Note: Descriptions are shown in the official language in which they were submitted.
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VASOPRESSIN AGONIST FORMULATION AND PROCESS
This application concerns new formulations for a class of tricyclic
vasopressin
agonist compounds, and pharmaceutically acceptable salts thereof, as well as
processes for manufacturing the formulations. The invention particularly
relates to
orally administered formulations of these compounds.
Background of the Invention
The art describes many methods of producing liquid or semi-solid
encapsulated pharmaceutical formulations. In Bull. Tech./Gattefosse Rep.
(1996), 89,
27-38, authors Shah et al. describe hard gelatin capsule technology,
particularly for
use in enhancing the bioavailability of poorly soluble or poorly absorbed
drugs.
U.S. Patent No. 4,620,974 (Hersh et al.) teaches a hard gelatin capsule
comprising a telescoping two-piece cap with a lubricant comprising a
polyethylene
glycol of a molecular weight between about 200 and about 900 present in
admixture
with the composition at a concentration of from about 0.5 to about 25 weight
percent.
WO 96/40071 (Lamberti) discloses methods and devices for producing
minimal volume capsules. WO 96/41622 (Tanner et al.) teaches suspensions
suitable
for encapsulation in gelatin capsules, particularly including a solid phase of
solid
particles and a liquid phase capable of suspending the solid phase.
U.S. Patent No. 5,641,512 (Cimiluca) teaches soft gelatin encapsulated
analgesics in which the shell contains a xanthine derivative, such as
caffeine.
U.S. Patent No. 4,578,391 (Kawata et al.) describes oily compositions for
antitumor agents comprising at least one sparingly oil soluble or water-
soluble
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antitumor drug, at least one fat or oil, and at least one solubilizing
adjuvant in an oily
vehicle, selected from crown ether, lecithin, polyethylene glycol, propylene
glycol,
vitamin E, polyoxyethylene alkylether, and sucrose esters of fatty acids.
EP 0 815 854 A1 discloses a substantially translucent, semi-solid fill
material
for a soft gelatin capsule, the semi-solid material being sufficiently viscous
that it
cannot be expelled from the capsule with a syringe at room temperature.
U.S. Patent No. 4,744,988 (Brox) teaches soft gelatin capsules comprising a
shell of gelatin, a softener and a filling of a polyethylene glycol and a low
polyhydric
alcohol and at least one active substance, characterized in that the shell
contains 4 to
40 percent sorbital or sorbitanes, at least half of the weight of polyethylene
glycol
used is a polyethylene glycol having a mean molecular weight of 600, and the
capsule
filling comprises up to 20% by weight of glycerol and/or 1,2-propylene glycol.
WO 95/19579 (Dhabhar) teaches a process for solubilizing difficulty soluble
pharmaceutical agents in a mixture of polyethylene glycol and propylene glycol
by
using a polyvinylpyrrolidone with a specific viscosity average molecular
weight of
from about 5,000 to about 25,000.
Summary of the Invention
This invention provides orally administerable formulations for tricyclic
vasopressin agonist compounds, or the pharmaceutically acceptable salts
thereof,
singularly or collectively optionally referred to herein as "active
ingredient", which
have the structure:
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~N
- G
N
r __
wherein:
A, B, E, G are, independently, CH or nitrogen;
D is, independently, C-W or nitrogen;
R' is alkanoyl of 2 to 7 carbon atoms, a group selected from CN, COOH,
CONHz,
H ~ - R9
or a moiety selected from the group:
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R3
S ~ N~ ~ N/ ~ \ N
N
\ -~R2 R2
> > ~N
(a) ~ (b) ~ (c) (d)
\ R2 \ R2 / R2 \
'~ ~' ~ \ >'
N- ' N- N ' ~ ~ '
N-
R4 ~Ra Ra/ ~Ra
) (~ (~ (b)
/N ~ ~ ~\ w / ~ /~
r V
> >
N-N ' ~N
Rah R2 R2
(j)
\~/N\ ~ / ~ ~ \
\ /~ ~ ~R2 or W
N- N , ~/ ~ i R2
Ra/ N= N
(m) (n) (o)
Rz, R3 and RS are, independently, hydrogen, straight chain alkyl of 1 to 6
carbon atoms, branched chain alkyl of 3 to 7 carbon atoms, cycloalkyl of 3 to
7
carbon atoms, or perfluoroalkyl of 1 to 6 carbons;
R4 is hydrogen, straight chain alkyl of 1 to 6 carbon atoms, branched chain
alkyl of 3 to 7 carbon atoms, cycloalkyl of 3 to 7 carbon atoms, alkoxyalkyl
of 2 to 7
carbon atoms, or an acyl substituent selected from the group consisting of
alkanoyl of
2 to 7 carbon atoms, alkenoyl of 3 to 7 carbon atoms, cycloalkanoyl of 3 to 7
carbon
atoms, amyl, or arylalkanoyl;
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X and Y are, independently, hydrogen, straight chain alkyl of 1 to 6 carbon
atoms, branched chain alkyl of 3 to 7 carbon atoms, cycloalkyl of 3 to 7
carbon
atoms, perfluoroalkyl of 1 to 6 carbons, alkoxyalkyl of 2 to 7 carbon atoms,
halogen
(including chlorine, bromine, fluorine, and iodine), alkoxy of 1 to 6 carbons,
hydroxy, CF3, or perfluoroalkyl of 2 to 6 carbons;
W is hydrogen, halogen (preferably chloro, bromo or iodo), alkyl, alkoxyalkyl
of 2 to 7 carbons, hydroxyalkyl of 1 to 6 carbons, or CHzNR6R';
R6 and R' are, independently, hydrogen, straight chain alkyl of 1 to 6 carbon
atoms, branched chain alkyl of 3 to 7 carbon atoms; or, taken together with
the
nitrogen atom of CHzNR6R', R6 and R' form a five or six membered ring
optionally
containing one or more additional heteroatoms such as, but not limited to,
those of the
group:
> >
bN ~ ~N , ~ or ~N~
~N ~ - VN -N
R$ is a straight chain alkyl of 1 to 6 carbon atoms
R9is independently hydrogen, trimethylsilyl or a straight chain alkyl of 1 to
6
carbon atoms;
or a pharmaceutically acceptable salt, ester or prodrug form thereof.
Among the more preferred active ingredient compounds of the formulations of
this invention are those of the formula:
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X
m I
Y~.A R 1
wherein:
A and B are, independently, CH or nitrogen;
D is C-W or nitrogen;
R' is alkanoyl of 2 to 7 carbon atoms or a group selected from
R3
2
\ ~ R5 \ ~ R2 ~ R
~'
N- N- N\
(a) R2 (b) R2 (e)~ Ra Ra
R2
or
R~N-N ~R4 R4~~N R2
(b)
R2, R3 and RS are, independently, hydrogen, straight chain alkyl of 1 to 6
carbon atoms, branched chain alkyl of 3 to 7 carbon atoms, cycloalkyl of 3 to
7
carbon atoms, or perfluoroalkyl of 1 to 6 carbons;
R', X, Y, W, R6, R' and Ra are as defined above;
or a pharmaceutically acceptable salt thereof.
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For the compounds defined above and referred to herein, unless otherwise
noted, aroyl groups include, for example, benzoyl, naphthoyl which can be
substituted independently with one or more substituents from the group of
hydrogen,
halogen, cyano, straight chain alkyl of 1 to 6 carbon atoms, branched chain
alkyl of 3
to 7 carbon atoms, alkoxy of 1 to 6 carbons, CF3, or phenyl.
Heteroaroyl groups herein refer to a carbonyl (radical) directly bonded to a
carbon atom of a five membered heterocyclic ring having one or two heteroatoms
selected from nitrogen, oxygen, sulfur, for example 2-thienoyl. The
heterocyclic ring
of the heteroaroyl groups may also include, but are not limited to, groups in
which the
aryl portion is a furan, pyrrole, 2H-pyrrole, imidazole, pyrazole,
isothiazole,
isoxazole, thiophene, pyrazoline, imidazolidine or pyrazolidine group. The
heteroaryl
groups herein can be substituted independently with one or more substituents
from
the group of hydrogen, halogen, cyano, straight chain alkyl of 1 to 6 carbon
atoms, or
branched chain alkyl of 3 to 7 carbon atoms.
The arylalkanoyl groups herein refer to a carbonyl group or radical directly
bonded to an alkyl group of 1 to 6 carbon atoms which is terminally
substituted by an
aryl group, for example, phenylacetic acid. The aryl group can be substituted
independently with one or more substituents from the group of hydrogen,
halogen,
cyano, straight chain alkyl of 1 to 6 carbon atoms, branched chain alkyl of 3
to 7
carbon atoms, alkoxy of 1 to 6 carbons, CF3, or phenyl or substituted phenyl
where
the substituents are selected from halogen, cyano, straight chain alkyl of 1
to 6 carbon
atoms, branched chain alkyl of 3 to 7 carbon atoms, alkoxy of 1 to 6 carbons,
CF3.
The halogens referred to herein may be selected from fluorine, chlorine,
bromine or iodine, unless otherwise specified.
It is understood by those practicing the art that the definition of the
compounds of formula (I), when R', Rz, R3, R', R5, R6, R', X, or Y contain
asymmetric
carbons, encompass all possible stereoisomers and mixtures thereof which
possess the
activity discussed below. In particular, it encompasses any optical isomers
and
diastereomers; as well as the racemic and resolved, enantiomerically pure R
and S
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_g_
stereoisomers; as well as other mixtures of the R and S stereoisomers and
pharmaceutically acceptable salts thereof, which possess the indicated
activity.
Optical isomers may be obtained in pure form by standard separation
techniques. It is
also understood that the definition of R', R2, R3, R°, R5, R6, R', X,
or Y of the
compounds of formula (I), encompasses all possible regioisomers, and mixtures
thereof which possess the activity discussed below. Such regioisomers may be
obtained pure by standard separation methods known to those skilled in the
art.
Also among the preferred groups of active ingredients in the formulations of
this invention are those in the subgroups:
a) compounds having the general formula:
w
~ I N
~\
B
A 1
R
wherein A, B, W, R', R2, R3, R4, R5, R6, R', Rg, R9, X, and Y are as defined
above;
b) compounds having the general formula:
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O
/N
N
r ..
wherein A, B, R', RZ, R3, R', R5, R9, X, and Y, are as defined above; and
c) compounds having the general formula:
~N~N
~N
X N
N
O
~\B
Y~ A t
wherein A, B, R', R2, R', R', R5, R9, X, and Y, are as defined above.
It is understood that subgroups a)-c), above, further include subgroups
wherein:
A and B are, independently, CH or nitrogen;
R' is alkanoyl of 2 to 7 carbon atoms or a group selected from
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R3
R5 ~ ~ ~ R2 ~ R2
R2 R2 R4
(e) ~' (~l
R2 ~ ~ R2
or
R4~~ N ~R4 R4/ ~ N R2
('~ (k)
R2, R3 and RS are, independently, hydrogen, straight chain alkyl of 1 to 6
carbon atoms, branched chain alkyl of 3 to 7 carbon atoms, cycloalkyl of 3 to
7
carbon atoms, or perfluoroalkyl of 1 to 6 carbons; and
R', X, Y, W, R6, R' and R8 are as defined above;
or a pharmaceutically acceptable salt thereof.
Particularly preferred among the compounds of group a), above, are those in
which W is H, A and B are each CH, and R' is the group of alkanoyl of 2 to 7
carbon
atoms or a group selected from the moieties (a), (b), (e), (f), (g), (h), (i)
or (k), listed
above.
The pharmaceutically acceptable salts include those derived from such organic
and inorganic acids as: lactic, citric, acetic, tartaric, succinic, malefic,
malonic,
hydrochloric, hydrobromic, phosphoric, nitric, sulfuric, methanesulfonic, and
similarly known acceptable acids.
Among the most preferred formulations of this invention are those described
herein having as the active ingredient [2-Chloro-4-(3-methyl-pyrazol-1-yl)-
phenyl-
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(5H, 11H)-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone, or
pharmaceutically
acceptable salts thereof, having the structure:
C
CH3
The formulations of this invention are useful in methods for treating in
humans or other mammals diseases, conditions or disorders in which vasopressin
agonist activity is desired. These methods of treatment include those for
diseases,
conditions or disorders which make it desirable to release factor VIII and von
Willebrand factor into the circulatory system, release tissue-type plasminogen
activator (t-PA) in the blood circulation, or affect the renal conservation of
water and
urine concentration. Such methods of treatment include, but are not limited
to,
treatments for diabetes insipidus, nocturnal enuresis, nocturia, urinary
incontinence,
or bleeding and coagulation disorders in humans or other mammals, including
hemophilia.
The methods herein for which these formulations are used include facilitation
in humans or other mammals of temporary delay of urination, which may also be
described as controlling or treating the inability to temporarily delay
urination,
whenever desirable. This method is understood to include treatments
facilitating the
temporary delay of urination which are separate from and not included in the
treatment of the conditions known as nocturnal enuresis and nocturia.
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In order to obtain consistency of administration, it is preferred that a
composition of the invention is in the form of a unit dose. Suitable unit dose
forms
preferably include tablets or capsules, though one skilled in the art will
understand
semi-solids or gels of this invention are also readily made and useful. Such
unit dose
forms may contain from 0.1 to 1000 mg of a compound of the invention and
preferably from 2 to 50 mg. Still further preferred unit dosage forms contain
1 to 25
mg, more preferably from 1 to 10 mg, of a compound of the present invention.
The
compounds of the present invention can be administered orally at a dose range
of
about 0.01 to 100 mg/kg or preferably at a dose range of 0.1 to 10 mg/kg. Such
compositions may be administered from 1 to 6 times a day, more usually from 1
to 4
times a day.
The compositions of the invention may be formulated with other conventional
carriers or excipients such as fillers, disintegrating agents, binders,
lubricants,
flavoring agents and the like.
The formulations of this invention comprise (by % w/w):
a) from about 1 % to about 20% of active ingredient, or a
pharmaceutically acceptable salt thereof, preferably from about 1% to about
16%
more preferably from about 5% to about 16% of this active ingredient;
b) from about 1% to about 18% of a surfactant component, preferably
from about 1% to about 5%, more preferably from about 5% to about 15% or from
about 5% to about 18%, most preferably from about 5% to about 10% or from
about
8% to about 12% of the surfactant component;
c) from about 50% to about 80% of a component of one or more
polyethylene glycols (PEG), preferably from about 55% to about 70% of one or
more
polyethylene glycols; and
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d) from about 1% to about 20%, preferably from about 5% to about 15%
and more preferably between about 8% and about 12%, of a component of:
i) one or more sucrose fatty acid esters; or
ii) a polyvinylpyrrolidone (PVP) with a K value between about 15
and 90, preferably with a K value of from about 17 as defined in USP/NF; or
iii) a combination of one or more sucrose fatty acid esters and a
PVP, as defined above.
The polyethylene glycol component may be comprised of one or more PEG
polymers, preferably commercially available PEG polymers between PEG 200 and
PEG 4,000, i.e. those PEG polymers having an average molecular weight between
about 190 and about 4800. More preferred are PEG polymers between average
molecular weights of from about 190 to about 3450, most preferably between
about
400 and 1540. Among the preferred PEG polymers are PEG 400, having an average
molecular weight between about 380 and about 420, and PEG 1,000, having an
average molecular weight between about 950 and about 1050. The ratio of high
and
low molecular weight PEG species within the PEG component is preferably from
about 2.5:1 to about 1:2.5, more preferably about 1:1. As an example, a
preferred
blend of PEG polymers within this invention would include a 1:1 blend of PEG
400
and PEG 1000. It may be preferable to choose a mixture of PEG components which
will have a melting point at or near the physiological temperature of the
mammal to
receive the formulation. Mixtures of final components which have a viscosity
range
of from about 140 to about 1500 centipoise at 37°C may be preferred,
more
preferably a range of from 300 to about 800 centipoise at 37°C.
The surfactants that may be used with the present formulations include, but
not limited to, polysorbate 20 (polyoxyethylene 20 sorbitan monolaurate),
Polysorbate 60, Polysorbate 40, polysorbate 80, Span 80 Sorbitan Oleate, a
product of
ICI Americas, Wilmington, Delaware, polysorbate 81, polysorbate 85,
polysorbate
120, bile acids and salts defined by Martindale The Extra Pharmacopoeia
Thirtieth
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Edition on page1341-1342 such as Sodium taurocholates, Sodium
deoxytaurocholates, Chenodeoxycholic acid, and ursodeoxycholic acid, and
pluronic
or poloxamers such as Pluronic F68, Pluronic L44, Pluronic L101, or
combinations of
one or more of the above. Polysorbate 80, by itself or in combination with one
or
more other surfactants, is preferred for use with this invention.
The sucrose fatty acid esters useful with this invention include those
commercially available and ast recognized esters useful for orally
administered
pharmaceutical compositions, including monoesters, diesters and triesters of
sucrose,
or mixtures or blends thereof. Specific examples of esters useful with this
invention
are sucrose monolaurate, sucrose monomyristate, sucrose monopalminate, sucrose
monostearate, sucrose distearate, sucrose tristearate, sucrose trimyristate,
and sucrose
tripalmitate, or combinations thereof.
In addition to these components, other enhancing or protective antioxidants or
preservatives may be added to the compositions of this invention, which may
account
for up to about 4% by weight of the formulation. Examples may include ascorbyl
palmitate, benzyl alcohol, butylated hydroxyanisole (BHA), butylated
hydroxytoluene
(BHT), etc. Examples of these components in the present formulations would
include BHA at a concentration from about 0.3% to about 2.5% (% w/w) and BHT
at
a concentration from about 0.005% to about 0.15% (% w/w), preferably with a
mixture of BHA and BHT within these ranges. Further embodiments comprise about
0.2% BHT.
A formulation of this invention utilizing one or more of these antioxidants or
preservatives comprises:
a) from about 1% to about 20% of active ingredient, or a
pharmaceutically acceptable salt thereof, preferably from about 1% to about
16%,
more preferably from 5% to about 16% of this active ingredient;
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b) from about 1% to about 18% of a surfactant component, preferably
from about 5% to about 15% of the surfactant component, more preferably from
about 5% to about 10% or from about 8% to about 12% of the surfactant
component;
c) from about 50% to about 80% of a component of one or more
polyethylene glycols (PEG), preferably from about 55% to about 70% of one or
more
polyethylene glycols;
d) from about 1% to about 20%, preferably about 5% to about 15%, of
one or more sucrose fatty acid esters or polyvinylpyrrolidone (PVP) with a K
value
between about 15 and 90, preferably with a K value of from about 17 as defined
in
USP/NF; and
e) from about 0.1% to about 4% of one or more preservatives or
antioxidants, for example from about 0.3% to about 2.5% (% w/w) BHA and/or
from
about 0.005% to about 0.15% (% w/w) BHT.
One preferred embodiment of this invention provides a pharmaceutical
formulation comprising:
a) from about 5% to about 16% of active ingredient;
b) from about 5% to about 10% of a surfactant component;
c) a component of from about 55% to about 70% of one or more
polyethylene glycols;
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d) from about 5% to about 15% of polyvinylpyrrolidone (PVP) with a K
value between about 15 and 90, preferably with a K value of from about 17 as
defined
in USP/NF; and
e) from about 0.3% to about 2.5% (% w/w) BHA and from about 0.005%
to about 0.15% (% w/w) BHT.
Preferably, the formulations of this invention are enclosed in a sealed
enclosure after manufacture, such as soft or hard gelatin capsules. The
formulations
of this invention may be created as a liquid or semi-liquid formulation and
introduced
into a capsule. Similarly, using an acceptable range of components and/or
temperatures, the formulation may be made as a gel or solid prior to
encapsulation.
Detailed Description of the Invention
The active compounds useful in the formulations of the present invention may
be prepared according to one of the general processes outlined below.
As shown in Scheme I, a tricyclic benzodiazepine of formula (1) is treated
with an appropriately substituted acetylaroyl (heteroaroyl) halide, preferably
an aroyl
(heteroaroyl) chloride of formula (2) in the presence of a base such as
pyridine or a
trialkylamine such as triethylamine, in an aprotic organic solvent such as
dichloromethane or tetrahydrofuran at temperatures from -40°C to
50°C to yield the
acylated derivative of formula (3). Treatment of (3) with a dialkylamide
dialkyl
acetal of formula (4) in an aprotic organic solvent such as dichloromethane at
temperatures ranging from 0°C to the reflux temperature of the solvent
yields the
enone of formula (5) according to the procedure of Lin et al., J. Het. Chem.,
14, 345
(1977). Treatment of (5) with hydroxylamine or a substituted hydrazine of
formula
(6) in acetic acid at temperatures ranging from ambient to the reflux
temperature of
the solvent yields the target compounds of formula (I) wherein A, B, D, E, G,
X, Y,
RZ and R4 are as defined above, and R' is an heterocyclic moiety selected from
the (f),
(g), or (j) group of heterocycles defined above.
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r~ o\ N
N-N
R2
(.))
The preferred substituted acetylaroyl (heteroaroyl) chlorides of formula (2)
of
Scheme I are conveniently prepared by treating the corresponding carboxylic
acids
with thionyl chloride at temperatures ranging from ambient to the reflux
temperature
of the solvent, or with oxalyl chloride in an aprotic solvent such as
dichloromethane
or tetrahydrofuran in the presence of a catalytic amount of dimethylformamide
at
temperatures ranging from 0°C to 40°C.
The preferred dialkylamide dialkylacetals are either available commercially,
or are known in the literature, or can be conveniently prepared according to
procedures analogous to those in the literature. Kantlehner, W. Chem. Ber.
105, 1340
( 1972).
The preferred tricyclic benzodiazepines of formula (1) are a 10,11-dihydro-
SH-pyrrolo[2,1-c][1,4]benzodiazepine (Albright et al., U.S. Patent No.
5,536,718,
issued July 16, 1996), a 10,11-dihydro-SH-pyrazole[5,1-c][1,4]benzodiazepine,
Cecchi, L. et. al., J. Het. Chem., 20, 871 (1983). and 10,11-dihydro-SH-
tetrazole[5,1-
c][1,4]benzodiazepine, Klaubert, D.H., J. Het. Chem., 22, 333 (1985).
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SCHEME I
Ø ~ X
-G
X \ N ~ Y ~A~O N
CH3 O
H > Y
A
(2) J= acylating moiety
(3)
i~
X i / G X i / N -G
R4-~2 (
R2 Oalkyl O' ~ or NH20H O
(alkyl)2N~Oalkyl Y ~A I O > Y ~ ~ 1
N(alkyl)2 A R
H
R2 (I)
An alternate process for the preparation of intermediates of formula (3) is
illustrated in the following Scheme II.
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SCHEME II
X-
Y
~~Y ~~ X ~ ~ G
A~B ~ A~B / H (1)
Br Br
Br
(~) J= COOH J= acy(lgjing moiety
HC= R9 , H2S04
pyridine, TEA
(Ph3P)ZPd(E)~ (II) S04
Cu(I)I R9
O
(10) (3)
Thus, a tricyclic benzodiazepine of formula (1) is treated with an
appropriately substituted bromo aroyl (heteroaroyl) halide, preferably an amyl
(heteroaroyl) chloride of formula (8) in the presence of an organic base such
as
pyridine or a trialkylamine such as triethylamine in an aprotic organic
solvent such as
dichloromethane or tetrahydrofuran at temperatures from -40°C to
50°C to yield the
acylated intermediate of formula (9). The intermediate (9) is subsequently
coupled
with a mono substituted terminal acetylene such as trimethylsilyl or a
straight chain
alkyl of 1 to 6 carbon atoms, in the presence of pyridine and a catalyst such
as
bis(triphenylphosphine) palladium (II) chloride and copper (I) iodide in an
organic
base such as triethylamine as the solvent, in a sealed pressure tube at
temperatures
ranging from ambient to 100°C essentially according to the procedure of
Martinez et
al., J. Med. Chem., 35, 620 (1992). The resulting acetylene intermediate of
formula
(10) is then hydrated by treatment with 1% sulfuric acid in an aprotic organic
solvent
such as tetrahydrofuran saturated with mercury (II) sulfate at ambient
temperature
essentially according to the procedure of Reed et al., J. Org Chem. , 52, 3491
(1987)
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to provide the desired acyl compound of formula (3) wherein A, B, D, E, G, X,
and
Y, are as defined above and R9 is hydrogen or a straight chain alkyl of 1 to 6
carbon
atoms. Alternatively, compound 9 where R9 is trimethyl is treated with
tetrabutylbutylammonium fluoride in an ether solvent such as tetrahydrofuran
to
afford compound (10) where R9 is hydrogen.
The preferred acylating agents of formula (8) of Scheme II are conveniently
prepared by treating an appropriately substituted aryl (heteroaryl) carboxylic
acid of
formula (7) with thionyl chloride at temperatures ranging from ambient to the
reflux
temperature of the solvent, or with oxalyl chloride in an aprotic organic
solvent such
as dichloromethane or tetrahydrofuran in the presence of a catalytic amount of
dimethylformamide at temperatures ranging from 0°C to 40°C.
The protected acetylene intermediates of Scheme II are either available
commercially, or are known in the art, or can be readily prepared by
procedures
analogous to those in the art.
As shown in Scheme III, the intermediate acetyl compounds (3) of Scheme I
can be prepared also by the Stille coupling of a bromo aryl (heteroaryl)
compound of
formula (9) of Scheme II with a (a-ethoxyvinyl)trialkyltin, preferably a (a-
ethoxyvinyl)tributylltin, in the presence of a catalytic amount of
bis(triphenylphosphine) palladium(II) chloride in an aprotic organic solvent
such as
toluene at temperatures ranging from ambient to the reflux temperature of the
solvent,
essentially according to the procedure of Kosugi et al., Bull. Chem. Soc.
Jpn., 60, 767
(1987).
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Scheme III
N' D~ E
N~D~E X ~ \ G
i \
X ~ ~G R3Sn OEt ~ ~ N
/ /N
(Ph3P)2Pd(II)C12
toluene Y
Y ~ ~ A ~ CHs
'A ~ Br
(9) (3)
The preparation of the acetyl compound (3) can also be accomplished via a
palladium-catalyzed arylation of a vinyl alkyl ether such as vinyl butylether,
with the
aryl halide intermediate of formula (9) according to the procedure of Cabri et
al.,
Tetrahedron Lett., 32, 1753 (1991).
The (a-alkoxyvinyl)trialkyltin intermediates of Scheme III are either
available
commercially, or are known in the art, or can be readily prepared by
procedures
analogous to those in the art.
In the case where R' in Scheme I is hydrogen, the heterocyclic nitrogen can be
alkylated or acylated according to the reactions outlined in Scheme IV.
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D~ E
SCHEME IV ~ N~ ~~
Xi
B
Y ' I
'°' I \ R2
(I Ra= H) . ~ N _
1. Alkylating or acylating agent
Base
2. Separation
of regioisomers
I ND.E ~,D.E
X i \ N i
X //r ~~G
/ ~/
1~B ~ /
Y B
Y ~I
I \ R2 A ~ R2
N,
(I) R1- f R4 (I) R1- g Ra~N'N
Ra = alkyl or acyl residue
Thus, the pyrazole compound of formula (I, R4 is H) is alkylated by treatment
with a strong base such as sodium or potassium hydride and an alkylating agent
such
as an alkyl halide, preferably an alkyl chloride (bromide or iodide) in an
aprotic
organic solvent such as dimethylformamide or tetrahydrofuran at temperatures
ranging from 0°C to 80°C to yield compound (I, R'= (f) or (g))
wherein A, B, D, E,
G, X, Y, and RZ are as defined above, and R4 is an alkyl or acyl moiety.
Alternatively, compound (I) is acylated by treatment with a carboxylic acid
halide,
preferably a chloride, or a carboxylic acid anhydride in the presence of an
amine base
such as pyridine or a trialkylamine, preferably triethylamine, in an aprotic
organic
solvent such as dichloromethane or tetrahydrofuran with no additional solvent
when
pyridine is used as the base, at temperatures ranging from -40°C to
ambient to yield
compound (I) wherein A, B, D, E, G, X, Y and RZ are as defined above, and R4
is an
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alkyl or acyl moiety. The alkylation or acylation of a compound of formula (I,
R' is
H) leads to a mixture of regioisomers wherein Rz is hydrogen and R' is an
heterocyclic moiety selected either from the (f) or (g) group of heterocycles
defined
above and illustrated below, respectively.
R2
R2
N' N
R4~ N
O
The compounds of general formula (I) of Scheme I wherein A and B are
carbon, Rz is H, and R' is an heterocyclic moiety selected from the (g) group
of
heterocycles defined above, can be prepared according to the general process
outlined
in Scheme V.
SCHEME V
R ~ , R'
J/ ~/
Y ~~ HC= C , TEA Y ~ Rs mCPBA
Br > C°~C~~N-R~ CH2C1 ~
(Ph3P)zPd(II)CI2
J= COOCH3
(11, A and B= carbon)) Cu(I)I (12) J= COOCH3
/ Rs
/ MeOH Y ~ I i
Y~ I ' \ \ N~R~
60°C
C~I~-R'
J= COOCH3 O (13) J= COOCH3
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a Y~ I Y Y\
R NHNH' ~ a/ N N/ +
N' ~Ra
(14) J= COOCH3 (15) J= COOCI~
(major) (minor)
Separation
MeOH
aq. NaOH
J
/ Y ~
Y
\ ~ --
R~ N~ N Ra/ N' N
(16) J= COOH (1~) J= acylating moiety
,D~
N E
G
N
-G N
N
O /
(1) Y ~ R1
(I)
Thus, an appropriately substituted haloaryl (heteroaryl) carboxylic acid
ester,
preferably a bromo (or iodo) methylester of formula (11) is coupled with a
dialkylamino propyne, preferably 1-dimethylamino propyne, in the presence of a
catalyst such as bis(triphenylphosphine) palladium(II) chloride and copper(I)
iodide
in an organic base such as triethylamine as the solvent and at temperatures
ranging
from ambient to 80°C essentially according to the procedures of Alami
et al.,
Tetrahedron Lett., 34, 6403 (1993), and of Sanogashira et al., Tetrahedron
Lett.,
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4467 (1975) to provide the substituted acetylene intermediate of general
formula (12).
The intermediate (12) is subsequently converted into its N-oxide by treatment
with an
oxidizing agent using any of a number of standard oxidative procedures
(Albini, A.,
Synthesis, 263 (1993) or with dioxirane reagents (Murray, R.W., Chem. Rev.,
1187
( 1989), in an aprotic organic solvent such as dichloromethane at temperatures
below
ambient. The intermediate N-oxide is not isolated but is rearranged in situ to
an
enone of general formula (13) by treatment with, preferably with heating, a
hydroxylic solvent, including any solvent or combination of solvents composed
of or
containing water, any C,-Cg straight chain or branched chain alkyl alcohol,
ethylene
glycol, polyethylene glycol, 1,2-propylene diol, polypropylene glycol,
glycerol, 2-
methoxyethanol, 2-ethoxyethanol, 2,2,2-trifluoroethanol, benzyl alcohol,
phenol, or
any equivalent solvent that contains one or more free hydroxyl (-OH)
substituent(s)
that is known to those skilled in the art.
Solvent systems containing one or more cosolvents, along with one or more
solvents may also be used for this process of rearranging the N-oxide to the
desired
enaminone. The cosolvents referred to herein may be defined as a diluent of
the main
solvents) and can be selected from: hydrocarbons such as pentane, hexane or
heptane; aromatic hydrocarbon such as benzene, toluene or xylene; ethers such
as
diethyl ether, tetrahydrofuran, dioxane or dimethoxy ethane; chlorinated
hydrocarbons such as dichloromethane, chloroform, dichloroethane, or
tetrachloroethane; or other common solvents such as ethyl acetate, N,N-
dimethylformamide, N,N-dimethylacetamide, acetonitrile, dimethylsulfoxide,
acetone, or the like.
The conversion of the amine N-oxide into an enaminone may be
accomplished by introducing the amine N-oxide into a suitable hydroxylic
solvent,
preferably with stirring, at or between about room or ambient temperature and
about
the reflux temperature of the solvent. In other instances the introduction of
the amine
N-oxide to a hydroxylic solvent, preferably with stirring, may be accomplished
in the
presence of an acceptable catalyst, such as a palladium(II) catalyst or a
copper (I)
catalyst, at or between room temperature and the reflux temperature of the
solvent.
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This procedure provides a novel synthesis of enaminone compounds from
propargylic amines or their N-oxides in hydroxylic solvents, which influence
the
ultimate outcome of the reaction. This new method of enaminone synthesis
provides
a convenient alternative to existing methods and further extends the range of
starting
materials that can be converted into enaminone products.
Although the precise mechanism by which a propargylic amine N-oxide is
converted into an enaminone product has not been rigorously determined, it
likely
resembles two known processes; the thermal [2,3]-sigmatropic rearrangement of
propargylic amine N-oxides (Craig, et. al., Tetrahedron Lett., 4025, 1979;
Hallstrom,
et. al., Tetrahedron Lett., 667, 1980; Khuthier, A-H, et. al., J. Chem. Soc.
Chem.
Commun., 9, 1979) and the conversion of certain isoxazoles into enaminones
(Liguori, et. al., Tetrahedron, 44, 1255, 1988).
Treatment of (13) with a substituted hydrazine (6) in acetic acid at
temperatures ranging from ambient to reflux leads to a mixture of
regioisomeric
compounds of general formulas (14) and (15) in a variable ratio. The major
isomer of
formula (14) is separated by means of chromatography and/or crystallization
and
subsequently hydrolyzed to the desired carboxylic acid of formula (16).
The intermediate (16) is then converted into an acylating species, preferably
an acid chloride (bromide or iodide) or a mixed anhydride of formula (17) by
procedures analogous to those described hereinbefore. The acylating agent (17)
is
then used to acylate a tricyclic benzodiazepine of formula (1) by any of the
procedures described hereinbefore to yield the desired compound of formula
(I),
wherein A, B are CH and D, E, G, X, Y, and R4 are as defined above, Rz is
hydrogen
and R' is a heterocyclic moiety selected from the (g) group of heterocycles
illustrated
below.
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~R2
R~ IN'- rN
(g)
Likewise, treatment of (13) with an unsubstituted hydrazine (6, R4 is H) in
acetic acid at temperatures ranging from ambient to the reflux temperature of
the
solvent yields the intermediate pyrazole ester of formula (18). In this case
the
heterocyclic nitrogen can be alkylated or acylated as shown in Scheme VI to
provide
compounds of formula (I) wherein Rz is hydrogen, and R' is an heterocyclic
moiety
selected from the (f) group of heterocycles defined above.
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SCHEME VI
R6 NH2NH2
_ I > Y
Y ~A ~ N~ 7 (6~ 4- ~A
R R _ H) N_ ~
O N
H
(13) J= COOCH3 (18, R2= H) J= COOCH3
Alkylating or / /
acylating agent Y
> ~A I ~ + A
Base
N-N 4,N_
R4 R
(15) J= COOCH3 (14) J= COOCH3
(maj or) (minor)
Separation
MeOH, aq. NaOH
Y ~ Y
A ~ ~~ > ->
N- N
Ra
(19) J= COOH (20) J= acylating moiety
0.
I \
i \ G X i /
X ~ / N
N
H
O / (I)
(1) Y
~A R~
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Thus, the intermediate ester of formula (18) is alkylated by treatment with a
strong base such as sodium or potassium hydride and an alkylating agent such
as an
alkyl halide, preferably an alkyl chloride (bromide or iodide), in an aprotic
solvent
such as dimethylformamide or tetrahydrofuran at temperatures ranging from
0°C to
80°C to yield a mixture of regioisomers of formulas (14) and (15) in a
variable ratio.
The major regioisomer of formula (15) is separated by chromatography andlor
crystallization and subsequently hydrolyzed to the desired carboxylic acid of
formula
(19), which is then converted into an acylating agent, preferably an acyl
chloride or a
mixed anhydride by procedures analogous to those described hereinbefore. The
acylating species of formula (20) is then used to acylate a tricyclic
benzodiazepine of
formula (1) to yield the desired compound of formula (I), wherein A, B, D, E,
G, X,
Y, and R4 are as defined above, Rz is hydrogen, and R' is a heterocyclic
moiety
selected from the (f) group of heterocycles defined above.
R2
,N~
N Ra
Compounds of general formula (I) wherein R' is an heterocyclic moiety
selected from the (h) group of R' heterocycles defined above, can be prepared
as
outlined in Scheme VII.
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SCHEME VII
H3 ~ 1) Ra~2
B O
Y~ Ai
Y 'q ,~ hi 2) KMnOa ~ NN
OOH Ra
J= COOH
(21) (22)
i \ N i \ N.
wB X , / N~G X ~ / N~G
Y 'A~ ~ N
N,
(1) O~ (I)
Ra >~
Y ~I
~A~R~
(23) J= acylating moiety
An appropriately substituted malondialdehyde of formula (21) is treated first
with a hydrazine in acetic acid at temperatures ranging from ambient to the
reflux
temperature of the solvent and the intermediate pyrazole is then oxidized with
potassium permanganate in a basic aqueous solution at temperatures ranging
from
ambient to the reflux temperature of the solvent to yield a carboxylic acid
intermediate of formula (22). The acid (22) is converted into an acylating
agent,
preferably an acid chloride (bromide or iodide) or a mixed anhydride by
procedures
analogous to those described hereinbefore. The acylating agent of formula (23)
is
finally reacted with a tricyclic benzodiazepine of formula (1) to yield
compounds of
general formula (I) wherein A, B, D, E, G, X, Y, and R° are as defined
above, and R'
is a heterocyclic moiety selected from the (h) group of heterocycles defined
above.
N
~N~
Ra
(h)
In the case where R4 in Scheme VII is hydrogen, the heterocyclic nitrogen can
be alkylated or acylated according to the procedures outlined hereinbefore.
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The preferred malondialdehydes of formula (21) and the hydrazines of
Scheme VII are either available commercially, or are known in the art, or can
be
readily prepared by procedures analogous to those in the literature for known
compounds, such as those of Knorr et al., J. Org. Chem., 49, 1288 (1984) and
Coppola et al., J. Het. Chem., 11, 51 (1974).
An alternative preparation of the intermediate carboxylic acids of formula
(22)
of Scheme VII wherein Y is as defined above and R4 is other than hydrogen, is
outlined in Scheme VIII.
SCHEME VIII
Br ~ J
N /~
~N Y''p,~ N H
J= COOCH3
(24) R4 (2~
n-BuLi
Sn(R)3 Cl NaNOz, HCl
~ KI, I2
J
(R~Sn ~B
N Y'~y
N J= COOCH3
(2~ R4 (2~
(Ph3P)4Pd(0), Cu(I~
DMF
J~ B J ~ ~ B
aq. NaOH
w ~ ~v
~NN MeOH A L -NN
Ra ~Ra
(2~ J=COOCH~ (22) J=COOH
The organotin reagent of formula (25) is reacted in a Stille coupling reaction
with an appropriately substituted aryl (heteroaryl) halide, preferably a
bromide or
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iodide of formula (28) in the presence of a catalyst such as
tetrakis(triphenylphosphine)palladium (0) and copper (I) iodide in an organic
aprotic
solvent such as dimethylformamide at temperatures ranging from ambient to
150°C,
essentially according to procedures analogous to those found in Farina et al.,
J. Ors.
Chem., 59, 5905 (1994). Basic hydrolysis of the resulting ester of formula
(26) with
sodium or lithium hydroxide in aqueous alcohol or tetrahydrofuran at
temperatures
ranging from ambient to the reflux temperature of the solvent yields the
desired
carboxylic acids of formula (22).
In turn, the organotin reagents of formula (25) wherein the R groups are
preferably alkyl groups, are conveniently prepared by metallation of a 4-bromo
N-
alkylpyrazole of formula (24) with a trialkyltin halide, preferably a
tributyltin
chloride (or bromide) in the presence of a metallating agent such as an
alkyllithium
such as n-butyl lithium, sec-butyl lithium, or tert-butyllithium in an aprotic
organic
solvent such as diethylether at temperatures ranging from -40°C to
ambient according
to procedures analogous to those found in Martina et al., Synthesis, 8, 613
(1991).
The preferred N-alkyl substituted 4-bromo pyrazoles of formula (24) are
conveniently prepared from 4-bromo pyrazole by alkylation with an alkyl
halide,
preferably an alkyl chloride (bromide or iodide) in the presence of a strong
base such
as lithium, sodium or potassium hydride in an aprotic organic solvent such as
dimethylformamide or tetrahydrofuran at temperatures ranging from 0°C
to 80°C.
Alternatively, alkylation of 4-bromopyrazole can be carried out with an
alkylating
agent mentioned above, and a strong alkaline base such as lithium, sodium or
potassium hydroxide in the presence of a phase transfer catalyst (Jones, R.A.
Aldrichimica ACTA, 9(3), 35, 1976) such as benzyldimethyltetradecylammonium
chloride, or benzyltrimethylammonium chloride.
The preferred aryl (heteroaryl) iodides of formula (28) are conveniently
prepared by diazotization of the corresponding substituted anilines of formula
(27)
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followed by reaction of the corresponding diazonium salt with iodine and
potassium
iodide in aqueous acidic medium essentially according to the procedures of
Street et
al., J. Med. Chem., 36, 1529 (1993) and of Coffen et al., J. Org. Chem., 49,
296
(1984).
An alternative preparation of compounds of general formula (I) is outlined in
Scheme IX.
SCHEME IX
Y~ ~ I \ N ~~ E
X ~G
N~ ~ E \A F(CI)
X ~ / N~G
O'
(29) J= acylating moiety Y
(1) 'A ~ F (CI)
N ~a E
1. R1-H (31) X ~ / G
Base
2. separation of O / I
regioisomers Y ' ~ ( )
A R~
(30)
A tricyclic benzodiazepine of formula (1) is treated with an appropriately
substituted haloaroyl (heteroaroyl) halide, preferably a fluoro aroyl or a
fluoro (or
chloro) heteroaroyl chloride of formula (29), in the presence of a base such
as
triethylamine or diisopropylethylamine in an aprotic organic solvent such as
dichloromethane or tetrahydrofuran at temperatures from -40°C to the
reflux
temperature of the solvent to yield the acylated derivative (30).
Alternatively, the acylating species can be a mixed anhydride of the
carboxylic acid described above, such as that prepared by reaction 2,4,6-
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trichlorobenzoyl chloride in a solvent such as dichloromethane according to
the
procedure of Inanaga et al., Bull. Chem. Soc. Jpn, 52, 1989 (1979). Treatment
of
said mixed anhydride of general formula (29) with the tricyclic benzodiazepine
of
formula (1) in a solvent such as dichloromethane and in the presence of an
organic
base such as 4-dimethylaminopyridine at temperatures ranging from 0°C
to the reflux
temperature of the solvent, yields the intermediate acylated derivative (30)
of Scheme
IX.
A compound of formula (30) is then treated with the lithium, sodium or
potassium salt of an appropriately substituted heterocycle of formula (31) in
a polar
aprotic organic solvent such as dimethylformamide or tetrahydrofuran at
temperatures
ranging from ambient to the reflux temperature of the solvent to yield a
compound of
general formula (I), wherein A, B, D, E, G, X, Y, R2, R3, and RS are as
defined above,
and R' is a heterocyclic moiety selected from the group consisting of (a),
(b), (c), (d),
(1), (n) or (o) defined above.
~ N
v
\ ~ RS \ N/ \\ \ N/
~~ R2
N
(a) ~ (b) ~ (c)
N
R2 ANN~ ~~~R2 ~ ~R2
~N
(d) (1) (n) (o)
The condensation of the intermediate of formula (30) with the intermediate
salt of formula (31) leads to a variable ratio of regioisomers of general
formula (I)
which are separated by means of chromatography and/or crystallization.
The preferred substituted fluoro aroyl and fluoro (or chloro) heteroaroyl
chlorides of formula (29) are either available commercially, or are known in
the art,
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or can be readily prepared by procedures analogous to those in the literature
for the
known compounds.
The lithium, sodium or potassium salts of the heterocycles of formula (31) are
prepared by treatment of said heterocycle with a strong base such as lithium
hydride,
sodium hydride, potassium hydride or a metal alkoxide at temperatures ranging
from
-40°C to ambient in an aprotic organic solvent such as
dimethylformamide or
tetrahydrof uran.
Alternatively, the compounds of general formula (I) described in Scheme IX
can be prepared according to the process outlined in Scheme X.
SCHEME X
1. R1-H (31)
J\
B (COCI)z, MeOH, DMF ~ 'B Base
> Y/
Y ~A~ F(CI) or SOC12, DMF, MeOH A F(CI) 2, separation of
or pTSA, MeOH regioisomers
(33) J= COOCH3
(32) J= COOH
Hydrolysis J
Y%T _, -~ Y/A~Ri
A ~ Ri NaOH, aq. MeOH
or LiOH, aq.THF
(34) J=COOCH3 (35) J=COOH
\ N. D_G X i / ~ G
X ~ / ~ N
N
J I \'B( H > G~ /
Y~A~R1 (1) Y
A R~
(36) J=acylating moiety
(I)
Thus, an appropriately substituted fluoroaryl or fluoro (or chloro)heteroaryl
carboxylic acid of formula (32) is esterified using methods known in the art
such as
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treatment with oxalyl chloride (or thionyl chloride) in an alcohol solvent
such as
methanol, in the presence of a catalytic amount of dimethylformamide; or by
condensation with methanol in the presence of an acid catalyst such as para-
toluenesulfonic acid at temperatures ranging from ambient to reflux.
The resulting ester of formula (33) is reacted with the lithium, sodium or
potassium salt of an appropriately substituted heterocycle of formula (31) in
a polar
aprotic organic solvent such as dimethylformamide at temperatures ranging from
ambient to 150°C, to yield an intermediate ester of formula (34). The
condensation
of (33) with (31) leads to a variable ratio of regioisomers of formula (34)
which are
separated by means of chromatography and/or crystallization.
Subsequent hydrolysis of the intermediate ester of formula (34) with an
aqueous base such as lithium, sodium or lithium hydroxide in methanol or
tetrahydrofuran affords the carboxylic acid of formula (35).
The intermediate carboxylic acid (35) is then converted into an acylating
agent preferably an acid chloride or a mixed anhydride of general formula (36)
using
any of the procedures described hereinbefore.
Subsequent reaction of the tricyclic benzodiazepine of formula (1) with the
intermediate acylating agent of formula (36) according to any of the
procedures
described hereinbefore yields the desired compounds of formula (I) of Scheme
IX.
Alternatively, the substituted carboxylic acids of formula (35) described in
Scheme X can be prepared according to the process outlined in Scheme XI.
SCHEME XI
N~ 1. Rl-H (31) N HOO
Y ~'~\ '~ Base Y~i H2S04 Y T
'A~ F(CI) 2, separation of ~A~ R~ A R1
regioisomers
(37, Y ~ CFA (38) (35)
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Thus, a fluoro aryl or fluoro (chloro)heteroaryl nitrile of formula (37) is
reacted with the lithium, sodium or potassium salt of a substituted
heterocycle of
formula (31) in an apolar aprotic solvent such as dimethylformamide at
temperatures
ranging from ambient to 150°C, to yield an intermediate of general
formula (38). The
reaction of (37) with (31) leads to a variable ratio of regioisomers of
formula (38)
which are separated by means of chromatography and/or crystallization.
Hydrolysis
of the intermediate nitriles of formula (38, Y~ CF3) is preferentially carried
out with
an inorganic acid such as sulfuric acid at temperatures ranging from ambient
to 60°C.
Alternatively, hydrolysis of the nitrile (38) can be carried out by heating in
ethanol in the presence of a strong alkaline base such as sodium hydroxide
with or
without a phase transfer catalyst (Jones, R.A. Aldrichimica Acta, 9(3), 35,
1976,)
such as benzyldimethyltetradecyl ammonium chloride.
The resulting carboxylic acids of formula (35) are then converted into the
desired compounds of formula (I) of Scheme IX by procedures analogous to those
described hereinbefore.
Alternatively, the substituted carboxylic acids of formula (35) of Scheme X
can be prepared according to the process outlined in Scheme XII by sequential
treatment of a nitrile of formula (38) wherein A and B are CH and where R' is
not
alkanoyl of 2 to 7 carbons, alkynyl, (b) or (d), with basic hydrogen peroxide
in
dimethylsulfoxide essentially according to the procedure of Katritzky et al.,
Synthesis, 949 (1989), followed by hydrolysis of the resulting amide of
formula (38)
preferably by treatment with dilute sulfuric acid and sodium nitrite according
to the
procedure of Hanes et al., Tetrahedron, 51, 7403 (1995).
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SCHEME XII
H NOC HOOC
NC 30% H20z 2 / 75% HZS04 /
Y / I > Y \ I ~ Y \ I ,
R
\ Ri DMSO, K2C03 R NaNO
(38) (39) (35)
Where R1 is not (b) or (d)
A preferred process for the preparation of the intermediate substituted
carboxylic acids of formula (35) of Scheme X wherein R' is a heterocyclic
moiety
selected from the (a) group of R' heterocycles defined above, is outlined in
Scheme
XIII.
SCHEME XIII
J
J~ Y ~~ ~B + SnClz, HCl
Y ' ~ H2S04, NaN02 A N v _ >
NH2 ~ ~ N
(40) J= COOCH3 (41) J= COOCH3
O R3
1. R2~P
J / Rs (47) J~ aq. NaOH J
Y ~ > Y ~ ~ > Y
A NHNH2 MeOH, H20 A R1 or LiOH, A R~
HCl aq. THF
2. crystallization
(42) J= COOCH3 (34) J= COOCH3 (35) J= COOH
~D, ~ ~ D- E
N
J X-', / ~G X ' ~ 'G
N N
Y I H
> ~A~R~ (1) > O
(36) J=acylating moiety Y
(I) ~A ~ Ri
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Diazotization of an appropriately substituted aniline of formula (40) followed
by reduction of the resulting diazonium salt of formula (41) with tin (II)
chloride in
concentrated hydrochloric acid according to the procedure of Street et al., J.
Med.
Chem., 36, 1529 (1993) provides the intermediate hydrazine hydrochloride salt
of
formula (42). Subsequent condensation of (42) with an aldehyde derivative of
formula (47), wherein RZ is as defined above, R3 and RS is H, and P is
dialkylacetal)
such as acetylacetaldehyde dimethyl acetal, or a ketone of formula (47),
wherein R2,
R3 and RS are as defined above, and P is =O or (O-alkyl)z in a solvent such as
aqueous methanol at temperatures ranging from ambient to 100°C provides
after
crystallization, the desired intermediate ester of formula (34, R' is (a) and
RS is H),
which is then converted to the compound of formula (I) as outlined in Scheme X
above.
R3
y ~ R5
R2
(a)
When Y is OCH3 the compounds of general formula (I) of Scheme I can be
conveniently demethylated as outlined in Scheme XIV.
SCHEME XIV
\ N.D. E .D. E
X ~ -G ~ \ N
i / ~ X -G
'/
N
~ BBr3, CH2C12
O-' \ > ~
/ Oi \
Y ~A~R1 Y
A R
(I) Y= OCH3
(I) Y= OH
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Thus, the reaction of compound (I) wherein Y is OCH3 with boron tribromide
in an organic solvent, such as dichloromethane, yields the corresponding
phenol of
formula (I) wherein Y is OH, and A, B, D, E, G, X, RZ and R3 are as defined
above
and R' is an heterocyclic moiety selected from the group (a) of heterocycles
defined
above and illustrated below.
(a) ~
Compounds in which R' contains three heteroatoms are prepared according to
Scheme XV.
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SCHEME XV
n_
X-
,0.
R2 a~ kyi
-G
~N 1) ~ ~ (43) ;y1)2 Oalkyl
H Y ~A~CN >
> (4)
(1) J= acylating moiety
NH2
2) H2S04 (44)
~~ E
\ I ~~ E
G ~ \ I
N R4~2 (() X ~ / G
> N
O / or NH20H
O
Y ~A O Y
N' 'N(alkyl)2 ( ) A R
(45)
R
Thus, a tricyclic benzodiazepine of formula (1) is treated with an
appropriately substituted cyano amyl (heteroaroyl) halide, preferably an aroyl
(heteroaroyl) chloride of formula (43) in the presence of a base in an aprotic
organic
solvent such as dichloromethane or tetrahydrofuran at temperatures ranging
from -
40°C to 80°C to yield an intermediate nitrile of formula (46,
Scheme XVI) which in
turn, is hydrolyzed to an amide intermediate of general formula (44) with an
inorganic acid such as sulfuric acid at ambient temperature to 50 °C.
Treatment of the
amide (44) with a dialkyl amide dialkyl acetal of formula (4) in an aprotic
organic
solvent such as dichloromethane or tetrahydrofuran at temperatures ranging
from 0°C
to 80°C yields the intermediate of formula (45). Treatment of (45) with
hydroxylamine or a hydrazine of formula (6) in acetic acid at temperatures
ranging
from ambient to reflux yields the desired target compounds of formula (I)
wherein A,
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B, D, E, G, X, Y, RZ and R' are as defined above, and R' is an heterocyclic
moiety
selected from the (e), (i) and (k) group of heterocycles defined above.
R2 / R2 O\ N
~~-N / N-N
R4 R2
Another preferred process for the preparation of the intermediate amide of
formula (44), see Scheme XV, wherein A and B are CH and D is not CH is
outlined
in Scheme XVI and consists of treating a nitrite of formula (46) with basic
hydrogen
peroxide in dimethylsulfoxide essentially according to the procedure of
Katritzky et
al., Synthesis, 949 (1989).
SCHEME XVI \ N ~~ E
\ X ~ -G
Y~ / /
E \1~ N
N ~ CN
X ~ ~G > _
J- acylating moiety O Y
CN
(43)
(1 D~ CH) (46)
N ~' E
X ~ ~G
30% H2O2 N
O
DMSO, K2C03 Y ,
~O
(44 D~ CH) NH2
The preferred process to prepare compounds of general formula (I) in which
R' contains four heteroatoms and R4 is hydrogen is outlined in Scheme XVII.
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SCHEME XVII
.D. E
\ N.D. E X ~ \ N ~G
X ~ 'G NaN3, NH4Cl '
DMF N
N
O-' \~ O
Y r' B Y
(46) 'A ~ CN (I) A R'
Treatment of the nitrile intermediate of formula (46) of Scheme XVI with
sodium azide and ammonium chloride in an aprotic organic solvent such as
dimethylformamide at temperatures ranging from ambient to the reflux
temperature
of the solvent yields the desired compounds of formula (I) wherein A, B, D, E,
G, X,
and Y, are as defined above, R4 is hydrogen and R' is an heterocyclic moiety
selected
from the group (m) of heterocycles defined above.
N~
N- N
(m)
The compounds of general formula (I) wherein D is CW and W is hydrogen,
can undergo Mannich condensation as shown in Scheme XVIII.
SCHEME XVIII
N E
N D'E X ' \ -G
X ~ G ~ /
N
N HOAc, aq, CH20
' O
O Y / R6R~NH Y
i
i (47) A R
A R
(I, D= CH) (I, W= CH2NR6R~ )
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Thus, reaction of compounds of formula (I, D is CH) with either aqueous
formaldehyde or paraformaldehyde, a substituted amine of formula (47), and
glacial
acetic acid in an alcohol solvent such as methanol at temperatures ranging
from
ambient to reflux yields the corresponding Mannich bases of general formula
(I),
wherein A, B, E, G, X, Y, R2, R3, R5, R6 and R' are as defined above; D is CW;
W is a
dialkylaminoalkyl residue preferably a dimethylaminomethyl residue, and R' is
an
heterocyclic moiety selected from the (a), (c), (e), (f), (g), (h), (i), (j),
(k), (1), (m), (n)
and (o) group of heterocycles defined above.
Likewise, the compounds of general formula (I) wherein D is CH can undergo
halogenation as shown in Scheme XIX.
SCHEME XIX
w
G
i~ G
N-Halosuccinimide N
N
CH2C12 O /
O / Y
Y ~ ~ A R~
A R'
(I, D= CH) (I W = halogen)
Thus, reaction of (I, D is CH) with a N-halosuccinimide such as N-chloro
(bromo or iodo)succinimide in a polar aprotic organic solvent such as
dichloromethane at temperatures ranging from -80°C to ambient yields
the
corresponding halogenated derivatives of general formula (I), wherein A, B, E,
G, X,
RZ, R3 and RS are as defined above, D is CW, W is a halogen such as chlorine
(bromine or iodine), and R' is an heterocyclic moiety selected from the (a),
(c), (e),
(~, (g), (h), (i), (j), (k), (1), (m), (n) and (o) group of heterocycles
defined above.
The subject compounds of the present invention were tested for biological
activity according to the following procedures.
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Vasopressin VZ Agonist Effects of Test Compounds in Normal Conscious
Water-Loaded Rats:
Male or female normotensive Sprague-Dawley rats (Charles River
Laboratories, Inc., Kingston, NY) of 350-500 g body weight were supplied with
standard rodent diet (Purina Rodent Lab. Chow 5001) and water ad libitum. On
the
day of test, rats were placed individually into metabolic cages equipped with
devices
to separate the feces from the urine and containers for collection of urine.
Test
compound or reference agent was given at an oral dose of 10 mg/kg in a volume
of 10
ml /kg. The vehicle used was 20% dimethylsulfoxide (DMSO) in 2.5% preboiled
corn starch. Thirty minutes after dosing the test compound, rats were gavaged
with
water at 30 ml /kg into the stomach using a feeding needle. During the test,
rats were
not provided with water or food. Urine was collected for four hours after
dosing of
the test compound. At the end of four hours, urine volume was measured.
Urinary
osmolality was determined using a Fiske One-Ten Osmometer (Fiske Associates,
Norwood, MA, 02062) or an Advanced CRYOMATIC Osmometer, Model 3C2
(Advanced Instruments, Norwood, MA). Determinations of Na', K" and Cl- ion
were
carned out using ion specific electrodes in a Beckman SYNCHRON EL-ISE
Electrolyte System analyzer. The urinary osmolality should increase
proportionally.
In the screening test, two rats were used for each compound. If the difference
in the
urine volume of the two rats was greater than 50%, a third rat was used.
Vas~ressin V2 Agonist Effects of Test Compounds in Normal Conscious
Homozy~~ous Brattleboro Rats with Central Diabetes Insipidus
Male or female homozygous Brattleboro rats (Harlan Sprague Dawley, Inc.,
Indianapolis, IN) of 250-350 g body weight were supplied with standard rodent
diet
(Purina Rodent Lab. Chow 5001) and water ad libitum. On the day of test, rats
were
placed individually into metabolic cages equipped with devices to separate the
feces
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from the urine and containers for collection of urine. Test compound or
reference
agent was given at an oral dose of 1 to 10 mg/kg in a volume of 10 ml /kg. The
vehicle used was 20% dimethylsulfoxide (DMSO) in 2.5% preboiled corn starch.
During the test, rats were provided with water ad libitum. Urine was collected
for six
hours after dosing of the test compound. At the end of six hours, urine volume
was
measured. Urinary osmolality was determined using a Fiske One-Ten Osmometer
(Fiske Associates, Norwood, MA, 02062) or an Advanced CRYOMATIC
Osmometer, Model 3C2 (Advanced Instruments, Norwood, MA). Determinations of
Na', K' and Cl- ion were carried out using ion specific electrodes in a
Beckman
SYNCHRON EL-ISE Electrolyte System analyzer. This animal model was mainly
used for evaluation of potency and duration of action of the active compounds.
The
results of this study are shown in Table I.
Example # Urine Volume Osmolality Rat Type '
(% decrease)a (% Increase)"
2 80% (1 mg/kg) 306% (1 mg/kg)CD
3 58% 240% CD
4 57% 225% CD
5 56% 231 % CD
6 5 8 % 270% CD
7 13% 137% CD
9A 70% 325% CD
9B 21 % 168% CD
11 70% 285% CD
12 69% 330% CD
13 50% 229% CD
14 86% 406% CD
15 47% 38% CD
16 88% 400% CD
18 52% 214% CD
25% (1 mg/kg) 152% (1 mg/kg)CD
21 49% 181 % CD
22 80% 322% CD
24 47% 159% CD
87% 979% CD
26 54% 279% CD
27 76% 183% CD
28 75% 37% CD
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29 66% 305% CD
30 81 % 334% BB
31 72% 298% CD
32 77% 373% CD
33 68% 362% CD
34 76% 407% BB
35 63% 308% CD
36 66% 164% BB
37 71 % 370% CD
38 66% 256% BB
39 69% 253% CD
40 46 % 183 % CD
41 69% 240% CD
49 74% 221 % BB
50 53% 223% CD
51 72% CD
52 66% 261 % CD
55 80% 164% CD
57 77% 288% CD
58 49% 324% CD
59 80% 607% CD
60 54% 165% CD
61 59% 245% CD
62 22% 150% CD
63 27% 214% CD
64 79% 349% CD
71 84% 264% CD
77 13% 90% CD
78 21% 115% CD
79 38% 123% CD
81 82% 490% CD
83 85% 442% CD
84 56% 291 % CD
85 76% 436% CD
86 5% 86% CD
87 71 % 214% CD
88 68% 226% CD
90 61 % 413 % CD
91 22% 69% CD
92 69% 454% CD
95 68% 300% CD
97 3% 106% CD
99 43 % 205 % CD
100 24% 248% CD
101 76% 376% CD
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107 31 % 125 % CD
108 30% 145% CD
109 21 % 95 % CD
115 66% 229% CD
116 66% 256% CD
117 68% 311% CD
120A 66% 269% CD
120B 67% 272% CD
121 22% 155% CD
123 88% 663% CD
°Percent decrease in urine volume vs. control at 10 mg per kg, unless
otherwise stated.
°Change in osmolality as percent of control at 10 mg/kg, unless
otherwise stated.
°Rat model used: Sprague-Dawley (CD) or Brattleboro (BB).
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The following examples are presented to illustrate rather than limit the scope
of the invention.
EXAMPLE 1
(4-Fluoro-2-tritluoromethyl-phenyl)-(5H,11H-pyrrolo[2,1-c] [1,4]benzodiazepin-
10-yl)-methanone
Oxalyl chloride (2.0 g) was added to a suspension of 4-fluoro-2-
trifluoromethylbenzoic acid (2.0 g) in dichloromethane (25 ml). Two drops of
dimethylformamide were added and the mixture was stirred for 18 hours at room
temperature. The resultant solution was evaporated to dryness to give the
crude acid
chloride. This was redissolved in dichloromethane and filtered. Evaporation of
this
material gave a liquid which was then redissolved in hexane, filtered, and
evaporated
to yield the acid chloride as a pale yellow viscous liquid, which was used
without
further purification.
The acid chloride (2.26 g) in dichloromethane (25 ml) was added portionwise
to a mixture of 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]benzodiazepine (1.66 g),
dichloromethane (10 ml), and diisopropylethylamine (1.30 g), cooled in an ice
bath.
After remaining at room temperature for 18 hours, the reaction mixture was
washed
with water and saturated aqueous sodium bicarbonate. The dichloromethane
solution
was dried over anhydrous sodium sulfate and filtered through a short column of
hydrous sodium magnesium silicate and further eluted with several volumes of
dichloromethane. The combined organic phase was concentrated on a hot plate
with
the gradual addition of hexane until crystallization occurred. After cooling,
the
crystals were collected by filtration to yield 2.57 g of the title compound,
m.p. 154
155 °C.
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EXAMPLE 2
[4-(3-Methyl-pyrazol-1-yl)-2-trifluoromethyl-phenyl] (5H,11H-pyrrolo[2,1-c]-
[1,4]benzodiazepin-10-yl)-methanone
60% Sodium hydride in oil (0.15 g) was washed with hexane and dry
dimethylformamide (25 ml) was added, followed by 3-methylpyrazole (0.25 g).
After hydrogen evolution ceased, (4-fluoro-2-trifluoromethyl-phenyl)(5H,11H-
pyrrolo[2,1-c][1,4]benzodiaxepin-10-yl)-methanone (1.0 g) was added. The
reaction
mixture was heated in a sand bath at 110 °C for 15 hours. The reaction
mixture was
poured onto ice and saturated saline solution was added. The precipitate was
collected by filtration. The crude reaction product was dissolved in
dichloromethane
and filtered through a short column of hydrous sodium magnesium silicate and
further
eluted with several additional volumes of dichloromethane. The combined
organic
phase was concentrated on a hot plate with the gradual addition of hexane.
After
cooling, the crystals were collected by filtration to yield 0.77 g of a crude
product.
Further purification by additional filtration through a short column of
hydrous sodium
magnesium silicate, followed by the addition of hexane, yielded the title
compound as
a crystalline solid (0.66 g), m.p. 194-195 °C.
EXAMPLE 3
[4-(4-Methyl-pyrazol-1-yl)-2-tritluoromethyl-phenyl]-(5H,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 2, employing (4-fluoro-2-trifluoromethyl-
phenyl)(SH,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.8 g), 60%
sodium hydride in oil (0.15 g), 4-methylpyrazole (0.20 g) and
dimethylformamide
(25 ml), the product (0.47 g) was obtained as a colorless amorphous solid, MS,
m/z:
437.3 (M+H)+, 873.2 (2M+H)+.
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EXAMPLE 4
(4-Pyrazol-1-yl-2-tritluoromethyl-phenyl)-(5H,11H-pyrrolo[2,1-
c] [1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 2, employing (4-fluoro-2-trifluoromethyl-phenyl)-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.0 g), 60% sodium
hydride in oil (0.20 g), pyrazole (0.25 g) and dimethylformamide (35 ml). The
product (0.62 g) was obtained as a colorless amorphous solid, MS, m/z: 423.2
(M+H)+, 445.2 (M+Na)+, 845.3 (2M+H)+.
EXAMPLE 5
[4-(3-Cyclopropyl-pyrazol-1-yl)-2-trifluoromethy-phenyl]-(SH,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 2, employing (4-fluoro-2-trifluoromethyl-phenyl)-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.42 g), 60% sodium
hydride in oil (0.20 g), 3-cyclopropylpyrazole (0.43 g) and dimethylformamide
(50
ml), the product (1.22 g) was obtained as a crystalline solid, m.p. 163-164
°C.
EXAMPLE 6
[4-(4-Methyl-imidazol-1-yl)-2-trifluoromethyl-phenyl]-(5H,11H-pyrrolo[2,1-
c] [1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 2, employing (4-fluoro-2-trifluoromethyl-phenyl)-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.0 g), 60% sodium
hydride in oil (0.20 g), 4-methylimidazole (0.25 g) and dimethylformamide (25
ml),
the title compound (0.66 g) was obtained as an amorphous solid. MS, m/z: 437.2
(M+H)+, 873.2 (2M+H)+.
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EXAMPLE 7
(5H,11H-Pyrrolo[2,1-c] [1,4]benzodiazepin-10-yl)-(4-[1,2,4]triazol-1-yl-2-
trifluoromethylphenyl)-methanone
In the manner of Example 2, employing (4-fluoro-2-trifluoromethyl-phenyl)-
(SH,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.0 g),
60°Io sodium
hydride in oil (0.20 g), 1,2,4-triazole (0.20 g), and dimethylformamide (25
ml), the
title compound (0.36 g) was obtained as a colorless amorphous solid, MS, m/z:
424.2
(M+H)+, 847.3 (2M+H)+
EXAMPLE 8
(2-Chloro-4-fluorophenyl)-(5H,11H-pyrrolo[2,1-c] [1,4]benzodiazepin-10-yl)-
methanone
Oxalyl chloride (2.60 g) was.added to a suspension of 2-chloro-4-fluorobenzoic
acid
(3.44 g) in dichloromethane (50 ml). Two drops of dimethylformamide were added
and the mixture was stirred for 18 hours at room temperature. The resultant
solution
was evaporated to give the crude 2-chloro-4-fluorobenzoyl chloride as a
viscous oil
(3.72 g).
The crude 2-chloro-4-fluorobenzoyl chloride (3.68 g) in dichloromethane (25
ml) was added portionwise to a stirred, ice cooled solution of 10,11-dihydro-
SH-
pyrrolo[2,1-c][1,4]benzodiazepine (2.76 g), diisopropylethylamine (2.47 g) and
dichloromethane (50 ml). After 18 hours at room temperature, the reaction
mixture
was washed with water and saturated aqueous sodium bicarbonate. The
dichloromethane solution was dried with anhydrous sodium sulfate and filtered
through a short column of hydrous sodium magnesium silicate and further eluted
with
several volumes of dichloromethane. The combined organic phase was
concentrated
on a hot plate with the gradual addition of hexane until crystallization
occurred. After
cooling, the crystals were collected by filtration to yield the title compound
(3.85 g),
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m.p. 110-112 °C.
EXAMPLE 9
[2-Chloro-4-(3-methyl-pyrazol-1-yl)-phenyl]-(SH,11H)-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone (Isomer A)
and
[2-Chloro-4-(5-methyl-pyrazol-1-yl)-phenyl]-(SH,11H)-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone (Isomer B)
Method 1: To 60% sodium hydride in oil (0.3 g, degreased with hexane)
in dimethylformamide (25 ml) was added 3-methylpyrazole (0.55 g). When the
hydrogen evolution subsided, (2-chloro-4-fluorophenyl)-(SH,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone (1.70 g) was added. The reaction mixture
was heated for 18 hours in a sand bath (internal temperature 125 °C).
The reaction
mixture was then poured onto ice and further diluted with a saturated saline
solution.
The precipitated solid was recovered by filtration. The crude product was
dissolved
in dichloromethane, dried over anhydrous sodium sulfate, and then filtered
through a
short column of hydrous sodium magnesium silicate and further eluted with
several
volumes of dichloromethane. The combined eluate was refluxed on a hot plate
with
the gradual addition of hexane until an opaque solution was observed. On
cooling an
amorphous solid was obtained. On subjecting this material to a second column
of
hydrous sodium magnesium silicate and evaporation of the solvent in vacuo gave
a
mixture of regioisomers 9A and 9B in approximately a 9:1 ratio as an amorphous
glass (1.11 g), MS, m/z: 403.2 (M+H)+.
Method 2: To a pre-cooled, stirred suspension of hexane-washed 60%
sodium hydride (3.00 g) in dry dimethylformamide (250 ml) was added dropwise
under nitrogen 3-methylpyrazole (5.50 g) at 0°C. The mixture was warmed
to room
temperature. After gas evolution ceased, 2-chloro-4-fluorophenyl)-(SH,11H-
pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (17.0 g) was added as a
solid,
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and the mixture heated to 130°C for one hour. The reaction mixture was
poured into
ice water, a precipitate collected by filtration, and air-dried. The
precipitate was
dissolved in dichloromethane, dried over anhydrous sodium sulfate, and
filtered
through a short column of silica gel, eluting with ethyl acetate. The combined
filtrate
was evaporated in vacuo to a residual foam (18.5 g). Purification and
separation of
regioisomers by low pressure column chromatography on silica gel eluting with
a
gradient mixture of ethyl acetate-hexane (10:90 to 25:75), yielded two
purified
regioisomers:
Isomer A, [2-chloro-4-(3-methyl-pyrazol-1-yl)-phenyl]-(5H,11H)-
pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (13.5 g), as a colorless
amorphous solid; MS (EI), m/z: 402 (M)+. A sample (0.5 g) was crystallized
from
diethyl ether, followed by recrystallization from ethanol to yield regioisomer
A
(0.275 g) as a colorless, crystalline solid, m.p. 141-143 °C;
Isomer B, [2-chloro-4-(5-methyl-pyrazol-1-yl)-phenyl]-(5H,11H)-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone (1.93 g) as a colorless amorphous solid.
A
sample was crystallized from diethyl ether, followed by recrystallization from
methanol to yield regioisomer B as colorless, needles (1.4 g), m.p. 160-163
~C; MS
(EI), m/z: 402 (M)+, MS (+FAB), m/z: 403 (M+H)+ .
EXAMPLE 10
[2-Chloro-4-(3-methyl-pyrazol-1-yl)-phenyl]-(SH,11H)-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone
Step a) 2-Chloro-4-(3-methylpyrazol-1-yl)benzonitrile: To a cooled (0
°C)
suspension of sodium hydride (60% in oil; 2.0 g) in dimethylformamide (50 ml)
was
added 3-methylpyrazole (3.39 g) in portions. After hydrogen gas evolution
ceased, 2-
chloro-4-fluorobenzonitrile (5.17 g) was added and the mixture was stirred at
room
temperature for 18 hours. The mixture was poured onto ice, diluted with brine,
and
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the resulting precipitate was collected by filtration. The crude product was
dissolved
in dichloromethane, filtered through a column of anhydrous sodium magnesium
silicate, and crystallized by the addition of hexane. Recrystallization from
ethanol
gave 4.42 g of product, m.p. 148-150 °C.
Step b) 2-Chloro-4-(3-methylpyrazol-1-yl)benzamide: A suspension of 2-
chloro-4-(3-methylpyrazol-1-yl)benzonitrile (4.35 g) from step a in dimethyl
sulfoxide (20 ml) containing potassium carbonate (0.40 g) was cooled in an ice
bath.
Hydrogen peroxide (30%, 2.4 ml) was added and the mixture was warmed to room
temperature over 1 hour. The resultant precipitate was recovered by filtration
and
recrystallized from ethanol to yield 2.44 g of product as fine needles, m.p.
159-160
°C; MS, m/z: 235.9 (M+H)+.
Step c) 2-Chloro-4-(3-methylpyrazol-1-yl)benzoic Acid: A solution of 2-
chloro-4-(3-methylpyrazol-1-yl)benzamide (1.09 g) from step b in aqueous 75%
sulfuric acid (25 ml) was cooled in an ice bath and sodium nitrite (1.73 g)
was added.
The mixture was warmed to room temperature over 1 h and poured onto ice. The
precipitate was collected by filtration and used directly in the next
reaction.
Step d) [2-Chloro-4-(3-methyl-pyrazol-1-yl)-phenyl]-(5H-10,11-dihydro-
pyrrolo[2,1-c][1,4] benzodiazepin-10-yl)-methanone: A mixture of 2-chloro-4-(3-
methylpyrazol-1-yl)benzoic acid (0.69 g), dichloromethane (25 ml) from step c
,
oxalyl chloride (1.0 g), and 1 drop of dimethylformamide was stirred at room
temperature for 18 hour. The mixture was concentrated, taken up in
dichloromethane
(25 ml), and added to a mixture of 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]-
benzodiazepine (0.51 g) in dichloromethane (25 ml) containing
diisopropylethylamine (0.76 g). The mixture was stirred at room temperature
for 18
h and washed with saturated aqueous sodium bicarbonate solution. The organic
layer
was dried over anhydrous sodium sulfate and filtered through a short column of
anhydrous sodium magnesium silicate. The solution was concentrated and the
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resultant material was crystallized from diethyl ether to give 0.67 g of
product, m.p.
137-138 °C; MS, m/z: 403.2 (M+H)+, 805.8 (2M+H)+.
EXAMPLE 11
[2-Chloro-4-(4-methyl-pyrazol-1-yl)-phenyl-(5H,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 9's Method 1, employing (2-chloro-4-
fluorophenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.0
g),
60% sodium hydride in oil (0.3 g, degreased with hexane), 4-methylpyrazole
(0.48 g)
and dimethylformamide (25 ml), the title compound (0.74 g) was obtained as an
amorphous solid, MS, m/z: 403.2 (M+H)+, 425.2 (M+Na)+, 805.3 (2M+H)+.
EXAMPLE 12
[2-Chloro-4-(4-methyl-imidazol-1-yl)-phenyl]-(5H,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 9's Method 1, employing (2-chloro-4-
fluorophenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.0
g),
60% sodium hydride in oil (0.3 g, degreased with hexane), 4-methylimidazole
(0.48
g), and dimethylformamide (25 ml), the title compound (0.38 g) was obtained as
an
amorphous solid, MS, m/z: 403.3 (M+H)+.
EXAMPLE 13
[2-Chloro-4-(3-trifluoromethyl-pyrazol-1-yl)-phenyl]-(5H,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 9's Method 1, employing (2-chloro-4-
fluorophenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.8
g),
60% sodium hydride in oil (0.25 g, degreased with hexane), 3-
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trifluoromethylpyrazole (0.61 g) and dimethylformamide (25 ml), the title
compound
was obtained as an amorphous solid, MS, m/z: 457.2 (M+H)+.
EXAMPLE 14
[2-Chloro-4-(1,2,4-triazol-1-yl)-phenyl]-(SH,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 9's Method 1, employing (2-chloro-4-
fluorophenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.7
g),
60% sodium hydride in oil (0.5 g, degreased with hexane), 1,2,4-triazole (0.70
g) and
dimethylformamide (50 ml), the title compound (0.51 g) was obtained as an
amorphous solid, MS, m/z: 390.3 (M+H)+, 779.3 (2M+H)+.
EXAMPLE 15
(2-Chloro-4-pyrrol-1-yl-phenyl)-(SH,11H-pyrrolo[2,1-c] [1,4]benzodiazepin-10-
yl)-methanone
In the manner of Example 9's Method 1, employing (2-chloro-4-
fluorophenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.7
g),
60% sodium hydride in oil (0.3 g, degreased with hexane), pyrrole (0.42 g) and
dimethylformamide (25 ml), the title compound (0.60 g) was obtained as an
amorphous solid, MS, m/z: 388.2 (M+H)+.
EXAMPLE 16
(2-Chloro-4-pyrazol-1-yl-phenyl)(5H,11H-Pyrrolo[2,1-c] [1,4]benzodiazepin-10-
yl)-methanone
In the manner of Example 9's Method 1, employing (2-chloro-4-
fluorophenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.0
g),
60% sodium hydride in oil (0.2 g, degreased with hexane), pyrazole (0.20 g)
and
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dimethylformamide (25 ml), the title compound was obtained as an amorphous
solid,
MS, m/z: 389.2 (M+H)+, 777.1 (2M+H)+.
EXAMPLE 17
[2-Chloro-4-(1H-imidazol-1-yl)-phenyl)-(5H,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 9's Method 1, employing (2-chloro-4-
fluorophenyl)-(SH,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (2.0
g),
60% sodium hydride in oil (0.50 g, degreased with hexane), imidazole (0.50 g)
and
dimethylformamide (25 ml), the title compound (0.57 g) was obtained as a tan
amorphous solid, MS, m/z: 389 (M+H)+.
EXAMPLE 18
[2-Chloro-4-(3-methylpyrazol-1-yl)-phenyl]-(3-methyl-5H,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone
Step a) 1-(SH, 11H-Pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-2,2,2-trifluoro-
ethanone: To an ice cooled solution of 10,11-dihydro-SH-pyrrolo[2,1-c][1,4]-
benzodiazepine (5.62 g) and diisopropylethylamine (4.0 g) in dichloromethane
(75
ml) was added dropwise trifluoroacetic anhydride (7.0 g) in dichloromethane.
The
mixture was stirred at room temperature for 18 hours, and washed with water
and
saturated aqueous sodium bicarbonate. The organic phase was dried over
anhydrous
sodium sulfate and filtered through a short column of anhydrous sodium
magnesium
silicate. The combined organic phase was concentrated on a hot plate with the
gradual
addition of hexane until crystallization occurred. After cooling, the crystals
were
collected by filtration to yield 7.70 g of product as fine needles, m.p. 134-
135 °C, MS
m/z: 281 (M+H)+.
Step b) 1-(3-Dimethylaminomethyl-SH,11H-pyrrolo[2,1-c][1,4]benzo-
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diazepin-10-yl)-2,2,2-trifluoroethanone. A mixture of 1-(5H, 11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-2,2,2-trifluoroethanone (2.80 g), ) from step a) ,
bis-
dimethylaminomethane (2.04 g), paraformaldehyde (2.70 g) and acetic acid (1.20
g)
in a mixture of tetrahydrofuran (50 ml) and methanol (50 ml) and was stirred
at room
temperature for 18 hours. The mixture was concentrated in vacuo , water was
added,
and the aqueous mixture was extracted with dichloromethane. The combined
extracts
were dried over anhydrous sodium sulfate and filtered through a short column
of
anhydrous sodium magnesium silicate. The solution was concentrated in vacuo
and
the residue was crystallized from hexane to yield 2.05 g of the product as a
colorless
solid m.p. 109-110 °C, MS m/z: 338.3 (M+H)+.
Step c) Trimethyl-(10-trifluoroacetyl-10,11-dihydro-5H-pyrrolo[2,1-
c][1,4]benzodiazepin-3-yl-methyl)-ammonium iodide: A mixture of 1-(3-
dimethylaminomethyl-5H,11H-pyrrolo[2,1-c] [ 1,4]benzodiazepin-10-yl)-2,2,2-
trifluoroethanone (1.83 g) from step b) and iodomethane (1.0 g) in
dichloromethane
(20 ml) was stirred at room temperature for 18 hours. Diethyl ether was added
and
the resulting precipitate was collected by filtration to give 2.54 g of
product as a
colorless solid, m.p. 140-155 °C (dec).
Step d) 10,11-Dihydro-3-methyl-5H-pyrrolo[2,1-c][1,4]benzodiazepine:
Sodium borohydride (2.6 g) was added in two portions to a refluxing mixture of
trimethyl-( 10-trifluoroacetyl-10,11-dihydro-5H-pyrrolo[2,1-c] [
1,4]benzodiazepin-3-
yl-methyl)-ammonium iodide (2.60 g) from step c) in ethanol. After 4 hours,
the
mixture was concentrated in vacuo. Water was added to the residue and the
mixture
was extracted with dichloromethane. The combined extracts were dried over
anhydrous sodium sulfate and filtered through a short column of anhydrous
sodium
magnesium silicate. The solution was concentrated in vacuo and the residue was
crystallized from hexane to yield 1.14 g of product, m.p. 150-151 °C,
MS m/z: 199.1
(M+H)+.
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Step e) [2-Chloro-4-(3-methylpyrazol-1-yl)-phenyl]-(3-methyl-SH,11H-
pyrrolo[2,1-c][1,4]-benzodiazepin-10-yl)-methanone: A mixture of 2-chloro-4-(3-
methylpyrazol-1-yl)-benzoic acid (0.18 g) from step d) , oxalyl chloride (0.18
g) and
one drop of dimethylformamide in dichloromethane (10 ml), was stirred at room
temperature for 18 hours. The mixture was concentrated in vacuo, and the
residue
was redissolved in dichloromethane and reconcentrated in vacuo to yield 2-
chloro-4-
(3-methyl-pyrazol-1-yl)-benzoyl chloride. A slurry of the acid chloride in
dichloromethane (25 ml) was added dropwise to a solution of 10,11-dihydro-3-
methyl-5H-pyrrolo[2,1-c][1,4]benzodiazepine (0.12 g) and diisopropylethylamine
(0.10 g) in dichloromethane (25 ml). The mixture was stirred at room
temperature for
18 h, and washed with water and saturated aqueous sodium bicarbonate. The
organic
phase was dried over anhydrous sodium sulfate and filtered through a short
column of
anhydrous sodium magnesium silicate. The solution was concentrated in vacuo
and
triturated with diethyl ether to yield 0.115 g of product as colorless
crystals, m.p. 178-
180 °C, MS m/z: 417.3 (M+H)+, 833.3 (2M+H)+.
EXAMPLE 19
(2-Chloro-4-trifluoromethyl-pyrimidin-5-yl)-(5H,11H-pyrrolo[2,1-
c] [1,4] benzodiazepin-10-yl)-methanone
2-Chloro-4-(trifluoromethyl)pyrimidine-5-carbonyl chloride (2.57 g) was
added gradually to an ice-cooled solution of 10,11-dihydro-5H-pyrrolo[2,1-c] [
1,4]-
benzodiazepine, (1.84 g) and diisopropylethylamine (1.37 g) in dichloromethane
(50
ml). After stirring at room temperature for 18 hours, the reaction mixture was
washed with water and saturated aqueous sodium bicarbonate. The
dichloromethane
solution was dried over anhydrous sodium sulfate and filtered through a short
column
of hydrous sodium magnesium silicate and further eluted with several volumes
of
dichloromethane. The combined organic phase was concentrated on a hot plate
with
the gradual addition of hexane until crystallization occurred. After cooling,
the
crystals were collected by filtration to yield the title compound (3.22 g),
m.p. 221-223
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°C.
EXAMPLE 20
[2-(3-Methyl-pyrazol-1-yl)-4-trifluoromethyl-pyrimidin-5-yl]-(SH,11H-
pyrrolo[2,1-c] [1,4]benzodiazepin-10-yl)-methanone
To 60% sodium hydride in oil (0.15 g, degreased with hexane) in
dimethylformamide (25 ml) was added 3-methylpyrazole (0.25 g). When the
hydrogen evolution subsided, (2-chloro-4-trifluoromethyl-pyrimidin-5-yl)-
(SH,11H-
pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.98 g) was added. The
reaction
mixture was heated for 18 hours in a sand bath (internal temperature 110
°C). The
mixture was then poured onto ice and further diluted with a saturated saline
solution.
The precipitate was filtered, redissolved in dichloromethane and dried over
anhydrous
sodium sulfate. Purification was aided by filtration through a short column of
hydrous sodium magnesium silicate and further elution with several volumes of
dichloromethane. The combined eluate was concentrated on a hot plate with the
gradual addition of hexane until crystallization occurred. After cooling, the
solid was
collected by filtration to yield the title compound (0.54 g) as colorless
crystals, m.p.
202-204 °C.
EXAMPLE 21
[2-(4-Methyl-pyrazol-1-yl)-4-trifluoromethyl-pyrimidin-5-yl]-(SH,11H-
pyrrolo[2,1-c] [1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 9's Method 1, employing (2-chloro-4-
trifluoromethyl-pyrimidin-5-yl)-(SH,11H-pyrrolo[2,l-c][1,4]benzodiazepin-10-
yl)-
methanone (0.98 g), 60% sodium hydride in oil (0.15 g), 4-methylpyrazole (0.42
g)
and dimethylformamide (25 ml), the title compound (0.73 g) was obtained as a
crystalline solid, m.p. 214-217 °C.
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EXAMPLE 22
1-[4-(SH,11H-Pyrrolo[2,1-c] [1,4]benzodiazepine-10-carbonyl)-phenyl]-ethanone
4-Acetylbenzoic acid (5.0 g) and thionyl chloride (10 ml) were heated on a
steam bath under argon for 0.75 hour, and the volatile material was removed
under
reduced pressure. Toluene was added and the volatiles were removed again to
give
the crude acid chloride as a red-orange oil. This compound tended to solidify
and
was used as such for further transformations.
The acid chloride (4.56 g) in dichloromethane (25 ml) was added portionwise
to an ice-cooled solution of 10,11-dihydro-5H-pyrrolo[2,1-c] [ 1,4]-
benzodiazepine
(3.68 g) and diisopropylethylamine (3.25 g) in dichloromethane (100 ml). After
stirring at room temperature for 18 hours, the reaction mixture was washed
with
water and saturated aqueous sodium bicarbonate. The dichloromethane solution
was
dried over anhydrous sodium sulfate and filtered through a short column of
hydrous
sodium magnesium silicate eluting with several volumes of dichloromethane. The
combined organic phase was concentrated on a hot plate with the gradual
addition of
hexane until crystallization occurred. After cooling, the crystals were
collected by
filtration to yield the title compound (1.75 g), m.p. 135-137 °C.
EXAMPLE 23
3-Dimethylamino-1-[4-(SH,11H-pyrrolo[2,1-c] [1,4] benzodiazepine-10-carbonyl)-
phenyl]-2-propen-1-one
A reaction mixture of 1-[4-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepine-10-
carbonyl)-phenyl]-ethanone (1.40 g), t-butoxy-bis-dimethylaminomethane (5.0
ml)
and dichloromethane (10 ml) was stirred for 18 hours. The red-orange
precipitate
was filtered to yield the title compound (1.22 g), m.p. 203-205 °C.
Additional
product (0.18 g) was isolated from the reaction mixture by concentration.
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EXAMPLE 24
[4-(1H-Pyrazol-3-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c] [1,4]benzodiazepin-
10-yl)-methanone
A reaction mixture of 3-dimethylamino-1-[4-(5H,11H-pyrrolo[2,1-
c][1,4]benzodiazepine-10-carbonyl)-phenyl]-2-prop-1-one (1.0 g), anhydrous
hydrazine (0.20 g), and glacial acetic acid (20 ml) was refluxed for 7 hours
and
evaporated to dryness. The crude residue was dissolved in dichloromethane,
washed
with a saturated aqueous sodium bicarbonate solution and dried over anhydrous
sodium sulfate. The solution was filtered through a short column of hydrous
sodium
magnesium silicate, eluting with several volumes of dichloromethane. The
combined
organic phase was concentrated on a hot plate with the gradual addition of
hexane
until crystallization occurred. After cooling, the crystals were collected by
filtration.
The column procedure was repeated to yield the title compound (0.65 g), m.p.
219-
221 °C.
EXAMPLE 25
[4-(1-Methyl-1H-pyrazolyl-3-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c] [1,4]-
benzodiazepin-10-yl)-methanone
To a mixture of 60% sodium hydride in oil (0.35 g, degreased with hexane)
and dimethylformamide (20 ml) was added [4-(1H-pyrazol-3-yl)-phenyl]-(5H,11H-
pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.98 g) followed in a few
minutes by iodomethane (0.50 g). The reaction mixture was stirred for 18 hours
at
room temperature and then poured into water and extracted with
dichloromethane.
After drying, the organic layer was filtered through a short column of hydrous
sodium
magnesium silicate, eluting with several volumes of dichloromethane. The
combined
organic phase was concentrated on a hot plate with the gradual addition of
hexane
until crystallization occurred. After cooling, the crystals were collected by
filtration
to yield the title compound (0.70 g), m.p. 194-195 °C.
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EXAMPLE 26
[4-(1-Ethyl-1H-pyrazol-3-yl)-phenyl]-(SH,11H-pyrrolo[2,1-c] [1,4]-
benzodiazepin-10-yl)-methanone
In the manner of Example 25, employing [4-(1H-pyrazol-3-yl)-phenyl]-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.0 g), 60% sodium
hydride in oil (0.27 g), dimethylformamide (25 ml), and ethyl iodide (0.87 g);
the title
compound (0.69 g) was obtained as a crystalline solid, m.p. 180-183 °C.
EXAMPLE 27
[4-(1-Propyl-1H-pyrazol-3-yl)-phenyl]-(SH,11H-pyrrolo[2,1-c][1,4]-
benzodiazepin-10-yl)-methanone
In the manner of Example 25, employing [4-(1H-pyrazol-3-yl)-phenyl]-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.98 g), 60% sodium
hydride in oil (0.30 g), dimethylformamide (25 ml), and 1-iodopropane (0.60
g), the
title compound (0.32 g) was obtained as a crystalline solid, m.p. 159-161
°C.
EXAMPLE 28
[4-(1-Butyl-1H-pyrazol-3-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c][1,4]-
benzodiazepin-10-yl)-methanone
In the manner of Example 25, employing [4-(1H-pyrazol-3-yl)-phenyl]-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.98 g),
60°Io sodium
hydride in oil (0.30 g), dimethylformamide (25 ml), and 1-iodobutane (0.60 g),
the
title compound (0.32 g) was obtained as a crystalline solid, m.p. 122-123
°C.
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EXAMPLE 29
[4-(1-methoxymethyl-1H-pyrazol-3-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c] [1,4]-
benzodiazepin-10-yl)-methanone
In the manner of Example 25, employing [4-(1H-pyrazol-3-yl)-phenyl]-
(5H,11H-pyrrolo[2,1-cJ[1,4]benzodiazepin-10-yl)-methanone (1.0 g), 60% sodium
hydride in oil (0.15 g), dimethylformamide (25 ml), and iodomethyl methyl
ether
(0.50 g), the title compound (0.26 g) was obtained as an amorphous solid, MS,
m/z:
399.2(M+H)+, 797.2 (2M+H)+.
EXAMPLE 30
1-{3-[4-(5H,11H-Pyrrolo[2,1-c] [1,4]benzodiazepine-10-carbonyl)-phenyl]-
pyrazol-1-yl}-ethanone
To a solution of [4-(1H-pyrazol-3-yl)-phenyl]-(5H,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone (0.50 g) in dry pyridine (10 ml) was
added
acetic anhydride (0.20 g). After stirring at room temperature for 18 hours the
reaction mixture was poured into water and the precipitate was collected by
filtration.
The crude product was dissolved in dichloromethane and dried over anhydrous
sodium sulfate. This solution was filtered through a short column of hydrous
sodium
magnesium silicate, eluting with several additional volumes of
dichloromethane. The
eluant was concentrated on a hot plate with the gradual addition of hexane
until
crystallization occurred. After cooling, the crystals were collected by
filtration to
yield the title compound (0.46 g), m.p. 192-194 °C.
EXAMPLE 31
1-{3-[4-(SH,11H-Pyrrolo[2,1-c] [1,4]benzodiazepine-10-carbonyl)-phenyl]-
pyrazol-1-yl}-propan-1-one
In the manner of Example 30, employing [4-(1H-pyrazol-3-yl)-phenyl]-
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(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.16 g) in dry
pyridine
(10 ml) and propionic anhydride (0.10 g), the title compound (0.17 g) was
obtained as
a crystalline solid, m.p. 150-152 °C.
EXAMPLE 32
[4-(1-Cyclopropanecarbonyl-1H-pyrazol-3-yl)-phenyl]-(SH,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone
To a solution of [4-(1H-pyrazol-3-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c][1,4]-
benzodiazepin-10-yl)-methanone (1.0 g) in dry pyridine (10 ml) was added
cyclopropanecarbonyl chloride (0.44 g). After stirring at room temperature for
18
hours the reaction mixture was poured into water and the precipitate was
collected by
filtration. The crude product was dissolved in dichloromethane and dried over
anhydrous sodium sulfate. This solution was filtered through a short column of
hydrous sodium magnesium silicate, eluting with several additional volumes of
dichloromethane. The eluant was concentrated on a hot plate with the gradual
addition of hexane until crystallization occurred. After cooling, the crystals
were
collected by filtration to yield the title compound (0.88 g) was obtained as a
crystalline solid, m.p. 197-199 °C.
EXAMPLE 33
1-{3-[4-(5H,11H-Pyrrolo[2,1-c][1,4]benzodiazepine-10-carbonyl)-phenyl]-
pyrazol-1-yl}-butan-1-one
In the manner of Example 32, employing [4-(1H-pyrazol-3-yl)-phenyl]-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.71 g) in dry
pyridine
(10 ml) and butyryl chloride (0.32 g), the title compound (0.54 g) was
obtained as a
solid, m.p. 105-110 °C; MS, m/z: 424 (M)+.
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EXAMPLE 34
(5H,11H-Pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-{4-[1-(thiophene-2-carbonyl)-
1H-pyrazoly-3-yl]phenyl}-methanone
In the manner of Example 32, employing [4-(1H-pyrazol-3-yl)-phenyl]-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.5 g) in dry
pyridine
(10 ml) and thiophene-2-carbonyl chloride (0.25 g), the title compound (0.41
g) was
obtained as a crystalline solid, m.p. 195-197 °C; MS, m/z: 464 (M)+.
EXAMPLE 35
{4-[1-(5-Fluoro-2-methyl-benzoyl)-1H-pyrazol-3-yl]-phenyl}-(5H,11H-
pyrrolo[2,1-c] [1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 32, employing [4-(1H-pyrazol-3-yl)-phenyl]-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.35 g) in dry
pyridine
( 10 ml) and 2-methyl-5-fluorobenzoyl chloride (0.22 g), the title compound
(0.11 g)
was obtained as an amorphous pale yellow solid, MS, m/z: 490 (M)+.
EXAMPLE 36
{4-[1-(2-Methyl-benzoyl)-1H-pyrazol-3-yl]-phenyl}-(5H,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 32, employing [4-(1H-pyrazol-3-yl)-phenyl]-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.71 g) in dry
pyridine
(20 ml) and o-toluyl chloride (0.39 g), the title compound (0.59 g) was
obtained as a
crystalline solid, m.p. 170-173 °C; MS, m/z: 472 (M)+.
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EXAMPLE 37
{4-[1-(2-Chloro-4-fluoro-benzoyl)-1H-pyrazol-3-yl]-phenyl}-(5H,11H-
pyrrolo[2,1-c] [1,4]benzodiazepin-10-yl}methanone
Portionwise, 2-chloro-4-fluorobenzoyl chloride (0.82 g) was added to a
solution of [4-(1H-pyrazol-3-yl)-phenyl]-(5H,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-
10-yl)-methanone (1.0 g) and diisopropylamine (0.55g) in dichloromethane (25
ml)
which was cooled in an ice bath. The reaction mixture was allowed to stir at
room
temperature overnight. The reaction mixture was washed with water and
saturated
sodium bicarbonate and dried over anhydrous sodium sulfate. The
dichloromethane
solution was passed through a short column of hydrous sodium magnesium
silicate,
eluting several additional volumes of dichloromethane. The eluant was
evaporated to
dryness to yield 1.06 g of the product as a solid, m.p. 150-157 °C; MS,
m/z: 510
(M)+.
EXAMPLE 38
{4-[1-(2,4-Dichloro-benzoyl)-1H-pyrazol-3-yl]-phenyl}-(5H,11H-pyrrolo[2,1-
c] [1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 32, employing [4-(1H-pyrazol-3-yl)-phenyl]-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.71 g) in dry
pyridine
(20 ml) and 2,4-dichlorobenzoyl chloride (0.52 g), the title compound (0.66 g)
was
obtained as a crystalline solid, m.p. 180-182 °C; MS, m/z: 528 (M)+.
EXAMPLE 39
2-(2,4-Dichloro-phenyl)-1-{3-[4-(SH,11H-pyrrolo[2,1-c] [1,4]benzodiazepine-10-
carbonyl)-phenyl]-pyrazol-1-yl}-ethanone
In the manner of Example 32, employing [4-(1H-pyrazol-3-yl)-phenyl]-
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(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.71 g) in dry
pyridine
(25 ml) and 2,4-dichlorophenylacetyl chloride (0.56 g), the title compound
(0.20 g)
was obtained as a crystalline solid, m.p. 130-140 °C, resolidifies,
m.p. 180-182 °C.
EXAMPLE 40
{4-[1-(Biphenyl-2-carbonyl)-1H-pyrazol-3-yl]-phenyl}-(5H,11H-pyrrolo[2,1-
c] [1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 32, employing [4-(1H-pyrazol-3-yl)-phenyl]-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.71 g) in dry
pyridine
(20 ml) and 2-biphenylcarbonyl chloride (0.65 g), the title compound (0.49 g)
was
obtained as an amorphous solid, MS, m/z: 534 (M)+.
EXAMPLE 41
{4-[1-(4'-Trifluoromethyl-biphenyl-2-carbonyl)-1H-pyrazol-3-yl]-phenyl}-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 32, employing [4-(1H-pyrazol-3-yl)-phenyl]-
(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (0.71 g) in dry
pyridine
(20 ml) and 4'-trifluoromethyl-2-biphenylcarbonyl chloride (0.71 g), the title
compound (0.59 g) was obtained as an amorphous solid, MS, m/z: 602 (M)+.
EXAMPLE 42
3-Dimethylamino-1-[4-(5H,11H-pyrrolo[2,1-c] [1,4]benzodiazepine-10-carbonyl)-
phenyl]-2-buten-1-one
A mixture of 1-[4-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepine-10-carbonyl)-
phenyl]-ethanone (2.0 g) and dimethylacetamide dimethylacetal (15 ml) was
refluxed
in an inert atmosphere for 15 hours and the volatiles were removed at reduced
pressure. The crude solid was dissolved in dichloromethane and filtered
through a
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short column of hydrous sodium magnesium silicate followed by several volumes
of
dichloromethane. The combined eluant was concentrated and hexane was gradually
added until crystallization occurred. The cooled solution was filtered to
recover the
title compound (1.03 g) as a crystalline solid, m.p. 183-185 °C.
EXAMPLE 43
[4-(5-Methyl-1H-pyrazol-3-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c][1,4]-
benzodiazepin-10-yl)-methanone
Anhydrous hydrazine (0.10 g) was added to a solution of 3-Dimethylamino-1-
[4-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepine-10-carbonyl)-phenyl]-2-buten-1-
one
(0.50 g) in glacial acetic acid (25 ml). The reaction mixture was refluxed for
18
hours and then concentrated under vacuum. The solid was extracted with
dichloromethane and washed with saturated aqueous sodium bicarbonate solution.
The dichloromethane solution was dried over anhydrous sodium sulfate and
filtered
through a short column of hydrous sodium magnesium silicate and further eluted
with
several volumes of dichloromethane. The combined organic phase was
concentrated
on a steam bath with the gradual addition of hexane to give an opaque
solution. After
cooling the amorphous solid was recovered by filtration to yield the product
(0.33 g),
MS, m/z: 368 (M)+.
EXAMPLE 44
4-(5H,11H-Pyrrolo[2,1-c][1,4]benzodiazepine-10-carbonyl)-benzonitrile
4-Cyanobenzoic acid (5.0 g) and thionyl chloride (5.0 ml) were heated on a
steam bath for one hour, and all of the volatiles were removed at reduced
pressure.
Hexane was added and the crude crystalline acid chloride (5.30 g) was
recovered by
filtration, and used without further purification.
To a reaction mixture of 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]-
benzodiazepine (3.68 g), dichloromethane (100 ml), and diisopropylethylamine
(2.80
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g) was added 4-cyanobenzoyl chloride (2.97 g). After remaining at room
temperature
for 18 hours, the reaction mixture was washed with water and saturated aqueous
sodium bicarbonate solution. The dichloromethane solution was dried over
anhydrous sodium sulfate and filtered through a short column of hydrous sodium
magnesium silicate and further eluted with several volumes of dichloromethane.
The
combined organic phase was concentrated on a hot plate with the gradual
addition of
hexane until crystallization occurred. After cooling, the crystals were
collected by
filtration to yield the title compound (5.05) g, m.p. 184-186 °C.
EXAMPLE 45
4-(SH,11H-Pyrrolo[2,1-c][1,4]benzodiazepine-10-carbonyl)-benzamide
4-(5H,11H-Pyrrolo[2,1-c][1,4]benzodiazepine-10-carbonyl)-benzonitrile (0.5
g) from Example 44 was added to concentrated sulfuric acid (5 ml) and the
mixture
was stirred for 18 hours at room temperature to yield a bright yellow
solution. The
solution was poured onto ice and made basic with the addition of concentrated
ammonium hydroxide. The resultant solid was filtered, dissolved in
dichloromethane, and filtered through a short column of hydrous sodium
magnesium
silicate and further eluted with several volumes of dichloromethane. The
combined
organic phase was concentrated on a hot plate with the gradual addition of
hexane
until crystallization occurred.. After cooling, the crystals were collected by
filtration
to yield the title compound (5.05 g), m.p. 226-228 °C.
EXAMPLE 46
N-(Dimethylaminomethylene)-4-(5H,11H-pyrrolo[2,1-c] [1,4]benzodiazepine-10-
carbonyl)-benzamide
A mixture of 4-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepine-10-carbonyl)-
benzamide (1.25 g) from Example 45 and dimethylformamide dimethylacetal (20
ml)
was refluxed for 4 hours and the volatiles removed in vacuo to give a solid.
The solid
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was dissolved in dichloromethane and filtered through a short column of
hydrous
sodium magnesium silicate and further eluted with several volumes of
dichloromethane. The combined organic phase was concentrated on a steam bath
with the gradual addition of hexane until crystallization occurred. After
cooling, the
crystals were collected by filtration to yield the title compound (1.40 g),
m.p. 232-234
°C.
EXAMPLE 47
N-(1-Dimethylaminoethylene)-4-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepine-10-
carbonyl)-benzamide
A mixture of 4-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepine-10-carbonyl)
benzamide (1.24 g) from Example 45 and dimethylacetamide dimethylacetal (5.0
ml)
was heated on a steam bath for 4 hours. On cooling for 18 hours a crystalline
solid
precipitated which was recovered by filtration. The solid was washed with
hexane to
yield the product (1.54 g), m.p. 210-212 °C; MS, m/z: 400 (M)+.
EXAMPLE 48
(SH,11H-Pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-[4-(2H-[1,2,4]triazol-3-yl)-
phenyl]-methanone
A mixture of N-(dimethylaminomethylene)-4-(5H,11H-pyrrolo[2,1-c][1,4]-
benzodiazepine-10-carbonyl)-benzamide (1.0 g) from Example 46, glacial acetic
acid
(15 ml), and anhydrous hydrazine (0.16 g) was refluxed for 15 hours and the
volatiles
removed in vacuo. Saturated aqueous sodium bicarbonate solution was added and
the
resultant solid was recovered by filtration. The solid was refluxed for 4
hours and the
volatiles removed in vacuo to give a solid. The solid was dissolved in
dichloromethane and filtered through a short column of hydrous sodium
magnesium
silicate and further eluted with several volumes of dichloromethane. The
combined
organic phase was concentrated on a steam bath with the gradual addition of
hexane
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until crystallization occurred. After cooling, the crystals were collected by
filtration
to yield the title compound (0.39 g), m.p. 225-227 °C; MS, m/z: 355
(M)+.
EXAMPLE 49
[4-(2-Methyl-2H-[1,2,4]triazol-3-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c] [1,4]-
benzodiazepin-10-yl)-methanone
In the same manner as Example 48, employing N-(dimethylaminomethylene)-
4-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepine-10-carbonyl)-benzamide (1.56 g)
from
Example 46 in glacial acetic acid (75 ml) and methylhydrazine (0.32 g), the
title
compound (0.10 g) was obtained as a solid, m.p. 155-158 °C; MS, m/z:
369 (M)+.
EXAMPLE 50
[4-(5-Methyl-2H-[1,2,4]triazol-3-yl)-phenyl]-(SH,11H-pyrrolo[2,1-c] [1,4]-
benzodiazepin-10-yl)-methanone
In the same manner as Example 48, employing N-(1-dimethylaminoethylene)-
4-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepine-10-carbonyl)-benzamide (1.00 g)
from
Example 47 in glacial acetic acid (75 ml) and anhydrous hydrazine (0.25 g),
the title
compound (0.20 g) was obtained as an amorphous solid, MS, m/z: 369 (M)+.
EXAMPLE 51
[4-(2,5-Dimethyl-2H-[1,2,4]triazol-3-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c]-
[1,4]benzodiazepin-10-yl)-methanone
In the manner of Example 48, employing N-(1-dimethylaminoethylene)-4-(5H,11H-
pyrrolo[2,1-c][1,4]benzodiazepine-10-carbonyl)-benzamide (1.18 g) from Example
47 in glacial acetic acid (75 ml) and methylhydrazine (0.30 g), the title
compound
(0.33 g) was obtained as a solid, m.p. 193-195 °C; MS, m/z: 383 (M)+.
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EXAMPLE 52
[4-(3-Methyl[1,2,4]oxadiazol-5-yl)-phenyl]-(5H,11H-pyrrolo-[2,1-c][1,4]-
benzodiazepin-10-yl)-methanone
A solution of N-(1-dimethylaminoethylene)-4-(SH,11H-pyrrolo[2,1-c][1,4]-
benzodiazepine-10-carbonyl)-benzamide (1.15 g) from Example 47 in glacial
acetic
acid (50 ml) containing hydroxylamine hydrochloride (0.40 g) and potassium
acetate
( 1.0 g) was refluxed for 2 hours. All volatiles were removed under reduced
pressure
and a saturated aqueous solution of sodium bicarbonate was added. The mixture
was
extracted with dichloromethane and the extracts were dried over anhydrous
sodium
sulfate. The solution was filtered through a short column of hydrous sodium
magnesium silicate and further eluted with several volumes of dichloromethane.
The
combined organic phase was concentrated on a steam bath with the gradual
addition
of hexane until crystallization occurred. After cooling, the crystals were
collected by
filtration to yield the title compound (0.38 g), m.p. 177-179 °C; MS,
m/z: 371.3
(M)+, 741.3 (2M)+.
EXAMPLE 53
1-Methyl-4-(4-methylphenyl)-1H-pyrazole
A mixture of 2-(4-methylphenyl)-malondialdehyde (3.05 g), absolute ethanol
(40 ml), and methylhydrazine (1.09 g) was stirred at room temperature for 18
hours
and the volatiles removed at room temperature. Water was added and the mixture
was extracted with dichloromethane. After drying over anhydrous sodium
sulfate, the
solution was filtered through a short column of hydrous sodium magnesium
silicate
and further eluted with several volumes of dichloromethane. The combined
organic
phase was concentrated on a steam bath with the gradual addition of hexane
until
crystallization occurred. After cooling, the crystals were collected by
filtration to
yield the title compound (2.91 g), m.p. 107-108 °C.
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EXAMPLE 54
4-(1-Methyl-1H-pyrazol-4-yl)-benzoic acid
A mixture of 1-methyl-4-(4-methylphenyl)-1H-pyrazole (1.70 g), potassium
permanganate (9.70 g), and 1 N sodium hydroxide (100 ml) was refluxed for 18
hours. The suspension was filtered through diatomaceous earth and cooled. The
aqueous solution was extracted with dichloromethane which was discarded. The
aqueous solution was acidified to pH 5.5. The resultant precipitate was
difficult to
filter and was thus extracted with dichloromethane. After evaporation of the
solvent,
the resulting solid was recrystallized from acetone to yield the title
compound (0.60
g), m.p. 274-275 °C; MS m/z: 202 (M)+.
EXAMPLE 55
[4-(1-Methyl-1H-pyrazol-4-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c] [1,4]-
benzodiazepin-10-yl)-methanone
Oxalyl chloride (0.30 g) was added to a suspension of 4-(1-methyl-1H-
pyrazol-4-yl)-benzoic acid (0.46 g) in dichloromethane (25 ml). Two drops of
dimethylformamide were added and the mixture was stirred for 18 hours at room
temperature. The resultant solution was evaporated to dryness to yield the
crude acid
chloride (0.57 g), which was utilized without further purification.
The acid chloride was added to a solution of 10,11-dihydro-SH-pyrrolo[2,1-
c][1,4]-benzodiazepine (0.37 g) and diisopropylethylamine (0.58 g) in
dichloromethane (50 ml). After 18 hours at room temperature, the reaction
mixture
was washed with water and saturated aqueous sodium bicarbonate. The
dichloromethane solution was dried over anhydrous sodium sulfate and filtered
through a short column of hydrous sodium magnesium silicate and further eluted
with
several volumes of dichloromethane. The combined organic phase was
concentrated
on a hot plate with the gradual addition of hexane until crystallization
occurred. After
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cooling, the crystals were collected by filtration to yield the title compound
(0.38 g),
m.p. 200-201 °C; MS m/z: 368 (M)+.
EXAMPLE 56
6-(1-Methyl-1H-pyrazol-4-yl)-pyridine-3-carboxylic acid
A suspension of 6-(1-formyl-2-hydroxyvinyl)pyridine-3-carboxylic acid (1.93
g) (Eastman Chemicals) in absolute ethanol (50 ml) and methylhydrazine (0.50
g)
was stirred for 18 hours at room temperature. The reaction mixture was
filtered to
give the product (1.30 g). The filtrate was evaporated to give a solid which
was
recrystallized from ethyl acetate to give an analytical sample of the title
compound
(0.55 g), m.p. 262-264 °C.
EXAMPLE 57
[6-(1-Methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-(5H,11H-pyrrolo[2,1-c][1,4]-
benzodiazepin-10-yl)-methanone
A suspension of 6-(1-methyl-1H-pyrazol-4-yl)pyridine-3-carboxylic acid
(0.48 g) in thionyl chloride (5.0 ml) was stirred at room temperature for 2
hours. The
volatile material was removed under reduced pressure to afford 6-(1-methyl-1H-
pyrazol-4-yl)pyridine-3-carbonyl chloride as a solid, which was utilized
without
further purification.
A solution of 10,11-dihydro-SH-pyrrolo[2,1-c][1,4]benzodiazepine (0.37 g)
and diisopropylethylamine (0.61 g) in dichloromethane (25 ml) was cooled to 0
~C
and a solution of 6-(1-methyl-1H-pyrazol-4-yl)pyridine-3-carbonyl chloride in
dichloromethane (25 ml) was added portionwise. After 18 hours at room
temperature, the reaction mixture was washed with water and a saturated
aqueous
sodium bicarbonate solution. The dichloromethane solution was dried over
anhydrous sodium sulfate and filtered through a short column of hydrous sodium
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magnesium silicate and further eluted with several volumes of dichloromethane.
The
combined organic phase was concentrated on a hot plate with the gradual
addition of
hexane until crystallization occurred. After cooling, the crystals were
collected by
filtration to yield the title compound (0.31 g), m.p. 173-175 °C; MS,
m/z: 370.3
(M+H)'.
EXAMPLE 58
[4-(Pyrazol-1-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c] [1,4]benzodiazepin-10-yl)-
methanone
To a suspension of 4-(pyrazol-1-yl)benzoic acid (1.56 g) in dichloromethane
(25 ml) was added oxalyl chloride (1.04 g) and one drop of dimethylformamide.
The
mixture was stirred at room temperature for 18 hours to yield a clear
solution. The
volatile material was removed under reduced pressure to afford 4-(pyrazol-1-
yl)benzoyl chloride as a pale yellow solid (1.58 g), which was utilized
without further
purification.
The 4-(pyrazol-1-yl)benzoyl chloride (0.75 g) was added to a solution of
10,11-dihydro-5H-pyrrolo[2,1-c][1,4]benzodiazepine (0.61 g) and
diisopropylethylamine (0.47 g) in dichloromethane (25 ml). After 18 hours at
room
temperature, the reaction mixture was washed with water and a saturated
aqueous
sodium bicarbonate solution. The dichloromethane solution was dried over
anhydrous sodium sulfate and filtered through a short column of hydrous sodium
magnesium silicate and further eluted with several volumes of dichloromethane.
The
combined organic phase was concentrated on a hot plate with the gradual
addition of
hexane until crystallization occurred. After cooling, the crystals were
collected by
filtration to yield the title compound (0.90 g), m.p. 179-181 °C.
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EXAMPLE 59
[4-(3-Methyl-pyrazol-1-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c] [1,4]-benzodiazepin-
10-yl)-methanone
To a suspension of 4-(3-methylpyrazol-1-yl)benzoic acid (1.84 g) in
dichloromethane (25 ml) was added oxalyl chloride (1.16 g) and one drop of
dimethylformamide. The mixture was stirred at room temperature for 18 hours
and
the volatile material was removed under reduced pressure. Dichloromethane was
added, the solution filtered, and the solvent evaporated under reduced
pressure to
yield 4-(3-methylpyrazol-1-yl)benzoyl chloride as a yellow oil (1.76 g), which
was
utilized without further purification.
The 4-(3-methylpyrazol-1-yl)benzoyl chloride was added to an ice-cooled
solution of 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]benzodiazepine (0.55 g) and
diisopropylethylamine (0.44 g) in dichloromethane (25 ml). After stirring at
room
temperature for 18 hours, the reaction mixture was washed with water and a
saturated
aqueous sodium bicarbonate solution. The dichloromethane solution was dried
over
anhydrous sodium sulfate and filtered through a short column of hydrous sodium
magnesium silicate and further eluted with several volumes of dichloromethane.
The
combined organic phase was concentrated on a hot plate with the gradual
addition of
hexane until crystallization occurred. After cooling, the title compound (0.90
g) was
obtained as an amorphous solid, MS, mJz: 369 (M+H)+.
EXAMPLE 60
[4-(4-Methyl-pyrazol-1-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c] [1,4]-benzodiazepin-
10-yl)-methanone
To a suspension of 4-(4-methylpyrazol-1-yl)benzoic acid (0.75 g) in
dichloromethane (15 ml) was added oxalyl chloride (0.50 g) and one drop of
dimethylformamide. The mixture was stirred at room temperature for 18 hours
and
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the volatile material was removed under reduced pressure. The residue was
dissolved
in hexane and filtered through diatomaceous earth. Evaporation of the solvent
in
vacuo yielded 4-(4-methylpyrazol-1-yl)benzoyl chloride (0.77 g), which was
used
without further purification.
The 4-(4-methylpyrazol-1-yl)benzoyl chloride (0.72 g) was added to an ice-
cooled solution of 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]benzodiazepine (0.60 g)
and
diisopropylethylamine (0.48 g) in dichloromethane (25 ml). After stirring at
room
temperature for 18 hours, the reaction mixture was washed with water and
saturated
aqueous sodium bicarbonate. The dichloromethane solution was dried over
anhydrous sodium sulfate and filtered through a short column of hydrous sodium
magnesium silicate and further eluted with several volumes of dichloromethane.
The
combined organic phase was concentrated on a hot plate with the gradual
addition of
hexane until crystallization occurred. After cooling, the crystals were
collected by
filtration to yield the title compound (0.75 g), m.p. 179-181 °C; MS
m/z: 369 (M+H)+.
EXAMPLE 61
[4-(3,5-Dimethyl-pyrazol-1-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c] [1,4]-
benzodiazepin-10-yl)-methanone
To a suspension of 4-(3,5-dimethylpyrazol-1-yl)benzoic acid (1.34 g) in
dichloromethane (25 ml) was added oxalyl chloride (1.0 g) and one drop of
dimethylformamide. The mixture was stirred at room temperature for 18 hours
and
the volatile material was removed under reduced pressure. The residue was
dissolved
in hexane and filtered through diatomaceous earth. Evaporation of the solvent
in
vacuo yielded 4-(3,5-dimethylpyrazol-1-yl)benzoyl chloride (0.80 g), which was
used
without further purification.
The 4-(3,5-dimethylpyrazol-1-yl)benzoyl chloride (0.75 g) was added to an
ice-cooled solution of 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]benzodiazepine
(0.55 g)
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and diisopropylethylamine (0.42 g) in dichloromethane (25 ml). After stirring
at room
temperature for 18 hours, the reaction mixture was washed with water and
saturated
aqueous sodium bicarbonate. The dichloromethane solution was dried over
anhydrous sodium sulfate and filtered through a short column of hydrous sodium
magnesium silicate and further eluted with several volumes of dichloromethane.
The
combined organic phase was concentrated on a hot plate with the gradual
addition of
hexane until crystallization occurred. After cooling, the title compound (0.79
g) was
obtained as an amorphous solid, MS m/z: 383 (M+H)+.
EXAMPLE 62
(5H,11H-Pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-[4-(3-trifluoromethyl-pyrazol-
1-yl)-phenyl]-methanone
A suspension of 4-(3-trifluoromethylpyrazolyl-1-yl)benzoic acid (1.45 g) in
thionyl chloride (5.0 ml) was heated at reflux for 3 hours. The volatile
material was
removed under reduced pressure, the residue dissolved in dichloromethane, and
filtered through diatomaceous earth. Evaporation of the solvent in vacuo
yielded 4-(3-
trifluoromethylpyrazol-1-yl)benzoyl chloride (1.45 g), which was used without
further purification.
To a solution of 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]-benzodiazepine (0.88
g) and diisopropylethylamine (0.66 g) in dichloromethane (50 ml) was added the
4
(3-trifluoromethylpyrazol-1-yl)benzoyl chloride (1.40 g). After stirnng at
room
temperature for 18 hours, the reaction mixture was washed with water and
saturated
aqueous sodium bicarbonate. The dichloromethane solution was dried over
anhydrous sodium sulfate and filtered through a short column of hydrous sodium
magnesium silicate and further eluted with several volumes of dichloromethane.
The
combined organic phase was concentrated on a hot plate with the gradual
addition of
hexane until crystallization occurred. After cooling, the crystals were
collected by
filtration to yield the title compound (1.70 g), m.p. ~166-167 °C; MS,
m/z: 423.3
(M+H)+, 845.4 (2M+H)+.
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EXAMPLE 63
[4-(Imidazol-1-yl)-phenyl]-(SH,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-
methanone
A suspension of 4-(imidazol-1-yl)benzoic acid (0.90 g) in thionyl chloride
(2.0 ml) was heated on a steam bath under argon for one hour. Evaporation of
the
volatile material under reduced pressure afforded a residue which crystallized
upon
the addition of hexane to yield the 4-(imidazol-1-yl)benzoyl chloride as the
hydrochloride salt (1.17 g), m.p. 242-247 °C.
To a solution of 10,11-dihydro-SH-pyrrolo[2,1-c][1,4]-benzodiazepine (0.75),
diisopropylethylamine (1.20 g), and 4-dimethylaminopyridine (0.1 g) in
dichloromethane (50 ml) was added 4-(imidazol-1-yl)benzoyl chloride
hydrochloride
(1.12 g). After stirnng at room temperature for 18 hours, the reaction mixture
was
washed with water and saturated aqueous sodium bicarbonate. The
dichloromethane
solution was dried over anhydrous sodium sulfate, filtered through a short
column of
hydrous sodium magnesium silicate, and further eluted with several volumes of
dichloromethane. The combined organic phase was concentrated on a hot plate
with
the gradual addition of hexane until crystallization occurred. After cooling,
the
crystals were collected by filtration to yield the title compound (0.57 g),
m.p. 171-172
°C; MS, m/z: 354 (M+H)'.
EXAMPLE 64
[4-(4-Methyl-imidazol-1-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c] [1,4]-benzodiazepin-
10-yl)-methanone
To a suspension of 4-(4-methylimidazol-1-yl)benzoic acid (0.80 g) in
dichloromethane (25 ml) was added oxalyl chloride (0.50 g) and one drop of
dimethylformamide. The mixture was stirred at room temperature for 18 hours
and
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the volatile material was removed under reduced pressure to yield 4-(4-
methylimidazol-1-yl)benzoyl chloride (1.02 g), which was utilized without
further
purification.
The 4-(4-methylimidazol-1-yl)benzoyl chloride (0.99 g) was added to an ice-
cooled solution of 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]benzodiazepine (0.64 g)
and
diisopropylethylamine (0.60 g) in dichloromethane (25 ml). After stirnng at
room
temperature for 18 hours, the reaction mixture was washed with water and
saturated
aqueous sodium bicarbonate. The dichloromethane solution was dried over
anhydrous sodium sulfate, filtered through a short column of hydrous sodium
magnesium silicate, and further eluted with several volumes of
dichloromethane. The
combined organic phase was concentrated on a hot plate with the gradual
addition of
hexane until crystallization occurred. After cooling, the title compound (0.52
g) was
obtained as a solid, m.p. 140-145 °C; MS, m/z: 369 (M+H)+.
EXAMPLE 65
4-Bromo-2-chloro-benzoic acid, methyl ester
Thionyl chloride (1.64 ml) was added dropwise to a suspension of 4-bromo-2-
chlorobenzoic acid (6.92 g) in methanol, and heated to 60 °C for 2
hours. The
solvent was removed in vacuo, the residue redissolved in ethyl acetate, and
washed
sequentially with 0.5 N sodium hydroxide (2x), water, and brine. The organic
phase
was dried over anhydrous sodium sulfate, and the solvent removed in vacuo to
afford
the title compound (7.8 g). 'H NMR (300 MHz), (DMSO-db) 8: 3.87 (s,3H), 7.68-
7.9
(m, 3H).
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EXAMPLE 66
2-Chloro-4-(3-dimethylamino-propyn-1-yl)benzoic acid, methyl ester
To a stirred solution of 4-bromo-2-chlorobenzoic acid, methyl ester (18.69 g)
in triethylamine (110 ml), was added 1-dimethylamino-2-propyne (12.1 ml),
bis(triphenylphosphine)palladium(II) chloride (1.26 g), and copper(I) iodide
(0.136
g). The mixture was heated slowly to 60 °C, and the temperature
maintained for one
hour. The reaction was cooled to room temperature, filtered through
diatomaceous
earth, and the collected solid washed with ethyl acetate. The solvent was
removed in
vacuo, the resulting residue redissolved in ethyl acetate, and washed with
water (3x).
The combined organic extract was dried over anhydrous sodium sulfate, and the
solvent removed in vacuo to give a crude product. The crude product was
purified by
column chromatography on silica gel (225 g), eluting with 40% ethyl
acetate/hexane.
After removing the solvent in vacuo, the title compound was obtained as a
viscous oil
(17.7 g), MS (+FAB), m/z: 252 (M+H)+.
EXAMPLE 67
2-Chloro-4-(3-dimethylamino-2-propen-1-on-1-yl)-benzoic acid, methyl ester
Gradually, 3-chloroperoxybenzoic acid (10.76 g) was added to a solution of 2-
chloro-4-(3-dimethylamino-propyn-lyl)-benzoic acid, methyl ester (15.07 g in
dichloromethane (40 ml), at a rate to maintain the reaction temperature at -20
~C.
The mixture was stirred for 10-15 minutes. The resulting N-oxide was purified
by
chromatography on Activity Grade I basic alumina (215 g), eluting with
10°Io
methanol/dichloromethane. The solvent was evaporated in vacuo between 12 to 18
°C. The resulting residue was dissolved in methanol (100 ml) and heated
at 60-65 °C
with stirnng for 18 hours. After removing the solvent in vacuo, and the
product was
purified by column chromatography on silica gel (190 g), eluting with 70%
ethyl
acetate/hexane. Trituration with diethyl ether containing some hexane afforded
the
title compound as a solid (5.68 g), m.p. 92-96 °C.
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EXAMPLE 68
2-Chloro-4-(1H-pyrazol-3-yl)-benzoic acid, methyl ester
To a suspension of 2-chloro-4-(3-dimethylamino-2-propen-1-on-1-yl)-benzoic
acid, methyl ester (13.67g) in ethanol (53 ml) was added hydrazine
monohydrochloride (7.0 g). The mixture was heated in an oil bath at 75-80
°C for
one hour. The solvent was removed in vacuo. The resulting residue was
dissolved in
ethyl acetate, washed with water and brine, dried over anhydrous sodium
sulfate, and
the solvent removed in vacuo to yield the title compound as a crude solid (12
g). A
purified sample had a melting point of 130-131 °C.
EXAMPLE 69
2-Chloro-4-(1-methyl-1H-pyrazol-3-yl)-benzoic acid, methyl ester
To a suspension of hexane washed sodium hydride (3.05 g, 60% dispersion) in
dimethylformamide (6 ml) under nitrogen was added a solution of 2-chloro-4-(1H-
pyrazol-3y1)-benzoic acid, methyl ester (12.0 g) in dimethylformamide (30 ml)
over a
period of 15 minutes. The mixture was stirred at room temperature for 30
minutes.
Iodomethane (9.5 ml) was added dropwise over 15 minutes. The mixture was
allowed to stir at room temperature for 45 minutes. Additional iodomethane
(5.16
ml) was added, and the reaction stirred another 75 minutes. The reaction was
diluted
with a small quantity of water, and concentrated in vacuo. The residue was
diluted
with water (500 ml) and extracted with a small quantity of ethyl acetate (5x).
The
combined organic phase was evaporated in vacuo to afford a crude product (
13.48 g).
The crude product was purified by column chromatography on silica gel (195 g)
eluting with 15% ethyl acetate/hexane to afford the pure 1-methyl regioisomer
(4.29
g), followed by a mixture of the 1-methyl and 2-methyl regioisomers (4.6 g).
The
mixture of isomers was triturated with hexane three times to give an
additional
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sample of the pure 1-methyl regioisomer (2.55 g), m.p. 66.5-67 °C; MS
(+FAB), m/z:
251 (M+H)+.
EXAMPLE 70
2-Chloro-4-(1-methyl-1H-pyrazol-3-yl)-benzoic acid
To a solution of 2-chloro-4-(1-methyl-1H-pyrazol-3-yl)-benzoic acid, methyl
ester (6.85 g) in methanol (32 ml) was added 2.5 N sodium hydroxide solution
(15.3
ml). The reaction was heated to 50 °C for one hour. The solvent was
removed in
vacuo, and the residue dissolved in water (250 ml), cooled in an ice bath, and
acidified with 2N hydrochloric acid (24 ml). The resulting precipitate was
filtered
and dried to give a colorless solid (6.3 g) m.p. 232-233 °C; MS (+FAB),
m/z: 236
(M+H)+.
EXAMPLE 71
[2-Chloro-4-(1-methyl-1H-pyrazol-3-yl)-phenyl]-(SH,11H-pyrrolo[2,1-c]-
[1,4]benzodiazepine-10-yl)-methanone
Well powdered 2-chloro-4-(1-methyl-1H-pyrazol-3-yl)-benzoic acid (6.3 g)
and dimethylformamide (2.16 ml) were suspended under nitrogen in a mixture of
tetrahydrofuran (70 ml) and dichlormethane (15 ml). A solution of oxalyl
chloride
(2.43 ml) in dichloromethane (5 ml) was added dropwise, and the reaction
stirred for
one hour. The resulting suspension of 2-chloro-4-(1-methyl-1H-pyrazol-3-yl)-
benzoyl
chloride was utilized without further purification.
To a suspension of 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]benzodiazepine (4.93
g) in dichloromethane (15 ml) was added diisopropylethylamine (7 ml). The
suspension of the freshly prepared acid chloride was gradually added over 15
minutes
under a positive flow of nitrogen. The slightly warm reaction mixture was
stirred
under nitrogen for 50 minutes. After stirnng one hour, the mixture was
concentrated
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in vacuo. The residue was dissolved in dichloromethane, washed with water, 5%
sodium bicarbonate, and water. The organic phase was washed with brine, dried
over
anhydrous sodium sulfate, and the solvent removed in vacuo to give a crude
product
(10.95 g). The crude product was purified by column chromatography on silica
gel
(200 g), loading the column with 25% ethyl acetate/hexane. Less polar
impurities
were eluted with 25-30% ethyl acetate/hexane. The product was eluted with 30-
40%
ethyl acetate/hexane to afford a pure sample (7.42 g); which, after seeding
with
crystals, was triturated with diethyl ether containing some hexane for 24
hours.
Filtration afforded the title compound as a crystalline solid (6.88 g), m.p.
148.5-150
~C; MS (EI), m/z: 402 (M)+.
EXAMPLE 72
2-chloro-4-(2-methyl-1H-pyrazol-3-yl)-benzoic acid, methyl ester
The title compound was prepared in the same manner as described in Example
68, employing methyl-2-chloro-4-(3-dimethylamino-2-propene-1-one)-benzoate
(0.8
g) and methylhydrazine (0.319 ml). The major 2-methyl regioisomer was isolated
by
column chromatography on silica gel, 'H NMR (300 MHz), (DMSO-db) b: 3.87 (s,
3H), 3.89 (s, 3H), 6.58 (d, 1H), 7.5 (d, 1H) 7.62-7.93 (m, 3H).
EXAMPLE 73
2-Chloro-4-(2-methyl-1H-pyrazol-3-yl)-benzoic acid
The title compound was prepared in the same manner as described in Example
70, employing 2-chloro-4-(2-methyl-1H-pyrazol-3yl)-benzoic acid, methyl ester
(0.464 g) and 2.5N sodium hydroxide (1.04 ml). 'H NMR (300 MHz), (DMSO-db) 8:
3.89 (s, 3H), 6.56 (d, 1H), 7.49 (d, 1H), 7.59-7.90 (m, 3H)
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EXAMPLE 74
[2-Chloro-4-(2-methyl-1H-pyrazol-3-yl)-pheny]-(5H,11H-pyrrolo[2,1-c]
[1,4]benzodiazepine-10-yl)-methanone
The title compound was prepared in the same manner as described in Example
71, Employing 2-chloro-4-(2-methyl-1H-pyrazol-3y1)-benzoic acid (3.98 g)
yielded
the corresponding acid chloride, and acylation with 10,11-dihydro-5H-
pyrrolo[2,1-
c][1,4]benzodiazepine (0.293 g) yielded the title compound as a foam, m.p. 78-
79
°C; MS (EI), m/z: 402 (M)+.
EXAMPLE 75
2-Chloro-4-cyanobenzoic acid, methyl ester
2-chloro-4-aminobenzoic acid, methyl ester (13.95 g) was suspended in a
mixture of water (65 ml) and concentrated hydrochloric acid (15.7 ml). After
stirring
at room temperature for 10 minutes, the suspension was cooled to 0 °C.
A solution of
sodium nitrite (5.71 g) in water (37 ml) was gradually added over 20 minutes,
maintaining a reaction temperature of 0 °C. After stirnng at 0
°C for 35 minutes, the
reaction mixture was partially neutralized by the addition of solid sodium
carbonate
(3.16 g) to afford a cold solution of the diazonium salt.
To a pre-cooled solution of copper(I) cyanide (8.4 g) and sodium cyanide
(9.19 g) in water (112 ml) was gradually added the above solution of diazonium
salt
over a 45-50 minute period. The diazonium salt solution was maintained at 0
°C
during the addition. The resulting mixture was stirred for 18 hours at room
temperature. A precipitate was filtered, air-dried, dissolved in ethyl acetate
(250 ml),
and filtered to remove insoluble matter. The organic phase was dried over
anhydrous
magnesium sulfate, and the solvent removed in vacuo to afford a crude product
as a
brown solid (13.2 g). The crude product was purified by column chromatography
on
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silica gel (250 g), eluting with 5-10% ethyl acetate/hexane to yield the title
compound
(10.9 g) as a solid, m.p. 90-92 °C; MS (EI), m/z: 195 (M)+.
EXAMPLE 76
2-Chloro-4-cyanobenzoic acid
To a stirred solution of 2-chloro-4-cyanobenzoic acid, methyl ester (24.3 g)
in
methanol (150 ml) was added 2.5N sodium hydroxide (54.5 ml). After stirring at
room temperature. for 45 minutes, the solvent was removed in vacuo. The
residue
was dissolved in water, cooled in an ice bath, and made acidic with 2N
hydrochloric
acid (14 ml). The resulting precipitate was filtered and dried in vacuo to
yield the title
compound as a solid (22.55g) m.p. 154-158 °C.
EXAMPLE 77
3-Chloro-4-(SH,11H-pyrrolo[2,1-c] [1,4]benzodiazepine-10-carbonyl)-
benzonitrile
To a cooled suspension of 2-chloro-4-cyanobenzoic acid (9.1 g) in a mixture
of dichloromethane (40 ml) and dimethylformamide (3.88 ml) was added dropwise
a
solution of oxalyl chloride (4.6 ml) in dichloromethane (10 ml) at 0
°C. The stirred
reaction was allowed to warm to room temperature over a one hour period. A
cloudy
solution of 2-chloro-4-cyanobenzoyl chloride was utilized without further
purification.
To a stirred suspension of 10,11-dihydro-5H-pyrrolo[2,1-
c][1,4]benzodiazepine (7.32 g) and diisopropylethylamine (13.6 ml) in
dichloromethane (35 ml) was added under nitrogen the cloudy solution of 2-
chloro-4-
cyanobenzoyl chloride. After one hour at room temperature, the mixture was
diluted
with dichloromethane and washed sequentially with water, 5% sodium
bicarbonate,
and 50% saturated brine, After drying over anhydrous sodium sulfate, the
solvent
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was removed in vacuo to afford a crude product ( 18.0 g). Purification by
column
chromatography on silica gel (250 g), eluting with 20% ethyl acetate/hexane,
followed by 25% ethyl acetate/hexane, yielded the title compound (13.56 g) as
a
straw yellow foam, MS (EI), m/z: 347 (M)+.
EXAMPLE 78
3-Chloro-4-(5H,11H-Pyrrolo[2,1-c] [1,4]benzodiazepine-10-carbonyl)-benzoic
acid
To a suspension of 3-chloro-4-(SH,11H-pyrrolo-[2,1-c][1.4]-benzodiazepine-
10-carbonyl)-benzonitrile (90.72 g) in ethanol was added 10 N sodium hydroxide
(1.02 ml) and the mixture heated under reflux for two hours. The solvent was
removed in vacuo, the residue dissolved in water, and made acidic with 2 N
hydrochloric acid (4.7 ml). The resulting precipitate was extracted with ethyl
acetate,
and the organic phase dried over anhydrous sodium sulfate. After removing the
solvent in vacuo, a foam was triturated with diethyl ether for 18 hours and
filtered to
give a crude product (0.69g). The crude product was purified by treatment with
activated charcoal in methanol. Crystallization from methanol/ether afforded
the title
compound as a purified solid (0.29 g), m.p. 198-199 °C; MS (EI), m/z:
366 (M)+.
EXAMPLE 79
3-Chloro-4-(5H,11H-pyrrolo[2,1-c] [1,4]benzodiazepine-10-carbonyl)-benzamide
Concentrated sulfuric acid (70 ml) was added to 3-chloro-4-(SH,11H-pyrrolo-
[2,1-c][1.4]benzodiazepine-10-carbonyl)-benzonitrile (12.85 g). The mixture
was
stirred at 60 °C for 3 hours, followed by stirring at room temperature
for 18 hours.
The reaction mixture was poured over ice and neutralized at 0 °C,
with 30%
ammonium hydroxide (184 ml). The resulting suspension was extracted with ethyl
acetate. The aqueous mixture was filtered, and reextracted with ethyl acetate.
The
combined organic phase was dried over anhydrous sodium sulfate, and the
solvent
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removed in vacuo. The residue was triturated with a mixture of diethyl ether
(50-60
ml) and a small quantity of ethyl acetate. Filtration of the precipitate
afforded the
title compound as a crystalline solid (10.44 g), m.p. 211-212 °C; MS
(EI), m/z: 365
(M)'.
EXAMPLE 80
N-(1-Dimethylaminoethylene)-3-chloro-4-(SH,11H-pyrrolo[2,1-c] [1,4]-
benzodiazepine-10-carbonyl)-benzamide
A suspension of 3-chloro-4-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepine-10-
carbonyl)-benzamide (5.48 g) and dimethylacetamide dimethyl acetal (10.97 ml)
was
heated at 90 °C for 20 minutes. The excess reagent was removed under
reduced
pressure, and the title compound utilized without further purification, MS
(EI), m/z:
434 (M)'.
EXAMPLE 81
[2-Choro-4-(5-methyl-2H-[1,2,4]triazol-3-yl)phenyl]-(SH,11H-pyrrolo[2,1-
c] [1,4]benzodiazepine-10-carbonyl)-methanone
To a solution of N-(1-dimethylaminoethylene)-3-chloro-4-(5H,11H-
pyrrolo[2,1-c][1,4]-benzodiazepine-10-carbonyl)-benzamide (3.01 g) in acetic
acid (4
ml) was added a solution of anhydrous hydrazine (0.435 ml) in acetic acid (4
ml).
The reaction mixture was stirred at between 85-90 °C for 45
minutes. After
removing the acetic acid in vacuo, the reaction mixture was diluted with water
(35-40
ml), neutralized to pH 7.0 with aqueous sodium bicarbonate, and extracted with
ethyl
acetate. The organic extract was washed with brine, dried over anhydrous
sodium
sulfate, and the solvent removed in vacuo to afford a crude product (2.68 g).
Purification of the crude product by column chromatography on silica gel (45
g),
eluting with 70°Io ethyl acetate/hexane, afforded a purified product
(2.5g), which,
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after trituration with diethyl ether, yielded the title compound as a solid (2
g), m.p.
211-212 °C; MS (EI), m/z: 403 (M)'.
EXAMPLE 82
N-(Dimethylaminomethylene)-3-chloro-4-(5H,11H-pyrrolo[2,1-c][1,4]
benzodiazepine-10-carbonyl)-benzamide
The title compound was prepared in the same manner as described in Example
80, employing 3-chloro-4-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepine-10-
carbonyl)-
benzamide (1.83 g) and dimethylformamide dimethylacetal (5.3 ml), MS (EI),
m/z::420 (M)'.
EXAMPLE 83
[2-Chloro-4-(2H-1,2,4-triazol-3-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c][1,4]-
benzodiazepin-10-yl)-methanone
The title compound was prepared in the same manner as described in Example
81, employing N-(dimethylaminomethylene)-3-chloro-4-(SH,11H-pyrrolo[2,1-
c[1,4]benzo-diazepine-10-carbonyl)-benzamide (2.53 g) and hydrazine (0.38 ml),
m.p. 174-177 °C; MS (EI), m/z: 389 (M)+.
EXAMPLE 84
[2-Chloro-4-(2-methyl-2H-[1,2,4]triazol-3-yl)-phenyl]-(5H,11H-pyrrolo [2,1-
c][1,4]benzodiazepin-10-carbonyl)-methanone
The title compound was prepared in the same manner as described in Example
48, using N-(dimethylaminomethylene)-3-chloro-4-(5H,11H-pyrrolo[2,1-
c[1,4]benzodiazepin-10-carbonyl)-benzamide (0.572 g), and methylhydrazine
(0.149
ml). m.p. 141-143°C. MS (EI): 403 (M)+.
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EXAMPLE 85
4-[(2,5-Dimethyl-2H-[1,2,4]triazol-3-yl)-2-chloro-phenyl]-(5H,11H-pyrrolo[2,1-
c] [1,4]benzodiazepin-10-carbonyl)-methanone
The title compound was prepared in the same manner as described in Example
48, using N-(1-dimethylaminoethylene)-3-chloro-4-(5H,11H-pyrrolo[2,1-c][1,4]-
benzodiazepin-10-carbonyl)-benzamide (0.51 g) and methylhydrazine (0.125 ml).
m.p. 197-199°C. MS (EI): 417 (M)+.
EXAMPLE 86
[2-Chloro-4-(1H-tetrazol-5-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c][1,4]
benzodiazepin-10-yl)- methanone
To a solution of 3-chloro-4-(5H,11H-pyrrolo[2,1-c][1,4]-benzodiazepine-10-
carbonyl)-benzonitrile (0.348 g) in dimethylformamide (2 ml) was added sodium
azide (0.078 g) and ammonium chloride (0.065 g). The mixture was heated to
100°C
for 18 hours.
Most of the dimethylformamide was removed in vacuo. The residue dissolved
in water (approximately 8 ml) and basified to pH 9.0 with 2.5N sodium
hydroxide
(0.6 ml) and extracted with ethyl acetate. The aqueous extract was acidified
with 2N
hydrochloric acid (1.1 ml), reextracted with ethyl acetate, dried over
anhydrous
sodium sulfate and the solvent removed in vacuo to give crude product (0.350g)
as an
oil. The oily product was triturated with diethyl ether, filtered through acid
treated
silica gel, and eluted with 40% ethyl acetate/hexane to give purer sample.
This was
further triturated with diethyl ether, and filtered to give a sample (0.88g)
m.p. 218-
220°C. MS (+FAB) 391 (M+H)+.
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EXAMPLE 87
[2-Chloro-4-(3-methyl-pyrazol-1-yl)-phenyl]-(3-dimethylaminomethyl-5H,11H-
pyrrolo[2,1-c] [1,4]benzodiazepin-10-yl)-methanone
To a stirred solution of [2-chloro-4-(3-methyl-pyrazol-1-yl)-phenyl]-(5H,11H-
pyrrolo-[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.61 g), N, N, N', N',
tetramethyldiamino-methane (0.82 g), and glacial acetic acid (0.48 g) in
methanol (25
ml) was added a solution of 37°Io aqueous formaldehyde (4 ml). The
mixture was
warmed to 40 °C for 10 minutes. After stirring one hour at room
temperature, the
reaction was concentrated in vacuo, redissolved in dichloromethane, and
extracted
sequentially with aqueous sodium bicarbonate and water (4x). The organic phase
was
dried over anhydrous sodium sulfate, and filtered through a plug of silica
gel, eluted
with ethyl acetate. Evaporation of the solvent in vacuo afforded an oil, which
on
trituration with hexane yielded 0.36 g of the title compound as a colorless
powder,
m.p. 100-102 °C; MS (+FAB), m/z: 482 (M+Na)+, 460 (M+H)'.
EXAMPLE 88
(3-Bromo-5H,11H-pyrrolo[2,1-c] [1,4]benzodiazepin-10-yl)-
[2-chloro-4-(3-methyl-pyrazol-1-yl)-phenyl]-methanone
To a stirred pre-cooled solution of [2-chloro-4-(3-methyl-pyrazol-1-yl)-
phenyl]-(5H, 11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.61 g) in
dichloromethane (25 ml) was added solid N-bromosuccinimide (0.712 g) over 10
minutes at -78 °C. The reaction was allowed to warm to -40 °C
over thirty minutes.
The mixture was diluted with dichloromethane, and extracted sequentially with
saturated aqueous sodium bicarbonate (2 x 100 ml) and water (100 ml). The
organic
phase was dried over anhydrous sodium sulfate, filtered through a plug of
silica gel,
and evaporated in vacuo to a residue. Crystallization from diethyl ether
yielded 1.47
g of the title compound as a colorless solid, m.p. 148-149 °C (dec); MS
(EI), m/z:
480 (M)+..
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EXAMPLE 89
(4-Bromo-2-chlorophenyl)-(SH,11H-pyrrolo[2,1-c] [1,4]benzodiazepin-10-yl)-
methanone
Dimethylformamide (1 drop) was added to a solution of 4-bromo-2-
chlorobenzoic acid (2.20 g) in anhydrous tetrahydrofuran (20 ml). Oxalyl
chloride
(1.46 g) was added and the mixture was warmed to reflux. The resultant
solution was
cooled to ambient temperature before being evaporated to dryness to give crude
4-
bromo-2-chlorobenzoyl chloride as a gold viscous liquid, which was used
without
further purification.
To a mixture of 10,11-dihydro-SH-pyrrolo[2,1-c][1,4]-benzodiazepine (1.44
g) and triethylamine (0.95 g) in dichloromethane (40 ml), cooled in an ice
bath, was
added dropwise a solution of 4-bromo-2-chlorobenzoyl chloride (2.42 g) in
dichloromethane (20 ml). The cooling bath was removed and after stirring for
22
hours, the reaction mixture was washed sequentially with water, saturated
aqueous
sodium bicarbonate, 0.5 N hydrochloric acid and water. The dichloromethane
solution was dried over anhydrous sodium sulfate, filtered, then evaporated in
vacuo
to dryness to yield an off-white foam. Purification by flash chromatography on
silica
gel eluting with hexane-ethyl acetate (2:1) resulted in a white foam (3.02 g),
m.p. 77-
80 °C, MS m/z: 400 (M)+.
EXAMPLE 90
[2-Bromo-4-(3-methyl-pyrazol-1-yl)-phenyl]-(5H,11H)-pyrrolo[2,1-c]
[1,4]benzodiazepin-10-yl)-methanone
Step a) 4-Fluoro-2-bromobenzoyl chloride: Dimethylformamide (2 drops)
was added to a solution of 4-fluoro-2-bromobenzoic acid (4.91 g) in anhydrous
tetrahydrofuran (55 ml). Oxalyl chloride (3.41 g) was added and the mixture
was
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warmed to reflux. The resultant solution was cooled to room temperature,
evaporated
in vacuo to give the crude acid chloride as a gold viscous liquid, which was
used
without further purification.
Step b) (4-Fluoro-2-bromophenyl)-(5H,11H-pyrrolo[2,1-c][l,4Jbenzo-
diazepin-10-yl)-methanone: A solution of 4-fluoro-2-bromobenzoyl chloride
(5.32 g)
from step a), in dichloromethane (35 ml), was added dropwise to a solution of
10,11-
dihydro-5H-pyrrolo[2,1-c][1,4]-benzodiazepine (3.44 g) and triethylamine (2.27
g) in
dichloromethane (80 ml) and cooled in an ice bath. The cooling bath was
removed
and after stirnng for 16 hours, the reaction mixture was washed sequentially
with
water, saturated aqueous sodium bicarbonate and saturated aqueous sodium
chloride.
The dichloromethane solution was dried over anhydrous magnesium sulfate,
filtered
and evaporated in vacuo to give a pale purple foam. Purification by flash
chromatography on silica gel eluting with hexane-ethyl acetate (1:1) resulted
in the
intermediate (4-fluoro-2-bromophenyl)-(5H,11H-pyrrolo[2,1-c] [
1,4]benzodiazepin-
10-yl)-methanone as a tan foam (6.91 g), MS m/z: 384 (M)+. This material was
used
without further purification in the next step.
Step c) [2-Bromo-4-(3-methyl-pyrazol-1-yl)-phenyl]-(5H,11H)-pyrrolo[2,1-
c] [1,4]benzodiazepin-10-yl)-methanone: A dispersion of 60% sodium hydride in
oil
(0.20 g) was washed with hexane, and then suspended in dimethylformamide ( 15
ml).
To this suspension was added 3-methylpyrazole (0.41 g). When hydrogen gas
evolution subsided, (4-fluoro-2-bromophenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzo-
diazepin-10-yl)-methanone (1.74 g) from step b) was added. The reaction
mixture
was heated to 130 °C for 6 hours. After the reaction mixture was cooled
to room
temperature, poured into a 50% saturated aqueous sodium chloride solution and
extracted with ethyl acetate. The ethyl acetate solution was dried over
anhydrous
magnesium sulfate, filtered and evaporated in vacuo to afford a brown oil.
Purification by flash chromatography on silica gel eluting with hexane-ethyl
acetate
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(1:1) gave a colorless solid (0.75 g). Recrystallization from methanol gave an
off-
white crystalline solid (0.53 g), m.p. 141-142.5°C, MS m/z: 446 (M)'.
EXAMPLE 91
(2,4-Ditluoro-phenyl)-(5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-
methanone
Step a) 2,4-Difluorobenzoyl chloride: A suspension of 2,4-difluorobenzoic
acid (3.6 g) containing a few drops of dimethylformamide in dichloromethane
(40
ml) was treated dropwise under nitrogen with oxalyl chloride (2.4 ml). After
gas
evolution subsided, the reaction mixture was refluxed for an additional 15
minutes.
The solution was evaporated to dryness in vacuo and the residue was utilized
without
further purification.
Step b) (2,4-Difluorophenyl)-(SH,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-
yl)-methanone: To a solution of the crude 2,4-difluorobenzoyl chloride acid
chloride
of Step a in dichloromethane under nitrogen was added solid 10,11-dihydro-SH-
pyrrolo[2,1-c][1,4]benzodiazepine amine (2.0 g) and diisopropylethylamine (3.4
ml).
The reaction mixture turned yellow-orange. After stirnng at room temperature
for 10
the reaction mixture was washed with water, 1 N hydrochloric acid, 1 N sodium
hydroxide and brine. The organic phase was dried over anhydrous sodium sulfate
and
evaporated to dryness to give a brown solid. The crude product was purified by
column chromatography on silica gel (Merck-60) with 20% ethyl acetate -hexane
to
provide 2.9 g of the title compound as a white foam.
MS (EI, m/z): 324 (M)+.
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EXAMPLE 92
[2-Fluoro-4-(3-methyl-pyrazol-1-yl)-phenyl]-(5H,11H-pyrrolo[2,1-c][1,4]-
benzodiazepin-10-yl)-methanone
A suspension of hexane washed 60% sodium hydride (0.31 g) in dry
dimethylformamide was treated dropwise with 3-methylpyrazole (0.62 ml) under
nitrogen at room temperature. Stirring was continued until the gas evolution
subsided
(10 minutes). In one portion (2,4-difluorophenyl)-(SH,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone (2.5 g) from step b) of Example 91 was
added and stirring was continued until a clear solution was attained. The
mixture was
heated in a preheated oil bath at 130°C for one hour. After cooling,
the mixture was
partitioned between water and ethyl acetate. The organic extracts were dried
over
anhydrous sodium sulfate and evaporated to dryness. The crude product was
purified
by flash column chromatography on silica gel (Merck-60) eluting with 20% ethyl
acetate -hexane to yield 0.82 g of the title product as a foam which was
crystallized
by sonication from ethanol/hexane, m.p. 192-193 °C. MS (EI) m/z: 386
(M)+.
EXAMPLE 93
Methyl 4-(3-methyl-pyrazol-1-yl)-2-trifluoromethyl-benzoate
Step a) Methyl 4-fluoro-2-trifluoromethylbenzoate: A suspension of 4-
fluoro-2-trifluoromethylbenzoic acid (25.6 g) and a few drops of
~dimethylformamide
in dichloromethane (250 ml) was treated dropwise under nitrogen with oxalyl
chloride (11.3 ml). After gas evolution subsided, the reaction mixture was
refluxed
for an additional 15 minutes. The reaction was cooled and methanol (50 ml) was
added. After stirnng for 2 hours, the reaction was concentrated in vacuo, and
the
residue was partitioned between dichloromethane and water. The organic phase
was
washed with saturated sodium bicarbonate, dried over anhydrous sodium sulfate,
and
evaporated to dryness to give 18.0 g of the title compound as a golden oil.
MS, (EI)
m/z: 222 (M)+ .
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The aqueous layer was acidified with 2 N hydrochloric acid to give a colorless
solid which was collected by filtration to give 7.5 g of the starting benzoic
acid.
Step b) Methyl 4-(3-methyl-pyrazol-1-yl)-2-trifluoromethyl-benzoate: A
suspension of hexane washed 60% sodium hydride (3.85 g) in dry
dimethylformamide (150 ml) was treated with the dropwise addition of a
solution of
3-methylpyrazole (7.75 ml) in dimethylformamide (50 ml) under nitrogen at room
temperature. Stirring was continued until the gas evolution subsided (10
minutes). A
solution of methyl 4-fluoro-2-trifluoromethylbenzoate (17.8 g) from step a) in
dimethylformamide (50 ml) was added dropwise to the clear solution. After
stirring
for 30 min. at room temperature the reaction was quenched with saturated
ammonium
chloride and extracted with ethyl acetate. The organic extracts (3x) were
dried over
anhydrous sodium sulfate and evaporated to dryness. The crude product was
purified
by flash column chromatography on silica gel (Merck 60) with a dichloromethane-
hexane gradient (50°Io - 75%) to give 13.6 g of the title product as a
colorless solid.
m.p. 59-61°C MS (EI, m/z): 284 (M)+.
EXAMPLE 94
4-(3-Methyl-pyrazol-1-yl)-2-trifluoromethyl-benzoic acid
Methyl 4-(3-methyl-pyrazol-1-yl)-2-trifluoromethyl-benzoate (1.19 g) from
Example 93, step b) was dissolved in methanol (10 ml) and a solution of 2.5 N
sodium hydroxide (3.3 ml) was added. The reaction was heated at reflux for 90
minutes, cooled to room temperature and concentrated in vacuo to dryness. The
residue was partitioned between ethyl acetate and 1 N hydrochloric acid. The
combined organic extracts were dried over anhydrous sodium sulfate and
concentrated in vacuo to give 1.14 g of the title compound as a colorless
solid. MS
(FAB) m/z: 271 (M+H)+ .
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EXAMPLE 95
[4-(3-Methylpyrazol-1-yl)-2-trifluoromethylphenyl]-(SH,11H-pyrazolo[5,1-c]-
[1,4]benzodiazepin-10-yl)-methanone
A solution of 4-(3-methyl-pyrazol-1-yl)-2-trifluoromethylbenzoic (0.26g)
from Example 94 in tetrahydrofuran (5 ml) was treated with dimethylformamide
(0.020 ml) followed by oxalyl chloride (0.090 ml). The solution was stirred at
room
temperature until gas evolution stopped and then the solution was warmed to
reflux
for 10 minutes. The sample was cooled to room temperature, concentrated to a
solid
and the solid was dissolved in tetrahydrofuran (25 mL). this solution was
added to a
solution (5H-10,11-dihydropyrazolo [5,1-c][1,4]benzodiazepine (0.143 g) and
triethylamine (0.150 ml) in tetrahydrofuran (20 ml). The solution was stirred
overnight at room temperature. A precipitate formed. The sample was diluted
with
dichloromethane to dissolve the precipitate and then the sample was
concentrated in
vacuo to about 1/3 of the original volume. The sample was partitioned between
dichloromethane and saturated aqueous ammonium chloride. The sample was
extracted with dichloromethane and the organic layers were pooled, dried over
anhydrous sodium sulfate, filtered and concentrated to an oil. The oil was
flash
chromatographed on silica gel using a gradient of 40% ethyl acetate/hexanes to
100°Io
ethyl acetate affording the title compound as a foam (0.30 g). A portion of
this
material was recrystallized from acetone/ hexanes to give heavy plates m.p.
100-
102°C, MS m/z: 437 (M)+.
EXAMPLE 96
2-Chloro-4-(3-methyl-1H-pyrazol-1-yl)-benzoic acid methyl ester and 2-chloro-
4-(5-methyl-1H-pyrazol-1-yl)-benzoic acid methyl ester
A suspension of hexane washed potassium hydride (0.424 g) in
dimethylformamide (5 ml) was treated in one portion with 3-methyl pyrazole
(0.85
ml) while stirnng. After the gas evolution ceased, 2-chloro-4-fluorobenzoic
acid
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methyl ester (2.0 g, 10.6) was added to the clear solution and heated at
130°C for 15
minutes. The reaction mixture was cooled to room temperature and partitioned
between ethyl acetate and brine. The organic phase was washed with water,
brine,
and dried over anhydrous sodium sulfate. Removal of solvent in vacuo afforded
2.2
g of a yellow oil. (Note: 20% hydrolysis of the ester was detected by analysis
of the
NMR spectrum of the crude product). The desired regioisomer 2-chloro-4-(3-
methyl-
1H-pyrazol-1-yl)-benzoic acid methyl ester was isolated from the other isomer
(described below) by flash column chromatography on silica gel (Merck 60)
eluting
with dichloromethane-hexane 2:1) to give 1.55 g of the title compound as a
colorless
solid. MS (EI m/z: 250/252 (M)+.
The 5-regioisomer, namely 2-chloro-4-(5-methyl-1H-pyrazol-1-yl)-benzoic
acid methyl ester was isolated from the above flash column chromatography on
silica
gel (Merck 60) by further eluting with dichloromethane-hexane 2:1 to give 0.20
g of
the product as a colorless solid. MS (EI), m/z: 250/252 (M)+.
EXAMPLE 97
2-Chloro-4-(3-methyl-1H-pyrazol-1-yl)-benzoic acid
A solution of 2-chloro-4-(3-methyl-1H-pyrazol-1-yl)-benzoic acid methyl
ester (1.42 g) from Example 96 and 6 ml of 1 M aqueous lithium hydroxide in
tetrahydrofuran (20 ml) was stirred for 18 hours at room temperature. The
reaction
mixture was partitioned between ethyl acetate and 1 N hydrochloric acid. The
organic layer was washed with water, brine and was dried over anhydrous sodium
sulfate. Evaporation of the solvent in vacuo afforded 1.05 g of the title
compound
as a colorless solid. m.p. 192-193°C. MS (EI), m/z: 236/238 (M)+.
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EXAMPLE 98
(2,6-Dichloropyridin-3-yl)(5H,11H-Pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-
methanone
A solution of 2,6-dichloronicotinic acid (3.84 g), oxalyl chloride (2.0 g),
and
1 drop of dimethylformamide in dichloromethane (25 ml), was stirred at room
temperature for 18 hours. The solution was concentrated in vacuo to give 3.50
g of
2,6-dichloronicotinyl chloride which was added portionwise in dichloromethane
(25
ml) to an ice cooled solution of 10,11-dihydro-SH-pyrrolo[2,1-c] [ 1,4]-
benzodiazepine
(2.15 g) and diisopropylethylamine (2.03 g) in dichloromethane (50 ml). The
mixture was stirred at room temperature for 18 hours and washed with saturated
aqueous sodium bicarbonate. The organic phase was dried over anhydrous sodium
sulfate and filtered through a short column of anhydrous sodium magnesium
silicate.
The combined organic phase was concentrated on a hot plate with the gradual
addition of hexane until crystallization occurred. After cooling, the crystals
were
collected by filtration to yield 2.65 g of the title as an amorphous solid.
m.p. 115-130
°C. MS, m/z: 358.1 (M+H)+.
EXAMPLE 99
(2-Chloro-6-pyrazol-1-yl-pyridin-3-yl)-(5H,11H-pyrrolo[2,1-c][1,4]
benzodiazepin-10-yl)-methanone
To a suspension of 60% sodium hydride in oil (0.1 g) in dimethylformamide
(25 ml) was added dropwise pyrazole (0.15 g). After hydrogen gas evolution
ceased,
(2,6-dichloropyridin-3-yl)(5H,11H-pyrrolo[2,1-c] [ 1,4]benzodiazepin-10-yl)-
methanone (0.67 g) was added and the reaction mixture was heated in a sand
bath at
110 °C for 18 hours. The mixture was poured onto ice, diluted with
brine, and
extracted with dichloromethane. The combined extracts were dried over
anhydrous
sodium sulfate and filtered through a short column of anhydrous sodium
magnesium
silicate. The solution was concentrated in vacuo and triturated with diethyl
ether to
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give 0.18 g of the title compound as a colorless solid, m.p. 133-135
°C. MS m/z:
390.8 (M+H)+, 779.1 (2M+H)+.
EXAMPLE 100
[2-Chloro-6-(3-methylpyrazol-1-yl)-pyridin-3-yl]-(5H,11H-pyrrolo[2,1-c]
[1,4]benzodiazepin-10-yl)-methanone
To a suspension of 60% sodium hydride in oil (0.1 g) in dimethylfonnamide
(25 ml) was added dropwise 3-methylpyrazole (0.15 g). After hydrogen gas
evolution ceased, (2,6-dichloropyridin-3-yl)(SH,11H-pyrrolo[2,1-c][1,4]benzo-
diazepin-10-yl)-methanone (0.67 g) was added and the reaction mixture was
heated in
a sand bath at 110 °C) for 18 hours. The mixture was poured onto ice,
diluted with
brine, and extracted with dichloromethane. The combined extracts were fined
over
anhydrous sodium sulfate and filtered through a short column of anhydrous
sodium
magnesium silicate. The crude product was purified by preparative hplc
(Dynamax
c60 silica cartridge) eluting with 40% ethyl acetate in hexanes to give 0.21 g
of
colorless crystals, m.p. 171-172 °C. MS, m/z: 404.2 (M+H)+, 807.1
(2M+H)+.
EXAMPLE 101
[2-Chloro-6-(4-methylpyrazol-1-yl)-pyridin-3-yl](5H,11H- pyrrolo[2,1-c]
[1,4]benzodiazepin-10-yl)-methanone
To a suspension of 60% sodium hydride in oil (0.1 g) in dimethylfonnamide
(25 ml) was added dropwise 3-methylpyrazole (0.45 g). After hydrogen gas
evolution ceased, (2,6-dichloropyridin-3-yl)(SH,11H-pyrrolo[2,1-c][1,4]benzo-
diazepin-10-yl)-methanone, (1.79 g) was added and the reaction mixture was
heated
in a sand bath at 110 °C for 18 hours. The mixture was poured onto ice,
diluted with
brine, and extracted with dichloromethane. The combined extracts were fined
over
anhydrous sodium sulfate and filtered through a short column of anhydrous
sodium
magnesium silicate. The crude product was purified by preparative hplc
(Dynamax
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c60 silica cartridge) eluting with 40% ethyl acetate in hexanes to give 0.26 g
of
colorless crystals, m.p. 155-156 °C, MS, m/z: 404.2 (M+H)+, 807.0
(2M+H)+.
EXAMPLE 102
[2-Chloro-4-(3-methyl-1,2,4-triazol-1-yl)-phenyl](5H,11H- pyrrolo[2,1-c]-
[1,4]benzodiazepin-10-yl)-methanone
To a suspension of 60% sodium hydride in oil (0.3 g) in dimethylformamide
(50 ml) was added dropwise 3-methyl-1,2,4-triazole (0.45 g). After hydrogen
gas
evolution ceased, 2-chloro-4-fluorophenyl-(5H,11H-pyrrolo[2,1-c][1,4]benzo-
diazepine-10-yl)-methanone (1.70 g) was added and the reaction mixture was
heated
in a sand bath at 110 °C for 18 hours. The mixture was poured onto ice,
diluted with
brine, and extracted with dichloromethane. The combined extracts were dried
over
anhydrous sodium sulfate and filtered through a short column of anhydrous
sodium
magnesium silicate. The solution was concentrated in vacuo and the residue was
triturated with diethyl ether to give 1.25 g of the title compound as
colorless crystals,
m.p. 191-193 °C, MS m/z: 404.1 (M+H)'.
EXAMPLE 103
[4-(3-Methyl-1,2,4-triazol-lyl)-2-trifluoromethyl-phenyl](5H,11H-pyrrolo[2,1-
c][1,4]benzodiazepin-10-yl)-methanone
To a suspension of 60% sodium hydride in oil (0.3 g) in dimethylformamide
(50 ml) was added dropwise 3-methyl-1,2,4-triazole (0.45 g). After hydrogen
gas
evolution ceased, 4-fluoro-2-trifluoromethyl-phenyl-(5H,11H-pyrrolo[2,1-
c][1,4]benzodiazepine-10-yl)-methanone (1.76 g) was added and the reaction
mixture
was heated in a sand bath at 110 °C for 18 hours. The mixture was
poured onto ice,
diluted with brine, and extracted with dichloromethane. The combined extracts
were
dried over anhydrous sodium sulfate and filtered through a short column of
anhydrous
sodium magnesium silicate. The solution was concentrated in vacuo and the
residue
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was triturated with diethyl ether to give 0.81 g of the title compound as
colorless
crystals, m.p. 148-150 °C, MS m/z: 438.2 (M+H)+, 875.8 (2M+H)+.
EXAMPLE 104
4-Hydrazino-2-methoxybenzoic acid, methyl ester, hydrochloride (1:1), hydrate
(2:1)
A stirred suspension of 4-amino-2-methoxybenzoic acid, methyl ester (21.74
g) in concentrated hydrochloric acid (110 ml), which was cooled to -10
°C, was
treated with a precooled solution of sodium nitrite (8.5 g) in water (45 ml)
at a rate
required to maintain a reaction temperature less than 0 °C. After the
addition was
complete, the reaction mixture was stirred at -2 °C for 10 minutes. The
cloudy,
orange solution was added dropwise to a vigorously stirred precooled solution
of tin
(II) chloride dihydrate (101 g) in concentrated hydrochloric acid (67 ml) at -
10 °C.
The rate of addition was controlled to maintain a reaction temperature less
than -5 °C.
After the addition was complete, the cream colored suspension was warmed to
room
temperature and a solid was filtered. The solid was washed with diethyl ether
and
dried over anhydrous sodium sulfate to yield 52 g of a crude product. The
crude
product (20 g) was partitioned between aqueous 2.5 N sodium hydroxide and
dichloromethane. The organic phase was filtered through diatomaceous earth,
washed with brine, and dried over anhydrous magnesium sulfate. Filtration and
evaporation of the solvent in vacuo afforded a cream colored solid (7.1 g),
which
upon treatment with one equivalent of an anhydrous hydrogen chloride solution
in
diethyl ether afforded the title compound as the monohydrochloride salt, m.p.
76-79
°C, MS, m/z: 197 (M+H)+.
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EXAMPLE 105
2-Methoxy-4-(3-methyl-pyrazol-1-yl)-benzoic acid, methyl ester
To a stirred solution of 4-hydrazino-2-methoxybenzoic acid, methyl ester
hydrochloride (0.88 g) from Example 104 and one drop of concentrated
hydrochloric
acid in a 1:1 water/methanol (10 ml) mixture was added acetylacetaldehyde
dimethylacetal (0.53 g). The reaction was heated to 90 °C for 5
minutes. The
reaction was concentrated in vacuo and partitioned between 1 N sodium
hydroxide
(10 ml) and ethyl acetate (50 ml). The organic phase was removed and washed
with
brine, dried over anhydrous magnesium sulfate and filtered. Evaporation of the
solvent in vacuo afforded a brown oil which was combined with a previous lot
(0.54
g) and recrystallized three times from diisopropyl ether to give 2-methoxy-4-
(3-
methyl-pyrazol-1-yl)-benzoic acid, methyl ester (0.5 g). m.p. 167-
169°C, MS, m/z:
246 (M)+.
EXAMPLE 106
2-Methoxy-4-(3-methyl-pyrazol-1-yl)-benzoic acid
A solution of 2-methoxy-4-(3-methyl-pyrazol-1-yl)-benzoic acid methyl ester
(0.5 g) from Example 105 in tetrahydrofuran (2.5 ml) was treated with 1 N
lithium
hydroxide (2.13 ml) at room temperature. After 14 hours the solvent was
removed in
vacuo and the title compound precipitated by the addition at 0 °C of 1N
hydrochloric
acid. After drying under vacuum 0.42g of the title compound was obtained as a
solid. MS, m/z: 232 (M)+~
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EXAMPLE 107
[2-Methoxy-4-(3-methyl-pyrazol-1-yl)-phenyl] (5H,11H-pyrrolo[2,1-c] [1,4]-
benzodiazepin-10-yl) methanone
Oxalyl chloride (0.17 ml) was added to a stirred solution of 2-methoxy-4-(3-
methyl-pyrazol-1-yl)-benzoic acid (0.41 g) from Example 106 and
dimethylformamide (0.004 ml) in anhydrous tetrahydrofuran (10 ml). The
reaction
was heated at 35 °C -for ten minutes. The resulting solution was
evaporated in vacuo
to yield the crude 2-methoxy-4-(3-methyl-pyrazol-1-yl)-benzoic acid carbonyl
chloride. Following coevaporation with dichloromethane the acid chloride was
dissolved in dichloromethane (10 ml) and 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]-
benzodiazepine (0.31 g) added. Diisopropylethylamine (0.37 ml) was added and
the
reaction stirred at room temperature for 2 hours. The reaction was diluted
with
dichloromethane and washed with water followed by 1N hydrochloric acid. The
organic phase was washed with brine, dried over anhydrous magnesium sulfate
and
concentrated to dryness in vacuo. The solid residue was purified by flash
column
chromatography on silica gel eluting with hexane /ethyl acetate (2/1) to give
0.35 g
of the title compound as a colorless solid, m.p. 92-94 °C.
EXAMPLE 108
(3-Dimethylaminomethyl-SH,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)[2-
methoxy-4-(3-methyl-pyrazol-1-yl)-phenyl]-methanone
To a stirred solution of [2-methoxy-4-(3-methyl-pyrazol-1-yl)-
phenyl](5H,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl) methanone from Example
107 (0.57 g) in warm methanol (10 ml) was added N,N,N,N'-tetramethyl-
diaminomethane (0.392 ml) and acetic acid (0.164 ml). Following the addition
of
aqueous 37% formalin solution (2.9 ml) the reaction was stirred for fifteen
minutes.
The mixture was concentrated in vacuo and partitioned between dichloromethane
and
sodium hydrogen carbonate. The organic phase was removed, washed with brine
and
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dried over anhydrous magnesium sulfate. The solution was filtered and the
solvent
removed in vacuo. The residue was purified by flash column chromatography on
silica eluting with chloroform/methanol (50/1) to afford a. solid.
Recrystallisation of
the solid from acetone gave the title compound as a colorless solid, m.p. 196-
198 °C.
EXAMPLE 109
[2-Hydroxy-4-(3-methyl-pyrazol-1-yl] (SH,11H-pyrrolo[2,1-c] [1,4]-
benzodiazepin-10-yl)-methanone
[2-Methoxy-4-(3-methyl-pyrazol-1-yl)-phenyl] (5H,11H-pyrrolo [2,1-c] [ 1,4]-
benzodiazepin-10-yl) methanone (0.82 g) from Example 107 was dissolved in
dichloromethane (20 ml) and cooled to -78 °C. Boron tribromide (6.2 ml)
was added
and the reaction stirred at 0 °C for five minutes. Ammonium hydroxide
(15 ml) was
added and extracted with dichloromethane. The organic phase was washed with
brine
and dried over anhydrous magnesium sulfate. The solid was removed by
filtration
and the solvent removed in vacuo. The residue was purified by flash column
chromatography on silica pressure eluting with hexane/ethyl acetate (3/1 then
2/1) to
afford 0.19g of the title compound as a colorless solid, m.p. 134-136
°C.
EXAMPLE 110
2-Chloro-4-iodo-benzoic acid, methyl ester
4-Amino-2-methoxy-benzoic acid methyl ester (22.97 g) was cooled to an
internal temperature of -10 °C in concentrated hydrochloric acid (110
ml) and stirred
as a suspension. A precooled solution of sodium nitrite (98.71 g) in water (45
ml) was
added to this mixture, at such a rate so as to maintain a reaction temperature
of less
than 0 °C. After stirring for 25 minutes at 0 °C the reaction
was treated with a
solution of potassium iodide (24.44 g) and iodine (18.37 g) in water (50 ml)
at such a
rate so as to maintain a reaction temperature of less than -4°C. Ethyl
acetate (100 ml)
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was added during the addition and the dark mixture was stirred at 0 °C
for one hour.
The organic layer was diluted with ethyl acetate and washed well with
saturated
sodium thiosulfate solution. The resulting orange solution was washed with
brine and
dried over anhydrous magnesium sulfate. The solvent was evaporated in vacuo to
yield an oil which was purified by suction filtration through silica gel
eluting with
hexane/ethyl acetate (50/1). The resulting purified oil solidified on cooling
to give
33.71 g of the title compound. MS, m/z: 296 (M)+~
EXAMPLE 111
4-Bromo-1-methyl-1H-pyrazole
To a suspension of prewashed (tetrahydrofuran) 60% sodium hydride in oil
(11.67 g) in tetrahydrofuran (200 ml) was added dropwise a solution of 4-
bromopyrazole (39.77 g) in tetrahydrofuran (50 ml). The solution was stirred
at room
temperature for two hours. Excess iodomethane (33 ml) in tetrahydrofuran (50
ml)
was added at such a rate as to maintain a slight increase in temperature. The
reaction
was stirred further for two hours. The solvent was removed in vacuo and the
residue
stirred in diethyl ether. A precipitate was removed by suction filtration and
washed
with diethyl ether. The combined organic phase was evaporated in vacuo to give
42.22g of the title compound as an oil. MS, m/z: 160 (M)+.
EXAMPLE 112
1-Methyl-4-tributylstannyl-1H-pyrazole
To a precooled (<-10°C internal temperature) solution of 1.6M n-butyl
lithium
in hexanes (100 ml) in anhydrous diethyl ether (100 ml) under argon was added
a
solution of 4-bromo-1-methyl-1H-pyrazole (23.42 g) from Example 111 in diethyl
ether (50 ml) at a rate to maintain the temperature. The reaction was allowed
to stir
for a further 20 minutes before tributyltin chloride (43.4 ml) was added in
diethyl
ether (50 ml). The reaction temperature was allowed to rise to 20 °C.
The reaction
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was diluted with diethyl ether and the insoluble material removed by suction
filtration. Evaporation of the solvent in vacuo afforded 56 g of the title
compound as
an oil. MS, m/z: 373 [M+H]+~ Residual amounts of tin residues were removed
from
the oil by distillation using a kugelrohr apparatus under high vacuum at 170
°C.
EXAMPLE 113
2-Chloro-4-(1-methyl-1H-pyrazol-4-yl)-benzoic acid, methyl ester
An argon degassed dimethylformamide solution (70 ml) of 2-chloro-4-iodo-
benzoic acid methyl ester (25.4 g) pyrazole from Example 110, 1-methyl-4-
tributylstannyl-1H- (31.77 g), tetrakis(triphenylphosphine)palladium (0) (1.8
g) and
catalytic copper (I) iodide was heated at 80 °C for 7 hours. The
solvent was removed
in vacuo and the residue adsorbed onto silica gel. Purification by suction
filtration
through a pad of silica gel eluting sequentially with hexane and followed by
hexane/ethyl acetate (2/1) afforded after evaporation of the solvent a solid
residue
which was recrystallised from diisopropyl ether to give 7.82 g. of the title
compound
MS, m/z 250 (M)+.
EXAMPLE 114
2-Chloro-4-(1-methyl-1H-pyrazol-4-yl)-benzoic acid
To a solution of 2-chloro-4-(1-methyl-1H-pyrazol-4-yl)-benzoic acid, methyl
ester (6.25 ) from Example 113 in methanol (80 ml) was added 1N sodium
hydroxide
(30 ml). The reaction was heated under reflux for one hour. The volume of
solvent
was reduced in vacuo by three quarters and the residue treated with 2N
hydrochloric
acid at 0 °C. A precipitate was filtered and dried in vacuo to yield
5.84 g the title
compound, MS m/z: 237 [M+H]+~
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EXAMPLE 115
[2-Chloro-4-(1-methyl-1H-pyrazol-4-yl)phenyl) (5H,11H-pyrrolo[2,1-c][1,4]-
benzodiazepin-10-yl)-methanone
Oxalyl chloride (0.49 ml) was added to a solution of 2-chloro-4-(1-methyl-
1H-pyrazol-4-yl)-benzoic acid (0.41 g) from Example 114 and dimethylformamide
(0.012 ml) in anhydrous tetrahydrofuran (20 ml). The reaction was heated at 35
°C
for ten minutes. The resulting solution was evaporated in vacuo to dryness to
yield
the crude 2-chloro-4-(1-methyl-1H-pyrazol-4-yl)-benzoic acid carbonyl
chloride.
Following co-evaporation with anhydrous methylene chloride, the acid chloride
was
dissolved in dichloromethane (20 ml) followed by the addition of 10,11-dihydro-
5H-
pyrrolo[2,1-c][1,4]-benzodiazepine (0.888 g) and diisopropylethylamine (1.06
ml).
The resulting solution was stirred at room temperature for 2 hours. The
reaction was
diluted with dichloromethane and washed with water followed by 1N hydrochloric
acid. The organic phase was washed with brine and dried over anhydrous
magnesium
sulfate. The dichloromethane was filtered and concentrated in vacuo to
dryness. .
The residue was purified by flash column chromatography on silica pressure
eluting
with hexane/ethyl acetate (2/1) to afford 1.4 g of the title compound as a
colorless
solid m.p. 105-109 °C.
EXAMPLE 116
[2-Chloro-4-(3-methyl-pyrazol-1-yl)-phenyl]-(4H,lOH-pyrazolo[5,1-c] [1,4]-
benzodiazepin-5-yl)-methanone
To solution of 2-chloro-4-(3-methyl-pyrazol-1-yl)-benzoyl chloride (0.214 g)
Example 18 Step a in dichloromethane (10 ml) was added 5H-10,11-
dihydropyrazolo[5,1-c][1,4] benzodiazepine (0.153 g) and diisopropylethylamine
(0.173 ml). The reaction was stirred at room temperature for 2 hours. The
reaction
was diluted with dichloromethane and washed with water followed by 1N
hydrochloric acid. The organic phase was washed with brine and dried over
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anhydrous magnesium sulfate. The dichloromethane solution was filtered and
concentrated in vacuo to dryness. The residue was purified by flash column
chromatography on silica pressure eluting with hexane/ethyl acetate (1/1) to
afford
0.3 g of the title compound as a colorless solid, m.p. 187-188 °C.
EXAMPLE 117
2-Chloro-4-(3-methyl-pyrazol-1-yl)-phenyl]-(5,10-dihydro-4H-tetrazolo[5,1-
c][1,4]benzodiazepin-5-yl)-methanone
To a solution of 2-chloro-4-(3-methyl-pyrazol-1-yl)-benzoyl chloride (0.18 g)
Example 18 Step a in dichloromethane (10 ml) was added 10,11-dihydro-5H-
tetrazole[5,1-c][1,4]benzodiazepine (0.13 g) and diisopropylethylamine (0.145
ml).
The reaction was stirred at room temperature for 2 hours. The reaction was
diluted
with dichloromethane and washed with water followed by 1N hydrochloric acid.
The organic phase was washed with brine and dried over anhydrous magnesium
sulfate. The dichloromethane solution was filtered and concentrated in vacuo
to
dryness. The residue was purified by flash column chromatography on silica
eluting
with hexane/ethyl acetate (1/1) to give 0.14 g of the title compound as a
colorless
solid. m.p. 110-114°C.
EXAMPLE 118
1-[4-(4H,lOH-pyrazolo[5,1-c][1,4]benzodiazepine-5-carbonyl) phenyl]-ethanone
A mixture of 5,10-dihydro-4H-pyrazolo[5,1-c][1,4]benzodiazepine ( 0.555 g),
4-acetylbenzoyl chloride (0.657 g) and N,N-diisopropylethylamine (0.464 g) in
dichloromethane (15 ml) was stirred at room temperature for 4 hours. The
mixture is
poured into water and extracted with dichloromethane. The dichloromethane
extract
was washed with saturated sodium hydrogen carbonate, water and brine and dried
over anhydrous sodium sulfate. The extract is filtered through a thin pad of
hydrous
magnesium silicate and the filter pad washed with dichloromethane. The
filtrate is
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concentrated in vacuo to give 1.53 g of yellow solid. Trituration of the solid
with
ethyl acetate gave 0.747 g of the title compound as a glass, m.p. 201-
210°C.
The mother liquors from the trituration were evaporated and the residue (0.30
g) was
chromatographed on thick layer silica gel plates (200 micron) using hexane-
ethyl
acetate (1:1) as solvent. The solid is triturated with ethyl acetate and
combined with
the 0.747 g of initially isolated product. The combined solids were
precipitated from
a mixture of dichloromethane -hexane to give 0.73 g of product as a glass .
EXAMPLE 119
1-[4-(4H,lOH-pyrazolo[5,1-c][1,4]benzodiazepine-5-carbonyl)phenyl]-3-
(dimethylamino)-prop-2-en-1-one
A mixture of 1-[4-(4H,lOH-pyrazolo[5,1-c][1,4]benzodiazepine-5-carbonyl)
phenyl]ethanone (0.73 g), tert-butoxybis-[dimethylamino]methane (0.964 g) in
dichloromethane (10 ml) was stirred at room temperature for 2 days. The
mixture is
concentrated in vacuo and the residue crystallized from dichloromethane-hexane
to
give 0.65 g of the title compound as yellow crystals, m.p. 225-230°C.
EXAMPLE 120
[4-(1-Methyl-1H-pyrazol-3-yl)phenyl](4H,lOH-pyrazolo[5,1c][1,4]-
benzodiazepin-5-yl) methanone (Isomer A)
and
[4-(2-Methyl-1H-pyrazol-3-yl)phenyl](4H,lOH-pyrazolo[5,1c][1,4]-
benzodiazepin-5-yl) methanone (Isomer B)
A mixture of 1-[4-(4H,lOH-pyrazolo[5,1-c][1,4]benzodiazepine-5-
carbonyl)phenyl]-3-(dimethylamino)-prop-2-en-1-one (0.83 g), hydrazine (0.198
g)and acetic acid (0.336 g) in 10 ml of ethanol is refluxed for 4 hours. The
volatiles
were removed in vacuo and the residue dissolved in dichloromethane. The
solution
is washed with water, 1N sodium hydrogen carbonate, water and brine and dried
over
anhydrous sodium sulfate. The solution is filtered through a thin pad of
hydrous
magnesium silicate and the filter pad washed with ethyl acetate. The filtrate
is
concentrated in vacuo to give 0.56 g of light yellow solid. The solid was
chromatographed on thick layer silica gel plates (200 microns) with ethyl
acetate as
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solvent to give 0.35 g of white solid as a mixture of A and B (1:4). Multiple
fractional crystallizations from ethyl acetate gives 89 mg of crystals, m.p.
155-156°C
as a mixture of A and B (9:1) and 65 mg of a glass as a mixture of A and B
(1:6)
EXAMPLE 121
1-[4-(SH,11H-pyrrolo[2,1-c] [1,4]benzodiazepine-10-carbonyl)-3-chlorophenyl]-
ethanone
Step a) Triethylamine (8.80 ml) was added to a solution of (4-bromo-2-
chlorophenyl)-(SH,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (2.37
g)
in pyridine (1.80 ml), in a 20 ml Carnus tube. The resultant solution was
purged with
nitrogen for 25 minutes then (trimethylsilyl)acetylene (1.67 ml),
bis(triphenyl-
phosphine)palladium(II) chloride (0.08 g) and copper(I) iodide (0.01 g) were
added.
The tube was filled with nitrogen-purged triethylamine, sealed and heated on
an oil
bath at 90 °C for 80 hours. The solution was cooled to room
temperature, the solvent
evaporated in vacuo, and the residue partitioned between dichloromethane and
water.
The dichloromethane extract was dried over anhydrous sodium sulfate, filtered,
and
evaporated in vacuo to a brown foam. Purification by flash chromatography on
silica
gel eluting with hexane-ethyl acetate (1:1) resulted in the intermediate
acetylene as an
off-white foam (2.11 g), MS m/z: 418 (M)+. This material was used without
further
purification in the next step.
Step b) A solution of 1 % sulfuric acid in tetrahydrofuran was saturated with
mercury
(II) sulfate. The intermediate acetylene (1.0O g) in tetrahydrofuran (5 ml)
was stirred
for 50 hours with 30 ml of the aforementioned mercury (II) sulfate-
tetrahydrofuran
solution. An additional amount of mercury (II) sulfate (0.01 g) and water 0.3
ml
was added. After stirnng for 120 hours, the reaction mixture was poured into
water
and extracted with dichloromethane. The dichloromethane solution was washed
sequentially with saturated aqueous sodium bicarbonate and water. The
dichloromethane solution was dried over anhydrous magnesium sulfate, filtered,
and
evaporated to dryness to yield a brown solid. Purification by flash
chromatography
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on silica gel eluting with hexane-ethyl acetate (1:1) gave a white solid (0.30
g), mp
98-100°C, MS m/z: 364 (M)+.
EXAMPLE 122
1-[4-(SH,11H-pyrrolo[2,1-c] [1,4]benzodiazepine-10-carbonyl)-3-chlorophenyl]-
ethanone
Tributyl(ethoxyvinyl)tin (1.17 g) was added to a solution of (4-bromo-2-
chlorophenyl)-(SH,11H-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone (1.24
g)
in toluene (10 ml). The resultant solution was purged with nitrogen for 10
minutes,
then bis(triphenylphosphine)palladium (II) chloride (0.11 g) was added. The
reaction mixture was heated to reflux for 24 hours. The solution was cooled to
room
temperature and 5% aqueous hydrochloric acid (10 ml) was added. . After
stirnng for
one hour, the mixture was filtered through a pad of diatomaceous. Diethyl
ether (5
ml) was added to the filtrate, and the resulting mixture was extracted with
ethyl
acetate. The organic layer was dried over anhydrous magnesium sulfate,
filtered, and
evaporated in vacuo to yield a brown glass. Purification by flash
chromatography on
silica gel eluting with hexane-ethyl acetate (1:1) resulted in a white solid
(0.30 g),
MS, m/z: 364 (M)+.
EXAMPLE 123
[2-Chloro-4-(3-methyl-4-ethynyl-phenyl) (5H,11H-pyrrolo[2,1-c][1,4]-
benzodiazepin-10-yl)-methanone
Treatment of the intermediate acetylene of Example 121 step A with a 1M
solution of tetrabutylammonium fluoride in tetrahydrofuran at room temperature
provided upon solvent removal an 84% yield the title compound as an orange -
yellow solid, m.p. 84-86°C, MS, m/z: 346 (M)+.
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Process for Preparing Pharmaceutical Formulations
This invention also includes methods for producing the formulations disclosed
herein. A process of this invention comprises the steps of:
a) combining, preferably with mixing or stirring, the PEG and surfactant
components to create a first carrier mixture;
b) raising the temperature of the first carrier mixture to a range of from
about 75°C to about 95°C, preferably from about 80°C to
about 90°C;
c) adding the active ingredient to create a first pharmaceutical
composition mixture;
d) stirring the first pharmaceutical composition mixture, preferably with
heating, until the first pharmaceutical composition mixture is clear,
preferably at a
temperature from about 115°C to about 170°C, preferably to a
temperature from
about 130°C to about 150°C, more preferably at a temperature
from about 135°C to
about 145°C;
e) cooling the first pharmaceutical composition, if necessary, to a
temperature of from about 75°C to about 95°C, preferably from
about 80°C to about
90°C;
g) adding the amount of sucrose fatty acid esters) and/or povidone to
create a final pharmaceutical composition mixture, preferably with stirring
until the
final pharmaceutical composition mixture is clear.
In cases wherein optional antioxidants or preservatives are used, such as BHA,
BHT, etc., the following steps may be employed:
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a) combining, preferably with mixing or stirnng, the PEG component
(such as a mixture of PEG 400 and PEG 1000) and the surfactant component (such
as
Polysorbate 80) to create a first carrier mixture;
b) raising the temperature of the first carrier mixture to a range of from
about 75°C to about 95°C, preferably from about 80°C to
about 90°C;
c) adding to the first carrier mixture optional antioxidant or preservative
components to create a second carrier mixture, which is then stirred or
otherwise
mixed until the second Garner mixture is a clear solution;
d) adding the active ingredient to create a first pharmaceutical
composition mixture;
e) stirring the first pharmaceutical composition mixture, preferably with
heating, until the first pharmaceutical composition mixture is clear,
preferably at a
temperature from about 130°C to about 150°C, more preferably at
a temperature from
about 135°C to about 145°C;
f) optionally cooling the first pharmaceutical composition to a
temperature of from about 75°C to about 95°C, preferably from
about 80°C to about
90°C;
g) adding the amount of sucrose fatty acid esters) and/or povidone to
create a final pharmaceutical composition mixture, preferably with stirnng
until the
final pharmaceutical composition mixture is clear.
One skilled in the art will understand the viscosity and form of the final
formulation may be manipulated with components within the scope of this
invention
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and temperature ranges during processing. For instance, a fluid or semi-solid
composition may be produced with the more fluid PEG, surfactant and PVP
species
within the scope of this invention. More gel-like, viscous or semi-solid
compositions
may be produced with combinations of higher molecular weight PEG components
and PVP components having higher K values. Moreover, the components may be
cooled below their melting point if milling or other processing of the final
composition is desired. To create a more pelletized initial composition, a
fluid
composition of this invention may be sprayed onto a cooled Teflon-coated
surface
to form small solid spheres, which may be individually coated or collected for
further
processing.
Specific non-limiting examples of formulations within the scope of this
invention are provided below.
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Example 1
50 mg/capsule: Oral Formulation at 10 % Drug Loading
In place of the polysorbate 80 in this formulation of Example l, other
polysorbate series such as Tween 20, 40 and 60 can also be used, alone or in
combination with each other and/or polysorbate 80.
(% w/w) per capsule 20,000
(mg) capsule batch (g)
active ingredient 10.42 50.00 1000.00
Inactive Ingredients:
PEG 1000, NF 30.96 148.61 2,972.16
Povidone USP K-17 10.00 48.00 960.00
Polysorbate 80, NF 10.00 48.00 960.00
B~~ ~' 0.09 0.42 8.32
B~, ~ 0.87 4.16 83.2
PEG 400, NFZ Q.S. to Q.S. to Q.S. to 9,600
100 480.00
1. Weigh the Polysorbate 80, PEG 400, and PEG 1000 into a suitable
mixing vessel, stir using a top mounted mixer, and warm to 85 ~ 5 °C.
2. Add BHT and BHA to the mixing vessel, very slowly to prevent
formation of lumps. Continue stirring at 85 ~ 5 °C, until a clear
solution was
formed.
3. Add active ingredient to the vessel at 85 ~ 5 °C , very slowly to
prevent formation of lumps. Slowly raise the temperature to 125 ~ 5 °C,
and stir
until the active ingredient dissolves completely.
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4. Cool the solution in step 4. to 60 ~ 5 °C.
5. Add Povidone, USP, K-17 (Plasdone C-15, ISP) slowly to step 5, to
prevent the formation of lumps.
Let the solution warm up to 85 ~ 5 °C. Stir until the solution becomes
clear.
6. Encapsulate 480 mg of the finished solution (in step 10) into size 1
capsules at 38 ~ 5 °C using Hoflinger and Karg (H&K) 800L encapsulator
machine.
During encapsulation cool the body of capsule using cool Nitrogen, which was
passed
through dry ice.
7. Band seal the capsules with gelatin solution.
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Example 2
50 mg/cansule: Oral Formulation at 10 % Drug Loading
In place of surfactant used in this formulation (poloxamer 188), other
polymers in the series such as Pluronic L44, Pluronic L101 can also be used.
(% w/w) per capsule 20,000
(mg) capsule batch
(g)
active ingredient 10.42 50.00 1000.00
Inactive Ingredients:
Povidone USP K-17 10.00 48.00 960.00
(Plasdone C-15, ISP)
Poloxamer 188, NF 12.00 57.60 1152.00
BHT NF 0.09 0.42 8.32
BHA NF 0.87 4.16 83.20
PEG 400 NF Q. S. to Q.S. to Q.S. to
100 480.00 9600 g
This formulation is manufactured the same as that of the formula of Example
1 (50 mg/capsule) with the exception that 12 % of poloxamer was used in place
of the
polysorbate 80 in this formulation. The encapsulation weight is 480 mg.
Example 3
50 mg/ca~sule
Example 3
provides a formulation with a combination of two or more surfactants.
(% w/w) per capsule (mg) 20,000 capsule
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batch (g)
active ingredient 10.64 51.07 1,021.44
Inactive Ingredient:
PEG 1000, NF 28.60 137.28 2,745.60
Povidone USP K-17 10.00 48.00 960.00
(Plasdone C-15, ISP)
Polysorbate 40, NF 5.00 24.00 480.00
Poloxamer 188, NF 10.00 48.00
BHT, NF 0.09 0.43 8.64
BHA, NF 0.87 4.18 83.52
PEG 400, NF Q.S. to Q.S. to 480.00Q.S. to
100
9600.00
The formulation of Example 3 is manufactured the same as that of Example 1
(SO mg/capsule) with the exception that two surfactants, polysorbate 40 and
poloxamer 188 were added in step 1 along with PEG 400 and PEG 1000. The
encapsulation weight is 480 mg.
Example 4
25 m capsule: Oral Formulation at 5 % Drub Loading
(% w/w) per capsule 20,000
(mg) capsule
batch (g)
active ingredient 5.49 25.00 500.00
Inactive Ingredient:
PEG 1000, NF 32.66 148.61 2,972.16
Povidone, USP K-17 10.55 48.00 960.00
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(Plasdone C-15,ISP)
Polysorbate 80, NF 10.55 48.00 960.00
BHT, NF 0.09 0.42 8.32
BHA, NF 0.91 4.16 83.2
PEG 400, NFz Q.S. to 100 Q.S. to Q.S. to
455.00 9,100 g
The formulation of Example 4 is produced in the same manner as that of 50
mg/capsule, above, with the exception that the heating temperature to
solubilize the
active ingredient in step 3 is 115 ~ 5 °C, instead of 120 ~ 5
°C. The encapsulation
weight is 455 mg.
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Example 5
100 m~/capsule~ Oral Formulation at 15 % Drug Loading
(% w/w) per capsule 20,000 capsule
(mg) batch (g)
active ingredient 15.38 100.00 2,000.00
Inactive Ingredient:
PEG 1000, NF 28.98 188.35 3,767.05
Povidone USP K-17 10.00 65.00 1,300.00
(Plasdone C-15, ISP)3
Polysorbate 80, NF 9.45 61.39 1,227.91
0.08 0.53 10.64
0.82 5.32 106.42
PEG 400, NF Q.S. to Q.S. to Q.S. to 13,000.00
100 650.00
This formulation is produced with the same steps as the 50 mg/capsule, above,
with the exception that the heating temperature to solubilize the active
ingredient in
step 3 is 145 ~ 5 °C, instead of 120 ~ 5 °C. The encapsulation
weight is 650 mg in
size 0 hard gelatin capsule.
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Example 6
150 mg Active Ingredient in Size 00 Capsule
(% w/w) per capsule 20,000
(mg) capsule batch
(g)
active ingredient 16.48 149.97 2,999.36
Inactive Ingredients:
PEG 1000, NF 26.3 239.33 4,786.60
Povidone USP K-17 15 136.50 2,730.00
(Plasdone C-15,ISP)
Polysorbate 80, NF 9.32 84.81 1,696.24
B~, ~ 0.08 0.73 14.56
B~~ ~ 0.81 7.37 147.42
PEG 400, NF Q.S. to 100 Q.S. Q.S. to
to
910.00 18,200.00
This formulation of Example 6 is produced with the same steps as that of 50
mg/capsule with the exception of the heating temperature to solubilize the
active
ingredient in step 3 is 150 ~ 5 °C, instead of 145 ~ 5 °C. The
encapsulation weight is
910 mg in size 00 hard gelatin capsule.
The following specific Examples 7 through 11 shown in Table 1, below, can
be formulated as described above to create formulations of 10% active
ingredient
with varying concentrations of PEG 400, PEG 1000, two PVP components with
respective K values of 15 and 90, and a combination of BHA and BHT as an
adjuvant
component.
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$ Table 1
Example PEG PEG PVP PVP BHT BHA NATO Active
No. 400 1000 K15 K90 Ingrednt
(%) (%) (%) (%) (%) (%) (%) (%)
7 55.40 20.00 10.00 0.00 0.20 2.002.40 10.00
8 40.40 3 5 10.00 0.00 0.20 2.002.40 10.00
.00
9 75.40 0.00 5.00 5.00 0.20 2.002.40 10.00
65.40 10.00 0.00 10.00 0.20 2.002.40 10.00
11 40.40 35.00 5.00 5.00 0.20 2.002.40 10.00
Similarly, the following Examples 12 through 32 may be formulated by the
methods herein using PEG 400, PEG 1000, PVP with a K value of 17, active
10 ingredient, BHA and BHT as antioxidants or preservatives and the additional
components listed as "other".
Table 2
Ex. PEG PEG PVP Active BHA BHT Other Other
No. 400 1000 K-17 Ingred.
12 40.40 35.00 10.00 2.00 0.20 Sodium -
10.00
Taurocholate
2.40
13 75.40 - 5.00 10.21 2.00 0.20 Sodium PVP
Taurocholate K-90
2.40 5.00
14 42.59 35.00 10.00 10.21 2.00 0.20 - -
34.35 28.23 10.00 10.21 2.00 0.20 Poloxamer 188
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15.00
1642.59 20.00 10.21 2.00 0.20 Poloxamer 188 -
10.00
15.00
1737.10 30.49 10.21 2.00 0.20 Poloxamer 188
10.00
10.00
1835.72 29.36 10.21 2.00 0.20 Poloxamer 188 -
10.00
12.50
1934.35 28.23 10.21 2.00 0.20 Poloxamer 188 -
10.00
15.00
2037.10 30.49 10.21 2.00 0.20 Poloxamer 188 -
10.00
10.00
2134.35 28.23 10.21 2.00 0.20 Poloxamer 188 -
10.00
15.00
2235.72 29.36 10.21 2.00 0.20 Poloxamer 188 -
10.00
12.50
2336.86 30.30 10.64 2.00 0.20 Pluronic L44 -
10.00
10.00
2436.86 30.30 10.64 2.00 0.20 Pluronic L101 -
10.00
10.00
2539.61 32.55 10.64 2.00 0.20 Tween 40 -
10.00
5.00
2641.25 33.91 10.64 2.00 0.20 Tween 40 -
10.00
2.00
2739.61 32.55 10.64 2.00 0.20 Tween 20 -
10.00
5.00
2841.25 33.91 10.64 2.00 0.20 Tween 20 -
10.00
2.00
2934.12 28.04 10.64 2.00 0.20 Tween 40 Poloxamer
10.00
5.00 188
10.00
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30 36.86 30.30 10.00 10.64 2.00 0.20 Tween 40 -
10.00
31 36.86 30.30 10.00 10.64 2.00 0.20 Tween 80 -
10.00
32 34.12 28.04 10.00 10.64 2.00 0.20 Tween 80 Poloxamer
5.00 188
10.00
