Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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METHOD FOR TREATING COGNITIVE DYSFUNCTION
This invention provides a method of using 2-methyl-
4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine, for the treatment of cognitivedysfunction.
Alzheimer's Disease is a degenerative brain
disorder characterized clinically by progressive loss of
memory, cognition, reasoning, and judgment. Eventually, a
global defect develops that involves all aspects of higher
cortical function. Initiative ~;m;n;shes, and the patient
may become distractible. In addition to the decreased
cognitive function, specific disturbances of speech, motor
activity, and recognition of perceptions may be discernible.
Normal personality traits may become exaggerated or
caricatured. The initial affective change may be dominated
by irritability, with periods of anger and violence. The
patient commonly exhibits impoverished thought process and
delusions. As the disease progresses uncontrollable
agitation commonly develops. To date, Alzheimer's Disease
has proven to be incurable.
Palliative treatments to alleviate the symptoms of
cognitive dysfunctions of disease processes such as
Alzheimer's Disease could improve the quality of life for
both the patient and the patient's caregiver. Such
treatments could m;n;mize or delay the emergence of symptoms
requiring hospitalization or institutional care. Therefore,
such treatments are desirable for both health economic
purposes and for the enhancement of the quality of life when
a cognitive dysfunction is present.
Further, although the cholinergic neurons
degenerate, it is characteristic of Alzheimer~s Disease that
the postsynaptic muscarinic receptors in the forebrain and
hippocampus still exist. Muscarinic cholinergic agonists are
therefore useful in the treatment of Alzheimer's Disease and
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in improving the cognitive functions of those afflicted with
Alzheimer~s Dis ease.
Surprisingly, and in accordance with this
invention, Applicants have discovered that the compound 2-
methyl-4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine can be useful for the treatment of a
cognitive dysfunction, particularly Alzheimer's Disease. The
compound 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][l,5]benzodiazepine is known and described in U.S. Patent
No. 5,229,382, herein incorporated by reference in its
entirety.
The presently claimed invention provides a method
for treating a cognitive dysfunction comprising administering
an effective amount of 2-methyl-4-(4-methyl-1-piperazinyl)-
lOH-thieno[2,3-b][1,5]benzodiazepine to a patient in need of
such treatment.
Further, the invention provides a method for the
palliative treatment of a cognitive dysfunction comprising
administering an effective amount of 2-methyl-4-(4-methyl-1-
piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine to a
patient in need of such treatment. The 2-methyl-4-(4-methyl-
l-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine compound
is particularly useful for the palliative treatment of
Alzheimer's Disease.
The 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
thieno[2,3-b][1,5]benzodiazepine compound is of the formula
~ NCH3
N~<
~N~CHa
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or an acid addition salt thereof. The free base of formula
(I) is 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine.
r 5 The substantially pure crystalline anhydrous Form I
2-methyl-4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine (Form I) has a typical X-ray powder
diffraction pattern substantially as follows, using a
Sieman~s D5000 diffractometer equipped with a copper
radiation source, wherein d represents the interplaner
spac1ng:
d I/I1
10.2689 100.00
8.577 7.96
7.4721 1.41
7.125 6.50
6.1459 3.12
6.071 5.12
5.4849 0.52
5.2181 6.86
5.1251 2.47
4.9874 7.41
4.7665 4.03
4.7158 6.80
4.4787 14.72
4.3307 1.48
4.2294 23.19
4.141 11.28
3.9873 9.01
3.7206 14.04
3.5645 2.27
3.5366 4.85
3.3828 3.47
3.2516 1.25
3.134 0.81
3.0848 0.45
3.0638 1.34
3.0111 3.51
2.8739 0.79
2.8102 1.47
2.7217 0.20
2.6432 1.26
2.6007 0.77
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Form II 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
thieno[2,3-b][1,5]benzodiazepine (Form II) has a typical X-
ray powder di~fraction pattern substantially as follows,
using a Sieman's D5000 diffractometer equipped with a copper
radiation source, wherein d represents the interplaner
spacing:
d I/Il
9.9463100.00
8.5579 15.18
8.2445 1.96
6.8862 14.73
6.3787 4.25
6.2439 5.21
5.5895 1.10
5.3055 0.95
4.9815 6.14
4.8333 68.37
4.7255 21.88
4.6286 3.82
4.533 17.83
4.4624 5.02
4.2915 9.19
4.2346 18.88
4.0855 17.29
3.8254 6.49
3.7489 10.64
3.6983 14.65
3.5817 3.04
3.5064 9.23
3.3392 4.67
3.2806 1.96
3.2138 2.52
3.1118 4.81
3.0507 1.96
2.948 2.40
2.8172 2.89
2.7589 2.27
2.6597 1.86
2.6336 1.10
2.5956 1.73
The x-ray powder dif~raction patterns set forth
herein were obtained with a copper K of wavelength = 1.541A.
The interplanar spacings in the column marked "d" are in
Angstroms. The typical relative intensities are in the
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W O 96/38151 PCT~US95106859
column marked "I/I1". The detector was a Kevex silicon
lithium solid state detector.
As used herein l'substantially pure" shall refer to
anhydrous Form I associated with < 5% Form II; and most
preferably it shall refer to < 2% Form II. It is further
preferred that llsubstantially pure~ shall refer to < 0.5%
non-Form I polymorph.
As used herein "substantially pure" shall refer to
anhydrous Form I associated with about < 5% Form II; and most
preferably it shall refer to about < 2% Form II. It is
further preferred that "substantially pure" shall refer to <
0.5% related substances. When the Form I polymorph is
formulated as a pharmaceutical composition, "substantially
pure~ shall preferably refer to about < 15% Form II
polymorph; more preferably, the term shall refer to about <
10% Form II polymorph when the Form I polymorph is formulated
as a pharmaceutical, and it is especially preferred that the
term shall refer to about <5% Form II polymorph when the
substantially pure substance is formulated.
As used herein, the term "2-methyl-4-(4-methyl-1-
piperazinyl)-lOH-thieno[2,3-b]tl,5]benzodiazepinel' refers to
a technical grade of 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
thienot2,3-b]tl,5]benzodiazePine when no specific solvate or
polymorph is named. Typically, the technical grade 2-methyl-
4-(4-methyl-1-piperazinyl)-lOH-thienot2,3-
b]tl,5]benzodiazePine contains less than about 5% undesired
related substances and may be a mixed polymorph. Such
technical grade 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
thienot2,3-b]tl,5]benzodiazepine usually contains less than
about 1% undesired related substances.
The term ~crude" refers to a form of 2-methyl-4-(4-
methyl-1-piperazinyl)-lOH-thienot2,3-b]tl,5]benzodiazepine
typically associated with undesired polymorph and/or greater
than about 5% undesired related substances. Such crude grade
2-methyl-4-(4-methyl-1-piperazinyl)-lOH-thienot2,3-
b]tl,5]benzodiazePine may contains less than about 1%
undesired related substances.
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As used herein, the term "m~mm~l~' shall refer to
the M~mm~l ia class of higher vertebrates. The term "m~mm~l "
includes, but is not limited to, a human. The term
~treating~ as used herein includes prophylaxis of the named
condition or amelioration or elimination of the condition
once it has been established.
As used herein, the term "palliative treatment~
shall refer to a treatment designed for the relie~ of
symptoms of cognitive dysfunction, rather than curing the
disease. Such symptoms of cognitive dysfunction may include,
but are not limited to progressive loss of memory, cognition,
reasoning, judgment, aspects of higher cortical function,
~;m; n; shed initiative, excessive distraction, speech, motor
activity, recognition of perceptions, exaggerated or
caricatured personality traits, irritability, excessive
anger, violence, uncontrollable agitation, and delusions.
The method of this invention is particularly useful for the
treatment of delusions and disturbance in perceptions from
such cognitive dysfunction. Further, the method of this
invention is especially desirable for the treatment of
excessive personality traits, irritability, excessive anger,
violence, and uncontrollable agitation.
The term "cognitive dysfunction~ shall refer to a
dysfunction in the ability of a subject to perceive, reason,
remember, and in the ability to acquire knowledge. Such
dysfunction may be associated wlth Alzheimer's Disease, AIDS,
and other central nervous system conditions which cause such
dysfunction.
The results of pharmacological studies show that 2-
methyl-4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine has muscarinic cholinergic receptor
activity. The compound is active at the dopamine D-l and D-2
receptors as indicated by an IC50 of less than 1 uM in the
3H-SCH233390 (Billard, et al. Life Sciences 35:1885 (1984))
and the 3H spiperone (Seeman et al Nature 216:717 (1976))
binding assays respectively. Further, the anhydrous Form I
compound is active at the 5-HT-2 receptor and 5-HTlC
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W O96/381Sl PCTrUS95/06859
receptor. The complex pharmacological profile of the
compound provides a medicament which can be useful for the
treatment of a cognitive dysfunction.
In vivo animal and clinical observations support
that 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine has a complex muscarinic receptor
subtype profile. For example, rats exposed to an overdose of
the compound surprisingly exhibited significant salivation.
Further, clinical subjects experienced pupilary constriction
rather than the expected pupilary dialation.
The compound 2-methyl-4-(4-methyl-1-
piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine can be
used for the methods of this invention, both in its ~ree
base and acid addition salt forms. The acid addition salts
are preferably the pharmaceutically acceptable, non-toxic
addition salts with suitable acids, such as those of
inorganic acids, for example hydrochloric, hydrobromic,
nitric, sulphuric or phosphoric acids, or of organic acids,
such as organic carboxylic acids, for example glycollic,
maleic, hydroxymaleic, fumaric, malic, tartaric, citric or
lactic acid, or organic sulphonic acids for example methane
sulphonic, ethane sulfonic, 2-hydroxyethane sulfonic, p-
toluene-sulfonic or naphthalene-2-sulfonic acid.
The compound 2-methyl-4-(4-methyl-1-
piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine can be
prepared using a process which comprises
(a) reacting N-methylpiperazine with a compound of
the formula
/
N C
3 0 ~ N ~CH3
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in which Q is a leaving group capable of being split of~,
or
(b) ring-closing a compound o~ the ~ormula
NCH3
~ /
NH2 C
~ N)~CHa
Appropriate reaction conditions and suitable values of
Q can readily be chosen for these processes.
In reaction (a) the radical Q can, for example, be an
amino group or a mono- or dialkyl-substituted amino group
(each alkyl substituent suitably containing 1 to 4 carbon
atoms), hydroxyl, thiol, or an alkoxy, alkylthio or
alkylsulphonyl group suitably containing 1 to 4 carbon
atoms, for example a methoxy or methylthio group, or a
haIogen atom, especially a chlorine atom. Preferably, Q is
amino (-NH2), hydroxyl or thiol, and amino is most
preferred. The reaction is preferably carried out at a
temperature of from 50~C to 200~C.
When Q is amino, the intermediate of formula (II) may
also exist in the imino form:
NH
Il
/NH--C\
and when Q is hydroxyl or thiol, the intermediates of
formula (II) may exist in their amide and thioamide ~orms:
O S
Il 11
/NH--C\ or NH--C\
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_g _
The amidine of formula (II) (Q is -NH2), can be in
salt form, for example a salt of a mineral acid such as the
hydrochloride, and can be reacted with N-methylpiperazine
in an organic solvent such as anisole, toluene,
dimethylformamide or dimethyl-sulphoxide, preferably at a
temperature range of 100 to 150~C.
The amidine is prepared by condensing a
thiophene compound of formula
NC~
H2N S CH3
with an ortho-halonitrobenzene, in the presence of a base,
for example sodium hydride, in a solvent such as
tetrahydrofuran or n-butyl lithium in tetrahydrofuran, or
potassium carbonate or lithium hydroxide in
dimethylsulphoxide or aqueous sodium hydroxide in
dimethylsulfoxide, or with a tetraalkyl-ammonium salt in a
two-phase system, to form a nitronitrile of formula:
NO2 NC
H ~
(IIa)
which can be simultaneously reduced and ring-closed to the
amidine of formula (II) employing, for example, stannous
chloride and hydrogen chloride in aqueous ethanol or,
alternatively by reduction with hydrogen and
palladium/carbon or ammonium polysulphide followed by acid-
catalysed ring closure. The intermediate of formula (IIa)
may be isolated using ammonium chloride (NHACl) or ammonium
acetate (NH40Ac).
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W O96/38151 PCTrUS95/06859
--10--
When Q is hydroxyl, reaction (a) is preferably
carried out in the presence of titanium tetrachloride which
has the ability to react with the N-methylpiperazine to
form a metal amine complex. Other metal chlorides such as
those of zirconium, hafnium or vanadium may also be
employed. The reaction can be carrled out in the presence
of an acid binding agent such as a tertiary amine, for
example, triethylamine.
Alternatively, the reaction can be carried out
using excess of N-methylpiperazine to act as an acid-
binding agent. A suitable organic solvent such as toluene
or chlorobenzene can be used as a reaction medium, although
the use of anisole is particularly desirable, at least as a
co-solvent, in view of its ability to form a soluble
complex with Tic14.
If desired, elevated temperatures, for example
up to 200~C, can be used to hasten the reaction and a
preferred temperature range for carrying out the reaction
is from 80~C to 120~C.
The intermediate amide of formula (II) (Q is
-OH) can be prepared from the corresponding amidine (Q is
-NH2) by alkaline hydrolysis, or can be derived from
compounds of formula
NH2 C02R
~ N ~ ~ S ~ CH3
in which R is an alleyl group, preferably Cl-4 alkyl, by
ring closure employing, for example, sodium methylsulfinyl
methanide in a suitable solvent such as dimethylsulfoxide.
Alternatively, the amide can be prepared by ring closure of
an amino-acid, employing for example dicyclo-
hexylcarbodiimide (DCC) in a suitable solvent such as
tetrahydrofuran. The amino-acid can be obtained for
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example from the above esters by basic hydrolysis using for
example sodium hydroxide in ethanol.
Thioamides of formula (II) (Q is -SH),
iminothio-ethers; iminoethers or iminohalides, or other
derivatives containing active Q radicals as specified
above, tend to be more reactive towards N-methylpiperazine
and can usually be reacted without the necessity for the
presence of TiC14, but otherwise employing the same
conditions of temperature and solvent.
The thioamide of formula (II) (Q is -SH) can be
prepared by treating a solution of the corresponding amide
in an anhydrous basic solvent, such as pyridine, with
phosphorous pentasulfide. Similarly, the amide can be
converted to the iminothioether, iminoether or iminohalide,
or other derivatives containing active Q radicals, by
treatment with conventional reagents such as for example in
the case of the iminochloride, phosphorous pentachloride.
The intermediate compounds of formula (II) in
which Q is a leaving group capable of being split off,
particularly those in which Q is -NH2, -OH or -SH and when
Q is -NH2 salts thereof, are novel compounds, and form a
further aspect of the present invention.
With regard to reaction (b) above, the compound
of formula (III) may be ring-closed by employing, for
example, titanium tetrachloride as catalyst and anisole as
solvent, and the reaction is preferably carried out at a
temperature of 100~C to 250~C, for example from 150~C to
200~C.
The intermediate compound of formula (III) is
preferably prepared n situ without isolation by reacting a
compound of formula
NH2 R02C
HN
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in which R is an alleyl group, preferably Cl-4 alkyl, with
N-methylpiperazine, by heating to a temperature of between
30~C and 120~C, for example about 100~C, in a suitable
solvent such as for example anisole, and employing Tic14 as
catalyst.
The compound of formula (IV) can be prepared
from the corresponding nitro compound of formula
NO2 R02C
~--H --
Such compounds of formula (V) in which R is an
allyl group, such as for example Cl-4 alkyl, are novel and
form a further aspect of the invention.
If convenient this nitro compound can be
converted to the amine of formula (IV) without isolation,
before reaction with N-methylpiperazine. Intermediate
compounds of formula (V) can be made by condensation of a
thiophene of formula
RO2~
H2N S CH3
with an ortho-halonitrobenzene, preferably ortho fluoro- or
chloro-nitrobenzene, in the presence of a base, for
example, (a) sodium hydride in a solvent such as for
example tetrahydrofuran and at a temperature of from -20~C
to 30~C, or (b) anhydrous potassium carbonate or lithium
hydroxide in a solvent such as dimethylsulfoxide at a
temperature of from 90~C to 120~C. The compound of formula
(V) is converted to that of formula (IV) by reduction, for
example catalytically, employing hydrogen and
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-13-
palladium/carbon, or chemically, employing stannous
chloride and hydrogen chloride in aqueous ethanol, or
ammonium polysulphide, or zinc in aqueous ammonium
chloride.
It will be appreciated that the compound of
formula (I) may be isolated ~er se or may be converted to
an acid addition salt using conventional methods.
The compound has an ICso of less than 1 mM in
the 3H-QNB binding assay described by Y~m~mllra, HI and
Snyder, SH in Proc. Nat. Acad. Sci. USA, 71, 1725 (1974)
indicating that it has muscarinic-cholinergic activity.
Alzheimer~s Disease is likely caused by up to 90%
degeneration of the muscarinic cholinergic neurons in
nucleus basalis, which is part of substantia innomin~ta,
and these neurons project to the frontal cortex and
hippocampus having a general stimulatory effect on the
cognitive functions of the forebrain and hippocampus.
Although the cholinergic neurons degenerate, the
postsynaptic muscarinic receptors in the forebrain and
hippocampus still exist. Therefore, such muscarinic
cholinergic agonists can be useful in the treatment of
Alzheimer's Disease.
A double-blind multicenter clinical trial was
designed to assess the safety and efficacy of 2-methyl-4-
(4-methyl-1-piperazinyl)-10H-thieno[2,3-
b][1,5]benzodiazepine in 237 elderly patients with
cognitive dysfunction, wherein the age of the patients was
greater than or equal to sixty-five (6S) years of age.
Patients were r~n~m'zed to 2-methyl-4-(4-methyl-1-
piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine or
placebo. Changes in behavioral manifestations were
measured using the BEHAVE-AD, BPRS, and CGI rating scales,
which are known and available to the skilled artisan.. The
results of the study suggest that 2-methyl-4-(4-methyl-1-
piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine can be
useful for the treatment of behavioral manifestations of
cognitive dysfunction.
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WO96/38151 PCTrUS95/~6B~9
-14- = =
The 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
thieno-[2,3-b][1,5]benzodiazepine compound is effective
over a wide dosage range, the actual dose administered
being dependent on the condition being treated. For
example, in the treatment of adult humans, dosages of from
about 0.25 to 50 mg, preferably from 1 to 30 mg, and most
preferably 1 to 20 mg per day may be used. A once a day
dosage is normally sufficient, although divided doses may
be administered. For treatment of cognitive dysfunction, a
dose range of from 1 to 30 mg, preferably 1 to 20 mg per
day is suitable. Radiolabelled 2-methyl-4-(4-methyl-1-
piperazinyl)-lOH-thieno-[2,3-b][1,5]benzodiazepine, can be
detected in the saliva and thus the compound can
potentially be monitored in patients to assess compliance.
A preferred formulation of the invention is a
solid oral formulation comprising from about 1 to about 20
mg or 1 to 10 mg of active anhydrous Form I 2-methyl-4-(4-
methyl-1-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine
as an effective amount of the active ingredient.
Most preferably, the solid oral formulation is
contained in packaging materials which protect the
formulation from moisture and light. For example, suitable
packaging materials include amber colored high density
polyethylene bottles, amber colored glass bottles, and other
containers made of a material which inhibits the passage of
light. Most preferably, the packaging will include a
desiccant pack. The container may be sealed with an aluminum
foil blister to provide the desired protection and maintain
product stability.
The 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
thieno-[2,3-b][1,5]benzodiazepine compound will normally be
- administered orally or by injection and, for this purpose,
it is usually employed in the form of a pharmaceutical
composition.
Accordingly, pharmaceutical compositions
comprising 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
thienot2,3-b][1,5]benzodiazepine, as active ingredient
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associated with a pharmaceutically acceptable carrier may
be prepared. In making the compositions of the invention
conventional techniques for the preparation of
pharmaceutical compositions may be used. For example, the
active ingredient will usually be mixed with a carrier, or
diluted by a carrier, or enclosed within a carrier which
may be in the form of a capsule, sachet, paper or other
container. When the carrier serves as a diluent, it may be
solid, semi-solid or liquid material which acts as a
vehicle, excipient or medium for the active ingredient. The
active ingredient can be adsorbed on a granular solid
container for example in a sachet. Some examples of
suitable carriers are lactose, dextrose, sucrose, sorbitol,
mannitol, starches, gum acacia, calcium phosphate,
alginates, tragacanth, gelatin, syrup, methyl cellulose,
methyl- and propyl-hydroxy-benzoate, talc, magnesium
stearate or mineral oil. The compositions of the invention
may, if desired, be formulated so as to provide quick,
sustained or delayed release of the active ingredient after
administration to the patient. For example, one such
pre~erred quick release formulation is described in U.S.
Patent Nos. 5,079,018, 5,039,540, 4,305,502, 4,758,598, and
4,371,516, hereby incorporated by reference. Such
formulation most preferably comprises 2-methyl-4-(4-methyl-
1-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine, water,
hydrolyzed gelatin, and mannitol.
Dep~n~;ng on the method of administration, the
compositions for the treatment of central nervous system
conditions may be formulated as tablets, capsules,
injection solutions for parenteral use, gel or suspension
for transdermal delivery, suspensions or elixirs for oral
use or suppositories. Preferably the compositions are
formulated in a unit dosage form, each dosage containing
from 0.25 to 100 mg, more usually 1 to 30 mg, of the active
ingredient. When a sustained release formulation is
desired, the unit dosage form may contain from 0.25 to 200
mg of the active ingredient. A preferred formulation of
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WO96/38151 PCTrUS95/06859
-16-
the invention is a capsule or tablet comprising 0.25 to 75
mg or 1 to 30 mg of active ingredient together with a
pharmaceutically acceptable carrier therefor. A further
preferred formulation is an injection which in unit dosage
form comprises 0.25 to 30 mg or 1 to 30 mg of active
ingredient tagether with a pharmaceutically acceptable
diluent therefor.
The materials for the present invention can be
purchased or prepared by a variety of procedures well known
to those of ordinary skill in the art. The 2-methyl-4-(4-
methyl-l-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine
compound can be prepared as described by Chakrabarti in U.S.
Patent No 5,229,382 ('382), herein incorporated by reference
in its entirety. It is most desirable to prepare a rapidly
dissolving formulation comprising substantially pure
crystalline Form I 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
thieno[2,3-b][1,5]benzodiazepine. Such substantially pure
crystalline Form I 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
thieno[2,3-b][1,5]benzodiazepine may be prepared using the
techniques described herein by the Preparation section herein
infra .
As used herein mixing steps may be accomplished
using common agitation methods such as stirring, shaking, and
the like. As used herein the phrase ~producing crystalline
product from the mixture" shall refer to crystallization from
the stated mixture of compound and solvent. Further, the
artisan recognizes that crystallization processes may include
seeding, chilling, scratching the glass of the reaction
vessel, and other such common techniques.
Compound characterization methods include, for
example, x-ray powder pattern analysis, thermogravimetric
analysis (TGA), differential ss~nning calorimetery (DSC),
titrametric analysis for water, and lH-NMR analysis for
solvent content.
The following examples are provided for purposes of
illustration and are not to be construed as limiting the
scope of the claimed invention.
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W O96/38151 PCTrUS95106859
Pre~aration
Crystalline Form II 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
thieno[2,3-b][1,5]benzodiazepine
A 10 gram sample of crude 2-methyl-4-(4-methyl-1-
piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine was
suspended in methylene chloride (lOO)gm and stirred at
ambient temperature (20-25~C) for a period of 1 hour. The
slurry was vacuum filtered and the filtrate was recovered.
The stirred filtrate was chilled to 0-5~C in an ice bath and
the solvent was slowly evaporated under a stream of nitrogen
to a thick paste. Approximately 3/4 of the solvent was
removed by evaporation. A quantity of prechilled methylene
chloride (30 gm, 0-5~C) was mixed into the thick paste. The
resulting slurry was vacuum filtered and allowed to air dry
on the filter. The isolated solid was further dried in a
vacuum oven at 50~C for a period of 30 minutes. Isolated:
4.8 gm. X-ray powder characterization: Form II + CH2Cl2
Solvate.
The isolated solid was redried in a vacuum oven at
50~C under a stream of nitrogen for a period of 30 hours.
Isolated: 4.5 gm X-ray powder characterization: Form II.
(described supra. )
Pre~aration 2
Form I 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][l,5]benzodiazepine
A sample of ethyl acetate which was saturated with
technical grade 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
thieno[2,3-b][1,5]benzodiazepine was contacted with Form II
2-methyl-4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine (0.3g), a seed of anhydrous Form I 2-
methyl-4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine and stirred at about 25~C for about 5
hours. The reaction product was isolated by vacuum
CA 02219902 1997-10-29
WO 961381Sl PCTrUS95/06859
-18-
~iltration and dried under ambient conditions. Yield: 0.25
g. X-ray powder analysis indicated that the product was
anhydrous Form I 2-methyl-4-(4-methyl-1-piperazinyl) -10H-
thieno[2,3-b][1,5]benzodiazepine.
PreDaratiQn 3
Technical Grade 2-methyl-4-(4-methyl-1-piperazinyl)-1 OH-
thieno[2,3-b][1,5]benzodiazepine
~~
Intermediate 1
In a suitable three neck flask the ~ollowing was added:
Dimethylsul~oxide (analytical): 6 volumes
Intermediate 1 : 75 g
N-Methylpiperazine (reagent) : 6 equivalents
Interm~iate 1 can be prepared using methods known to the
skilled artisan. For example, the preparation o~ the
Interm~iate 1 is taught in the '382 patent.
A sub-sur~ace nitrogen sparge line was added to remove the
Amm~i a formed during the reaction. The reaction was heated
to 120~C and maintained throughout the duration of the
reaction. The reactions were ~ollowed by HPLC until < 5% o~
the intermediate 1 was le~t unreacted. After the reaction
was complete, the mixture was allowed to cool slowly to 20~C
(about 2 hours). Each reaction mixture was then trans~erred
to an appropriate three neck round bottom ~lask and water
bath. To this solution with agitation was added 10 volumes
reagent grade methanol and the reaction was stirred at 20~C
CA 02219902 1997-10-29
W O96/38151 PCTrUS95/06859
--19--
~or 30 minutes. Three volumes of water was added slowly over
about 30 minutes. The reaction slurry was cooled to zero to
5~C and stirred for 30 minutes. The product was filtered and
the wet cake was washed with chilled methanol. The wet cake
was dried in vacuo at 45~C overnight. The product was
identified as technical 2-methyl-4-(4-methyl-1-piperazinyl)-
lOH-thieno[2,3-b][1,5]benzodiazepine.
Yield: 76.7%; Potency: 98.1%
The procedure of Preparation 3 was repeated substantially as
described above and provided a yield of 81% with a potency of
101 . 1% .
Pre~aration 4
Technical Grade 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
thieno[2,3-b] [1,5]benzodiazepine
Intermediate 1 ( supra ) was suspended in DMSO (3.2
vol.) and toluene (4.5 vol.). A portion (~0.65 vol.) of the
solvent was removed by distillation at 120-125~C. The
mixture was cooled to 110~C, N-methylpiperazine (NMP, 4.2
equiv.) was added and the mixture heated back to reflux (120-
125~C). Another portion (--1 vol.) of the solvent was removed
by distillation to dry the reaction mixture. A vigorous
reflux was desired to drive the reaction to completion (about
7 hrs.) by removing ammonia from the reaction. The product
was isolated by the slow addition of water (12.75 vol.) to
the cooled (10~C) reaction solution. The product was
collected by filtration and washed with chilled water (2
vol.). The crude 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-
4 thieno[2,3-b] [1,5]benzodiazepine was dried in vacuo at 60~C.
The product was recrystallized from hot toluene (5 vol.) to
give a technical grade 2-methyl-4-(4-methyl-1-piperazinyl)-
lOH-thieno[2,3-b] [1,5]benzodiazepine. After drying in vacuo
at 50~C, the technical grade 2-methyl-4-(4-methyl-1-
piperazinyl)-lOH-thieno[2,3-b] [1,5]benzodiazepine was
recrystallized again from ethyl acetate (10 vol.)/toluene
CA 022l9902 l997-l0-29
W O96/38151 PCTAUS95/06859
-20-
(0.62 vol.)/methanol (3.1 vol.) to give 2-methyl-4-(4-methyl--
1-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine as a
methanol solvate. The methanol solvate upon drying at >50~C
was converted to an anhydrous technical grade 2-methyl-4-(4-
methyl-1-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine.
Pre~aration S
Form I from acetone
A 3.0 g sample of technical grade 2-methyl-4-(4-
methyl-1-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine
was suspended in acetone (30 g). The mixture was stirred and
heated to about 60~C. The mixture was maintained at about
60~C for about 30 minutes. The mixture was allowed to cool
to about 25~C. The resulting product was isolated using
vacuum filtration. The product was identified as Form I 2-
methyl-4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine using x-ray powder analysis. Yield:
0.8 g.
Pre~aration 6
Form I using tetrahydrofuran
An 8.0 g sample of technical grade 2-methyl-4-(4-
methyl-1-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine
was suspended in tetrahydrofuran (25 g). The mixture was
stirred and heated to about 60~C. The mixture was maintained
at about 60~C for about 30 minutes. The mixture was allowed
to cool to about 25~C. The resulting product was isolated
using vacuum filtration. The product was identified as Form
I 2-methyl-4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine using x-ray powder analysis. Yield:
1.3 g.
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Pre~aration 7
Form I using ethyl acetate
A 270 g sample of technical grade 2-methyl-4-(4-
methyl-1-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine
was suspended in ethyl acetate (2.7 L). The mixture was
heated to about 76~C and maintained at about 76~C ~or about 30
minutes. The mixture was allowed to cool to about 25~C. The
resulting product was isolated using vacuum ~iltration. The
product was identified as Form I 2-methyl-4-(4-methyl-1-
piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine using x-ray
powder analysis.
Yield: 197 g.
Pre~aration 8
Form I from t-butanol
A 1.0 g sample o~ technical grade 2-methyl-4-(4-
methyl-1-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine
was suspended in tert-butanol (30 g). The stirred mixture
was heated to about 60~C and maintained at about 60~C ~or
about 30 minutes. The mixture was allowed to cool to about
25~C. The resulting product was isolated using vacuum
~iltration. The product was identi~ied as Form I 2-methyl-4-
(4-methyl-1-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine
using x-ray powder analysis.
Yield: 0.3 g.
Pre~aration 9
Form I ~rom Slurry Conversion of Form II in Toluene
A 0.5 g sample o~ technical grade 2-methyl-4-(4-
methyl-1-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine
and a 0.5 g sample o~ Form II 2-methyl-4-(4-methyl-1-
piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine were
suspended in toluene (5 ml), presaturated with 2-methyl-4-(4-
CA 02219902 1997-10-29
WO 96138151 PCTrUS95/06859
methyl-l-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine.
The mixture was stirred in a sealed vial at about ambient
temperature ~or about 22 hours. The resulting product was
isolated using vacuum filtration and dried under vacuum at
about 45~C. The product was identi~ied as Form I 2-methyl-4-
(4-methyl-1-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine
using x-ray powder analysis.
EXAMPLE
A portion of the hydroxypropyl cellulose was
dissolved in puri~ied water to form a solution for
granulation. The r~m~; n; ng hydroxypropyl cellulose (total of
4.0% w/w final tablet weight), which was an extra ~ine grade,
was combined with the 2-methyl-4-(4-methyl-1-piperazinyl)-
lOH-thieno[2,3-b][1,5]benzodiazepine (1.18% w/w), lactose
(79.32% w/w) and a portion of the crospovidone (5% w/w) in a
high shear granulator. All ingredients were security sieved
prior ~o addition and dry blended in the graFlulator. This
mixture was then granulated with the hydroxypropyl cellulose
solution in the high shear granulator. The granulation was
wet sized using standard methods. The wet granulation was
then dried in a fluidized bed dryer and sized. The material
was then added to a tumble bin mixer.
The running powders consisting of microcrystalline cellulose
(granular) (10% w/w), magnesium stearate (0.5% w/w), and the
remainder of the crospovidone were added to the sized
granulation. The mixture was blended and compressed with the
appropriate tooling on tablet compression eauipment.
Subcoatina:
Hydroxypropyl methylcellulose (10% w/w) was mixed
with purified water to form a solution. Core tablets were
divided into approximately equal sections and spray coated
with the hydroxypropyl methylcellulose solution. The
operation was performed in a per~orated coating pan.
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W O961381S1 PCTrUS9~/06859
-23-
Coatina of Core Tablets:
Color Mixture White (hydroxypropyl methylcellulose,
polyethylene glycol, polysorbate 80, and titanium dioxide)
was mixed with purified water to form the coating suspension.
Subcoated tablets were divided into approximately equal
sections and spray coated with the coating suspension
described above. The operation was performed in a perforated
coating pan.
The coated tablets were lightly dusted with carnauba wax
and imprinted with appropriate identification.
EXAMPLE 2
The process substantially as described above in
Example 1 was repeated using the following ingredients to
provide pharmaceutically elegant tablet formulations
containing 1, 2.5, 5, 7.5, and 10 mg 2-methyl-4-(4-methyl-
l-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine,
respectively, per tablet:
1 ma 2-methyl-4-(4-methvl-1-~i~erazinyl)-lOH-thienor2,3-
blrl,5lbenzodiaze~ine ~e~ tablet:
Name~ of IngrnaiQnt~ Quantity
(mg/tabl~t)
Activ~ Ingr~ai~nt
2-methyl-4-(4-methyl-l- l.O
piperazinyl)-lOH-thieno[2,3-
b][l,5]benzodiazepine
OthQr Ingr~ai~lnt~
Lactose 67.43
Hydroxypropyl Cellulose 3.40
Crospovidone 4.25
Microcrystalline Cellulose 8.50
Magnesium Stearate 0.42
Subcoating
Hydroxypropyl l.70
Methylcellulose
Coating
Color Mixture White 3.47
Poli~hing
Carnauba Wax trace
Imprinting
Edible Blue Ink trace
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W 096/381S1 PCTrUS95106859
-24-
2-methvl- 4-(4 -methvl-1-~i~erazinvl)-lOH-thieno r 2,3-
bl~1,51benzodiaze~ine 2.5 ma Tablets
Names o~ IngrQaients Quantity
(mg/tablet )
Active Ingredient
2-methyl-4-(4-methyl-l- 2.5
piperazinyl)-lOH-thieno[2,3-
b][l,5]benzodiazepine
othQr Ingrodients
Lactose 102.15
Hydroxypropyl Cellulose 5.20
Crospovidone 6.50
Microcrystalline Cellulose 13.00
Magnesium Stearate 0.65
Subcoating
Hydroxypropyl 2.60
Methylcellulose
Coating
Color Mixture White 5.30
Poli~hing
Carnauba Wax trace
Imprinti~g
Edible Blue Ink trace
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W 096/38151 PCTrUS95/06859
2-methvl-4-(4-methvl-1-~iperazinvl)-lQH-thieno r 2,3-
blrl,51benzodiaze~ine 5.0 ma Tablets
A
Namo~ of Ingroai~nts Quantity
~ (mg/tablot)
Activo Ingr~dient
2-methyl-4-(4-methyl-l- 5.00
piperazinyl)-lOH-thieno[2,3-
b][l,5]benzodiazepine
Othor IngrQaionts
Lactose 156.00
Hydroxypropyl Cellulose 8.00
Crospovidone . lO.OO
Microcrystalline Cellulose 20.00
Magnesium Stearate l.OO
Subcoating
Hydroxypropyl 4.00
Methylcellulose
Coating
Color Mixture White 8.16
Polishing
Carnauba Wax trace
Imprinting
Edible Blue Ink trace
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-26-
2-methyl-4-(4-methyl-1-piperazinvl)-lOH-thieno r 2,3-
blrl,51benzodiaze~ine 7.5 m~ Tablets
N~me~ o~ Ingr~di~nts Qu~ntity
(mg/tabl~t)
Active Ingr~aient
2-methyl-4-(4-methyl-1- 7.50
piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine
Other Ingredi~nts
Lactose 234 00
Hydroxypropyl Cellulose 12.00
Crospovidone 15.00
Microcrystalline Cellulose 30.00
Magnesium Stearate 1.50
Subcoating
Hydroxypropyl 6.00
Methylcellulo~e
Coating
Color Mixture White 12.24
Polishing
Carnauba Wax trace
Imprinting
Edible Blue Ink trace
CA 02219902 1997-10-29
W 096/38151 PCTrUS9~/O~P~9
2-methyl-4-(4-methyl-1-~i~erazinvl)-1 OH- thieno r 2,3-
blrl.5lbenzodiaze~ine 10.0 ma Tablets
o
Namns of Ingreaisnts Quantity
(mg/tablQt )
Active Ingreaient
2-metnyi-4-(4-methyl-i- lû. ûû
piperazinyl)-lûH-thieno[2,3-
b][1,5]benzodiazepine
Other Ingreaients
Lactose 312.00
Hydroxypropyl Cellulose 16.00
Crospovidone 20 00
Microcrystalline Cellulose 40.0û
Magnesium Stearate 2.00
Subcoating
Hydroxypropyl 8.ûO
Methylcellulose
Coating
Color Mixture White 16.32
Polinhing
Carnauba Wax trace
Imprinting
Edible Blue Ink trace
EXAMPLE 4
Pul w le Formulation
A pul w le ~ormulation is prepared by blending the
active with silicone starch, and ~illing it into hard
gelatin capsules.
Per 300 mg capsule
Compound o~ the invention30.0 mg
Silicone 2.9 mg
- Starch ~lowable 267.1 mg
~ 15
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W O96/38151 PCTrUS95/06859
-28-
EXAMPLE 5
Tablet Formulation
A tablet formulation is made by granulating the active
with appropriate diluent, lubricant, disintegrant and
binder and compressing
Compound of the invention10.0 mg
Magnesium stearate 0.9 mg
Microcrystalline cellulose75.0 mg
Povidone 15.0 mg
Starch, directly compressible204.1 mg
EXAMPLE 6
Aqueous Injection Formulation
An aqueous injection of active is prepared as a
freeze-dried plug, for reconstitution in a suitable,
sterile diluent before use (to a total volume of 10 ml).
Compound of the invention is contacted with Mannitol N
Hydrochloric acid and/or N sodium hydroxide to adjust pH to
5-5.5.
Compound of the invention 20.0 mg
Mannitol 20.0 mg
N Hydrochloric acid and/or N
sodium hydroxide to adjust pH to
5-5.5.
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W O96/38151 PCTrUS95106859
-29-
EXAMPLE 7
Controlled Release IM Formulation
,,
A controlled release injection for intramuscular
injection is formed from a sterile suspension of micronised
active in an oleaginous vehicle.
Compound of the invention 50.0 mg
Aluminium stearate 0.04 mg
Sesame oil 2 ml
EXAMPLE 8
Capsule Formulation
A formulation is prepared by blending the active
with silicone starch and starch, and filling it into hard
gelatine capsules.
Per 300 mg capsule
Compound of the invention 2.5 mg
Starch flowable with 0.96%
silicone 220 222.5 mg
Starch flowable 75.0 mg
Ry~ le 9
2-methyl-4-(4-methyl-1-piperazinyl)-lOH-thieno[2,3-
b][l,5]benzodiazepine Granules
The granules were produced by blending the mannitol and
7 Hydroxymethyl propyl cellulose in a high shear mixer;
granulating with the aqueous suspension of 2-methyl-4-(4-
methyl-l-piperazinyl)-lOH-thieno[2,3-b][1,5]benzodiazepine
and polysorbate 20; wet sized and subsequently dried in a
fluid bed dryer. These are dry sized and reblended prior to
packaging.
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W O96/38151 PCTrUS95/06859
-30-
1 a. 250mg Sachets
INGREDIENT MG/SACHET
Active
2-methyl-4-(4-methyl-1- 250
piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine
Other Ingredients
Mannitol 234.97
Hydroxypropyl methyl 12.50
cellulose 3cps
Polysorbate 20 0.028
1 b. 750mg Sachets
INGREDIENT MG/SACHET
Active
2-methyl-4-(4-methyl-1- 750
piperazinyl)-lOH-thieno[2,3-
b][1,5]benzodiazepine
Other Ingredients
Mannitol 704.93
Hydroxypropyl methyl 37.49
cellulose 3cps
Polysorbate 20 0.08
lc. lOOOmg Sachets
INGREDIENT MG/SACHE~
Active
2-methyl-4-(4-methyl-1- 1000
piperazinyl)-lOH-thieno[2,3-
b]~1,5~benzodiazepine
Other Ingredients
Mannitol 939.90
Hydroxypropyl methyl 49.99
cellulose 3cps
Polysorbate 20 0.11
Such granules are most preferably contacted with an
acidic medium if a suspension or solution is desired.
