, 7.03; S,
5.38; H.sub.2O, 0.47; CH.sub.2Cl.sub.2, 4.94.
##STR00071##
[0088] Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-1-
1-methoxy-, (+/-)-. To a solution of (+/-)
cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-1-
1-methoxy-, methyl ester (100 mg, 0.177 mmol) in THF/Methanol
mixture (2.0 mL/2.0 mL), 2N NaOH solution (1.0 mL) was added. The
reaction mixture was heated at 90.degree. C. under microwave
conditions for 5 min. It was then concentrated, acidified with 1N
HCl solution and extracted with ethyl acetate (2.times.20 mL). The
organic layers were combined, dried (MgSO.sub.4), filtered and
concentrated. The residue was purified by preparative HPLC to
afford the desired product as a light yellow solid, (59 mg, 60%
yield). MS m/z 552 (MH.sup.+), Retention time: 3.850 min. 1H NMR
(300 MHz, MeOD) .delta. ppm 0.25 (m, 0.38H) 1.14-2.22 (m, 11.62H)
2.69-2.98 (m, 2H) 3.02 (s, 2.28H) 3.02 (s, 3.72H) 3.41 (d, J=15.00
Hz, 0.62H) 3.88 (s, 3H) 4.01 (d, J=15.00 Hz, 0.38H) 5.26 (d,
J=15.00 Hz, 0.38H) 5.45 (d, J=14.64 Hz, 0.62H) 6.94-7.02 (m, 1H)
7.13 (d, J=2.56 Hz, 0.38H) 7.21 (d, J=2.20 Hz, 0.62H) 7.26 (d,
J=8.42 Hz, 0.62H) 7.30 (d, J=8.78 Hz, 0.38H) 7.53 (dd, J=8.42, 1.46
Hz, 0.62H) 7.61 (dd, J=8.60, 1.65 Hz, 0.38H) 7.85 (d, J=8.42 Hz,
0.62H) 7.89 (d, J=8.42 Hz, 0.38H) 8.10 (s, 0.38H) 8.28 (d, J=1.46
Hz, 0.62H).
##STR00072##
[0089]
Cycloprop[d]indolo[2,1-a][2]benzazepine-1a,5(2H)-dicarboxamide,
8-cyclohexyl-N.sup.5-[(dimethylamino)sulfonyl]-1,12b-dihydro-N.sup.1a-[(2-
R,3S)-3-hydroxy-4,7,7-trimethylbicyclo[2.2.1]hept-2-yl]-11-methoxy-,
(1aR)-[partial]-. TBTU (437 mg, 1.36 mmol) and DIPEA (0.95 mL,
5.436 mmol) were added to a solution of (+/-)
cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-1-
1-methoxy- (500 mg, 0.906 mmol) in DMSO (20.0 mL). The reaction
mixture was stirred at rt for 15 min.
(2S,3R)-3-Amino-1,7,7-trimethylbicyclo[2.2.1]heptan-2-ol (280 mg,
1.36 mmol) was then added and the reaction mixture was stirred at
rt overnight. The reaction mixture was quenched with water and
acidified with 1N HCl solution. A brown solid separated which was
collected by filtration. This material was then fractionated by
Preparative HPLC under the following conditions. Column. Waters
Sunfire 19 mm.times.100 mm; Solvent A: 10% CH3CN-90% H2O-0.1% TFA;
Solvent B: 90% CH3CN-10% H2O-0.1% TFA; Program: Start with 65%
solvent B, initial hold time for 5 min, then gradually increase to
90% solvent B in 30 min with flow rate 25 mL/min. Load: 50-60
mg/run.
[0090]
Cycloprop[d]indolo[2,1-a][2]benzazepine-1a,5(2H)-dicarboxamide,
8-cyclohexyl-N.sup.5-[(dimethylamino)sulfonyl]-1,12b-dihydro-N.sup.1a-[(2-
R,3S)-3-hydroxy-4,7,7-trimethylbicyclo[2.2.1]hept-2-yl]-11-methoxy-,
(1aR)-[partial]- elutes before
Cycloprop[d]indolo[2,1-a][2]benzazepine-1a,5(2H)-dicarboxamide,
8-cyclohexyl-N.sup.5-[(dimethylamino)sulfonyl]-1,12b-dihydro-N.sup.1a-[(2-
R,3S)-3-hydroxy-4,7,7-trimethylbicyclo[2.2.1]hept-2-yl]-11-methoxy-,
(1aS)-[partial]- under the HPLC conditions described above. Product
obtained as a light yellow solid, 230 mg, 36% yield). MS m/703
(MH.sup.+), Retention time: 3.936 min. 1H NMR (500 MHz, MeOD)
.delta. ppm 0.14-0.24 (m, 2.64H) 0.51 (s, 2.46H) 0.72-2.21 (m,
20.9H) 2.49 (m, 0.18H) 2.62 (m, 0.82H) 2.85 (m, 0.18H) 2.96 (m,
0.82H) 3.03 (s, 6H) 3.39 (m, 0.82H) 3.49-3.58 (m, 1.64H) 3.71-3.80
(m, 0.36H) 3.90 (s, 3H) 4.17 (d, J=14.65 Hz, 0.18H) 5.06 (d,
J=14.65 Hz, 0.18H) 5.37 (d, J=14.95 Hz, 0.82H) 6.73 (d, J=5.49 Hz,
0.82H) 6.98-7.05 (m, 1H) 7.08 (d, J=4.58 Hz, 0.18H) 7.10 (d, J=2.44
Hz, 0.18H) 7.21 (d, J=2.44 Hz, 0.82H) 7.31 (d, J=8.55 Hz, 0.82H)
7.34 (d, J=8.55 Hz, 0.18H) 7.59-7.64 (m, 1H) 7.87-7.93 (m, 1H) 7.99
(s, 0.18H) 8.09 (d, J=1.22 Hz, 0.82 H).
##STR00073##
[0091]
Cycloprop[d]indolo[2,1-a][2]benzazepine-1a,5(2H)-dicarboxamide,
8-cyclohexyl-N.sup.5-[(dimethylamino)sulfonyl]-1,12b-dihydro-N.sup.1a-[(2-
R,3S)-3-hydroxy-4,7,7-trimethylbicyclo[2.2.1]hept-2-yl]-11-methoxy-,
(1aS)-[partial]-. TBTU (437 mg, 1.36 mmol) and DIPEA (0.95 mL,
5.436 mmol) were added to a solution of (+/-)
cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-1-
1-methoxy- (500 mg, 0.906 mmol) in DMSO (20.0 mL). The reaction
mixture was stirred at rt for 15 min. Then
(2S,3R)-3-amino-1,7,7-trimethylbicyclo[2.2.1]heptan-2-ol (280 mg,
1.36 mmol) was added, and the reaction mixture was stirred at rt
overnight. The reaction mixture was quenched with water and then
acidified with 1N HCl solution. A brown colored solid separated
that was collected by filtration. This material was then
fractionated by preparative HPLC under the following conditions.
Column: Waters Sunfire 19 mm.times.100 mm; Solvent A: 10% CH3CN-90%
H2O-0.1% TFA; Solvent B: 90% CH3CN-10% H2O-0.1% TFA; Program: Start
with 65% solvent B, initial hold time for 5 min, then gradually
increase to 90% solvent B in 30 min with flow rate 25 mL/min. Load:
50-60 mg/run.
[0092]
Cycloprop[d]indolo[2,1-a][2]benzazepine-1a,5(2H)-dicarboxamide,
8-cyclohexyl-N.sup.5-[(dimethylamino)sulfonyl]-1,12b-dihydro-N.sup.1a-[(2-
R,3S)-3-hydroxy-4,7,7-trimethylbicyclo[2.2.1]hept-2-yl]-11-methoxy-,
(1aS)-[partial]- elutes after
cycloprop[d]indolo[2,1-a][2]benzazepine-1a,5(2H)-dicarboxamide,
8-cyclohexyl-N.sup.5-[(dimethylamino)sulfonyl]-1,12b-dihydro-N.sup.1a-[(2-
R,3S)-3-hydroxy-4,7,7-trimethylbicyclo[2.2.1]hept-2-yl]-11-methoxy-,
(1aR)-[partial]- under the HPLC conditions described above. Product
obtained as a light yellow solid, 215 mg, 34% yield). MS m/703
(MH.sup.+), Retention time: 4.038 min. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 0.20 (m, 0.38H) 0.75 (s, 1.86H) 0.76 (s, 1.86H) 0.84
(s, 1.86H) 0.85 (s, 1.14H) 0.89-2.18 (m, 18.9H) 2.52 (m, 0.38H)
2.70 (m, 0.62H) 2.85 (m, 0.38H) 2.97 (m, 0.62H) 3.03 (s, 2.28H)
3.04 (s, 3.72H) 3.33-3.39 (m, 0.62H) 3.43-3.51 (m, 1.24H) 3.73-3.77
(m, 0.38H) 3.78-3.84 (m, 0.38H) 3.90 (s, 1.86H) 3.90 (s, 1.14H)
4.14 (d, J=14.65 Hz, 0.38H) 5.11 (d, J=14.65 Hz, 0.38H) 5.44 (d,
J=15.26 Hz, 0.62H) 6.68 (d, J=4.88 Hz, 0.62H) 6.96-7.03 (m, 1H)
7.07 (d, J=5.19 Hz, 0.38H) 7.12 (d, J=2.44 Hz, 0.38H) 7.23 (d,
J=2.14 Hz, 0.62H) 7.27 (d, J=8.54 Hz, 0.62H) 7.33 (d, J=8.54 Hz,
0.38H) 7.55 (dd, J=8.39, 1.68 Hz, 0.62H) 7.62 (dd, J=8.55, 1.53 Hz,
0.38H) 7.87 (d, J=8.54 Hz, 0.62H) 7.91 (d, J=8.55 Hz, 0.38H) 8.08
(d, J=1.22 Hz, 0.38H) 8.10 (d, J=1.22 Hz, 0.62H).
##STR00074##
[0093] Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-1-
1-methoxy-, (-)-. 10 N NaOH (2.0 mL, 20 mmol) solution and 4 mL of
water were added to a solution of
cycloprop[d]indolo[2,1-a][2]benzazepine-1a,5(2H)-dicarboxamide,
8-cyclohexyl-N.sup.5-[(dimethylamino)sulfonyl]-1,12b-dihydro-N.sup.1a-[(2-
R,3S)-3-hydroxy-4,7,7-trimethylbicyclo[2.2.1]hept-2-yl]-11-methoxy-,
(1aR)-[partial]- (160 mg, 0.228 mmol) in THF/MeOH (7 mL/7 mL). The
reaction mixture was heated at 120.degree. C. under microwave
conditions for 1 hr. It was then concentrated, acidified with conc.
HCl solution and extracted with ethyl acetate twice (2.times.30
mL). The organic layers were combined, dried (MgSO.sub.4), filtered
and concentrated in vacuo to an orange oil. The crude product was
then purified by Prep. HPLC column to afford the product a light
yellow solid, (80 mg, 64% yield). Average specific rotation
-130.85.degree.; Solvent MeOH; Wavelength 589 nm; 50 cm cell. MS
m/552 (MH.sup.+), Retention time: 3.760 min. .sup.1H NMR (500 MHz,
MeOD) .delta. ppm 0.27 (m, 0.38H) 1.14-2.22 (m, 11.62H) 2.76 (m,
0.38H) 2.80-2.92 (m, 1H) 2.92-3.09 (m, 6.62H) 3.45 (d, J=14.95 Hz,
0.62H) 3.90 (s, 1.86H) 3.91 (s, 1.14H) 4.04 (d, J=15.26 Hz, 0.38H)
5.28 (d, J=15.26 Hz, 0.38H) 5.47 (d, J=15.26 Hz, 0.62H) 6.95-7.05
(m, 1H) 7.15 (d, J=2.75 Hz, 0.38H) 7.23 (d, J=1.83 Hz, 0.62H) 7.28
(d, J=8.55 Hz, 0.62H) 7.33 (d, J=8.54 Hz, 0.38H) 7.54 (dd, J=8.39,
1.68 Hz, 0.62H) 7.63 (dd, J=8.55, 1.53 Hz, 0.38H) 7.86 (d, J=8.55
Hz, 0.62H) 7.91 (d, J=8.55 Hz, 0.38H) 8.11 (d, J=1.22 Hz, 0.62H)
8.29 (d, J=1.22 Hz, 0.38H).
##STR00075##
[0094] Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-5-[[[(dimethylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-1-
1-methoxy-, (+)-. 10 N NaOH (1.8 mL, 18 mmol) solution and 4 mL of
water were added to a solution of
cycloprop[d]indolo[2,1-a][2]benzazepine-1a,5(2H)-dicarboxamide,
8-cyclohexyl-N.sup.5-[(dimethylamino)sulfonyl]-1,12b-dihydro-N.sup.1a-[(2-
R,3S)-3-hydroxy-4,7,7-trimethylbicyclo[2.2.1]hept-2-yl]-11-methoxy-,
(1aS)-[partial]- (130 mg, 0.185 mmol) in bTHF/MeOH (7 mL/7 mL). The
reaction mixture was heated at 120.degree. C. under microwave
conditions for 1 hr. It was concentrated, acidified with conc. HCl
solution and extracted with ethyl acetate twice (2.times.30 mL).
The organic layers were combined, dried (MgSO.sub.4), filtered and
concentrated in vacuo to give an orange oil. The crude product was
then purified by Prep. HPLC column to afford the product as a light
yellow solid, (68 mg, 67% yield). Average specific
rotation+174.73.degree.; Solvent MeOH; Wavelength 589 nm; 50 cm
cell MS m/552 (MH.sup.+), Retention time: 3.773 min. .sup.1H NMR
(500 MHz, MeOD) .delta. ppm 0.27 (m, 0.38H) 1.14-2.22 (m, 11.62H)
2.76 (m, 0.38H) 2.80-2.92 (m, 1H) 2.92-3.09 (m, 6.62H) 3.45 (d,
J=14.95 Hz, 0.62H) 3.90 (s, 1.86H) 3.91 (s, 1.14H) 4.04 (d, J=15.26
Hz, 0.38H) 5.28 (d, J=15.26 Hz, 0.38H) 5.47 (d, J=15.26 Hz, 0.62H)
6.95-7.05 (m, 1H) 7.15 (d, J=2.75 Hz, 0.38H) 7.23 (d, J=1.83 Hz,
0.62H) 7.28 (d, J=8.55 Hz, 0.62H) 7.33 (d, J=8.54 Hz, 0.38H) 7.54
(dd, J=8.39, 1.68 Hz, 0.62H) 7.63 (dd, J=8.55, 1.53 Hz, 0.38H) 7.86
(d, J=8.55 Hz, 0.62H) 7.91 (d, J=8.55 Hz, 0.38H) 8.11 (d, J=1.22
Hz, 0.62H) 8.29 (d, J=1.22 Hz, 0.38H).
##STR00076##
[0095] 1H-Indole-6-carboxylic acid, 2-bromo-3-cyclohexyl-,
1,1-dimethylethyl ester. To a mechanically stirred solution of
2-bromo-3-cyclohexyl-1H-indole-6-carboxylic acid (80 g, 0.24 m) in
dry methylene dichloride (1.2 L) and THF (100 mL) were added
activated molecular sieves (4A, 80 g) and silver carbonate (275 g,
0.99 m). The reaction mixture was cooled to 0.degree. C. and
t-Butyl bromide (142 g, 1.04 m) was added drop wise. The mixture
was stirred overnight at rt and monitored by TLC (Hexane-Ethyl
acetate 80:20, R.sub.f (Product)=0.7). If any bromo acid was left
unconverted a further 10% of silver carbonate was added and
stirring was continued for an addition 2-4 h. On completion, the
reaction mixture was filtered through a thin bed of celite. The
filtrand was washed with methylene dichloride (500 mL). The
combined filtrates were concentrated in-vacuo, and the crude
product thus obtained was purified by silica gel chromatography:
(230-400 mesh, eluted with a gradient of ethyl acetate in pet ether
0-2%). Homogeneous fractions were combined and evaporated under
reduced pressure to give 80 g (85%) of the title compound. HPLC:
90.1% (RT=6.56 min), Column: C18 BDS, (50.times.4.6 mm), Mobile
Phase: Gradient of 0.1% TFA in water: ACN
(30.fwdarw.100.fwdarw.30), Flow rate 0.8 mL/min. LCMS: 99.8%
(RT=4.44 min), Column: Geneis, C18 50.times.4.6 mm Mobile Phase:
Gradient of 0.1% Formic acid in water: ACN
(70.fwdarw.95.fwdarw.70), Flow rate: 0.8 mL/min; M-1=376.5; .sup.1H
NMR CDCl.sub.3) (400 MHz) .delta. 1.37-1.40 (m, 3H, cyc.Hexyl),
1.62 (s, 9H, t-Bu), 1.80-1.94 (two sets of m, 3H, & 4H
respectively, cyc.Hexyl part), 2.81 (m, 1H, CH of cyc.Hexyl-
benzylic), 7.70-7.75 (m, 2H, Indole-H.sub.4&5), 8.04 (s, 1H,
Indole-H.sub.7), 8.52 (s, 1H, Indole-NH).
##STR00077##
[0096] 1H-Indole-6-carboxylic acid,
3-cyclohexyl-2-(2-formyl-4-methoxyphenyl)-, 1,1-dimethylethyl
ester. tert-Butyl 2-bromo-3-cyclohexyl-1H-indole-6-carboxylate (72
g, 0.19 m) was dissolved in a 1:1 mixture of toluene and ethanol
(720 mL) and degasified. LiCl (23.9 g, 0.51 m) was then added,
followed by sodium carbonate (720 mL, 1.0 M solution degasified
separately,) and Pd-tetrakis (13.1 g, 0.011 m). After stirring for
0.25 h, 2-formyl-4-methoxyphenylboronic acid (41.1 g, 0.22 m) was
added and the reaction mixture was heated to 85.degree. C. for 4 h.
The reaction was then monitored by TLC, (Hexane-Ethyl acetate
80:20, R.sub.f (Product)=0.55). On completion, the reaction mixture
was cooled to rt and water (1.0 L) was added followed by ethyl
acetate (1.0 L). The organic layer was washed with brine, and dried
and concentrated under vacuum to afford the title compound as a
yellow solid. Yield 75 g (74%). HPLC: 99.7% (RT=6.30 min), Column:
C18 BDS (4.6.times.50 mm), SC-307, Mobile Phase: Gradient of 0.1%
TFA in water: ACN (30.fwdarw.100.fwdarw.30), Flow rate 0.8 mL/min.
LCMS: 98.0% (RT=5.28 min), Column: Geneis, C18 (50.times.4.6 mm),
Mobile Phase: Gradient of 0.1% Formic acid in water: ACN
(70.fwdarw.95.fwdarw.70), Flow rate: 0.8 mL/min; M-1=432.2; .sup.1H
NMR (DMSO-d.sub.6) (400 MHz) .delta. 1.40-1.48 (m, 3H, cyc.Hexyl),
1.57 (s, 9H, t-Bu), 1.84-1.90 (m, 7H, cyc.Hexyl part), 3.09 (m, 1H,
CH of cyc.Hexyl- benzylic), 3.84 (s, 3H, OCH.sub.3), 6.55 (d, J=4
Hz, 1H, aryl H.sub.2'), 7.06 (d, 1H, aryl H.sub.3'), 7.08 (s, 1H,
aryl H.sub.6'), 7.23 (d, 1H, Indole-H.sub.5), 7.53 (d, J=8 Hz, 1H,
Indole-H.sub.4), 7.70-7.75 (m, 2H, NH+Indole-H.sub.7), 8.06 (s, 1H,
CHO).
##STR00078##
[0097] 7H-Indolo[2,1-a][2]benzazepine-6,10-dicarboxylic acid,
13-cyclohexyl-, 10-(1,1-dimethylethyl) 6-methyl ester. tert-Butyl
3-cyclohexyl-2-(2-formyl-4-methoxyphenyl)-1H-indole-6-carboxylate
(62.5 g, 0.144 m) was dissolved in dry DMF (1.2 L) and stirred
mechanically. Cesium carbonate (84 g, 0.17 m) and methyl
2-(dimethoxyphosphoryl)acrylate (65-70% GC pure, 56.2 g, 0.18 m)
were then added and the reaction mixture was heated to 65.degree.
C. for 4 h, and the reaction was monitored by TLC (Hexane-Ethyl
acetate 80:20, R.sub.f (Product)=0.7). On completion, the mixture
was cooled to rt, then quenched with water (1.0 L). A yellow solid
precipitated, which was collected by filtration and air dried. This
material was then slurried in methanol, filtered, and dried under
vacuum to give the product as a yellow powder, (70 g, 90%). HPLC:
99.1% (RT=6.45 min), Column: C18 BDS (4.6.times.50 mm), Mobile
Phase: Gradient of 0.1% TFA in water: ACN
(30.fwdarw.100.fwdarw.30), Flow rate 0.8 mL/min. LCMS: 100%
(RT=7.00 min), Column: Geneis, C18 (50.times.4.6 mm), Mobile Phase:
Gradient of 0.1% Formic acid in water: ACN
(70.fwdarw.95.fwdarw.70), Flow rate: 0.8 mL/min; M+1=502.2; .sup.1H
NMR (CDCl.sub.3) (400 MHz) .delta. 1.10-1.30 (m, 3H, cyc.Hexyl),
1.64 (s, 9H, t-Bu), 1.77-2.07 (m, 7H, cyc.Hexyl part), 2.80 (m, 1H,
CH of cyc.Hexyl- benzylic), 3.84 (s, 3H, OCH.sub.3), 3.93 (s, 3H,
COOCH.sub.3), 4.15 & 5.65 (two br. peak., 1H each, allylic
CH.sub.2), 6.95 (s, 1H, aryl H.sub.6'), 7.01 (d, 1H, aryl
H.sub.2'), 7.53 (d, J=8 Hz, 1H, aryl H.sub.3'), 7.70 (d, J=4 Hz,
1H, Indole-H.sub.5), 7.84 (s+d, 2H, olefinic H+ Indole-H.sub.4),
8.24 (s, 1H, indole-H.sub.7); .sup.13C NMR (CDCl.sub.3) (100.0 MHz)
.delta. 166.92, 165.71, 158.96, 142.28, 136.47, 13.50, 134.61,
132.43, 132.01, 129.73, 124.78, 124.68, 120.33, 119.39, 119.04,
115.62, 115.05, 111.27, 80.27, 55.49, 52.50, 39.09, 36.81, 33.40,
28.38, 27.15, 26.28.
##STR00079##
[0098] 2-Propenoic acid, 2-(dimethoxyphosphinyl)-, methyl ester. To
a 5 L four necked round bottom flask equipped with a mechanical
stirrer, a condenser, a temperature controller and a N2 inlet, was
charged paraformaldehyde (40.5 g, 1.35 mol), MeOH (2 L) and
piperidine (2 mL). The reaction mixture was heated to reflux under
N2 for 3 h. After cooling to 50.degree. C.,
2-(dimethoxyphosphoryl)acetate (150 g, 0.824 mol) was added in one
portion. The reaction mixture was continued to reflux for 18 h.
After cooling to rt, the reaction solution was concentrated in
vacuo to give a clear colorless oil. The above oil was dissolved in
dry toluene (1 L) in a 3 L four necked round bottom flask equipped
a temperature controller, a N.sub.2 inlet, a magnetic stirrer and a
Dean-Stark apparatus. To the solution was added TsOH.H.sub.2O (5.2
g). The reaction mixture was then refluxed azeotropically to remove
methanol for 18 h. After cooling to rt, the solution was
concentrated in vacuo to give a yellow oil which was vacuum
distilled at 150-155.degree. C./0.2 mmHg to afford the product as a
colorless oil (135.0 g). Purity, 90% based on .sup.1H NMR. .sup.1H
NMR (CDCl3, 300 MHz) .delta. 7.0 (dd, J=42.4 and 1.5 Hz, 1H), 6.73
(dd, J=20.5 and 1.8 Hz, 1H), 3.80 (s, 6H), 3.76 (s, 3H).
##STR00080##
[0099]
Cycloprop[d]indolo[2,1-a][2]benzazepine-1a,5(2H)-dicarboxylic acid,
8-cyclohexyl-1,12b-dihydro-11-methoxy-, 5-(1,1-dimethylethyl)
1a-methyl ester, (+/-). Sodium hydride (96 mg, 4 mmol) was added to
a stirred suspension of trimethylsulfoxonium chloride (567 mg, 4.4
mmol) in anhydrous DMSO (10 mL) under nitrogen. The resultant
mixture was stirred at rt for 30-45 min and then neat
7H-indolo[2,1-a][2]benzazepine-6,10-dicarboxylic acid,
13-cyclohexyl-3-methoxy-, 10-(1,1-dimethylethyl) 6-methyl ester
(1.0, 2 mmol) was added in small portions. The suspension was
diluted with DMSO (5 mL) and heated at 50.degree. C. for 3-4 h. The
reaction mixture was allowed to cool to rt and water was added. A
solid separated, which was collected by filtration and washed with
water and then air dried overnight to afford 1.15 g of crude
product. This material was purified by flash column chromatography
(silica gel, 3% MeOH in DCM) to provide pure title compound (0.96
g): LC/MS: Retention time 3.816 min; m/e 516 (MH.sup.+). .sup.1H
NMR (400 MHz, CDCl.sub.3): The product was observed to exist as
inter-converting rotamers, as evidenced from the compound's NMR
spectrum.
[0100] The following procedure is an example of a method to effect
the resolution of racemic
cycloprop[d]indolo[2,1-a][2]benzazepine-1a,5(2H)-dicarboxylic acid,
8-cyclohexyl-1,12b-dihydro-11-methoxy-, 5-(1,1-dimethylethyl)
1a-methyl ester, (+/-). A sample of
cycloprop[d]indolo[2,1-a][2]benzazepine-1a,5(2H)-dicarboxylic acid,
8-cyclohexyl-1,12b-dihydro-11-methoxy-, 5-(1,1-dimethylethyl)
1a-methyl ester, (+/-)-was dissolved in a mixture of isopropanol
and acetonitrile (8:2) to give a final concentration of 20 mg/mL.
This mixture was injected on a preparative chiral SFC
chromatography system using the following conditions: Chiralcel
OJ-H column, 4.6.times.250 mm, 5 .mu.m; Mobile Phase: 8% MeOH in
CO.sub.2; Temp: 35.degree. C.; Flow rate: 2 mL/min for 16 min; UV
monitored @ 260 nm; Injection: 5 .mu.L of .about.20.0 mg/mL in
IPA:ACN (8:2).
##STR00081##
[0101]
Cycloprop[d]indolo[2,1-a][2]benzazepine-1a,5(2H)-dicarboxylic acid,
8-cyclohexyl-1,12b-dihydro-11-methoxy-, 1a-methyl ester, (+/-)-.
TFA (5 mL) was added to a solution of (+/-)
8-Cyclohexyl-1,1a,2,12b-tetrahydro-11-methoxy-1a-(methoxycarbonyl)-cyclop-
rop[d]indolo[2,1-a][2]benzazepine-5-carboxylic acid, tert-butyl
ester (515 mg, 1 mmol) in anhydrous DCM (10 mL). The resultant
solution was stirred at rt for approximately 8 to 12 hr. The
reaction was then evaporated to dryness to afford the title
compound (0.47 g, 100%). LC/MS: Retention time 2.245 min; m/e 460
(MH.sup.+). .sup.1H NMR (400 MHz, CDCl.sub.3): From the compounds
NMR spectrum, the product was observed to exist as a mixture of
interconverting rotamers.
##STR00082##
[0102] Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-1,12b-dihydro-11-methoxy-5-[[[(methylamino)sulfonyl]amino]ca-
rbonyl]-, methyl ester. A solution of
8-Cyclohexyl-1,1a,2,12b-tetrahydro-11-methoxy-1a-(methoxycarbonyl)-cyclop-
rop[d]indolo[2,1-a][2]benzazepine-5-carboxylic acid (140 mg, 0.31
mmol) and CDI (64 mg, 0.40 mmol) in THF (3 mL) was stirred for 1 hr
at 60.degree. C. N-methylsulfamide (68 mg, 0.62 mmol) and DBU (71.6
mg, 0.47 mmol) were added and the mixture was stirred at 60.degree.
C. overnight. The reaction was then poured into cold water,
acidified with dilute hydrochloric acid and extracted into ethyl
acetate. The extracts were washed sequentially with dilute
hydrochloric acid (0.1 N), and brine, and then dried (anhy. sodium
sulfate), filtered and evaporated to provide the title compound as
a brown solid. ESI-MS m/e 552 (MH.sup.+). This material was used
without further purification.
##STR00083##
[0103] Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-1,12b-dihydro-11-methoxy-5-[[[(methylamino)sulfonyl]amino]ca-
rbonyl]-. Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-5-[[[(methylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-11--
methoxy-, methyl ester was dissolved in THF, MeOH mixture (2 mL, 2
mL). 2.5 M NaOH (aq.) (1.2 mL, 3 mmol) was then added and the
reaction was shaken at 22.degree. C. for 2 hr. The solution was
then neutralized with 1M HCl (aq.) (3 mL) and concentrated to
remove the organic solvents. The residue was slurried with H.sub.2O
and the solids were collected by filtration, washed with H.sub.2O
and dried to yield compound the title compound (160 mg, 0.30 mmol).
ESI-MS m/e 538 (MH.sup.+). This material was used without further
purification.
##STR00084##
[0104] Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-5-[[[(benzylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-11--
(methoxy)-12-(methoxy)-, methyl ester, (+/-)-. A solution of (+/-)
8-cyclohexyl-1,1a,2,12b-tetrahydro-11-methoxy-1a-(methoxycarbonyl)-cyclop-
rop[d]indolo[2,1-a][2]benzazepine-5-carboxylic acid (200 mg, 0.44
mmol) and CDI (92 mg, 0.57 mmol) in THF (5 mL) was stirred for 1 hr
at 60.degree. C. N-benzylsulfamide (164 mg, 0.88 mmol) and DBU (100
mg, 0.66 mmol) were then added and the resultant mixture was
stirred at 60.degree. C. overnight. The reaction was then poured
into cold water, acidified with dilute hydrochloric acid and
extracted into ethyl acetate. The organic phase was washed
hydrochloric acid (0.1 N), brine and dried (sodium sulfate) and
evaporated in vacuo to provide the title compound as a brown solid.
ESI-MS m/e 628 (MH.sup.+).
##STR00085##
[0105] Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-1,12b-dihydro-11-methoxy-5-[[[[(phenylmethyl)amino]sulfonyl]-
amino]carbonyl]-, (+/-)-. The title compound was prepared using a
similar procedure to that described for
cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid,
8-cyclohexyl-5-[[[(methylamino)sulfonyl]amino]carbonyl]-1,12b-dihydro-11--
methoxy-cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid starting from (+/-)
8-cyclohexyl-1,1a,2,12b-tetrahydro-11-methoxy-1a-(methoxycarbonyl)-cyclop-
rop[d]indolo[2,1-a][2]benzazepine-5-carboxylic acid. ESI-MS m/e 613
(MH.sup.+), .sup.1H NMR (500 MHz, MeOD) .delta. ppm 1.22-2.20 (m,
13H) 3.27-3.31 (m, 1 H) 3.47 (d, J=14.95 Hz, 0.6H) 3.92 (d, J=2.44
Hz, 3H) 4.04 (d, 0.4H) 4.31 (d, J=2.75 Hz, 2H) 5.24 (d, 0.4H) 5.48
(d, 0.6H) 7.02 (d, 1H) 7.17 (d, J=2.75 Hz, 1H) 7.19-7.35 (m, 5H)
7.39 (t, J=7.48 Hz, 2H) 7.45-7.52 (m, 1H) 7.80 (d, J=1.53 Hz, 0.4H)
7.85 (dd, J=8.39, 6.87 Hz, 1H) 8.22 (d, J=1.53 Hz, 0.6H).
##STR00086##
[0106] Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-5-[[(cyclopropylsulfonyl)amino]carbonyl]-1,12b-dihydro-11-me-
thoxy-, (+/-)-. A mixture of (+/-)
8-cyclohexyl-1,1a,2,12b-tetrahydro-11-methoxy-1a-(methoxycarbonyl)-cyclop-
rop[d]indolo[2,1-a][2]benzazepine-5-carboxylic acid (1 equiv), and
carbonyldiimidazole (1.5 equiv) in anhydrous THF was heated at
50.degree. C. for 30 min and allowed to cool to rt. Then 1 equiv of
cyclopropanesulfonamide and 1,8-diazabicyclo[5.4.0]undec-7-ene (2
equiv) were added consecutively. The resultant mixture was stirred
at rt overnight. After acidic aqueous workup, the isolated crude
product was purified by prep. HPLC. The intermediate ester was then
hydrolyzed using 1N NaOH in THF-MeOH to afford the title compound.
LC/MS: Retention time: 2.030 min; m/e 549 (MH.sup.+). .sup.1H NMR
(400 MHz, CDCl.sub.3): The product was observed to exist as
inter-converting rotamers, as evidenced from the compound's NMR
spectrum.
[0107] Intermediates 27-38 use the experimental procedures that
follow until otherwise noted. LCMS data: Stop time: Gradient time+1
minute; Starting conc: 0% B unless otherwise noted; Ending conc:
100% B unless otherwise noted; Eluent A: 5% CH.sub.3CN/95% H.sub.2O
with 10 mM NH.sub.4OAc (for columns A, D and E); 10% MeOH/90%
H.sub.2O with 0.1% TFA (for columns B and C); Eluent B: 95%
CH.sub.3CN/5% H.sub.2O with 10 mM NH.sub.4OAc (for columns A, D and
E); 90% MeOH/10% H.sub.2O with 0.1% TFA (for columns B and C);
Column A: Phenomenex 10.mu. 4.6.times.50 mm C18; Column B:
Phenomenex C18 10.mu. 3.0.times.50 mm; Column C. Phenomenex
4.6.times.50 mm C18 10.mu.; Column D: Phenomenex Lina C18 5.mu.
3.0.times.50 mm; Column E: Phenomenex 5.mu. 4.6.times.50 mm C18;
Preparative HPLC data: Conditions for H.sub.2O/CH.sub.3CN with 10
mM NH.sub.4OAc buffer; Gradient: Linear over 20 min. unless
otherwise noted; Starting conc: 15% B unless otherwise noted;
Ending conc: 100% B; Eluent A: 5% CH.sub.3CN/95% H.sub.2O with 10
mM NH.sub.4OAc; Eluent B:95% CH.sub.3CN/5% H.sub.2O with 10 mM
NH.sub.4OAc; Column: Sunfire Prep C.sub.18 OBD 5.mu. 30.times.100
mm; Conditions for H.sub.2O/MeOH with 0.1% TFA buffer; Gradient:
Linear over 20 min. unless otherwise noted; Starting conc: 30% B
unless otherwise noted; Ending conc: 100% B; Eluent A: 10% MeOH/90%
H.sub.2O with 0.1% TFA; Eluent B: 90% MeOH/10% H.sub.2O with 0.1%
TFA; Column: phenomenex 21.times.100 mmC18 H.sub.2O.
##STR00087##
[0108] Cycloprop[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid,
8-cyclohexyl-5-[[(cyclopropylsulfonyl)amino]carbonyl]-1,12b-dihydro-11-me-
thoxy-, (+/-)-. A mixture of (+/-)
8-cyclohexyl-1,1a,2,12b-tetrahydro-11-methoxy-1a-(methoxycarbonyl)-cyclop-
rop[d]indolo[2,1-a][2]benzazepine-5-carboxylic acid (1 equiv), and
carbonyldiimidazole (1.5 equiv) in anhydrous THF was heated at
50.degree. C. for 30 min and allowed to cool to rt. Then 1 equiv of
cyclopropanesulfonamide and 1,8-diazabicyclo[5.4.0]undec-7-ene (2
equiv) were added consecutively. The resultant mixture was stirred
at rt overnight. After acidic aqueous workup, the isolated crude
product was purified by prep. HPLC. The intermediate ester was then
hydrolyzed using 1N NaOH in THF-MeOH to afford the title compound.
LC/MS: Retention time: 2.030 min; m/e 549 (MH.sup.+). .sup.1H NMR
(400 MHz, CDCl.sub.3): The product was observed to exist as
inter-converting rotamers, as evidenced from the compound's NMR
spectrum.
##STR00088##
[0109]
13-Cyclohexyl-3-methoxy-6-(methoxycarbonyl)-7H-indolo[2,1-a][2]benz-
azepine-10-carboxylic acid. Trifluoroacetic acid (30 mL) was added
dropwise to a stirring slurry of 10-tert-butyl 6-methyl
13-cyclohexyl-3-methoxy-7H-indolo[2,1-a][2]benzazepine-6,10-dicarboxylate
(10 g, 20 mmol) in dichloroethane (30 mL) under N.sub.2. The clear
dark green solution was stirred at rt for 2.5 h, concentrated to
dryness and stirred with EtOAc (100 mL) overnight. The solids were
collected by filtration, washed with EtOAc and Et.sub.2O to yield
13-cyclohexyl-3-methoxy-6-(methoxycarbonyl)-7H-indolo[2,1-a][2]benzazepin-
e-10-carboxylic acid (8.35 g, 18.8 mmol, 94%) was as a yellow solid
which was used without further purification. 1H NMR (300 MHz,
CDCl.sub.3) .delta. 1.13-2.16 (m, 10H), 2.74-2.88 (m, 1H), 3.84 (s,
3H), 3.89 (s, 3H), 4.06-4.29 (m, 1H), 5.54-5.76 (m, 1H), 6.98 (d,
J=2.6 Hz, 1H), 7.08 (dd, J=8.4, 2.6 Hz, 1H), 7.52 (d, J=8.4 Hz,
1H), 7.78 (dd, J=8.8, 1.1 Hz, 1H), 7.80 (s, 1H), 7.86 (d, J=8.8 Hz,
1H), 8.34 (d, J=1.1 Hz, 1H). LCMS: m/e 446 (M+H).sup.+, ret time
3.21 min, column B, 4 minute gradient.
##STR00089##
[0110] Methyl
13-cyclohexyl-10-((cyclopropylsulfonyl)carbamoyl)-3-methoxy-7H-indolo[2,1-
-a][2]benzazepine-6-carboxylate. 1,1'-Carbonyldiimidazole (1.82 g,
11.2 mmol) was added to a slurry of
13-cyclohexyl-3-methoxy-6-(methoxycarbonyl)-7H-indolo[2,1-a][2]benzazepin-
e-10-carboxylic acid (3.85 g, 8.65 mmol) in THF (15 mL). The
reaction mixture was heated at 60.degree. C. for 1.5 h, cooled to
rt, treated with cyclopropanesulfonamide (1.36 g, 11.2 mmol),
stirred 10 min and then treated with the dropwise addition of a
solution of DBU (2.0 mL, 13 mmol) in THF (3 mL). The reaction
mixture was stirred at rt overnight, diluted with EtOAc (100 mL)
and washed with H.sub.2O (.about.30 mL), 1N HCl (aq.) (2.times.50
mL) and brine (.about.30 mL). The combined aqueous layers were
extracted with EtOAc (100 mL) and the organic layer was washed with
1N HCl (aq.) (.about.50 mL). The combined organic layers were
washed with brine (.about.30 mL), dried (MgSO.sub.4), filtered and
concentrated. The residue was stirred with Et.sub.2O (.about.100
mL) for 2 h and the solids were collected by filtration, rinsed
with Et.sub.2O and dried to yield methyl
13-cyclohexyl-10-((cyclopropylsulfonyl)carbamoyl)-3-methoxy-7H-ind-
olo[2,1-a][2]benzazepine-6-carboxylate (4.24 g, 7.73 mmol, 89%) as
a pale yellow solid which was used without further purification. 1H
NMR (300 MHz, CDCl.sub.3) .delta. 1.08-2.13 (m, 14H), 2.73-2.87 (m,
1H), 3.13-3.24 (m, 1H), 3.82 (s, 3H), 3.89 (s, 3H), 4.04-4.27 (m,
1H), 5.50-5.71 (m, 1H), 6.98 (d, J=2.6 Hz, 1H), 7.08 (dd, J=8.8,
2.6 Hz, 1H), 7.44 (dd, J=8.4, 1.1 Hz, 1H), 7.50 (d, J=8.8 Hz, 1H),
7.80 (s, 1H), 7.86 (d, J=8.4 Hz, 1H), 8.11 (br s, 1H), 8.78 (br s,
1H). LCMS: m/e 549 (M+H).sup.+, ret time 3.79 min, column B, 4
minute gradient.
##STR00090##
[0111]
13-Cyclohexyl-10-((cyclopropylsulfonyl)carbamoyl)-3-methoxy-7H-indo-
lo[2,1-a][2]benzazepine-6-carboxylic acid. Methyl
13-cyclohexyl-10-((cyclopropylsulfonyl)carbamoyl)-3-methoxy-7H-indolo[2,1-
-a][2]benzazepine-6-carboxylate (1.0 g, 1.8 mmol) was dissolved
into MeOH//THF (1:1, 24 mL) and treated with 1M aqueous NaOH (5
mL). The reaction mixture was stirred and heated at 60.degree. C.
for 1.5 h and cooled to rt. The clear solution was neutralized with
1M aqueous HCl (5 mL) and concentrated to remove organic solvents.
The resultant solids were collected by filtration, washed with
H.sub.2O and dried under vacuum to yield
13-cyclohexyl-10-((cyclopropylsulfonyl)carbamoyl)-3-methoxy-7H-i-
ndolo[2,1-a][2]benzazepine-6-carboxylic acid (1.0 g, 1.7 mmol, 94%)
as a bright yellow solid (with 0.75 equiv. of THF) which was used
without further purification. 1H NMR (300 MHz, CD.sub.3OD) .delta.
1.11-2.24 (m, 17H, 3H from THF), 2.81-2.96 (m, 1H), 3.17-3.28 (m,
1H), 3.69-3.79 (m, 3H, from THF), 3.94 (s, 3H), 4.07-4.33 (m, 1H),
5.55-5.81 (m, 1H), 7.14-7.24 (m, 2H), 7.55-7.64 (m, 2H), 7.88-7.94
(m, 2H), 8.20 (br s, 1H). LCMS: m/e 535 (M+H).sup.+, ret time 3.73
min, column B, 4 minute gradient.
##STR00091##
[0112] Methyl
8-cyclohexyl-5-((cyclopropylsulfonyl)carbamoyl)-11-methoxy-1,12b-dihydroc-
yclopropa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylate. To
slurry of sodium hydride (60% dispersion in mineral oil, 370 mg,
9.2 mmol) in DMSO (8 mL) stirring under N.sub.2 was added
trimethylsulfoxonium iodide (2.03 g, 9.2 mmol). The reaction
mixture was stirred for 45 min and then methyl
13-cyclohexyl-10-((cyclopropylsulfonyl)carbamoyl)-3-methoxy-7H-indolo[2,1-
-a][2]benzazepine-6-carboxylate (2.2 g, 4.0 mmol) in DMSO (5 mL)
was added (flask rinsed with DMSO (2.times.3 mL)). The reaction
mixture was stirred 1 h, poured into 0.25N HCl (100 mL), and
extracted with EtOAc (150 mL). The organic layer was washed with
brine (20 mL) and the combined aqueous layers were extracted with
EtOAc (100 mL). The combine organic layers were washed with brine
(.about.20 mL), dried (MgSO.sub.4), filtered and concentrated to
dryness. The residue was stirred with EtOAc/Et.sub.2O (1:3, 50 mL)
and the solids were removed by filtration. The motherliquor was
concentrated and dried under high vacuum to yield methyl
8-cyclohexyl-5-((cyclopropylsulfonyl)carbamoyl)-11-methoxy-1,12b-dihydroc-
yclopropa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylate (1.92 g,
3.4 mmol, 85%) as a yellow solid which was used without further
purification. Presents as a .about.2:1 mixture of rotamers or
atrope isomers. 1H NMR (300 MHz, CD.sub.3OD) .delta. 0.19-0.26 (m,
0.4H), 0.78-2.19 (m, 15.6H), 2.64-3.02 (m, 2H), 3.16-3.28 (m, 1H),
3.41 (d, J=15.0 Hz, 0.6H), 3.51 (s, 1.8H), 3.80 (s, 1.2H), 3.88 (s,
3H), 4.00 (d, J=15.0 Hz, 0.4H), 5.22 (d, J=15.0 Hz, 0.4H), 5.42 (d,
J=15.0 Hz, 0.6H), 6.93-7.01 (m, 1H), 7.12 (d, J=2.6 Hz, 0.4H), 7.19
(d, J=2.6 Hz, 0.6H), 7.25 (d, J=8.8 Hz, 0.6H), 7.29 (d, J=8.8 Hz,
0.4H), 7.55 (dd, J=8.8, 1.5 Hz, 0.6H), 7.63 (dd, J=8.8, 1.5 Hz,
0.4H), 7.85 (d, J=8.8 Hz, 0.6H), 7.88 (d, J=8.8 Hz, 0.4H), 8.08 (d,
J=1.5 Hz, 0.4H), 8.31 (d, J=1.5 Hz, 0.6H). LCMS: m/e 563
(M+H).sup.+, ret time 3.75 min, column B, 4 minute gradient.
##STR00092##
[0113]
8-Cyclohexyl-5-((cyclopropylsulfonyl)carbamoyl)-11-methoxy-1,12b-di-
hydrocyclopropa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic
acid. Methyl
8-cyclohexyl-5-((cyclopropylsulfonyl)carbamoyl)-11-methoxy-1,12b-d-
ihydrocyclopropa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylate
(1.92 g, 3.41 mmol) was dissolved into MeOH//THF (1:1, 40 mL) and
treated with 1M aqueous NaOH (8 mL). The reaction mixture was
stirred and heated at 60.degree. C. for 2 h and cooled to rt. The
clear solution was neutralized with 1M aqueous HCl (8 mL) and
concentrated to remove organic solvents. The resultant solids were
collected by filtration, washed with H.sub.2O and dried under
vacuum to yield
8-cyclohexyl-5-((cyclopropylsulfonyl)carbamoyl)-11-methoxy-1,12b-dihydroc-
yclopropa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid
(1.66 g, 3.03 mmol, 89%) as a yellow powder which was used without
further purification. Presents as a 1:1 mixture of rotamers or
atrope isomers. 1H NMR (300 MHz, CDCl.sub.3) .delta. 0.32 (t, J=6.2
Hz, 0.5H), 0.71-2.12 (m, 15.5H), 2.61-2.94 (m, 2H), 3.16-3.27 (m,
1H), 3.41 (d, J=15.0 Hz, 0.5H), 3.82 (s, 1.5H), 3.86 (s, 1.5H),
3.99 (d, J=15.0 Hz, 0.5H), 5.28 (d, J=15.0 Hz, 0.5H), 5.49 (d,
J=15.0 Hz, 0.5H), 6.85 (dd, J=8.4, 2.6 Hz, 0.5H), 6.91 (dd, J=8.4,
2.6 Hz, 0.5H), 6.96 (d, J=2.6 Hz, 0.5H), 7.08 (d, J=2.6 Hz, 0.5H),
7.19 (d, J=8.4 Hz, 0.5H), 7.24 (d, J=8.4 Hz, 0.5H), 7.61 (d, J=8.4
Hz, 0.5H), 7.67 (d, J=8.4 Hz, 0.5H), 7.83 (d, J=8.4 Hz, 0.5H), 7.85
(d, J=8.4 Hz, 0.5H), 8.06 (s, 0.5H), 8.35 (s, 0.5H), 9.31-10.35 (m,
1H). LCMS: m/e 547 (M-H).sup.-, ret time 2.06 min, column A, 4
minute gradient.
##STR00093##
[0114] Methyl
13-cyclohexyl-10-((isopropylsulfonyl)carbamoyl)-3-methoxy-7H-indolo[2,1-a-
][2]benzazepine-6-carboxylate. 1,1'-Carbonyldiimidazole (262 mg,
1.62 mmol) was added to a slurry of
13-cyclohexyl-3-methoxy-6-(methoxycarbonyl)-7H-indolo[2,1-a][2]benzazepin-
e-10-carboxylic acid (603 mg, 1.36 mmol) in THF (3 mL). The
reaction mixture was heated at 60.degree. C. for 1.5 h, cooled to
rt, treated with propane-2-sulfonamide (200 mg, 1.62 mmol), stirred
10 min and then treated with the dropwise addition of a solution of
DBU (0.27 mL, 1.8 mmol) in THF (0.75 mL). The reaction mixture was
stirred at rt overnight, diluted with EtOAc (15 mL) and washed with
H.sub.2O (.about.5 mL), 1N HCl (aq.) (2.times.10 mL) and brine
(.about.5 mL). The combined aqueous layers were extracted with
EtOAc (15 mL) and the organic layer was washed with 1N HCl (aq.)
(.about.10 mL). The combined organic layers were washed with brine
(.about.5 mL), dried (MgSO.sub.4), filtered and concentrated. The
residue was stirred with Et.sub.2O (.about.15 mL) for 2 h and the
solids were collected by filtration, rinsed with Et.sub.2O and
dried to yield methyl
13-cyclohexyl-10-((isopropylsulfonyl)carbamoyl)-3-methoxy-7H-indolo[2,1-a-
][2]benzazepine-6-carboxylate (640 mg, 1.2 mmol, 85%) as a bright
yellow solid which was used without further purification. 1H NMR
(300 MHz, CDCl.sub.3) .delta. 1.12-2.13 (m, 10H), 1.47 (d, J=7.0
Hz, 6H), 2.73-2.86 (m, 1H), 3.82 (s, 3H), 3.89 (s, 3H), 4.06-4.26
(m, 1H), 4.09 (septet, J=7.0 Hz, 1H), 5.51-5.71 (m, 1H), 6.98 (d,
J=2.6 Hz, 1H), 7.08 (dd, J=8.4, 2.6 Hz, 1H), 7.44 (dd, J=8.4, 1.5
Hz, 1H), 7.50 (d, J=8.4 Hz, 1H), 7.80 (s, 1H), 7.87 (d, J=8.4 Hz,
1H), 8.10 (d, J=1.5 Hz, 1H), 8.57 (s, 1H). LCMS: m/e 551
(M+H).sup.+, ret time 3.87 min, column B, 4 minute gradient.
##STR00094##
[0115] Methyl
8-cyclohexyl-5-((isopropylsulfonyl)carbamoyl)-11-methoxy-1,12b-dihydrocyc-
lopropa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylate. To slurry
of sodium hydride (60% dispersion in mineral oil, 97 mg, 2.4 mmol)
in DMSO (2 mL) stirring under N.sub.2 was added
trimethylsulfoxonium iodide (530 g, 2.4 mmol). The reaction mixture
was stirred for 45 min and then methyl
13-cyclohexyl-10-((isopropylsulfonyl)carbamoyl)-3-methoxy-7H-indolo[2,1-a-
][2]benzazepine-6-carboxylate (578 g, 1.05 mmol) in DMSO (1.5 mL)
was added (flask rinsed with DMSO (2.times.0.75 mL)). The reaction
mixture was stirred 1 h, poured into 0.25N HCl (25 mL), and
extracted with EtOAc (40 mL). The organic layer was washed with
brine (10 ml) and the combined aqueous layers were extracted with
EtOAc (25 mL). The combine organic layers were washed with brine
(.about.10 mL), dried (MgSO.sub.4), filtered and concentrated to
dryness. The residue was stirred with EtOAc/Et.sub.2O (1:4, 10 mL)
and the solids were removed by filtration. The motherliquor was
concentrated and dried under high vacuum to yield methyl
8-cyclohexyl-5-((isopropylsulfonyl)carbamoyl)-11-methoxy-1,12b-dih-
ydrocyclopropa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylate
(620 mg, 1.0 mmol, quant.) as a yellow solid which was used without
further purification. Presents as a .about.2:1 mixture of rotamers
or atrope isomers. 1H NMR (300 MHz, CDCl.sub.3) .delta. 0.32-0.39
(m, 0.4H), 0.77-2.09 (m, 17.6H), 2.60-2.96 (m, 2H), 3.41 (d, J=15.0
Hz, 0.6H), 3.53 (s, 1.8H), 3.79 (s, 1.2H), 3.87 (s, 3H), 4.02-4.14
(m, 1.4H), 5.14 (d, J=15.0 Hz, 0.4H), 5.39 (d, J=15.0 Hz, 0.6H),
6.89 (dd, J=8.4, 2.6 Hz, 0.4H), 6.91 (dd, J=8.4, 2.6 Hz, 0.6H),
7.00 (d, J=2.6 Hz, 0.4H), 7.11 (d, J=2.6 Hz, 0.6H), 7.23 (d, J=8.4
Hz, 0.6H), 7.25 (d, J=8.4 Hz, 0.4H), 7.38 (dd, J=8.4, 1.5 Hz,
0.6H), 7.43 (dd, J=8.4, 1.5 Hz, 0.4H), 7.83 (d, J=8.4 Hz, 0.6H),
7.86 (d, J=8.4 Hz, 0.4H), 7.96 (d, J=1.5 Hz, 0.4H), 8.20 (d, J=1.5
Hz, 0.6H), 8.39 (s, 0.4H), 8.43 (s, 0.6H). LCMS: m/e 563
(M-H).sup.-, ret time 3.00 min, column A, 4 minute gradient.
##STR00095##
[0116]
8-Cyclohexyl-5-((isopropylsulfonyl)carbamoyl)-11-methoxy-1,12b-dihy-
drocyclopropa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid.
Methyl
8-cyclohexyl-5-((isopropylsulfonyl)carbamoyl)-11-methoxy-1,12b-dihydrocyc-
lopropa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylate (606 mg,
1.07 mmol) was dissolved into MeOH//THF (1:1, 14 mL) and treated
with 1M aqueous NaOH (2.5 mL). The reaction mixture was stirred and
heated at 60.degree. C. for 2 h and cooled to rt. The clear
solution was neutralized with 1M aqueous HCl (2.5 mL) and
concentrated to remove organic solvents. The residue was stirred
with H.sub.2O (10 mL) overnight and the resultant solids were
collected by filtration, washed with H.sub.2O and dried under
vacuum to yield
8-cyclohexyl-5-((isopropylsulfonyl)carbamoyl)-11-methoxy-1,12b-dihydrocyc-
lopropa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid (530
mg, 0.96 mmol, 90%) as a bright yellow solid which was used without
further purification. Presents as a .about.2:1 mixture of rotamers
or atrope isomers. 1H NMR (300 MHz, CD.sub.3OD) .delta. 0.23-0.30
(m, 0.4H), 0.80-2.24 (m, 17.6H), 2.70-3.11 (m, 2H), 3.46 (d, J=15.0
Hz, 0.6H), 3.95 (s, 3H), 3.93-4.10 (m, 1.4H), 5.29 (d, J=15.0 Hz,
0.4H), 5.48 (d, J=15.0 Hz, 0.6H), 6.98-7.05 (m, 1H), 7.16 (d, J=2.6
Hz, 0.4H), 7.23 (d, J=2.6 Hz, 0.6H), 7.29 (d, J=8.8 Hz, 0.6H), 7.33
(d, J=8.8 Hz, 0.4H), 7.56 (dd, J=8.8, 1.5 Hz, 0.6H), 7.64 (dd,
J=8.4, 1.5 Hz, 0.4H), 7.87 (d, J 8.8 Hz, 0.6H), 7.92 (d, J=8.4 Hz,
0.4H), 8.13 (d, J=1.5 Hz, 0.4H), 8.31 (d, J=1.5 Hz, 0.6H). LCMS:
m/e 551 (M+H).sup.+, ret time 3.74 min, column B, 4 minute
gradient.
##STR00096##
[0117] Methyl
10-((aminosulfonyl)carbamoyl)-13-cyclohexyl-3-methoxy-7H-indolo[2,1-a][2]-
benzazepine-6-carboxylate. 1,1'-Carbonyldiimidazole (1.23 g, 7.60
mmol) was added to a slurry of
13-cyclohexyl-3-methoxy-6-(methoxycarbonyl)-7H-indolo[2,1-a][2]benzazepin-
e-10-carboxylic acid (2.6 g, 5.8 mmol) in THF (11 mL). The reaction
mixture was heated at 60.degree. C. for 1.5 h, cooled to rt,
treated with sulfamide (1.12 g, 11.7 mmol), stirred 10 min and then
treated with the dropwise addition of a solution of DBU (1.8 mL,
11.7 mmol) in THF (3 mL). The reaction mixture was stirred at rt
for 3 h, diluted with EtOAc (80 mL) and CH.sub.2Cl.sub.2 (100 mL)
and concentrated to dryness. The residue was diluted with
CH.sub.2Cl.sub.2 (100 mL) and washed with 1N HCl (aq.) (2.times.100
mL). The combined aqueous layers were extracted with
CH.sub.2Cl.sub.2 (100 mL) and the combined organic layers were
washed with 1/2 saturated brine (.about.50 mL), dried (MgSO.sub.4),
filtered and concentrated. The residue was stirred with Et.sub.2O
(.about.75 mL) for 1 h and the solids were collected by filtration,
rinsed with Et.sub.2O and dried to yield methyl
10-((aminosulfonyl)carbamoyl)-13-cyclohexyl-3-methoxy-7H-indolo[2,1-a][2]-
benzazepine-6-carboxylate (2.8 g, 5.3 mmol, 91%) as a bright yellow
solid which was used without further purification. 1H NMR (300 MHz,
CDCl.sub.3) .delta. 1.08-2.10 (m, 10H), 2.71-2.84 (m, 1H), 3.79 (s,
3H), 3.89 (s, 3H), 4.00-4.18 (m, 1H), 5.50-5.64 (m, 1H), 5.68 (s,
2H), 6.97 (d, J=2.6 Hz, 1H), 7.07 (dd, J=8.8, 2.6 Hz, 1H), 7.46
(dd, J=8.4, 1.5 Hz, 1H), 7.49 (d, J=8.8 Hz, 1H), 7.78 (s, 1H), 7.82
(d, J=8.4 Hz, 1H), 8.10 (br s, 1H), 9.49 (s, 1H). LCMS: m/e 524
(M+H).sup.+, ret time 3.60 min, column B, 4 minute gradient.
##STR00097##
[0118]
10-((Aminosulfonyl)carbamoyl)-13-cyclohexyl-3-methoxy-7H-indolo[2,1-
-a][2]benzazepine-6-carboxylic acid. Methyl
10-((aminosulfonyl)carbamoyl)-13-cyclohexyl-3-methoxy-7H-indolo[2,1-a][2]-
benzazepine-6-carboxylate (725 mg, 1.39 mmol) was dissolved into
MeOH//THF (1:1, 16 mL) and treated with 1M aqueous NaOH (3 mL). The
reaction mixture was stirred and heated at 60.degree. C. for 0.5 h
and cooled to rt. The reaction solution was diluted with
MeOH/H.sub.2O (2:1, 15 mL), neutralized with 1M aqueous HCl (3 mL)
and concentrated to remove organic solvents. The resultant solids
were collected by filtration, washed with H.sub.2O and dried under
vacuum to yield
10-((aminosulfonyl)carbamoyl)-13-cyclohexyl-3-methoxy-7H-indolo[2,1-a][2]-
benzazepine-6-carboxylic acid (650 g, 1.3 mmol, 92%) as a bright
yellow solid which was used without further purification. 1H NMR
(300 MHz, CDCl.sub.3) .delta. 1.16-2.22 (m, 10H), 2.82-2.96 (m,
1H), 3.94 (s, 3H), 4.07-4.29 (m, 1H), 5.57-5.80 (m, 1H), 7.14-7.23
(m, 2H), 7.55-7.63 (m, 2H), 7.88-7.94 (m 2H), 8.18 (s, 1H). LCMS:
m/e 510 (M+H).sup.+, ret time 2.85 min, column B, 4 minute
gradient.
##STR00098##
[0119] Methyl
5-((aminosulfonyl)carbamoyl)-8-cyclohexyl-11-methoxy-1,12b-dihydrocyclopr-
opa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylate. To slurry of
sodium hydride (60% dispersion in mineral oil, 350 mg, 8.8 mmol) in
DMSO (8 mL) stirring under N.sub.2 was added trimethylsulfoxonium
iodide (1.93 g, 8.8 mmol) in three portions. The reaction mixture
was stirred for 0.5 h and then methyl
10-((aminosulfonyl)carbamoyl)-13-cyclohexyl-3-methoxy-7H-indolo[2,1-a][2]-
benzazepine-6-carboxylate (2.0 g, 3.8 mmol) in DMSO (8 mL) was
added (flask rinsed with DMSO (2.times.2 mL)). The reaction mixture
was stirred 1 h, poured into 0.25N HCl (100 mL), and diluted with
CH.sub.2Cl.sub.2 (100 mL). The solution was filtered to collect
solids, and the organic layer of the motherliquor was separated and
concentrated to dryness. The residue was dissolved into EtOAc
(.about.150 mL) was washed with H.sub.2O (.about.50 mL) and brine
(.about.50 mL) dried (MgSO.sub.4), filtered and concentrated to
dryness. The residue was stirred with EtOAc/Et.sub.2O (4:1, 50 mL)
and the solids were collected by filtration and washed with EtOAc.
These solids were combined with the initially collected solids to
yield methyl 5-((amino
sulfonyl)carbamoyl)-8-cyclohexyl-11-methoxy-1,12b-dihydrocyclopropa[d]ind-
olo[2,1-a][2]benzazepine-1a(2H)-carboxylate (1.39 g, 2.6 mmol, 68%)
as a tan solid which was used without further purification.
Presents as a 1:1 mixture of rotamers or atrope isomers. 1H NMR
(300 MHz, DMSO-d.sub.6) .delta. 0.13-0.21 (m, 0.5H), 1.06-2.12 (m,
11.5H), 2.64-2.94 (m, 2H), 3.46 (s, 1.5H), 3.49 (d, J=15.0 Hz,
0.5H), 3.75 (s, 1.5H), 3.85 (s, 3H), 4.02 (d, J=15.0 Hz, 0.5H),
5.21 (d, J=15.0 Hz, 0.5H), 5.42 (d, J=15.0 Hz, 0.5H), 6.99-7.09 (m,
1H), 7.17-7.31 (m, 1H), 7.41 (s, 0.5H), 7.43 (s, 0.5H), 7.66-7.56
(m, 1H), 7.82 (d, J=8.4 Hz, 0.5H), 7.87 (d, J=8.8 Hz, 0.5H), 8.25
(s, 0.5H), 8.47 (s, 0.5H), 11.62 (s, 0.5H), 11.69 (s, 0.5H). LCMS:
m/e 538 (M+H).sup.+, ret time 3.56 min, column B, 4 minute
gradient.
##STR00099##
[0120]
5-((Aminosulfonyl)carbamoyl)-8-cyclohexyl-11-methoxy-1,12b-dihydroc-
yclopropa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid.
Methyl
5-((aminosulfonyl)carbamoyl)-8-cyclohexyl-11-methoxy-1,12b-dihydrocyclopr-
opa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylate (1.1 mg, 2.0
mmol) was dissolved into MeOH//THF (1:1, 24 mL) and treated with 1M
aqueous NaOH (5 mL). The reaction mixture was stirred and heated at
60.degree. C. for 2 h and cooled to rt. The clear solution was
neutralized with 1M aqueous HCl (5 mL) and concentrated to remove
organic solvents. The residue was stirred with H.sub.2O (10 mL) for
1 h and the resultant solids were collected by filtration, washed
with H.sub.2O and dried under vacuum to yield
5-((aminosulfonyl)carbamoyl)-8-cyclohexyl-11-methoxy-1,12b-dihydrocyclopr-
opa[d]indolo[2,1-a][2]benzazepine-1a(2H)-carboxylic acid (1.05 mg,
2.0 mmol, 98%) as a light yellow solid which was used without
further purification. Presents as a 1:1 mixture of rotamers or
atrope isomers. 1H NMR (300 MHz, DMSO-d.sub.6) .delta. 0.08-0.17
(m, 0.5H), 0.79-2.13 (m, 11.5H), 2.65-2.94 (m, 2H), 3.44 (d, J=14.6
Hz, 0.5H), 3.85 (s, 3H), 3.96 (d, J=14.6 Hz, 0.5H), 5.20 (d, J=14.6
Hz, 0.5H), 5.40 (d, J=14.6 Hz, 0.5H), 6.98-7.08 (m, 1H), 7.17-7.46
(m, 4H), 7.58 (d, J=8.1 Hz, 0.5H), 7.62 (d, J=8.1 Hz, 0.5H), 7.81
(d, J=8.8 Hz, 0.5H), 7.87 (d, J=8.8 Hz, 0.5H), 8.25 (s, 0.5H), 8.44
(s, 0.5H), 11.48-13.19 (m, 2H). LCMS: m/e 524 (M+H).sup.+, ret time
3.51 min, column B, 4 minute gradient.
[0121] Intermediates 40-44 use the experimental procedures that
follow until noted.
##STR00100##
[0122] Dry NaH (96 mg, 4 mmol) was added to a stirred suspension of
trimethylsulfoxonium chloride (567 mg, 4.4 mmol) in an. DMSO (10
mL) under nitrogen. The resultant mixture was stirred at rt for
30-45 min and then neat olefin (1.0, 2 mmol) was added in small
portions. The suspension was diluted with DMSO (5 mL) and heated at
50.degree. C. for 3-4 h. Reaction mixture was allowed to cool to rt
and water was added. Precipitated solid was filtered and washed
with water and then air dried overnight to afford 1.15 g of crude
product which was purified by flash column chromatography (silica
gel, 3% MeOH in DCM), to provide pure desired cyclopropyl compound
(0.96 g), as a off-white solid: LC/MS: Retention time 3.816 min;
m/e 516 (MH.sup.+). .sup.1H NMR (400 MHz, CDCl.sub.3): The product
was observed to exist as inter-converting rotamers.
##STR00101##
[0123] The tert-butyl ester (515 mg, 1 mmol) and TFA (5 mL) in an.
DCM (10 mL) was stirred at rt until hydrolysis is complete (8-12
hr). Excess TFA and DCM were evaporated to dryness to afford
desired acid (0.47 g, 100%) as a light beige solid. LC/MS:
Retention time 2.245 min; m/e 460 (MH.sup.+). .sup.1H NMR (400 MHz,
CDCl.sub.3): The product was observed to exist as inter-converting
rotamers.
##STR00102##
[0124] General procedure. A mixture of acid (1 equiv) and
carbonyldiimidazole (1.5 equiv) in an. THF was heated at 50.degree.
C. for 30 min and allowed to cool to rt. Then 1 equiv of either
sulfamide (R.dbd.NR.sub.2) or sulfonamide (R=alkyl or aryl) and DBU
(2 equiv) were added consecutively. The resultant mixture was
stirred at rt overnight. After acidic aqueous workup, isolated
crude product was purified by prep. HPLC to afford the product.
##STR00103##
[0125] Methyl ester moiety was hydrolyzed using 1N NaOH in THF-MeOH
to provide the corresponding acids.
##STR00104##
[0126] Acid derivatives (1 equiv) were combined with corresponding
amine (RRNH, 1.2 equiv), triethylamine (2-3 equiv) and TBTU (1.3
equiv) in an. DMF and stirred at rt for 1-2 h until completion of
the amide coupling. Isolated crude products were purified by prep.
HPLC to provide desired amides.
[0127] Intermediates 45-49 described below were analyzed by the
following LC/MS method: Analysis Conditions: Column:
PHENOMENNEX-LUNA 3.0.times.50 mm S10; Mobile Phase: (A) 10:90
methanol-water; (B) 90:10 methanol-water; Buffer: 0.1% TFA;
Gradient Range: 0-100% B; Gradient Time: 2 min; Flow Rate: 4
mL/min; Analysis Time: 3 min; Detection: Detector 1: UV at 220 nm;
Detector 2: MS (ESI+)/.
##STR00105##
[0128]
(+/-)-8-Cyclohexyl-1,1a,2,12b-tetrahydro-11-methoxy-1a-(methoxycarb-
onyl)-cycloprop[d]indolo[2,1-a][2]benzazepine-5-carboxylic acid,
tert-butyl ester. LC/MS: Retention time 3.816 min; m/e 516
(MH.sup.+). .sup.1H NMR (400 MHz, CDCl.sub.3): The product was
observed to exist as inter-converting rotamers.
##STR00106##
[0129] (+/-)-8-Cyclohexyl-1,1a,
2,12b-tetrahydro-11-methoxy-1a-(methoxycarbonyl)-cycloprop[d]indolo[2,1-a-
][2]benzazepine-5-carboxylic acid. Retention time 2.245 min; m/e
460 (MH.sup.+). .sup.1H NMR (400 MHz, CDCl.sub.3). The product was
observed to exist as inter-converting rotamers.
##STR00107##
[0130]
(+/-)-8-cyclohexyl-5-(morpholinosulfonylcarbamoyl)-1,1a,2,12b-tetra-
hydro-11-methoxy-cycloprop[d]indolo[2,1-a][2]benzazepine-1a-carboxylic
acid. The product was purified by prep HPLC and isolated as a beige
solid. LC/MS: Retention time: 1.968 min; m/e 460 (MH.sup.+).
.sup.1H NMR (400 MHz, CDCl.sub.3). The product was observed to
exist as inter-converting rotamers.
##STR00108##
[0131]
(+/-)-8-cyclohexyl-5-(4-methylpiperazin-1-ylsulfonylcarbamoyl)-1,1a-
,
2,12b-tetrahydro-11-methoxy-cycloprop[d]indolo[2,1-a][2]benzazepine-1a-c-
arboxylic acid. The product was purified by prep HPLC and isolated
in mono TFA salt form as a beige solid. LC/MS: Retention time:
1.687 min; m/e 607 (MH.sup.+). .sup.1H NMR (400 MHz, CDCl.sub.3).
The product was observed to exist as inter-converting rotamers.
##STR00109##
[0132] (+/-)-8-cyclohexyl-5-(cyclopropylsulfonylcarbamoyl)-1,1a,
2,12b-tetrahydro-11-methoxy-cycloprop[d]indolo[2,1-a][2]benzazepine-1a-ca-
rboxylic acid. LC/MS: Retention time: 2.030 min; m/e 549
(MH.sup.+). .sup.1H NMR (400 MHz, CDCl.sub.3): The product was
observed to exist as inter-converting rotamers.
[0133] Intermiates 50-60 were analyzed by the following LC/MS
method: Start % B: 0; Final % B: 100; Gradient time: 3 min; Stop
time: 4 min; Flow rate: 4 ml/min; Wavelenth: 220; Solvent A: 10%
MeOH/90% H.sub.2O/0.1% Trifluoroacetic Acid; Solvent B: 10%
H.sub.2O/90% MeOH/0.1% Trifluoroacetic Acid; Column. XBridge
4.6.times.50 mm S5.
##STR00110##
[0134] A mixture of the acid (1.3 g, 2.83 mmol) and CDI (0.64 g,
3.97 mmol) in THF (20 mL) was heated at 50.degree. C. for 0.5 h,
cooled down and added methylsulfonamide (0.4 g, 4.2 mmol) and DBU
(0.264 mL, 1.77 mmol). The mixture was stirred for 20 h and diluted
with EtOAc, washed with cold 1N HCl (2.times.), brine, dried
(MgSO4), removed the solvent and purified by flash (Biotage 40 M)
to afford the compound 1-2 (1.28 g, 85%) as a pale yellow solid.
LC-MS retention time: 3.51; MS m/z 537 (M+H). Compound 1-2 was
observed to exist as inter-converting rotamers. The major isomer: p
.sup.1H NMR (400 MHz, CHLOROFORM-D) .delta. ppm 1.11-2.17 (m, 12H),
2.84-2.98 (m, 2H), 3.43 (d, J=14.86 Hz, 1H), 3.49 (s, 3H), 3.55 (s,
3H), 3.89 (s, 3H), 5.40 (d, J=15.11 Hz, 1H), 6.91-6.96 (m, 1H),
7.13 (d, J=2.52 Hz, 1H), 7.22-7.27 (m, 1H), 7.39 (dd, J=8.31, 1.51
Hz, 1H), 7.85 (d, J=8.81 Hz, 1H), 8.23 (d, J=1.26 Hz, 1H), 8.75 (s,
1H).
##STR00111##
[0135] To a solution of the ester (1.28 g, 2.4 mmol) in THF (5 mL)
and MeOH (5 mL) was added NaOH (1N, 12 mL, 12 mmol). After being
stirred at room temperature for 3 h, the mixture was diluted with
EtOAc, washed with cold 1N HCl, brine, dried (MgSO4), and removed
the solvent in vacuo to afford the acid as a beige solid (1.20 g,
96%). LC-MS retention time: 3.46; MS m/z 523 (M+H). The acid was
observed to exist as inter-converting rotamers (.about.1/1) .sup.1H
NMR (400 MHz, CHLOROFORM-D).
##STR00112##
[0136] Typical general procedure for amine coupling: To a mixture
of the acid (0.060 g, 0.11 mmol) and a secondary/tertiary amine
containing diamine bishydrochloric acid salt (0.034 g, 0.17 mmol)
in DMC (1.5 mL) was added Et.sub.3N (0.096 mL, 0.69 mmol) and HBTU
(0.065 g, 0.17 mmol). The mixture was stirred at room temperature
for 0.5 h, diluted with MeOH, removed the solvent. The residue was
dissolved in methanol, filtered, and purified by prep-HPLC to
afford A TFA salt of an amide 1 (0.0378 g, 82%) as TFA salt which
was characterized by LC-MS and .sup.1H NMR.
##STR00113##
[0137] The product was prepared from the acid (0.47 g, 44%). LC-MS
retention time: 3.54; MS m/z 551 (M+H).
##STR00114##
[0138] The product was prepared (0.43 g, 94%). LC-MS retention
time: 3.49; MS m/z 537 (M+H).
##STR00115##
[0139] The product was prepared from the acid (0.96 g, 59%). LC-MS
retention time: 3.58; MS m/z 578 (M+H). compound was observed to
exist as inter-converting rotamers (3/4). The major isomer: .sup.1H
NMR (400 MHz, CHLOROFORM-D) .delta. ppm 1.16-1.59 (m, 4H), 1.72
(dd, J=9.44, 4.15 Hz, 3H), 1.88-2.12 (m, 4H), 2.24-2.36 (m, 2H),
2.75-2.97 (m, 2H), 3.44 (d, J=14.86 Hz, 1H), 3.56 (s, 3H), 3.89 (s,
3H), 4.09 (d, 1H), 4.24-4.37 (m, 4H), 5.41 (d, J=14.86 Hz, 1H),
6.92-6.96 (m, 1H), 7.13 (d, J=2.01 Hz, 1H), 7.24-7.30 (m, 1H), 7.39
(dd, J=8.31, 1.51 Hz, 1H), 7.84-7.88 (m, 1H), 8.24 (d, J=1.51 Hz,
1H).
##STR00116##
[0140] The product was prepared (0.93 g, 100%). LC-MS retention
time: 3.51; MS m/z 564 (M+H). Compound was observed to exist as
inter-converting rotamers (.about.3/4). The major isomer: .sup.1H
NMR (400 MHz) ppm 0.34-0.42 (m, 1H), 1.15-2.10 (m, 11H), 2.22-2.38
(m, 2H), 2.65-2.78 (m, 1H), 2.84-2.94 (m, J=3.02 Hz, 1H), 3.84 (s,
3H), 4.03 (d, J=15.11 Hz, 1H), 4.21-4.43 (m, 4H), 5.34 (d, J=14.86
Hz, 1H), 6.87 (dd, J=8.56, 2.77 Hz, 1H), 6.98 (d, J=2.52 Hz, 1H),
7.21 (d, J=8.31 Hz, 1 H), 7.69-7.75 (m, 1H), 7.86-7.90 (m, 1H),
8.13 (s, 1H).
##STR00117##
[0141] The product was prepared from the acid (0.109 g, 67%). LC-MS
retention time: 3.60; MS m/z 580 (M+H). Compound was observed to
exist as inter-converting rotamers (.about.5/4). The major isomer:
.sup.1H NMR (400 MHz) ppm 1.16-2.09 (m, 14H), 2.73-2.93 (m, 2H),
3.07 (s, 3H), 3.31-3.52 (m, 3H), 3.76 (s, 3H), 3.88 (s, 3H),
4.05-4.10 (m, 1H), 5.40 (d, J=15.11 Hz, 1H), 6.88-6.93 (m, 1H),
7.13 (d, J=2.27 Hz, 1H), 7.22-7.29 (m, 1H), 7.33-7.42 (m, 1H),
7.82-7.86 (m, 1H), 8.19 (d, J=1.51 Hz, 1H).
##STR00118##
[0142] The product was prepared (0.108 g, 100%). LC-MS retention
time: 3.55; MS m/z 566 (M+H).
##STR00119##
[0143] The product was prepared from the acid (0.127 g, 67%). LC-MS
retention time: 3.64; MS m/z 594 (M+H). Compound was observed to
exist as inter-converting rotamers: .sup.1H NMR (400 MHz) ppm
1.11-2.13 (m, 18H), 2.64 (dd, J=10.07, 6.80 Hz, 1H), 2.84-2.96 (m,
1H), 3.34-3.67 (m, 4H), 3.75 (s, 3H), 3.88 (s, 3H), 4.03-4.10 (m,
1H), 5.40 (d, J=15.36 Hz, 1H), 6.90-6.95 (m, 1H), 7.13 (d, J=2.01
Hz, 1 H), 7.21-7.29 (m, 1H), 7.33-7.39 (m, 1H), 7.83 (d, J=8.06 Hz,
1H), 8.20 (d, J=1.26 Hz, 1H).
##STR00120##
[0144] The product was prepared (0.126 g, 100%). LC-MS retention
time: 3.57; MS m/z 580 (M+H).
##STR00121##
[0145]
8-Cyclohexyl-11-methoxy-N-((1-methylcyclopropyl)sulfonyl)-1a-((3-me-
thyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahydrocyclop-
ropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide. CDI (26.4 mg,
0.163 mmol) was added to a stirring solution of
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid TFA (60 mg, 0.092 mmol) in THF (1 mL) and the reaction
was stirred at 60.degree. C. for 1 h and allowed to cool to rt.
Then 1-methylcyclopropane-1-sulfonamide (22 mg, 0.16 mmol) and DBU
(0.03 mL, 0.2 mmol) were added, and the reaction mixture was
stirred ON at rt. The reaction mixture was diluted with MeOH and
purified by preparative HPLC (H.sub.2O/CH.sub.3CN with 10 mM
NH.sub.4OAc buffer) to yield
8-cyclohexyl-11-methoxy-N-((1-methylcyclopropyl)sulfonyl)-1a-((3-methyl-3-
,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahydrocyclopropa[d-
]indolo[2,1-a][2]benzazepine-5-carboxamide (15.8 mg, 0.024 mmol,
26% yield) as a off white solid. The compound was isolated as a
mixture of enantiomers and presents as a 1:3 mixture of rotamers or
atrope isomers. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm 8.15
(s, 0.25H), 8.03 (s, 0.75H), 7.90 (d, J=8.8 Hz, 1H), 7.72-7.59 (m,
1H), 7.38-7.30 (m, 1H), 7.22 (s, 0.75H), 7.16 (s, 0.25H), 7.07-6.97
(m, 1H), 5.17 (d, J=15.0 Hz, 0.75H), 4.23-4.13 (m, 0.25H), 3.91 (s,
3H), 3.66 (d, J=15.0 Hz, 1H), 3.33-3.20 (m, 2H), 3.12-0.87 (m,
31.75H), 0.26-0.08 (m, 0.25H). LCMS: m/e=671 (M+H).sup.+, retention
time=2.33 min (Column=(3) phenomenex 10u C18 4.6.times.30 mm
Solvent A=5% CH.sub.3CN-95% H.sub.2O-10 mm Ammonium Acetate Solvent
B=95% CH.sub.3CN-5% H.sub.2O-10 mM Ammonium acetate, Start % B=0
Final % B=100 Gradient Time=4 min, Hold time=1 min, Flow Rate=4
mL/min)
##STR00122##
[0146]
8-Cyclohexyl-N-(isopropenylsulfonyl)-11-methoxy-1a-((3-methyl-3,8-d-
iazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,
2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
CDI (26.4 mg, 0.163 mmol) was added to a stirring solution of
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid TFA (60 mg, 0.092 mmol) in THF (1 mL) and the reaction
was stirred at 60.degree. C. for 1 h and allowed to cool to rt.
Then prop-1-ene-2-sulfonamide (20 mg, 0.16 mmol) and DBU (0.03 mL,
0.2 mmol) were added, and the reaction mixture was stirred ON at
rt. The reaction mixture was diluted with MeOH and purified by
preparative HPLC (H.sub.2O/CH.sub.3CN with 10 mM NH.sub.4OAc
buffer) to yield
8-cyclohexyl-N-(isopropenylsulfonyl)-11-methoxy-1a-((3-methyl-3,8-diazabi-
cyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,-
1-a][2]benzazepine-5-carboxamide (16.0 mg, 0.024 mmol, 26% yield)
as a white solid. The compound was isolated as a mixture of
enantiomers and presents as a 1:3 mixture of rotamers or atrope
isomers. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm 8.20 (s,
0.25H), 8.07 (s, 0.75H), 7.84-7.70 (m, 2H), 7.35-7.27 (m, 1H),
7.23-7.18 (m, 0.75H), 7.17-7.13 (m, 0.25H), 7.05-6.95 (m, 1H), 6.00
(s, 1H), 5.54 (s, 1H), 5.24-4.81 (m, 1H), 4.54-3.30 (m, 1H), 3.91
(s, 0.75H) 3.90 (s, 2.25H), 3.09-0.98 (m, 29.75H), 0.28-0.18 (m,
0.25H). LCMS: m/e=657 (M+H).sup.+, retention time=2.05 min
(Column=(3) phenomenex 10u C18 4.6.times.30 mm, Solvent A=5%
CH.sub.3CN-95% H.sub.2O-10 mm Ammonium Acetate Solvent B=95%
CH.sub.3CN-5% H.sub.2O-10 mM Ammonium acetate, Start % B=0 Final %
B=100 Gradient Time=4 min, Hold time=1 min, Flow Rate=4 mL/min)
##STR00123##
[0147] tert-Butyl
13-cyclohexyl-3-methoxy-64(3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbo-
nyl)-7H-indolo[2,1-a][2]benzazepine-10-carboxylate. HATU (680 mg,
1.8 mmol) was added to a stirring solution of
10-(tert-butoxycarbonyl)-13-cyclohexyl-3-methoxy-7H-indolo[2,1-a][2]benza-
zepine-6-carboxylic acid (670 mg, 1.37 mmol) and
3-methyl-3,8-diazabicyclo[3.2.1]octane di HCl salt (560 mg, 2.81
mmol) in DMF (6 mL) and TEA (1.2 mL, 8.2 mmol) and the reaction was
stirred for 30 min (complete by LCMS). The reaction mixture was
diluted with water (.about.35 mL) (precipitate formed) and stirred
ON. The precipitate was collected by filtration, flushed with water
and dried under high vacuum at 55.degree. C. to yield tert-butyl
13-cyclohexyl-3-methoxy-6-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carb-
onyl)-7H-indolo[2,1-a][2]benzazepine-10-carboxylate (775 mg, 1.30
mmol, 95% yield) as a light yellow solid. The material was used
without further purification. 1H NMR (300 MHz, CDCl.sub.3) .delta.
ppm 1.14-3.95 (m, 24H), 1.59 (s, 9H), 3.86 (s, 3H), 4.21-5.26 (m,
2H), 6.82 (s, 1H), 6.88 (d, J=2.6 Hz, 1H), 7.01 (dd, J=8.8, 2.6 Hz,
1H), 7.47 (d, J=8.8 Hz, 1H), 7.66 (dd, J=8.4, 1.1 Hz, 1H), 7.80 (d,
J=8.4 Hz, 1H), 8.02 (br s, 1H). LC-MS retention time: 3.72 min; m/z
596 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Phenomenex-Luna 10u C18 3.0.times.50
mm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 5
mL/min, a gradient of 100% solvent A/0% solvent B to 0% solvent
A/100% solvent B, a gradient time of 4 min, a hold time of 1 min,
and an analysis time of 5 min where solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a Micromass Platform for LC in electrospray mode.
##STR00124##
[0148] g. tert-Butyl
13-cyclohexyl-3-methoxy-6-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carb-
onyl)-7H-indolo[2,1-a][2]benzazepine-10-carboxylate (300 mg, 0.504
mmol) was dissolved into DCE (5 mL) and then TFA (700 .mu.l, 9.09
mmol) was added (reaction became green) and the reaction was
stirred at rt for 1 h (.about.70% conversion by LCMS). More TFA
(700 .mu.l, 9.09 mmol) was added and the reaction was stirred 1 h
(complete by LCMS). The reaction mixture was concentrated on a
rotary evaporator, diluted with diethyl ether and reconcentrated
twice to yield
13-cyclohexyl-3-methoxy-6-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carb-
onyl)-7H-indolo[2,1-a][2]benzazepine-10-carboxylic acid
trifluoroacetate (362 mg, 0.55 mmol, quant.) as a dark yellow
solid. Used without further purification. 1H NMR (300 MHz,
DMSO-d.sub.6) .delta. ppm 1.06-2.13 (m, 21H), 2.68-2.86 (m, 1H),
3.36-3.50 (m, 2H), 3.90 (s, 3H), 4.11-5.35 (m, 2H), 7.14 (s, 1H),
7.18-7.28 (m, 2H), 7.53 (d, J=8.4 Hz, 1H), 7.61 (dd, J=8.4, 1.1 Hz,
1H), 7.87 (d, J=8.4 Hz, 1H), 8.21 (br s, 1H), 9.55 (br s, 1H).
LC-MS retention time: 3.72 min; m/z 596 (MH+). LC-MS retention
time: 2.50 min; 538 m/z (MH-). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Phenomenex-Luna 10u
C18 4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% solvent A/0% solvent B to
0% solvent A/100% solvent B, a gradient time of 4 min, a hold time
of 1 min, and an analysis time of 5 min where solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray
mode.
##STR00125##
[0149]
13-Cyclohexyl-N-((1-(cyclopropylmethyl)cyclopropyl)sulfonyl)-3-meth-
oxy-6-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)
carbonyl)-7H-indolo[2,1-a][2]benzazepine-10-carboxamide. CDI (27
mg, 0.17 mmol) was added to a stirring solution of
13-cyclohexyl-3-methoxy-6-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carb-
onyl)-7H-indolo[2,1-a][2]benzazepine-10-carboxylic acid
trifluoroacetate (70 mg, 0.130 mmol)) in THF (0.5 mL) and the
reaction mixture was heated at 60.degree. C. for 1.5 h. The
reaction was cooled to rt and then
1-(cyclopropylmethyl)cyclopropane-1-sulfonamide (29.6 mg, 0.169
mmol) was added. The reaction was stirred 5 min. and then DBU
(0.029 mL, 0.195 mmol) was added. The reaction was stirred at rt
for 1 h, more DBU (0.029 mL, 0.195 mmol) was added and stirred at
rt ON (.about.75% conversion by LCMS). The reaction was diluted
with EtOAc (.about.2 mL) and washed with 1M aq. HCl (2.times.2 mL).
The organic layer was concentrated under a steam of nitrogen,
dissolved into MeOH (.about.3 mL) and purified by preparative HPLC
(Column: Xterra Prep MS C18 5u 30.times.100 mm, Eluent A: 5%
acetonitrile/water with 10 mM ammonium acetate, Eluent B: 95%
acetonitrile/water with 10 mM ammonium acetate, Flow Rate: 42
mL/min, linear gradient from 15% Eluent B to 100% Eluent B over 20
min) to yield
13-cyclohexyl-N-((1-(cyclopropylmethyl)cyclopropyl)sulfonyl)-3-methoxy-6--
((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-7H-indolo[2,1-a][2]be-
nzazepine-10-carboxamide (32 mg, 0.046 mmol, 35% yield) as a yellow
solid. 1H NMR (300 MHz, CDCl.sub.3) .delta. ppm 0.02-0.09 (m, 2H),
0.38-0.47 (m, 2H), 0.61-0.74 (m, 1H), 1.14-2.95 (m, 31H), 3.89 (s,
3H), 4.20-4.42 (m, 1H), 5.11-5.32 (m, 1H), 6.82 (s, 1H), 6.86-6.90
(m, 2H), 7.05 (dd, J=8.8, 2.6 Hz, 1H), 7.49 (d, J=8.8 Hz, 1H),
7.49-7.54 (m, 1H), 7.86 (d, J=8.4 Hz, 1H), 8.10 (br s, 1H). LC-MS
retention time: 3.46 min; m/z 697 (MH+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
Phenomenex-Luna 10u C18 3.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 5 mL/min, a gradient of 100%
solvent A/0% solvent B to 0% solvent A/100% solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a Micromass Platform for LC in
electrospray mode.
##STR00126##
[0150] tert-Butyl
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylate. Trimethylsulfoxonium iodide (375 mg, 1.69 mmol) was added
in three portions to s stirring slurry of a 60% NaH dispersion (68
mg, 1.7 mmol) in DMSO (1.5 mL) (foaming occurred). The reaction
mixture was stirred 20 min and then a solution of tert-butyl
13-cyclohexyl-3-methoxy-6-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carb-
onyl)-7H-indolo[2,1-a][2]benzazepine-10-carboxylate (438 mg, 0.735
mmol) in DMSO (2.5 mL) was added and the reaction was stirred for 1
h (no desired product by LCMS). The reaction mixture was heated at
90.degree. C. for 3 h (complete by LCMS), cooled to rt, quenched
with 0.25M aq. HCl (20 mL), and extracted with EtOAc (2.times.20
mL). The combined organic layers were washed with brine (20 mL),
dried (MgSO.sub.4), filtered and concentrated to yield 72009-057 as
an orange oil. The oil was used without further purification as
starting material in the preparation of
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid trifluoroacetate. The compound was isolated as a
mixture of enantiomers. LC-MS retention time: 3.68 min; m/z 610
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Phenomenex-Luna 10u C18 3.0.times.50
mm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 5
mL/min, a gradient of 100% solvent A/0% solvent B to 0% solvent
A/100% solvent B, a gradient time of 4 min, a hold time of 1 min,
and an analysis time of 5 min where solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a Micromass Platform for LC in electrospray mode.
##STR00127##
[0151]
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-
-5-carboxylic acid trifluoroacetate. The acid (448 mg, 0.735 mmol)
was dissolved into DCE (6 mL) and then TFA (1.5 mL, 19 mmol) was
added (reaction became dark red) and the reaction was stirred at rt
for 2 h (complete by LCMS). The reaction was concentrated on a
rotary evaporator, diluted twice with diethyl ether and
reconcentrated to an orange oil. The residue was slurried with
diethyl ether and the solids (360 mg yellow solid) were collected
by filtration and rinsed with hexanes. Addition solids (52 mg of a
yellow solid) were collected from the filtrate and rinsed with
hexanes. The combined solids were shown to be
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid trifluoroacetate (412 mg, 0.62 mmol, 84%). The
compound was isolated as a mixture of enantiomers and presents as a
1:5 mixture of rotamers or atrope isomers. For major isomer: 1H NMR
(300 MHz, CDCl.sub.3) .delta. ppm 0.81-1.01 (m, 2H), 1.13-2.65 (m,
18H), 2.62 (s, 3H) 2.69-3.70 (m, 2H), 3.60 (d, J=15.4 Hz, 1H), 3.87
(s, 3H), 4.36-5.30 (m, 3H), 6.95 (dd, J=8.8, 2.2 Hz, 1H), 7.09 (d,
J=2.2 Hz, 1H), 7.28 (d, J=8.8 Hz, 1H), 7.75 (d, J=8.4 Hz, 1H), 7.86
(d, J=8.4 Hz, 1H), 8.08 (s, 1H). LC-MS retention time: 3.24 min;
554 m/z (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Phenomenex-Luna 10u C18 3.0.times.50
mm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 5
mL/min, a gradient of 100% solvent A/0% solvent B to 0% solvent
A/100% solvent B, a gradient time of 4 min, a hold time of 1 min,
and an analysis time of 5 min where solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a Micromass Platform for LC in electrospray mode.
##STR00128##
[0152]
8-Cyclohexyl-N-((1-ethylcyclopropyl)sulfonyl)-11-methoxy-1a-((3-met-
hyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahydrocyclopr-
opa[d]indolo[2,1-a][2]benzazepine-5-carboxamide. CDI (21 mg, 0.13
mmol) was added to a stirring solution of
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid trifluoroacetate (56 mg, 0.10 mmol) in THF (0.5 mL)
and the reaction mixture was heated at 60.degree. C. for 1.5 h. The
reaction was cooled to rt and 1-ethylcyclopropane-1-sulfonamide (20
mg, 0.13 mmol) and then DBU (0.025 mL, 0.15 mmol as a 20% solution
in THF) were added. The reaction was stirred at rt for 1 h, more
DBU (.about.0.025 mL) was added and stirring continued for 2 h. The
reaction was heated at 60.degree. C. for 1 h, and then stirred ON
at rt (.about.25% conversion). Still more DBU (0.025 mL) and
1-ethylcyclopropane-1-sulfonamide (20 mg, 0.13 mmol) were added and
the reaction was allowed to stir at rt for 3 days. The reaction
solution was diluted with EtOAc (2 mL) and washed with 1M HCl (2
mL). The organic layer was concentrated to dryness with a stream of
nitrogen, dissolved into MeOH (1.5 mL), filtered and purified by
preparative HPLC (CH.sub.3CN/H.sub.2O with 10 mM NH.sub.4OAc) to
yield
8-cyclohexyl-N-((1-ethylcyclopropyl)sulfonyl)-11-methoxy-1a-((3-methyl-3,-
8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]-
indolo[2,1-a][2]benzazepine-5-carboxamide (5.3 mg, 7.7 .mu.mol, 8%
yield) as a yellow solid. The compound was isolated as a mixture of
enantiomers and presents as a 1:2 mixture of rotamers or atrope
isomers. 1H NMR (500 MHz, CDCl.sub.3) .delta. ppm -0.27--0.12 (m,
0.33H), 0.27-0.32 (m, 0.33H), 0.94-2.84 (m, 32.33H), 2.91-3.00 (m,
1H), 3.29-3.50 (m, 1H), 3.59 (d, J=15.3 Hz, 1H), 3.89 (s, 2H), 3.90
(s, 1H), 4.14 (d, J=14.7 Hz, 0.33H), 4.35-4.51 (m, 0.67H), 4.76 (d,
J=14.7 Hz, 0.33H), 5.18 (d, J=14.7 Hz, 0.67H), 6.92 (dd, J=8.2, 2.6
Hz, 0.33H), 6.96 (dd, J=8.6, 2.6 Hz, 0.67H), 7.01 (d, J=2.6 Hz,
0.33H), 7.12 (d, J=2.6 Hz, 0.67H), 7.29 (d, J=8.6 Hz, 0.67H), 7.29
(d, J=8.2 Hz, 0.33H), 7.49 (d, J=8.2 Hz, 0.33H), 7.54-7.61 (m,
0.67H), 7.87 (d, J=8.6 Hz, 0.67H), 7.88 (d, J=8.2 Hz, 0.33H), 7.98
(s, 1H). LC-MS retention time: 3.23 min; 685 m/z (MH.sup.+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Phenomenex-Luna 10u C18 3.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 5 mL/min, a
gradient of 100% solvent A/0% solvent B to 0% solvent A/100%
solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a Micromass Platform for LC in electrospray mode.
##STR00129##
[0153]
8-Cyclohexyl-N-((1-(cyclopropylmethyl)cyclopropyl)sulfonyl)-11-meth-
oxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tet-
rahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide. CDI
(24 mg, 0.15 mmol) was added to a stirring solution of
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid trifluoroacetate (64 mg, 0.12 mmol) in THF (0.5 mL)
the reaction mixture was heated at 60.degree. C. for 1.5 h. The
reaction was cooled to rt and
1-(cyclopropylmethyl)cyclopropane-1-sulfonamide (33 mg, 0.19 mmol)
and then DBU (0.09 mL, 0.6 mmol) were added and the reaction was
stirred at rt ON (.about.50% conversion by LCMS). The reaction
mixture was quenched with 1M HCl (aq), concentrated, dissolved into
MeOH (1.5 mL), filtered and purified by preparative HPLC
(CH.sub.3CN/H.sub.2O with 10 mM NH.sub.4OAc) to yield
8-cyclohexyl-N-((1-(cyclopropylmethyl)cyclopropyl)sulfonyl)-11-methoxy-1a-
-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahydr-
ocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide (14.2 mg,
0.020 mmol, 17% yield) as a light yellow solid. The compound was
isolated as a mixture of enantiomers and presents as a 1:3 mixture
of rotamers or atrope isomers. .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. ppm -0.33--0.14 (m, 0.25H), 0.01-0.11 (m, 2H), 0.20-0.29
(m, 0.25H), 0.38-0.47 (m, 2H), 0.61-3.44 (m, 32.5H), 3.56 (d,
J=14.6 Hz, 0.75H), 3.87 (s, 2.25H), 3.88 (s, 0.75H), 4.10 (d,
J=14.3 Hz, 0.25H), 4.24-4.54 (m, 1H), 4.73 (d, J=14.3 Hz, 0.25H),
5.16 (d, J=14.6 Hz, 0.75H), 6.87-6.97 (m, 1H), 6.99 (d, J=2.6 Hz,
0.25H), 7.10 (d, J=2.2 Hz, 0.75H), 7.27 (d, J=8.8 Hz, 0.75H), 7.28
(d, J=8.4 Hz, 0.25H), 7.48 (dd, J=8.8, 1.1 Hz, 0.25H), 7.59 (br d,
J=8.4 Hz, 0.75H), 7.85 (d, J=8.4 Hz, 0.75H), 7.85 (d, J=8.8 Hz,
0.25H), 7.95 (s, 0.75H), 7.97 (s, 0.25H). LC-MS retention time:
3.04 min; m/z 309 (MH-). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Phenomenex-Luna 10u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% solvent A/0% solvent B to
0% solvent A/100% solvent B, a gradient time of 4 min, a hold time
of 1 min, and an analysis time of 5 min where solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray
mode.
##STR00130##
[0154]
13-Cyclohexyl-N-((1-ethylcyclopropyl)sulfonyl)-3-methoxy-6-((3-meth-
yl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-7H-indolo[2,1-a][2]benzazepin-
e-10-carboxamide. CDI (80 mg, 0.49 mmol) was added to a solution of
tert-butyl
13-cyclohexyl-3-methoxy-6-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carb-
onyl)-7H-indolo[2,1-a][2]benzazepine-10-carboxylate TFA (200 mg,
0.30 mmol) in THF (1.5 mL) and the reaction mixture was heated at
60.degree. C. for 2 h. The reaction was cooled to rt and then 1/3
(.about.0.55 mL) of the reaction solution was added to a stirring
solution of 1-ethylcyclopropane-1-sulfonamide (40 mg, 0.27 mmol) in
DBU (0.20 mL, 1.3 mmol) and THF (0.20 mL). The reaction was stirred
at rt ON (.about.75% conversion by LCMS), concentrated to an oil,
quenched with 1N HCl (.about.1 mL) (prec. formed) and extracted
with EtOAc (2.times.1 mL) and DCM (2.times.1 mL). The combined
organics were concentrated to dryness, dissolved into MeOH (1.5 mL)
and purified by preparative HPLC (CH.sub.3CN/H.sub.2O with 10 mM
NH.sub.4OAc) to yield
13-cyclohexyl-N-((1-ethylcyclopropyl)sulfonyl)-3-methoxy-6-((3-methyl-3,8-
-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-7H-indolo[2,1-a][2]benzazepine-10-c-
arboxamide (30.9 mg, 0.046 mmol, 45% yield) as a yellow solid. 1H
NMR (300 MHz, CD.sub.3OD) .delta. ppm 0.97-2.35 (m, 29H), 2.56-2.73
(m, 1H), 2.80-2.94 (m, 1H), 3.38-3.58 (m, 1H), 3.94 (s, 3H),
4.29-4.64 (m, 2H), 5.14-5.31 (m, 1H), 7.05 (s, 1H), 7.09-7.20 (m,
2H), 7.57 (d, J=8.4 Hz, 1H), 7.65 (dd, J=8.4, 1.5 Hz, 1H), 7.92 (d,
J=8.4 Hz, 1H), 8.16 (br s, 1H). LC-MS retention time: 2.91 min; m/z
669 (MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Phenomenex-Luna 10u C18 4.6.times.50
mm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 5
mL/min, a gradient of 100% solvent A/0% solvent B to 0% solvent
A/100% solvent B, a gradient time of 4 min, a hold time of 1 min,
and an analysis time of 5 min where solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray
mode.
##STR00131##
[0155]
13-Cyclohexyl-N-(isopropylsulfonyl)-3-methoxy-6-((3-methyl-3,8-diaz-
abicyclo[3.2.1]oct-8-yl)carbonyl)-7H-indolo[2,1-a][2]benzazepine-10-carbox-
amide. CDI (80 mg, 0.49 mmol) was added to a solution of tert-butyl
13-cyclohexyl-3-methoxy-6-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carb-
onyl)-7H-indolo[2,1-a][2]benzazepine-10-carboxylate TFA (200 mg,
0.30 mmol) in THF (1.5 mL) and the reaction mixture was heated at
60.degree. C. for 2 h. The reaction was cooled to rt and then 1/3
(.about.0.55 mL) of the reaction solution was added to a stirring
solution of propane-2-sulfonamide (40 mg, 0.33 mmol) in DBU (0.20
mL, 1.3 mmol) and THF (0.20 mL). The reaction was stirred at rt ON
(complete by LCMS), concentrated to an oil, quenched with 1N HCl
(.about.1 mL) (prec. formed) and extracted with EtOAc (2.times.1
mL). The combined organics wee concentrated to dryness, dissolved
into MeOH (1.5 mL) and purified by preparative HPLC
(CH.sub.3CN/H.sub.2O with 10 mM NH.sub.4OAc) to yield
13-cyclohexyl-N-(isopropylsulfonyl)-3-methoxy-6-((3-methyl-3,8-diazabicyc-
lo[3.2.1]oct-8-yl)carbonyl)-7H-indolo[2,1-a][2]benzazepine-10-carboxamide
(28.5 mg, 0.044 mmol, 43% yield) as a yellow solid. 1H NMR (300
MHz, CD.sub.3OD) .delta. ppm 1.13-2.77 (m, 28H), 2.80-2.93 (m, 1H),
3.93 (s, 3H), 3.90-4.02 (m, 1H), 4.27-4.65 (m, 2H), 5.11-5.31 (m,
1H), 7.04 (s, 1H), 7.11 (d, J=2.6 Hz, 1H), 7.16 (dd, J=8.4, 2.6 Hz,
1H), 7.56 (d, J=8.4 Hz, 1H), 7.67 (dd, J=8.4, 1.5 Hz, 1H), 7.88 (d,
J=8.4 Hz, 1H), 8.18 (br s, 1H). LC-MS retention time: 2.55 min; m/z
643 (MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Phenomenex-Luna 10u C18 4.6.times.50
mm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 5
mL/min, a gradient of 100% solvent A/0% solvent B to 0% solvent
A/100% solvent B, a gradient time of 4 min, a hold time of 1 min,
and an analysis time of 5 min where solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray
mode.
##STR00132##
[0156]
13-Cyclohexyl-3-methoxy-6-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-y-
l)carbonyl)-N-((1-methyl-1H-imidazol-4-yl)sulfonyl)-7H-indolo[2,1-a][2]ben-
zazepine-10-carboxamide. tert-Butyl
13-cyclohexyl-3-methoxy-6-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carb-
onyl)-7H-indolo[2,1-a][2]benzazepine-10-carboxylate TFA (200 mg,
0.30 mmol) was dissolved into THF (1.5 mL) and stirred under
nitrogen. CDI (80 mg, 0.49 mmol) was added and the reaction mixture
was heated at 60.degree. C. for 2 h. The reaction was cooled to rt
and then 1/3 (.about.0.55 mL) of the reaction solution was added to
a stirring solution of 1-methyl-1H-imidazole-4-sulfonamide (60.0
mg, 0.37 mmol) in DBU (0.20 mL, 1.3 mmol) and THF (0.20 mL). The
reaction was stirred at rt ON (.about.75% conv. by LCMS),
concentrated to an oil, quenched with 1N HCl (.about.1 mL) (prec.
formed) and extracted with EtOAc (2.times.1 mL) and DCM (2.times.1
mL). The combined organics were concentrated, dissolved into MeOH
(1.5 mL) and purified by preparative HPLC (CH.sub.3CN/H.sub.2O with
10 mM NH.sub.4OAc) to yield
13-cyclohexyl-3-methoxy-6-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carb-
onyl)-N-((1-methyl-1H-imidazol-4-yl)sulfonyl)-7H-indolo[2,1-a][2]benzazepi-
ne-10-carboxamide (15.6 mg, 0.023 mmol, 22% yield) as a yellow
solid. 1H NMR (300 MHz, CD.sub.3OD) .delta. ppm 1.08-2.77 (m, 22H),
2.78-2.92 (m, 1H), 3.81 (s, 1H), 3.93 (s, 3H), 4.26-4.65 (m, 2H),
5.08-5.31 (m, 1H), 7.01 (s, 1H), 7.10 (d, J=2.6 Hz, 1H), 7.15 (dd,
J=8.4, 2.6 Hz, 1H), 7.55 (d, J=8.4 Hz, 1H), 7.65 (d, J=8.4 Hz, 1H),
7.67 (s, 1H), 7.81 (s, 1H), 7.83 (d, J=8.4 Hz, 1H), 8.17 (s, 1H).
LC-MS retention time: 2.15 min; m/z 681 (MH-). LC data was recorded
on a Shimadzu LC-10AS liquid chromatograph equipped with a
Phenomenex-Luna 10u C18 4.6.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 5 mL/min, a gradient of 100%
solvent A/0% solvent B to 0% solvent A/100% solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where solvent A was 5% acetonitrile/95% H.sub.2O/10 mM ammonium
acetate and solvent B was 5% H.sub.2O/95% acetonitrile/10 mM
ammonium acetate. MS data was determined using a Micromass Platform
for LC in electrospray mode.
##STR00133##
[0157] 8-Cyclohexyl-N-((3,5-dimethyl-4-isoxazolyl)
sulfonyl)-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbon-
yl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carbox-
amide.
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-
-5-carboxylic acid TFA (400 mg, 0.61 mmol) was dissolved into THF
(4 mL) and stirred under nitrogen. CDI (160 mg, 0.98 mmol) was
added and the reaction mixture was heated at 60.degree. C. for 2 h.
The reaction was cooled to rt and then 1/8 (.about.0.50 mL) of the
reaction solution was added to a stirring solution of
3,5-dimethylisoxazole-4-sulfonamide (30 mg, 0.17 mmol) in DBU (100
.mu.L, 0.66 mmol) and THF (0.100 mL). The reaction was stirred at
rt for 2 d (complete by LCMS) quenched with 1M HCl (aq) (0.75 mL)
(slight exotherm), diluted with MeOH and concentrated. The residue
was dissolved into MeOH and purified in one injection using
preparative HPLC (CH.sub.3CN/H.sub.2O with 10 mM NH.sub.4OAc) to
yield
8-cyclohexyl-N-((3,5-dimethyl-4-isoxazolyl)sulfonyl)-11-methoxy-1a-((3-me-
thyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahydrocyclop-
ropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide (33.7 mg, 0.047
mmol, 63% yield) as a yellow solid. The compound was isolated as a
mixture of enantiomers and presents as a 1:4 mixture of rotamers or
atrope isomers. 1H NMR (300 MHz, CDCl.sub.3) .delta. ppm 0.10-0.17
(m, 0.2H), 0.71-0.92 (m, 0.8H), 1.00-2.14 (m, 17H), 2.25-2.39 (m,
2H), 2.45 (s, 2.4H), 2.46 (s, 0.6H), 2.78 (s, 3H), 2.85-3.47 (m,
6H), 3.54 (d, J=15.4 Hz, 1H), 3.87 (s, 2.4H), 3.88 (s, 0.6H), 4.04
(d, J=15.0 Hz, 0.2H), 4.29-4.56 (m, 0.8H), 4.69 (d, J=15.0 Hz,
0.2H), 5.08-5.24 (m, 0.8H), 6.91 (dd, J=8.8, 2.6 Hz, 0.2H), 6.94
(dd, J=8.8, 2.6 Hz, 0.8H), 7.04 (d, J=2.6 Hz, 0.8H), 7.21-7.26 (m,
0.2H), 7.26 (d, J=8.8 Hz, 0.8H), 7.29 (d, J=8.8 Hz, 0.2H), 7.50
(dd, J=8.4, 1.5 Hz, 0.2H), 7.80 (dd, J=8.4, 1.1 Hz, 0.8H), 7.81 (d,
J=8.4 Hz, 0.8H), 7.82 (d, J=8.4 Hz, 0.2H), 7.89 (br s, 0.8H), 7.93
(br s, 0.2H). LC-MS retention time: 2.22 min; m/z 710 (MH-). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Phenomenex-Luna 10u C18 4.6.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 5 mL/min, a
gradient of 100% solvent A/0% solvent B to 0% solvent A/100%
solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where solvent A was 5% acetonitrile/95%
H.sub.2O/10 mM ammonium acetate and solvent B was 5% H.sub.2O/95%
acetonitrile/10 mM ammonium acetate. MS data was determined using a
Micromass Platform for LC in electrospray mode.
##STR00134##
[0158]
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-N-((1-methyl-1H-imidazol-4-yl) sulfonyl)-1,1a,
2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid, TFA (400 mg, 0.61 mmol) was dissolved into THF (4 mL)
and stirred under nitrogen. CDI (160 mg, 0.98 mmol) was added and
the reaction mixture was heated at 60.degree. C. for 2 h. The
reaction was cooled to rt and then 1/8 (.about.0.50 mL) of the
reaction solution was added to a stirring solution of
1-methyl-1H-imidazole-4-sulfonamide (30 mg, 0.19 mmol) in DBU (100
.mu.L, 0.66 mmol) and THF (0.100 mL). The reaction was stirred at
rt for 2 d (.about.60% conv. by LCMS) quenched with 1M HCl (aq)
(0.75 mL) (slight exotherm), diluted with MeOH and concentrated.
The residue was dissolved into MeOH (1.5 mL), filtered, and
purified in one injection using preparative HPLC
(CH.sub.3CN/H.sub.2O with 10 mM NH.sub.4OAc) to yield
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-N41-methyl-1H-imidazol-4-yl)sulfonyl)-1,1a,2,12b-tetrahydrocyclopro-
pa[d]indolo[2,1-a][2]benzazepine-5-carboxamide (21.8 mg, 0.031
mmol, 42% yield) as a yellow solid. The compound was isolated as a
mixture of enantiomers and presents as a 1:2 mixture of rotamers or
atrope isomers. 1H NMR (300 MHz, CDCl.sub.3) .delta. ppm
-0.39--0.56 (m, 0.33H), 0.12-0.21 (m, 0.33H), 0.74-3.68 (m,
25.33H), 3.65 (s, 3H), 3.85 (s, 2H), 3.86 (s, 1H), 4.02 (d, J=14.6
Hz, 0.33H), 4.18-4.59 (m, 0.67H), 4.72 (d, J=14.6 Hz, 0.33H), 5.11
(d, J=14.6 Hz, 0.67H), 5.96-6.21 (m, 1H), 6.87 (dd, J=8.8, 2.6 Hz,
0.33H), 6.91 (dd, J=8.8, 2.6 Hz, 0.67H), 6.95 (d, J=2.6 Hz, 0.33H),
7.06 (d, J=2.6 Hz, 0.67H), 7.19-7.27 (m, 1H), 7.41 (s, 1H), 7.56
(d, J=8.8 Hz, 0.33H), 7.61 (d, J=8.8 Hz, 0.67H), 7.68-7.78 (m, 2H),
7.96 (br s, 0.67H), 7.99 (br s, 0.33H). LC-MS retention time: 2.09
min; m/z 695 (MH-). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Phenomenex-Luna 10u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% solvent A/0% solvent B to
0% solvent A/100% solvent B, a gradient time of 4 min, a hold time
of 1 min, and an analysis time of 5 min where solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray
mode.
##STR00135##
[0159]
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-N-((5-methyl-2-pyridinyl)sulfonyl)-1,1a,2,12b-tetrahydrocyclo-
propa[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
8-Cyclohexyl-11-methoxy-1a((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carb-
onyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carb-
oxylic acid TFA (400 mg, 0.61 mmol) was dissolved into THF (4 mL)
and stirred under nitrogen. CDI (160 mg, 0.98 mmol) was added and
the reaction mixture was heated at 60.degree. C. for 2 h. The
reaction was cooled to rt and then 1/8 (.about.0.50 mL) of the
reaction solution was added to a stirring solution of
5-methylpyridine-2-sulfonamide (30 mg, 0.17 mmol) in DBU (100
.mu.L, 0.66 mmol) and THF (0.100 mL). The reaction was stirred 3 h
(complete by LCMS), quenched with 1M HCl (aq) (0.75 mL) (slight
exotherm was noted) and concentrated under a stream of nitrogen
over 2 d. The residue was dissolved into MeOH (.about.1 mL) and
purified in one injection by preparative HPLC (CH.sub.3CN/H.sub.2O
with 10 mM NH.sub.4OAc) to yield
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-N-((5-methyl-2-pyridinyl)sulfonyl)-1,1a,2,12b-tetrahydrocyclopropa[-
d]indolo[2,1-a][2]benzazepine-5-carboxamide (35.9 mg, 0.051 mmol,
68% yield) as a yellow solid. The compound was isolated as a
mixture of enantiomers and presents as a 1:2 mixture of rotamers or
atrope isomers. 1H NMR (300 MHz, CDCl.sub.3) .delta. ppm 0.19-0.27
(m, 0.33H), 0.95-3.11 (m, 24.67H), 2.45 (s, 3H), 3.59 (d, J=14.6
Hz, 1H), 3.90 (s, 2H), 3.91 (s, 1H), 4.02 (d, J=14.6 Hz, 0.33H),
4.15 (d, J=14.6 Hz, 0.33H), 4.64-4.26 (m, 1.67H), 5.12 (d, J=14.6
Hz, 0.67H), 6.95-7.03 (m, 1H) 7.15 (d, J=2.6 Hz, 0.33H), 7.19 (d,
J=2.6 Hz, 0.67H), 7.30 (d, J=8.4 Hz, 0.67H), 7.31 (d, J=8.8 Hz,
0.33H), 7.66-7.84 (m, 3H), 8.03-8.10 (m, 1.67H), 8.22 (s, 0.33H),
8.44 (s, 1H). LC-MS retention time: 2.19 min; m/z 706 (MH-). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Phenomenex-Luna 10u C18 4.6.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 5 mL/min, a
gradient of 100% solvent A/0% solvent B to 0% solvent A/100%
solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where solvent A was 5% acetonitrile/95%
H.sub.2O/10 mM ammonium acetate and solvent B was 5% H.sub.2O/95%
acetonitrile/10 mM ammonium acetate. MS data was determined using a
Micromass Platform for LC in electrospray mode.
##STR00136##
[0160]
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-N-(phenylsulfonyl)-1, 1a,
2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid TFA (400 mg, 0.61 mmol) was dissolved into THF (4 mL)
and stirred under nitrogen. CDI (160 mg, 0.98 mmol) was added and
the reaction mixture was heated at 60.degree. C. for 2 h. The
reaction was cooled to rt and then 1/8 (.about.0.50 mL) of the
reaction solution was added to a stirring solution of
benzenesulfonamide (30 mg, 0.19 mmol) in DBU (100 .mu.L, 0.66 mmol)
and THF (0.100 mL). The reaction was stirred at rt for 2 d
(complete by LCMS), quenched with 1M HCl (aq) (0.75 mL) (slight
exotherm), diluted with MeOH and concentrated to dryness. The
residue was dissolved into MeOH (1.5 mL) and purified in one
injection using preparative HPLC (CH.sub.3CN/H.sub.2O with 10 mM
NH.sub.4OAc) to yield
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-N-(phenylsulfonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2-
,1-a][2]benzazepine-5-carboxamide (27.6 mg, 0.040 mmol, 53% yield)
as a light yellow solid. The compound was isolated as a mixture of
enantiomers and presents as a 1:3 mixture of rotamers or atrope
isomers. 1H NMR (300 MHz, CDCl.sub.3) .delta. ppm 0.14-0.22 (m,
0.25H), 0.93-2.97 (m, 24.75H), 3.39-3.38 (m, 1.25H), 3.55 (d,
J=15.0 Hz, 0.75H), 3.87 (s, 3H), 3.99-4.11 (m, 0.25H), 4.20-4.11
(m, 1H), 5.13 (d, J=15.0 Hz, 0.75H), 6.87-6.98 (m, 1.25H), 7.09 (d,
J=2.2 Hz, 0.75H), 7.23-7.29 (m, 1H), 7.44-7.63 (m, 4H), 7.80 (d,
J=8.4 Hz, 1H), 7.90 (s, 1H), 8.11-8.19 (m, 2H). LC-MS retention
time: 2.32 min; m/z 691 (MH-). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Phenomenex-Luna 10u
C18 4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% solvent A/0% solvent B to
0% solvent A/100% solvent B, a gradient time of 4 min, a hold time
of 1 min, and an analysis time of 5 min where solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray
mode.
##STR00137##
[0161]
N-((4-Chloro-3-pyridinyl)sulfonyl)-8-cyclohexyl-11-methoxy-1a-((3-m-
ethyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahydrocyclo-
propa[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid TFA (400 mg, 0.61 mmol) was dissolved into THF (4 mL)
and stirred under nitrogen. CDI (160 mg, 0.98 mmol) was added and
the reaction mixture was heated at 60.degree. C. for 2 h. The
reaction was cooled to rt and then 1/8 (.about.0.50 mL) of the
reaction solution was added to a stirring solution of
4-chloropyridine-3-sulfonamide (40 mg, 0.21 mmol) in DBU (100
.mu.L, 0.66 mmol) and THF (0.100 mL). The reaction was stirred 3 h
(complete by LCMS) quenched with 1M HCl (aq) (0.75 mL) (slight
exotherm was noted) and concentrated under a stream of nitrogen
over 2 d. The residue was dissolved into MeOH (.about.1 mL) and
purified by preparative HPLC (CH.sub.3CN/H.sub.2O with 10 mM
NH.sub.4OAc) to yield
N-((4-chloro-3-pyridinyl)sulfonyl)-8-cyclohexyl-11-methoxy-1a-((3-methyl--
3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahydrocyclopropa[-
d]indolo[2,1-a][2]benzazepine-5-carboxamide (28.6 mg, 0.039 mmol,
52% yield) as a bright yellow solid. The compound was isolated as a
mixture of enantiomers and presented as a mixture of atrope isomers
or rotamers. Partial 1H NMR (300 MHz, CD.sub.3OD) .delta. 3.86 (s,
3H), 6.93 (d, J=8.8, 2.2 Hz, 1H), 7.08 (br s, 1H), 7.20-7.28 (m,
1H), 7.35 (d, J=5.1 Hz, 1H), 7.59 (d, J=8.4 Hz, 1H), 7.76 (d, J=8.4
Hz, 1H), 7.98 (s, 1H), 8.58 (d, J=5.1 Hz, 1H), 9.87 (s, 1H). LC-MS
retention time: 2.18 min; m/z 726 (MH-). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
Phenomenex-Luna 10u C18 4.6.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 5 mL/min, a gradient of 100%
solvent A/0% solvent B to 0% solvent A/100% solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where solvent A was 5% acetonitrile/95% H.sub.2O/10 mM ammonium
acetate and solvent B was 5% H.sub.2O/95% acetonitrile/10 mM
ammonium acetate. MS data was determined using a Micromass Platform
for LC in electrospray mode.
##STR00138##
[0162]
8-Cyclohexyl-11-methoxy-N-((2-methoxy-4-methylphenyl)sulfonyl)-1a-(-
(3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahydroc-
yclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid TFA (100 mg, 0.15 mmol) was dissolved into THF (1 mL)
and stirred under nitrogen. CDI (40 mg, 0.25 mmol) was added and
the reaction mixture was heated at 60.degree. C. for 2 h. The
reaction was cooled to rt and then 1/2 (.about.0.50 mL) of the
reaction solution was added to a stirring solution of
2-methoxy-4-methylbenzenesulfonamide (35 mg, 0.17 mmol) in DBU (100
.mu.L, 0.66 mmol) and THF (0.100 mL). The reaction was stirred at
rt for 1 d (complete by LCMS), quenched with 1M HCl (aq) (0.75 mL)
(slight exotherm), diluted with MeOH and concentrated. The residue
was diluted with MeOH (1.3 mL) and DMF (0.2 mL) and purified in one
injection using preparative HPLC (H.sub.2O/CH.sub.3CN, 10 mM
NH.sub.4OAc) to yield
8-cyclohexyl-11-methoxy-N-((2-methoxy-4-methylphenyl)sulfonyl)-1-
a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahyd-
rocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide (14.6 mg,
0.020 mmol, 27% yield) as an off-white solid. The compound was
isolated as a mixture of enantiomers and presents as a 1:4 mixture
of rotamers or atrope isomers. 1H NMR (300 MHz, CD.sub.3OD) .delta.
ppm 0.14-0.20 (m, 0.2H), 0.89-3.12 (m, 22.8H), 3.88 (s, 2H), 2.44
(s, 3H), 3.37 (s, 3H), 3.64 (d, J=15.0 Hz, 0.8H), 3.87-3.94 (m,
6H), 4.17 (d, J=15.4 Hz, 0.2H), 4.25-4.35 (m, 0.8H), 4.51-4.56 (m,
0.2H), 4.87-4.97 (m, 0.2H), 5.13 (d, J=15.4 Hz, 0.8H), 6.93-7.06
(m, 3H), 7.14-7.18 (m, 0.2H), 7.19-7.23 (m, 0.8H), 7.29-7.37 (m,
1H), 7.53-7.61 (m, 1H), 7.83-7.99 (m, 2.8H), 8.10 (s, 0.2H). LC-MS
retention time: 2.90 min; m/z 735 (MH-). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
Phenomenex-Luna 10u C18 4.6.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 5 mL/min, a gradient of 100%
solvent A/0% solvent B to 0% solvent A/100% solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where solvent A was 5% acetonitrile/95% H.sub.2O/10 mM ammonium
acetate and solvent B was 5% H.sub.2O/95% acetonitrile/10 mM
ammonium acetate. MS data was determined using a Micromass Platform
for LC in electrospray mode.
##STR00139##
[0163] 4-tert-Butylthiazole-2-sulfonamide. Sodium hypochlorite (15
mL, 28 mmol) (Reagent grade) (cooled internally with ice (.about.15
g)) was added dropwise to a vigorously stirring solution of
4-tert-butylthiazole-2-thiol (800 mg, 4.62 mmol) in DCM (25 mL) and
1M aq. HCl (25 mL, 25 mmol) cooled in an ice bath. The reaction was
stirred at 5.degree. C. for 10 min. and the layers were separated.
The organic layer was cooled in a flask with a dry ice and acetone
bath and ammonia bubbled into the solution over 5 minutes. The cold
bath was removed and the reaction was allowed to come to rt. The
crude reaction was concentrated, dissolved into MeOH and filter to
remove solids. The solution was concentrated to yield
4-tert-butylthiazole-2-sulfonamide as a waxy solid which was used
without further purification. LC-MS retention time: 1.85 min; m/z
219 (MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Phenomenex-Luna 10u C18 4.6.times.50
mm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 5
mL/min, a gradient of 100% solvent A/0% solvent B to 0% solvent
A/100% solvent B, a gradient time of 4 min, a hold time of 1 min,
and an analysis time of 5 min where solvent A was 5%
acetonitrile/95% H2O/10 mM ammonium acetate and solvent B was 5%
H2O/95% acetonitrile/10 mM ammonium acetate. MS data was determined
using a Micromass Platform for LC in electrospray mode.
##STR00140##
[0164]
N((4-tert-Butyl-1,3-thiazol-2-yl)sulfonyl)-8-cyclohexyl-11-methoxy--
1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,
1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide-
.
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)ca-
rbonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-ca-
rboxylic acid TFA (100 mg, 0.15 mmol) was dissolved into THF (1 mL)
and stirred under nitrogen. CDI (40 mg, 0.25 mmol) was added and
the reaction mixture was heated at 60.degree. C. for 2 h. The
reaction was cooled to rt and then 1/2 (.about.0.50 mL) of the
reaction solution was added to a stirring solution of
4-tert-butylthiazole-2-sulfonamide (35 mg, 0.16 mmol) in DBU (100
.mu.L, 0.66 mmol) and THF (0.100 mL). The reaction was stirred at
rt for 1 d (complete by LCMS), queched with 1M HCl (aq) (0.75 mL)
(slight exotherm), diluted with MeOH and concentrated. The residue
was diluted with MeOH (1.3 mL) and DMF (0.2 mL) and purified in one
injection using preparative HPLC (H.sub.2O/CH.sub.3CN, 10 mM
NH.sub.4OAc) to yield
N-((4-tert-butyl-1,3-thiazol-2-yl)sulfonyl)-8-cyclohexyl-11-meth-
oxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tet-
rahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide (9.0
mg, 0.012 mmol, 15% yield) as an off-white solid. LC-MS retention
time: 2.59 min; m/z 754 (MH-). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Phenomenex-Luna 10u
C18 4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% solvent A/0% solvent B to
0% solvent A/100% solvent B, a gradient time of 4 min, a hold time
of 1 min, and an analysis time of 5 min where solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray
mode.
##STR00141##
[0165] Quinoline-8-sulfonamide. Quinoline-8-sulfonyl chloride (1.0
g, 4.39 mmol) was slurried into DCM (20 mL) and cooled to
-60.degree. C. Then ammonia (.about.5 g, 294 mmol) was bubbled into
the reaction over 5 min (volume increased by .about.5 mL). The
reaction was stirred and allowed to come to rt overnight. The
residue was partitioned between water (50 mL) and EtOAc (50 mL) and
the organic layer was washed with brine (20 mL) and diluted with
hexanes (.about.50 mL). The solution was stirred 20 min and the
solids were collected by filtration to yield
quinoline-8-sulfonamide (432 mg, 2.07 mmol, 47% yield) as a light
yellow solid. 1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 7.25 (s,
2H), 7.73 (dd, J=8.3, 4.3 Hz, 1H), 7.76 (dd, J=8.6, 7.3 Hz, 1H),
8.28 (d, J=8.3 Hz, 1H), 8.31 (d, J=7.3 Hz, 1H), 8.56 (br d, J=8.6
Hz, 1H), 9.08 (dd, J=4.3, 0.9 Hz, 1H). LC-MS retention time: 1.27
min; m/z 209 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Phenomenex-Luna 10u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% solvent A/0% solvent B to
0% solvent A/100% solvent B, a gradient time of 4 min, a hold time
of 1 min, and an analysis time of 5 min where solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray
mode.
##STR00142##
[0166]
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-N-(8-quinolinylsulfonyl)-1, 1a,
2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid TFA (104 mg, 0.16 mmol) and CDI (40 mg, 0.25 mmol)
were dissolved into THF (1.0 mL), stirred under nitrogen and then
heated at 60.degree. C. for 2 h. The reaction was cooled to rt and
then 1/2 (.about.0.50 mL) of the reaction solution was added to a
stirring solution of quinoline-8-sulfonamide (40 mg, 0.19 mmol) in
DBU (0.10 mL) and THF (0.10 mL). The reaction was stirred at rt for
1 d, quenched with 1M aq. HCl (0.7 mL), diluted with MeOH and DMSO
(0.2 mL), and concentrated. The crude oil was diluted with MeOH (1
mL), filtered and purified with via preparative HPLC
(H.sub.2O/CH.sub.3CN, 10 mM NH.sub.4OAc) in a single injection to
yield
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-N-(8-quinolinylsulfonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2-
,1-a][2]benzazepine-5-carboxamide (11.2 mg, 0.015 mmol, 20% yield)
as a yellow solid. The compound was isolated as a mixture of
enantiomers and presents as a 1:2 mixture of rotamers or atrope
isomers. 1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 0.19-0.27 (m,
0.33H), 0.67-3.61 (m, 26.67H), 3.88 (s, 2H), 3.89 (s, 1H),
4.00-4.32 (m, 1H), 4.62-4.72 (m, 0.33H), 5.02-5.16 (m, 0.67H), 6.90
(dd, J=8.5, 2.4 Hz, 0.33H), 6.94 (dd, J=8.6, 2.5 Hz, 0.67H), 7.00
(br s, 0.33H), 7.10 (br, s, 0.67H), 7.19-7.27 (m, 1H), 7.39-7.55
(m, 2H), 7.66-7.83 (m, 2H), 7.90 (s, 1H), 8.01-8.11 (m, 1H),
8.15-8.27 (m, 1H), 8.71-8.77 (m, 1H), 8.93-9.09 (m, 1H). LC-MS
retention time: 2.57 min; m/z 742 (MH-). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
Phenomenex-Luna 10u C18 4.6.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 5 mL/min, a gradient of 100%
solvent A/0% solvent B to 0% solvent A/100% solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where solvent A was 5% acetonitrile/95% H.sub.2O/10 mM ammonium
acetate and solvent B was 5% H.sub.2O/95% acetonitrile/10 mM
ammonium acetate. MS data was determined using a Micromass Platform
for LC in electrospray mode.
##STR00143##
[0167]
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-N((2-pyridinylmethyl)sulfonyl)-1,1a,2,12b-tetrahydrocycloprop-
a[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid trifruoroacetate (40 mg, 0.060 mmol),
pyridin-2-ylmethanesulfonamide (25 mg, 0.15 mmol) and DMAP (7 mg,
0.060 mmol) were slurried into DCM (0.5 mL) and then EDC (20 mg,
0.10 mmol) was added. The vial was flushed with nitrogen, sealed
and the reaction was stirred at rt ON (complete by LCMS). The
reaction mixture was diluted with MeOH and purified by preparative
HPLC (CH.sub.3CN/H.sub.2O w/10 mM NH.sub.4OAc) in a single
injection to yield
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-N-((2-pyridinylmethyl)sulfonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]i-
ndolo[2,1-a][2]benzazepine-5-carboxamide (15.8 mg, 0.022 mmol, 37%
yield) as a yellow solid. The compound was isolated as a mixture of
enantiomers and presents as a .about.1:1 mixture or rotamers or
atrope isomers. 1H NMR (300 MHz, CDCl.sub.3) .delta. ppm 0.19-0.26
(m, 0.5H), 0.34-3.66 (m, 26.5H), 3.88 (s, 3H), 3.98-4.58 (m, 1.5H),
4.72 (d, J=14.3 Hz, 0.5H), 4.87-4.99 (m, 1.5H), 5.09 (d, J=14.6 Hz,
0.5H), 6.86-7.01 (m, 1.5H), 7.06-7.13 (m, 1H), 7.16-7.30 (m, 1.5H),
7.46-7.52 (m, 1H), 7.59-7.75 (m, 2H), 7.78-8.13 (m, 2H), 8.53 (d,
J=5.1 Hz, 1H). LC-MS retention time: 2.19 min; m/z 706 (MH-). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Phenomenex-Luna 10u C18 4.6.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 5 mL/min, a
gradient of 100% solvent A/0% solvent B to 0% solvent A/100%
solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where solvent A was 5% acetonitrile/95%
H.sub.2O/10 mM ammonium acetate and solvent B was 5% H.sub.2O/95%
acetonitrile/10 mM ammonium acetate. MS data was determined using a
Micromass Platform for LC in electrospray mode.
##STR00144##
[0168] 2,4-Dimethylthiazole-5-sulfonamide. Liquid ammonia was added
dropwise via a CO.sub.2/acetone condenser to a solution of
2,4-dimethylthiazole-5-sulfonyl chloride (260 mg, 1.228 mmol) in
THF (3 mL) at -78.degree. C. until the volume had approximately
doubled (.about.10 min) The reaction was stirred at -78.degree. C.
for 1 h and then allowed to slowly warm to rt. The reaction was
concentrated to dryness and the residue was diluted with
CH.sub.2Cl.sub.2. The slurry was filtered to remove the solids and
the solution was concentrated to yield
2,4-dimethylthiazole-5-sulfonamide (230 mg, 1.2 mmol, 97% yield) as
a light yellow solid. LC-MS retention time 0.43 min; m/z 191 (MH-).
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Phenomenex-Luna 10u C18 3.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 5 ml/min, a
gradient of 100% solvent A/0% solvent B to 0% solvent A/100%
solvent B, a gradient time of 3 min, a hold time of 1 min, and an
analysis time of 4 min where solvent A was 5% acetonitrile/95%
H.sub.2O/10 mM ammonium acetate and solvent B was 5% H.sub.2O/95%
acetonitrile/10 mM ammonium acetate. MS data was determined using a
Micromass Platform for LC in electrospray mode.
##STR00145##
[0169]
8-Cyclohexyl-N-((2,4-dimethyl-1,3-thiazol-5-yl)sulfonyl)-11-methoxy-
-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrah-
ydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid (40 mg, 0.072 mmol),
2,4-dimethylthiazole-5-sulfonamide (26 mg, 0.14 mmol) and DMAP (10
mg, 0.082 mmol) were slurried into CH.sub.2Cl.sub.2 (0.5 mL) and
then treated with EDC (20 mg, 0.10 mmol). The reaction was stirred
at rt ON. The reaction was concentrated with a stream of nitrogen,
dissolved into MeOH, filtered and purified by Prep HPLC
(CH.sub.3CN/H.sub.2O, w/10 mM NH.sub.4OAc, 15-100% over 20 mM) to
yield
8-cyclohexyl-N-((2,4-dimethyl-1,3-thiazol-5-yl)sulfonyl)-11-methoxy-
-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrah-
ydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide (25.4
mg, 0.035 mmol, 48% yield) as a yellow solid. LC-MS retention time
3.14 min; m/z 728 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Phenomenex-Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 ml/min, a gradient of 100% solvent A/0% solvent B to
0% solvent A/100% solvent B, a gradient time of 4 min, a hold time
of 1 min, and an analysis time of 5 mM where solvent A was 10%
MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a Micromass Platform for LC in electrospray mode.
##STR00146##
[0170] 1H-1,2,4-Triazole-3-sulfonamide. Liquid ammonia was added
dropwise via a CO.sub.2/acetone condenser to a solution of
1H-1,2,4-triazole-3-sulfonyl chloride (500 mg, 2.98 mmol) in THF (5
mL) at -78.degree. C. until the volume had approximately doubled
(.about.15 min) The reaction was stirred at -78.degree. C. for 1 h
and then allowed to slowly warm to rt. The reaction was
concentrated to dryness and the residue was diluted with DCM. The
slurry was filtered to remove the solids and the solution was
concentrated to yield 1H-1,2,4-triazole-3-sulfonamide (240 mg,
1.620 mmol, 54.3% yield) as an off white solid. 1H NMR (300 MHz,
CD.sub.3OD) .delta. ppm 8.60 (br s, 1H). LC-MS retention time 0.19
min; m/z 147 (MH-). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Phenomenex-Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 ml/min, a gradient of 100% solvent A/0% solvent B to
0% solvent A/100% solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray
mode.
##STR00147##
[0171]
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-N-(1H-1,2,4-triazol-3-ylsulfonyl)-1,1a,2,12b-tetrahydrocyclop-
ropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid (39 mg, 0.070 mmol), 1H-1,2,4-triazole-3-sulfonamide
(25 mg, 0.17 mmol) and DMAP (10 mg, 0.082 mmol) were slurried into
CH.sub.2Cl.sub.2 (0.5 mL) and then treated with EDC (20 mg, 0.104
mmol). The reaction was stirred at rt ON. The reaction was
concentrated with a stream of nitrogen, dissolved into MeOH,
filtered and purified by Prep HPLC (CH.sub.3CN/H.sub.2O, w/10 mM
NH.sub.4OAc, 15-100% over 20 min) to yield
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-N-(1H-1,2,4-triazol-3-ylsulfonyl)-1,1a,2,12b-tetrahydrocyclop-
ropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide (5.4 mg, 6.3
.mu.mol, 9% yield) as a yellow solid. LC-MS retention time 2.88
min; m/z 684 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Phenomenex-Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 ml/min, a gradient of 100% solvent A/0% solvent B to
0% solvent A/100% solvent B, a gradient time of 4 min, a hold time
of 1 min, and an analysis time of 5 min where solvent A was 10%
MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a Micromass Platform for LC in electrospray mode.
##STR00148##
[0172] Ethenesulfonamide. A solution of TEA (7.67 mL, 55.0 mmol) in
Et.sub.2O (50 ml) was added to a solution of 2-chloroethanesulfonyl
chloride (5.22 mL, 50 mmol) in Et.sub.2O (100 ml) at -24.degree. C.
(CCl.sub.4-dry ice bath) and the mixture was stirred and allowed to
warm to rt over 2 h. The reaction mixture was filtered to remove a
white precipitate and the filtrate was concentrated (.about.20 mL
of volume). NH.sub.3 was bubbled into the solution of
ethenesulfonyl chloride (1.266 g, 10.00 mmol) in Et.sub.2O (20 mL)
for 10 min and the white precipitate was removed by filtration. The
filtrate was concentrated to yield ethenesulfonamide (180 mg, 1.680
mmol, 16.80% yield) as a colorless gel which was used without
further purification. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm
6.80 (dd, J=16.5, 9.9 Hz, 1H), 6.16 (d, J=16.5 Hz, 1H), 5.87 (dd,
J=9.9 Hz, 1H). LC-MS retention time 0.17 min; m/z 108 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Phenomenex-Luna 10u C18 4.6.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 5 ml/min, a
gradient of 100% solvent A/0% solvent B to 0% solvent A/100%
solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 5 min where solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a Micromass Platform for LC in electrospray mode.
##STR00149##
[0173]
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-N-(vinylsulfonyl)-1,1a,
2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
Ethenesulfonamide (14.5 mg, 0.135 mmol) and DBU (0.03 ml, 0.2 mmol)
were added to a stirring solution of
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid (50 mg, 0.090 mmol) and CDI (22 mg, 0.14 mmol) in THF
(1 mL). The reaction mixture was stirred at 60.degree. C. for 1 h
and then at rt for 16 h. The reaction was diluted with MeOH,
filtered and purified by prep HPLC (H.sub.2O-MeOH with 0.1% TFA
buffer) to yield product
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-N-(vinylsulfonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2-
]benzazepine-5-carboxamide (35.2 mg, 0.055 mmol, 61% yield) as a
off white solid. The compound was isolated as a mixture of
enantiomers and presents as a 1:4 mixture of rotamers or atrope
isomers. 1H NMR (300 MHz, CD.sub.3OD) .delta. ppm 8.07 (s, 0.2H),
8.00 (s, 0.8H), 7.93 (d, J=8.4 Hz, 0.2H), 7.92 (d, J=8.4 Hz, 0.8H),
7.61 (d, J=8.4 Hz, 0.2H), 7.57 (d, J=8.4 Hz, 0.8H), 7.35 (d, J=8.8
Hz, 0.2H), 7.34 (d, J=8.4 Hz, 0.8H), 7.25-7.00 (m 3H), 6.49 (d,
J=16.8 Hz, 1H), 6.23 (d, J=9.9 Hz, 1H), 5.16 (d, J=15.4 Hz, 0.8H),
4.96 (d, J=15.0 Hz, 0.2H), 4.74-4.63 (m, 1H), 4.37-4.26 (m, 0.8H),
4.21 (d, J=15.0 Hz, 0.2H), 4.01-3.86 (m, 3H), 3.73-1.15 (m, 25.8H),
0.24-0.16 (m, 0.2H). LC-MS retention time 1.72 min; m/z 641 (MH-).
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Phenomenex-Luna 10u C18 4.6.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 5 ml/min, a
gradient of 100% solvent A/0% solvent B to 0% solvent A/100%
solvent B, a gradient time of 3 min, a hold time of 1 min, and an
analysis time of 4 min where solvent A was 5% acetonitrile/95%
H.sub.2O/10 mM ammonium acetate and solvent B was 5% H.sub.2O/95%
acetonitrile/10 mM ammonium acetate. MS data was determined using a
Micromass Platform for LC in electrospray mode.
##STR00150##
[0174] 2-Methylprop-1-ene-1-sulfonamide. t-Butyllithium (13 mL,
22.00 mmol) was added to a stirring solution of
1-bromo-2-methylprop-1-ene (2.0 mL, 20 mmol) in Et.sub.2O (100 mL)
at -70.degree. C. The reaction mixture was stirred at -70.degree.
C. under N.sub.2 for 1 h and then added dropwise to a stirring
solution of sulfuryl dichloride (3.2 mL, 40 mmol) in Et.sub.2O (100
mL) at 0.degree. C., (a white precipitate formed). The reaction
mixture was allowed to slowly warm to rt and stirred under N.sub.2
over 16 h. The reaction mixture was filtered and the filtrate
(still cloudy) was concentrated. The residue was dissolved into THF
(100 mL) to form a clear yellow solution. NH.sub.3 was bubbled into
this stirred solution of 2-methylprop-1-ene-1-sulfonyl chloride
(3.09 g, 20 mmol) in THF (100 mL) in a 250 ml 3 necked flask
equipped with cold finger (-70.degree. C.) over a period of 10
minutes (white precipitate formed, and a reflux of NH.sub.3
observed). After 1 h, the cold finger was removed and the reaction
was stirred at rt for 16 h. The reaction mixture was filtered to
remove the white precipitate, flushed with EtOAc. and concentrated
under vacuum to yield 2-methylprop-1-ene-1-sulfonamide (2.59 g,
19.2 mmol, 96% yield) as a clear yellow oil. LC-MS retention time
0.41 min; m/z 136 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Phenomenex-Luna 10u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 ml/min, a gradient of 100% solvent A/0% solvent B to
0% solvent A/100% solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 5 min where solvent A was 10%
MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a Micromass Platform for LC in electrospray mode.
##STR00151##
[0175]
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-N-((2-methyl-1-propen-1-yl)sulfonyl)-1,1a,2,12b-tetrahydrocyc-
lopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
2-Methylprop-1-ene-1-sulfonamide (18.3 mg, 0.135 mmol) and DBU
(0.03 ml, 0.2 mmol) were added to a stirring solution of
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid (50 mg, 0.090 mmol) and CDI (22 mg, 0.14 mmol) in THF
(1 mL). The reaction mixture was stirred at rt for 16 h. The
reaction was diluted with MeOH, filtered and purified by prep HPLC
(H.sub.2O-MeOH with 0.1% TFA buffer) to yield
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-N-((2-methyl-1-propen-1-yl)sulfonyl)-1,1a,2,12b-tetrahydrocycloprop-
a[d]indolo[2,1-a][2]benzazepine-5-carboxamide (13.8 mg, 0.020 mmol,
22% yield) as a white solid. The compound was isolated as a mixture
of enantiomers and presents as a 1:4 mixture of rotamers or atrope
isomers. 1H NMR (300 MHz, CD.sub.3OD) .delta. ppm 8.07 (s, 0.2H),
8.00 (s, 0.8H), 7.92 (d, J=8.4 Hz, 1H), 7.65-7.54 (m, 1H),
7.39-7.31 (m, 1H), 7.24-7.17 (m, 1H), 7.09-7.01 (m, 1H), 6.47 (s,
1H), 5.18 (d, J=15.4 Hz, 0.8H), 5.01-4.63 (m, 1.2H), 4.34-4.18 (m,
1H), 3.91 (s, 3H), 3.73-1.16 (m, 25.8H), 2.25 (s, 3H), 2.02 (s,
3H), 0.24-0.18 (m, 0.2H). LC-MS retention time 3.37 min; m/z 671
(MH.sup.+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Phenomenex-Luna 10u C18 4.6.times.50
mm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 5
ml/min, a gradient of 100% solvent A/0% solvent B to 0% solvent
A/100% solvent B, a gradient time of 4 min, a hold time of 1 min,
and an analysis time of 5 min where solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a Micromass Platform for LC in electrospray mode.
##STR00152##
[0176] Pyrimidine-2-sulfonamide. Sodium hypochlorite (30.9 mL, 60.0
mmol) was added dropwise with rapid stirring to a solution of
2-mercaptopyrimidine (1122 mg, 10 mmol) in CH.sub.2Cl.sub.2 (60 mL)
and 1N HCl (55.0 mL, 55.0 mmol) at 0.degree. C. After the addition
was complete, the mixture was stirred for 15 min at 0.degree. C.
and the phases where then separated. The organic layer was
transferred to a 250 mL 3-necked flask which was then chilled in an
ice bath and equipped with a cold finger (-70.degree. C.). NH.sub.3
was bubbled through the reaction mixture for 15 min and then the
reaction was slowly allowed to warm to rt and stirred 16 h. The
resulting white precipitate was separated by filtration and the
filtrate was concentrated under vacuum to yield
pyrimidine-2-sulfonamide (350 mg, 1.98 mmol, 20% yield) as a light
yellow solid. Additional product could be obtained from the
precipitate. 1H NMR (300 MHz, acetone-d.sub.6) .delta. ppm 9.00 (d,
J=4.9 Hz, 2H), 7.73 (t, J=4.9 Hz, 1H), 6.77 (br s, 2H). LC-MS
retention time 0.17 min; m/z 160 (MH+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
Phenomenex-Luna 10u C18 4.6.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 5 ml/min, a gradient of 100%
solvent A/0% solvent B to 0% solvent A/100% solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a Micromass Platform for LC in
electrospray mode.
##STR00153##
[0177]
8-Cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8--
yl)carbonyl)-N-(2-pyrimidinylsulfonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]i-
ndolo[2,1-a][2]benzazepine-5-carboxamide. EDC (26.0 mg, 0.135 mmol)
and DMAP (11 mg, 0.090 mmol) were added to a stirring solution of
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid (50 mg, 0.090 mmol) and pyrimidine-2-sulfonamide (21.6
mg, 0.135 mmol) in CH.sub.2Cl.sub.2 (1.5 mL) and the reaction
mixture was stirred at rt for 16 h. The reaction was diluted with
MeOH, filtered and purified by prep HPLC (H.sub.2O--CH.sub.3CN with
10 mM NH.sub.4OAc buffer) to yield
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-N-(2-pyrimidinylsulfonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[-
2,1-a][2]benzazepine-5-carboxamide (29.7 mg, 0.041 mmol, 45.0%
yield) as a light yellow solid. LC-MS retention time 1.71 min; m/z
693 (MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Phenomenex-Luna 10u C18 4.6.times.50
mm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 5
ml/min, a gradient of 100% solvent A/0% solvent B to 0% solvent
A/100% solvent B, a gradient time of 3 min, a hold time of 1 min,
and an analysis time of 4 min where solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray
mode.
##STR00154##
[0178] 4,6-Dimethylpyrimidine-2-sulfonamide. Sodium hypochlorite
(30.9 mL, 60.0 mmol) was added dropwise with rapid stirring to a
solution of 4,6-dimethylpyrimidine-2-thiol (1.40 g, 10.00 mmol) and
CaCl.sub.2 (14 g) in CH.sub.2Cl.sub.2(60 mL) and 1N HCl (55.0 mL,
55.0 mmol) at -23.degree. C. After the addition was complete, the
mixture was stirred for 15 min at -23.degree. C. and the phases
where separated. The organic layer was transferred to a 250 mL
3-necked flask which was then chilled to -23.degree. C. and
equipped with a cold finger (-70.degree. C.). NH.sub.3 was bubbled
through the reaction mixture for 15 min. and then the reaction was
slowly allowed to warm to rt and stirred 16 h. The resulting white
precipitate was separated by filtration and the filtrate was
concentrated under vacuum to yield
4,6-dimethylpyrimidine-2-sulfonamide (1.003 g, 5.36 mmol, 53.6%
yield) as white solid. 1H NMR (300 MHz, acetone-d.sub.6) .delta.
ppm 7.43 (s, 1H), 6.63 (br s, 2H), 2.53 (s, 6H). LC-MS retention
time 0.39 min; m/z 188 (MH.sup.+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
Phenomenex-Luna 10u C18 4.6.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 5 ml/min, a gradient of 100%
solvent A/0% solvent B to 0% solvent A/100% solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a Micromass Platform for LC in
electrospray mode.
##STR00155##
[0179]
8-Cyclohexyl-N-((4,6-dimethyl-2-pyrimidinyl)sulfonyl)-11-methoxy-1a-
-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahydr-
ocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide. EDC (26.0
mg, 0.135 mmol) and DMAP (11 mg, 0.090 mmol) were added to a
stirring solution of
8-cyclohexyl-11-methoxy-1a-((3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)car-
bonyl)-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-car-
boxylic acid (50 mg, 0.090 mmol) and
4,6-dimethylpyrimidine-2-sulfonamide (25.4 mg, 0.135 mmol) in
CH.sub.2Cl.sub.2 (1.5 mL) and the reaction mixture was stirred at
rt for 16 h. The reaction was diluted with MeOH and purified by
prep HPLC (H.sub.2O--CH.sub.3CN with 10 mM NH.sub.4OAc buffer) to
yield
8-cyclohexyl-N-((4,6-dimethyl-2-pyrimidinyl)sulfonyl)-11-methoxy-1a-((3-m-
ethyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl)-1,1a,2,12b-tetrahydrocyclo-
propa[d]indolo[2,1-a][2]benzazepine-5-carboxamide (24.3 mg, 0.032
mmol, 35.0% yield) as a light yellow solid. LC-MS retention time
1.25 min; m/z 721 (MH-). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Phenomenex-Luna 10u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 ml/min, a gradient of 100% solvent A/0% solvent B to
0% solvent A/100% solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray
mode.
[0180] Compounds in the following procedures were analyzed using
the following LC/MS method until noted: Analysis Conditions:
Column. PHENOMENNEX-LUNA 3.0.times.50 mm S10; Mobile Phase: (A)
10:90 methanol-water; (B) 90:10 methanol-water; Buffer: 0.1% TFA;
Gradient Range: 0-100% B; Gradient Time: 2 min; Flow Rate: 4
mL/min; Analysis Time: 3 min; Detection: Detector 1: UV at 220 nm;
Detector 2: MS (ESI+).
[0181] General procedure for the preparation of acylsulfonamide
derivatives. A mixture of acid (1 equiv) and carbonyldiimidazole
(1.5 equiv) in an. THF was heated at 50.degree. C. for 30 min and
allowed to cool to rt. Then, 1 equiv of sulfonamide (R=alkyl or
aryl) and DBU (2 equiv) were added consecutively. The resultant
mixture was stirred at rt overnight. After acidic aqueous workup,
isolated crude product was purified by prep. HPLC to afford
acylsulfonamide derivatives.
##STR00156##
[0182]
(+/-)-8-cyclohexyl-N-(1-benzylcyclopropane-1-sulfonyl)-1,1a,2,12b-t-
etrahydro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carbonyl-
)cycloprop[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The product
was purified by Prep HPLC and isolated as a TFA salt. LC-MS
retention time: 3.365 min; MS m/z (M+H) 747. The product was
observed to exist as inter-converting rotamers. 1H NMR (400 MHz,
CHLOROFORM-d) .delta. ppm 1.57 (25H, m), 2.74 (4H, m), 3.36 (3 H,
m), 3.64 (1H, d, J=15.11 Hz), 3.88 (3H, m), 4.41 (1H, br. s.), 5.16
(1H, m), 6.97 (1H, m), 7.11 (1H, m), 7.20 (6H, m), 7.30 (1H, m),
7.83 (2H, m).
##STR00157##
[0183]
(+/-)-8-cyclohexyl-N-(1-(pent-2-ynyl)cyclopropane-1-sulfonyl)-1,1a,-
2,12b-tetrahydro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-c-
arbonyl)cycloprop[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The
product was purified by Prep HPLC and isolated as a TFA salt. LC-MS
retention time: 3.334 min; MS m/z (M+H) 723. The product was
observed to exist as inter-converting rotamers. 1H NMR (400 MHz,
CHLOROFORM-d) .delta. ppm 3.62 (1H, d), 3.89 (3H, s), 4.46 (1H, m),
5.16 (1H, m), 6.98 (1H, dd, J=8.56, 2.52 Hz), 7.11 (1H, d, J=2.52
Hz), 7.30 (1 H, d, J=8.81 Hz), 7.55 (1H, m), 7.88 (1H, d, J=8.31
Hz), 8.03 (1H, s).
##STR00158##
[0184]
(+/-)-8-cyclohexyl-N-(1-(trimethylsilyl)cyclopropane-1-sulfonyl)-1,
1a, 2,
12b-tetrahydro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octa-
ne-8-carbonyl)cycloprop[d]indolo[2,1-a][2]benzazepine-5-carboxamide.
The product was purified by Prep HPLC and isolated as a TFA salt.
LC-MS retention time: 3.383 min; MS m/z (M+H) 729. The product was
observed to exist as inter-converting rotamers. 1H NMR (400 MHz,
CHLOROFORM-d) .delta. ppm 0.15 (9H, s), 1.09 (2H, m), 1.41 (7H, m),
1.80 (5H, m), 2.01 (7H, m), 2.49 (5H, m), 2.96 (2H, m), 3.61 (2H,
d, J=15.11 Hz), 3.89 (3H, s), 4.48 (2H, m), 5.16 (1H, m), 6.98 (1H,
dd, J=8.44, 2.64 Hz), 7.11 (1H, d, J=2.52 Hz), 7.28 (1H, d, J=8.81
Hz), 7.49 (1H, m), 7.90 (1H, d, J=8.31 Hz), 7.99 (1H, s).
##STR00159##
[0185]
(+/-)-8-cyclohexyl-N-(1-benzoylcyclopropane-1-sulfonyl)-1,1a,2,12b--
tetrahydro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carbony-
l)cycloprop[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The
product was purified by Prep HPLC and isolated as a TFA salt. LC-MS
retention time: 3.256 min; MS m/z (M+H) 761.
##STR00160##
[0186]
(+/-)-8-cyclohexyl-N-(2-cyanobenzenesulfonyl)-1,1a,2,12b-tetrahydro-
-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)cyclopro-
p[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The product was
purified by Prep HPLC and isolated as a TFA salt. LC-MS retention
time: 3.320 min; MS m/z (M+H) 733.
##STR00161##
[0187]
(+/-)-8-cyclohexyl-N-(cyclobutylmethanesulfonyl)-1,1a,2,12b-tetrahy-
dro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)cyclo-
prop[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The product was
purified by Prep HPLC and isolated as a TFA salt. LC-MS retention
time: 3.263 min; MS m/z (M+H) 685. The product was observed to
exist as inter-converting rotamers. 1H NMR (400 MHz, CHLOROFORM-d)
.delta. ppm 1.36 (5H, m), 1.99 (20H, m), 2.80 (5 H, m), 3.64 (5H,
m), 3.90 (3H, s), 4.40 (2H, m), 5.23 (1H, m), 6.97 (1H, m), 7.12
(1H, d, J=2.52 Hz), 7.27 (1H, m), 7.51 (1H, m), 7.90 (1H, m), 8.04
(1H, m).
##STR00162##
[0188]
(+/-)-8-cyclohexyl-N-(cycloprpylmethanesulfonyl)-1,1a,2,12b-tetrahy-
dro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)cyclo-
prop[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The product was
purified by Prep HPLC and isolated as a TFA salt. LC-MS retention
time: 3.170 min; MS m/z (M+H).671. Compound 22m was observed to
exist as inter-converting rotamers. 1H NMR (400 MHz, CHLOROFORM-d)
.delta. ppm 0.39 (2H, m), 0.67 (2H, m), 1.32 (6H, m), 1.65-3.22
(18H, m), 3.56 (5H, m), 3.89 (3H, s), 4.34 (2H, m), 5.20 (1H, m),
6.98 (1H, m), 7.11 (1H, d, J=2.27 Hz), 7.30 (1H, m), 7.66 (1H, m),
7.90 (1H, m), 8.10 (1H, m).
##STR00163##
[0189]
(+/-)-8-cyclohexyl-N-(2-methoxyethanesulfonyl)-1,1a,2,12b-tetrahydr-
o-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)cyclopr-
op[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The product was
purified by Prep HPLC and isolated as a TFA salt. LC-MS retention
time: 3.070 min; MS m/z (M+H) 675. The compound was observed to
exist as inter-converting rotamers. 1H NMR (400 MHz, CHLOROFORM-d)
.delta. ppm 1.58 (16H, m), 2.56 (2H, br. s.), 2.95 (3H, br. s.),
3.30 (3H, s), 3.59 (3H, m), 3.85 (8H, m), 4.43 (2H, m), 5.18 (2H,
br. s.), 6.97 (1H, m), 7.11 (1H, d, J=2.52 Hz), 7.30 (1H, m), 7.53
(1H, m), 7.91 (1H, m), 8.03 (1H, br. s.).
##STR00164##
[0190]
(+/-)-8-cyclohexyl-N-(phenylcarbamoyl)cyclopropylsulfonyl)-1,1a,2,1-
2b-tetrahydro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carb-
onyl)cycloprop[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The
product was purified by Prep HPLC and isolated as a TFA salt. LC-MS
retention time: 3.303 min; MS m/z (M+H) 776.
##STR00165##
[0191]
(+/-)-8-cyclohexyl-N-(1-allylcyclopropane-1-sulfonyl)-1,1a,2,12b-te-
trahydro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-
cycloprop[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The product
was purified by Prep HPLC and isolated as a TFA salt. LC-MS
retention time: 3.235 min; MS m/z (M+H) 697. The product was
observed to exist as inter-converting rotamers. 1H NMR (400 MHz,
CHLOROFORM-d) .delta. ppm 1.26 (6H, m), 1.98 (17H, m), 2.70 (6 H,
m), 3.10 (3H, m), 3.63 (1H, d, J=15.11 Hz), 3.89 (3H, s), 4.36 (2H,
m), 5.11 (2 H, m), 5.73 (1H, m), 6.99 (1H, m), 7.11 (1H, d, J=2.27
Hz), 7.29 (1H, d, J=8.56 Hz), 7.53 (1H, m), 7.89 (1H, m), 8.01 (1H,
s).
##STR00166##
[0192]
(+/-)-8-cyclohexyl-N-(phenylmethanesulfonyl)-1,1a,2,12b-tetrahydro--
11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)cycloprop-
[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The product was
purified by Prep HPLC and isolated as a TFA salt. LCMS method 1
(72007-001): LC-MS retention time: 3.253 min; MS m/z (M+H) 707. The
product was observed to exist as inter-converting rotamers. 1H NMR
(400 MHz, CHLOROFORM-d) .delta. ppm 1.35 (6 H, m), 1.96 (8H, m),
2.76 (12H, m), 3.32 (1H, br. s.), 3.60 (1H, d, J=15.11 Hz), 3.90
(3H, s), 4.21 (1H, m), 4.79 (1H, m), 4.88 (1H, m), 5.10 (0H, m),
6.97 (1H, dd, J=8.56, 2.52 Hz), 7.10 (1H, d, J=2.52 Hz), 7.32 (7H,
m), 7.66 (1H, d, J=8.06 Hz), 7.92 (1H, d, J=8.56 Hz).
##STR00167##
[0193]
(+/-)-8-cyclohexyl-N-((3,5-dichlorophenyl)methanesulfonyl)-1,1a,2,1-
2b-tetrahydro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carb-
onyl)cycloprop[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The
product was purified by Prep HPLC and isolated as a TFA salt. LC-MS
retention time: 3.405 min; MS m/z (M+H) 775.
##STR00168##
[0194] (+/-)-8-cyclohexyl-N((3-chlorophenyl)methanesulfonyl)-1,1a,
2,12b-tetrahydro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8--
carbonyl)cycloprop[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The
product was purified by Prep HPLC and isolated as a TFA salt. LC-MS
retention time: 3.3301 min; MS m/z (M+H) 741.
##STR00169##
[0195]
(+/-)-8-cyclohexyl-N-((methoxycarbonyl)cyclopropylsulfonyl)-1,1a,2,-
12b-tetrahydro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-car-
bonyl)cycloprop[d]indolo[2,1-a][2]benzazepine-5-carboxamide. The
product was purified by Prep HPLC and isolated as a TFA salt. LC-MS
retention time: 3.090 min; MS m/z (M+H) 715.
##STR00170##
[0196]
8-cyclohexyl-N-(2-methoxyethanesulfonyl)-1,1a,2,12b-tetrahydro-11-m-
ethoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)cycloprop[d]i-
ndolo[2,1-a][2]benzazepine-5-carboxamide. Step 1: To a solution of
t-butylester-ethylester (0.4 g, 0.776 mmol) in THF (8 ml) was added
1N NaOH (3.10 ml, 3.10 mmol) and MeOH (.about.5 ml) (2:20 PM). The
white mixture was stirred at room for 2 h. Diluted with EtOAc,
washed with, brine, dried (MgSO.sub.4), and removed the solvent in
vacuo to afford ester-acid as a white solid (0.38 g, 100%). LC-MS
retention time: 3.913 min; MS m/z (M+H) 502. Step 2: To a mixture
of ester-acid (0.38 g, 0.758 mmol) and
3-methyl-3,8-diazabicyclo[3.2.1]octane (0.181 g, 0.909 mmol) in DCM
(3 ml) was added TEA (0.5 ml, 3.59 mmol) and HBTU (0.345 g, 0.909
mmol). The mixture was stirred at room temperature for 2 h. Diluted
with EtOAc and washed with 1N NaOH, brine, dried (MgSO4), and
purified by Biotage 25M column afford ester-amide as a white foamy
solid (0.398 g, 86%). LC-MS retention time: 3.493 min; MS m/z (M+H)
610. Step 3: Ester-amide (0.398 g, 0.653 mmol) was dissolved in DCM
(2 mL) and added TFA (2 mL, 26.0 mmol) in ice cold bath. The
reaction mixture was stirred for 1.5 h at r.t., removed the solvent
to afford acid-amide as a brown solid (0.36 g, 100%). LC-MS
retention time: 3.186; MS m/z (M+H) 554. Step 4: Amide-amide was
purified and isolated as TFA salt. LC-MS retention time: 3.078 min;
MS m/z (M+H) 675, and was observed to exist as inter-converting
rotamers. 1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 1.28 (7H, m),
2.15 (22H, m), 3.28 (3H, s), 3.47 (1H, m), 3.86 (3H, m), 4.06 (2H,
br. s.), 5.24 (1 H, m), 6.96 (1H, m), 7.11 (1H, d, J=2.52 Hz), 7.29
(1H, m), 7.60 (1H, br. s.), 7.90 (1H, m), 8.10 (1H, m).
All samples for the procedures which follow were analyzed by the
LCMS method 1 described below:
[0197] LCMS method 1: Start % B: 0, Final % B: 100; Gradient time:
3 min; Stop time: 4 min; Flow rate: 4 ml/min; Wavelenth: 220;
Solvent A: 10% MeOH/90% H.sub.2O/0.1% Trifluoroacetic Acid; Solvent
B: 10% H.sub.2O/90% MeOH/0.1% Trifluoroacetic Acid; Column. Xbridge
4.6.times.50 mm S5.
##STR00171##
[0198]
(+/-)-8-cyclohexyl-N-[(2-fluorophenyl)methanesulfonyl]-1,1a,2,12b-t-
etrahydro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carbonyl-
)cycloprop[d]indolo[2,1-a][2]benzazepine-5-carboxamide(YT4)
Step 1: Preparation of Acylsulfonamide YT2
[0199] To a solution of compound YT1 (60 mg, 0.105 mmol),
(2-fluorophenyl)methanesulfonamide (39.6 mg, 0.209 mmol) and DMAP
(55 mg, 0.450 mmol) in DCM (1 ml) was added EDCI (40 mg, 0.209
mmol). The mixture was stirred at r.t. for three days. The reaction
mixture was purified by prep HPLC and repurified by Biotage 12M
(MeOH/DCM 0 to 10%) to afford the compound YT2 as a colorless glass
(66 mg, 100%). LC-MS retention time: 3.55 min; MS m/z (M+H)
631.
Step 2: Hydrolysis of methyl ester YT2 to acid YT3
[0200] To a solution of compound YT2 (66 mg, 0.102 mmol) in THF (3
ml) and MeOH (1 ml) was added NaOH (1 ml, 1.000 mmol). The mixture
was stirred at room for 2 h and then diluted with EtOAc, washed
with cold HCl (1N), brine, dried (MgSO4). Evaporation of solvents
in vacuum to afforded YT3 as a yellow solid (63 mg; 100%). LC-MS
retention time: 3.501 min; MS m/z (M+H) 617.
Step 3: Amide Coupling
[0201] To a mixture of compound YT3 (63 mg, 0.102 mmol) and
3-methyl-3,8-diazabicyclo[3.2.1]octane, 2HCl (20.34 mg, 0.102 mmol)
in DCM (1.5 ml) was added TEA (0.05 ml, 0.359 mmol) and HBTU (54.2
mg, 0.143 mmol). The mixture was stirred at room temperature for 2
h. Reaction was quenched with MeOH (1 ml) and evaporated to dryness
and the residue was dissolved in MeOH, filtered and purified by
Prep-HPLC to afford YT4 as a beige solid in mono TFA salt form
(30.8 mg; 35.9%). 1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm
7.88-7.97 (1H, m), 7.63 (1H, br. s.), 7.34-7.45 (1H, m), 7.20-7.32
(4H, m), 7.05-7.18 (2H, m), 6.96 (1H, dd, J=8.94, 2.64 Hz), 5.14
(1H, d, J=16.12 Hz), 4.95-5.04 (1H, m), 4.83-4.90 (1H, m), 3.88
(3H, s), 3.67-4.62 (2H, m), 3.63 (1H, d, J=14.86 Hz), 3.30-3.42
(1H, m), 2.13-3.08 (10H, m), 1.62-2.16 (8H, m), 0.95-1.52 (6H, m).
LC-MS retention time: 3.171 min; MS m/z (M+H) 725.
##STR00172##
[0202]
(+/-)-8-cyclohexyl-N-[1-(methoxycarbonyl)cyclopropanesulfonyl]-1,1a-
,2,12b-tetrahydro-11-methoxy-1a-(3-methyl-3,8-diazabicyclo[3.2.4]octane-8--
carbonyl)cycloprop[d]indolo[2,1-a][2]benzazepine-5-carboxamide(YT6).
To compound YT5 (240 mg, 0.359 mmol) and CDI (146 mg, 0.899 mmol)
was added THF (1 ml). The mixture was stirred at 50.degree. C. for
0.5 h and cooled down. Methyl 1-sulfamoylcyclopropanecarboxylate
(160 mg, 0.893 mmol) and DBU (0.217 ml, 1.438 mmol) were added. The
mixture was stirred for 3 days and diluted with EtOAc, washed with
satd. NaH2PO4, brine, dried (MgSO4), and purified by Biotage 25S
column (MeOH/DCM: 0 to 25%) to afford YT6 as a white solid (160 mg,
27%). LC-MS retention time: 3.171 min; MS m/z (M+H) 725.
* * * * *