U.S. patent application number 09/769740 was filed with the patent office on 2002-01-17 for pyrrolo {2,1-b}{1,3}benzothiazepines with atypical antipsychotic activity.
Invention is credited to Campiani, Giuseppe, Cesare, Maria Assunta Di, Minetti, Patrizia, Nacci, Vito.
Application Number | 20020006921 09/769740 |
Document ID | / |
Family ID | 11380538 |
Filed Date | 2002-01-17 |
United States Patent
Application |
20020006921 |
Kind Code |
A1 |
Campiani, Giuseppe ; et
al. |
January 17, 2002 |
Pyrrolo {2,1-b}{1,3}benzothiazepines with atypical antipsychotic
activity
Abstract
Polycondensated heterocycles with a
pyrrole[2,1-b][1,3]benzothiazepine structure of the following
formula (I) 1 where the groups are defined as in the description
are disclosed. As compared to known antipsychotic agents, these
compounds present substantial activity associated with a
simultaneous reduction in unwanted extrapyramidal symptoms. These
compounds can be formulated in pharmaceutical compositions for the
treatment of psychoses such as, for example, schizophrenia.
Inventors: |
Campiani, Giuseppe;
(Chianciano Terme, IT) ; Nacci, Vito; (Siena,
IT) ; Minetti, Patrizia; (Rome, IT) ; Cesare,
Maria Assunta Di; (Rome, IT) |
Correspondence
Address: |
NIXON & VANDERHYE P.C.
8th Floor
1100 North Glebe Road
Arlington
VA
22201
US
|
Family ID: |
11380538 |
Appl. No.: |
09/769740 |
Filed: |
January 26, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
09769740 |
Jan 26, 2001 |
|
|
|
PCT/IT99/00240 |
Jul 27, 1999 |
|
|
|
Current U.S.
Class: |
514/211.12 ;
540/547 |
Current CPC
Class: |
A61P 25/28 20180101;
A61P 25/22 20180101; A61P 25/18 20180101; A61P 25/00 20180101; A61P
25/24 20180101; C07D 513/04 20130101; A61P 43/00 20180101 |
Class at
Publication: |
514/211.12 ;
540/547 |
International
Class: |
A61K 031/554; C07D
497/02 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 28, 1998 |
IT |
MI98A001748 |
Claims
1. Compounds, in racemic form or as isolated optical isomers,
having formula (I): 10where: R=H, Cl, Br, F, I, C.sub.1-C.sub.4
alkoxy, C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkyl,
C.sub.5-C.sub.6 cycloalkyl; R.sub.1=dialkylamine,
4-alyl-1-piperazinyl, 4-hydroxyalkyl-1-piperanyl, 1-imidazolyl,
4-alkyl-1-piperidinyl R.sub.2=hydrogen, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.4 alkylthio and the pharmaceutically acceptable salts
thereof.
2. Compounds according to claim 1, where R represents chlorine or
hydrogen, R.sub.1 a 4-methyl-1-piperazinyl group; said R.sub.2
hydrogen.
3. Compounds according to claim 1, where R represents fluorine,
bromine or hydrogen, R.sub.1 a 4-methyl-1-piperaiznyl group; and
R.sub.2 hydrogen.
4.
(.+-.)-9-(4-methylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-b][1,3]benzot-
hiaze-pine.
5. (.+-.)-7-chloro-9-(4-methylpiperazin-1-yl)
-9,10-dihydropyrrolo[2,1-b][- 1,3]benzothiazepine.
6. (.+-.) -7-chloro-9-(4-methylpiperazin-1-yl) -9,10-dihydropyrrolo
[2,1-b][1,3]benzothiazepine.
7.
(.+-.)-7-fluoro-9-(4-methylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-b][1-
,3]benzothiazepine.
8.
(.+-.)-7-fluoro-9-(4-ethylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-b][1,-
3]benzothiazepine.
9.
(.+-.)-7-fluoro-9-(4-hyroxyethylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-
-b][1,3]benzothiazepine.
10.
(.+-.)-7-Bromo-9-(4-methylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-b][1-
,3]benzothiazepine.
11. Process for the preparation of the compounds of claims 1-10
comprising the cyclisationl reaction of a derivative containing a
phenyl group and a pyrrole group, and where the cyclisation leads
to the formation of a 1,3 thiazepine ring.
12. Process according to claim 11, where the result of said
cyclisation reaction is a pyrrolobenzothiazepinone, which is
transformed into a formula (I) derivative by substituting the keto
group on the thiazepine ring with a group selected from the
foregoing definitions of the radical R.sub.1.
13. Process according to claim 11, where said derivative containing
a phenyl group and a pyrrole group is a
1-[2-(methylsulphinyl)phenyl]-2-(py- rrol-1-yl)ethanone, and the
cyclisation reaction is carried out in the presence of
trifluoracetic anhydride and dimethylformamide.
14. Process according to claim 11, where said derivative containing
a phenyl group and a pyrrole group is an alkyl ester of
2-(phenylthio)pyrrolo-1-selenoacetic acid and the cyclisation
reaction is carried out in the presence of a crystalline complex of
triflate copper(I) and benzene.
15. Process for the resolution of racemic compounds of claims 1-10
into the corresponding optically active isomers, by means of
fractionated crystallisation of the diastereoisomeric salts
obtained by salification with an optically active acid.
16. Compounds of claims 1-10 for use in therapy in racemic form or
as single isolated optical isomers.
17. Pharmaceutical composition comprising, in racemic form or as
single isolated optical isomers, at least a compound of claims 1-10
in combination with pharmaceutically acceptable excipients and/or
vehicles, and optionally with other active ingredients which are
useful in the treatment of psychoses.
18. Pharmaceutical composition according to claim 17, in solid or
liquid form.
19. Use of the compounds of claims 1-10, in racemic form or as
single isolated optical isomers, in the preparation of an
antipsychotic medicinal agent for the treatment and prevention of
psychoses.
20. Use according to claim 19, where said psychosis takes the form
of schizophrenia, paranoid states, manic-depressive states,
affective disorders, social withdrawal, personality regression, or
hallucinations.
21. Use according to claim 19, in particular of the compounds of
claim 1, wherein R is hydrogen, fluorine, bromine, for the
treatment of the negative symptoms of schizophrenia involving the
emotional and cognitive spheres.
22. Use according to claim 21, where said negative symptom takes
the form of dementia.
Description
[0001] The present invention relates to the field of antipsychotic
drugs, in particular to polycondensated heterocycles with a
pyrrolo[2, 1-b][1,3]benzothiazepine structure.
STATE OF THE ART
[0002] The intervention of dopamine and dopaminergic neurons in the
pathology of a variety of psychiatric and neurological disorders
has been amply documented (Caine, D. B., Therapeutics and
Neurology; Blackwell Scientific Publications, Oxford 1980, p. 281).
In addition, it is also known that drugs which are active on
dopamine receptors may play an important role in the therapy of
such disorders; there is therefore considerable interest in the
effects of dopamine agonist and antagonist compounds on
dopaminergic is receptors, particularly with a view to their
therapeutic implications.
[0003] Chlorpromazine and aloperidol have long been the treatment
of choice for acute and chronic schizophrenia. It has been
postulated that these drugs relieve the positive symptoms of the
disease by blocking dopaminergic transmission in certain areas of
the brain. Chlorpromazine and aloperidol are defined as "typical
neuroleptic agents": their action is characterised by remission of
the symptoms of schizophrenia, accompanied, however, by unwanted
extrapyramidal collateral symptoms (motor disorders, catalepsy,
hyperprolactinaemia, etc.). The elimination of these adverse
effects therefore constitutes an important objective in the
development of new neuroleptic therapies.
[0004] Clinical trials have demonstrated that not only dopamine
antagonists but also 5-HT.sub.2 antagonist compounds are capable of
improving the symptoms of schizophrenia; in particular, it has been
observed that the co-administration of 5-HT.sub.2 antagonists and
"typical" antipsychotic agents reduces the incidence of
extrapyramidal symptoms as compared to treatment with neuroleptic
agents alone (Psychopharmacol., 99, 1989, S18-S27; Niemegeers et al
in 5-HT.sub.2 Receptor Antagonists in Schizophrenia, Racagni Ed.,
Elsevier Publishers, 1991, Vol 1, pp. 535-537).
[0005] Further developments in this sense have led to the
generation of drugs with a mixed antagonist component, i.e. which
are active on different receptors.
[0006] Clozapine
(8-chloro-11-(4-methyl-1-piperazinyl)-5H-dibenzo[b,e][1,4-
]diazepine) is an antipsychotic agent capable of simultaneously
antagonising dopamine on D.sub.2 receptors and serotonin on
5-HT.sub.2 receptors. This new action profile, called "atypical",
allows schizophrenia to be treated with a lower incidence of
extrapyramidal symptoms (J. Med. Chem., 39, 1996, pp.
1172-1188).
[0007] Unfortunately, the occurrence of cases of agranulocytosis
has limited the therapeutic use of this drug (Lancet. 1975, 2,
657).
[0008] Octoclothepin
(8-chloro-10-(4-methylpiperazino)-10,11-dihydrodibenz-
o[b,f]thiepin) is a compound partly endowed with "atypical"
activity. Its pharmacological activity has been studied in relation
to the optical isomers of this compound (J. Med. Chem., 1991, 34,
2023-2030): a slightly greater effect on schizophrenia by the (S)
form is unfortunately associated with a greater incidence of
extrapyramidal effects, so that its use has been withdrawn from
clinical trials. The (R) isomer presents a more "atypical" profile,
with fewer side effects, but also an inferior general potency.
Moreover, the two isomers prove to be endowed with the same
activity as 5-HT.sub.2 and D.sub.1 antagonists.
[0009] In view of the studies cited above, the need for
antipsychotic agents with substantial therapeutic activity and
without side effects remains unsatisfied. In particular, the search
continues for antipsychotic agents which present greater
neuroleptic activity and a lower incidence of extrapyramidal
effects.
[0010] It has now surprisingly been found that polycondensated
heterocycles with a pyrrolo[2,1-b][1,3]benzothiazepine are endowed
with an interesting pharmacological profile as antipsychotic
activity.
ABSTRACT OF THE INVENTION
[0011] The present invention describes polycondensated heterocycles
with a pyrrolo[2,]-b[1,3]benzothiazepine structure. As compared to
known antipsychotic agents, the compounds according to the
invention present substantial activity associated with a
simultaneous reduction in unwanted extrapyramidal symptoms. The
compounds object of the invention described herein can be
formulated in pharmaceutical compositions for the treatment of
psychoses such as, for example, schizophrenia.
[0012] Accordingly, it is an object of the present invention
polycondensated heterocycles with a
pyrrolo[2,1-b][1,3]benzothiazepine structure as disclosed in the
formula (I) below.
[0013] Another object of the present invention is a process for the
preparation of said compounds.
[0014] Still another object of the present invention is the use of
said compounds as medicaments, in particular as antipsychotic
agents, for the treatment of psychosis, such as schizophrenia,
paranoid states, manic-depressive states, affective disorders,
social withdrawal, personality regression, or hallucinations.
[0015] In its industrial aspects, the present invention provides
pharmaceutical compositions comprising at least a compound of the
invention.
DETAILED DESCRIPTION OF THE INVENTION
[0016] The invention described herein relates to new derivatives
with a neuroleptic action, corresponding to the following
structural formula (I): 2
[0017] where:
[0018] R.dbd.H, Cl, Br, F, I, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkyl, C.sub.5-C.sub.6
cycloalkyl;
[0019] R.sub.1=dialkylamine, 4-alkyl-1-piperazinyl,
4-hydroxyalkyl-1-piperazinyl, 1-imidazolyl,
4-alkyl-1-piperidinyl
[0020] R.sub.2=hydrogen, C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4
alkylthio; and the pharmaceutically acceptable salts thereof.
[0021] The formula (I) derivatives possess a chiral carbon atom in
position 9 on the benzothiazepine ring. The invention described
herein comprises both the formula (I) derivative in racemic form
and the single (R) and (S) isomers, separately.
[0022] In formula (I), R preferably represents bromine, chlorine,
fluorine or hydrogen, more preferably chlorine or fluorine; R.sub.1
is preferably a 4-methylpiperazinyl group; and R.sub.2 preferably
hydrogen.
[0023] Preferred derivatives according to the invention are the
products:
[0024]
(.+-.)-9-(4-methylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-b][1,3]be-
nzothiazepine, hereinafter referred to as (.+-.)-3a;
[0025]
(.+-.)-7-chloro-9-(4-methylpiperazin-1-y)-9,10-dihydropyrrolo[2,1-b-
][1,3]benzothiazepine, hereinafter referred to as (.+-.)-3b
(ST1455);
[0026]
(+)-7-chloro-9-(4-methylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-b][-
1,3]benzothiazepine, particularly preferred, hereinafter referred
to as (+)-3b (ST1460)
[0027]
(.+-.)-7-fluoro-9-(4-methylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1--
b][1,3]benzothiazepine, hereinafter referred to as (3c)
(ST1456);
[0028]
(.+-.)-7-fluoro-9-(4-ethylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-b-
][1,3]benzothiazepine (4c) (ST1457);
[0029]
(.+-.)-7-fluoro-9-[4-(2'-hyroxyethylpiperazin-1-yl]-9,10-dihydropyr-
rolo[2,1b][1,3]benzothiazepine (5c) (ST1458);
[0030] The invention described herein also relates to new,
effective methods of synthesis to obtain the new
pyrrolo[2,1-b][1,3]benzothiazepine structures. One of the problems
encountered was that of realising a cyclisation method that made it
possible to obtain the particular formula (I) tricyclic system with
high yields.
[0031] The various synthesis methods described herein include a
cyclisation reaction of a derivative comprising a phenyl group and
a pyrrole group, where the cyclisation leads to the formation of a
[1,3] thiazepine ring. The result of said cyclisation reaction is
preferably a pyrrolobenzothiazepinone, which can be transformed
into a formula (I) derivative by substituting the keto group on the
thiazepine ring with a group selected from the definitions given
above for the radical R.sub.1.
[0032] A process for the synthesis of formula (I) products is
illustrated in Scheme 1A in its essential steps, and in Scheme 1B
in detail. 3 4
[0033] With reference to Scheme 1B, the process involves the
reaction of 2-thiocyanate-pyrrole (11) with phenyl magnesium
bromide to form thioether (12). Thioether (12), subjected to
esterification and transacylation reactions, gives rise to the
derivative methylselenolate (15) comprising a phenyl group and a
pyrrole group. Derivative (15) is then subjected to a cyclisation
reaction, with formation of product (9a)
(pyrrolobenzothiazepinone). The cyclisation reaction is conducted
in the presence of a crystalline complex of triflate copper (I) and
benzene.
[0034] Lastly, product (9a) is transformed into a formula (I)
derivative by means of subsequent modifications of the keto group
on the thiazepine ring to obtain derivatives with R.sub.1 as
described above.
[0035] One preferred process for the synthesis of formula (I)
products is illustrated in Scheme 2A in its essential steps and in
Schemes 2B in detail. 5 6 7
[0036] With reference to Schemes 2B/1 and 2B/2, the process
involves the formation of the intermediate product (21a,b,c. SCHEME
2B/1; bromophenylethanone; 21a: R.dbd.H; 21b: R.dbd.Cl; 21c=F). As
far as the intermediate compound 21d is concerned, the synthetic
path is outlined in the Scheme C below.
[0037] The intermediate product (21a,b,c,d) is transformed into the
corresponding sulphoxide (23a,b,c,d). The latter, when subjected to
a cyclisation reaction, leads to the formation of the product
(9a,b,c,d) (pyrrolobenzothiazepinone). The cyclisation reaction is
conducted in the presence of trifluoracetic anhydride and
dimethylformamide.
[0038] This second synthesis method is preferred in that the
cyclisation reaction and consequent formation of the thiazepine
ring occur with distinctly greater yields.
[0039] Products (9a), (9b), (9c) and (9d), obtained with the
different synthesis methods described above are then transformed
into a formula (I) derivative by substituting the keto group with a
group selected from the definitions given above for the radical
R.sub.1. 8
[0040] It is then always possible to resolve the racemic products,
however they are obtained, into the two active isomers by means of
fractionated crystallisation of the diastereoisomeric salts
obtained by salification with an optically active acid, such as
tartaric acid, dibenzoyltartaric acid, camphoric acid, or
camphor-sulphonic acid
[0041] A detailed description of the above-mentioned synthesis
methods is presented in the experimental part.
[0042] A further subject of the invention described herein consists
in pharmaceutical compositions comprising formula (I) derivatives
in combination with pharmacologically acceptable excipients and
vehicles and optionally with additional active ingredients which
are useful in the treatment of psychoses.
[0043] Examples of such optional active ingredients are the
phenothiazines (e.g. chlorpromazine), the thiaxanthenes (e.g.
chlorprothixene, titothixene), the butyrophenones (e.g.
aloperidol), the dihydroindolones (e.g. molindone), the
dibenzoxazepines (e.g. loxapine), the Rauwolfia alkaloids, etc.
[0044] Formula (I) compounds can be formulated in solid, liquid or
semisolid pharmaceutical forms. Examples of liquid formulations are
injectable solutions or solutions for oral use, syrups, elixirs,
suspensions and emulsions. Examples of solid forms are tablets,
capsules, microcapsules, powders and granulates.
[0045] Formula (I) compounds are endowed with a pronounced
neuroleptic and antipsychotic activity. This enables them to be
used in the prevention and treatment of psychoses such as
schizophrenia, paranoid states, manic-depressive states, affective
disorders, social withdrawal, personality regression,
hallucinations, appetite disorders (anorexia) and related
disorders. Additional indications may be analgesia/anaesthesia,
neuroleptic anaesthesia, anxiety manifestations in the elderly, and
extrapyramidal disturbances. The invention described herein
therefore includes the use of formula (I) products in the
preparation of medicinal products which are useful for the
prevention and treatment of said disorders.
[0046] Some of the formula (I) products have an interesting
D.sub.3;D.sub.1 ratio, indicating them useful in the treatment of
the negative symptoms of schizophrenia involving the emotional and
cognitive spheres such as, for instance, dementia.
[0047] As documented in the experimental part, the "atypicity" of
the neuroleptic action of formula (I) derivatives makes it possible
to treat the above-mentioned pathologies effectively, at the same
time reducing to a minimum the extrapyramidal side effects normally
associated with the classic antipsychotic agents. The substantial
receptor activity that characterises these compounds also makes it
possible to considerably reduce the dose necessary to achieve a
therapeutic response, thus reducing toxicity and accumulation
phenomena. The reduction of the daily dose is an aspect of
particular interest in the treatment of chronic diseases such as
schizophrenia, which require prolonged exposure to the drug.
[0048] Formula (I) compounds can be administered over a dosage
range generally varying from 0.02 to 200 mg/kg, depending upon the
severity of the disease to be treated and its acute or chronic
component. Doses outside the range indicated are, however, possible
in particular conditions, under the supervision of a doctor.
[0049] The invention is now illustrated by means of the following
examples.
EXPERIMENTAL PART
1. Chemistry
[0050] 1.1 Synthetic Approaches
[0051] The synthesis of the new pyrrolo[2,1-b][1,3]benzothiazepine
structure was accomplished by adopting the two retrosynthetic
approaches described in Schemes 1A and 2A. The synthesis is based
on a cyclisation method to obtain the pyrrolobenzothiazepinone
intermediate products 9a,b. Scheme 1A gives our retrosynthetic
analysis of compound 9a. When pyrrolo was treated with copper
thiocyanate in methanol, thyocyanation occurred in a few minutes,
obtaining 2-thiocyanopyrrolo 11 with good yields. The Grignard
reaction with phenylmagnesium bromide (12), followed by alkylation
with ethyl bromroacetate, produced ester 13 with a high overall
yield.
[0052] Ester 13 may serve as a starting product in carrying out the
cyclisation to 9a, on the basis of an acylation reaction promoted
by coppery(I). In fact, dimethyl aluminium methylselenolate was
used to obtain selenoester 15. Like the thioesters, this
selenoester could be used in the carbon-carbon bond formation
reaction. Consequently, by exposure of 15 to the highly reactive
crystalline complex of triate copper(I) and benzene
[(CuOTf).sub.2PhH], the tricylic compound 9a was obtained together
with ketone 16 and other unidentified reaction products.
[0053] Another synthesis method, shown in Scheme 2A, was based on a
thionium ion intermediate. This made it possible to prepare the key
tricyclic intermediates 9a,b,c,d in conditions of Pummerer
rearrangement, starting from sulphoxide, which in turn were
prepared from 1-[2-(methythio)phenyl]ethanones. As shown in Scheme
2B/1, the key intermediate products 21a,b,c were prepared by
brominiation of the corresponding phenylethanones 18 and 20, which
in turn were prepared, respectively, by reaction of methyllithium
and the lithium salt of 2-(methylthio)benzoic acid 17, or from
(methylthio)benzene, 1-chloro- or 1-fluoro-4-(methylthio)benzene
19a,b,c, with a Friedels-Crafts reaction with acetic anhydride.
Subsequently (SCHEME 2B/2), the bromophenylethanones 21a,b,c,d were
transformed into the pyrrole derivatives 22a,b,c,d. Oxidation, for
example with sodium periodate or 3-chloroperbenzoic acid (MCPBA)
(23a,b,c,d), followed by exposure of these sulphoxides to
trifluoracetic anhycide produced the ketones 9a,b,c,d with a yield
of 40%. The mechanism proposed for the cyclisation stage is given
in Scheme 2B/3. 9
[0054] The "interrupted" Pummerer rearrangement started with
activation of the oxygen of the sulphoxide followed by attachment
of the pyrrole ring to the sulphur which shifted the trifluoracetic
ion. The sulphonium salt then underwent the shift of the methyl
group, generating the new heterocyclic system and
methyltrifluoroacetate. Starting from the ketones 9a,b,c,d (FIG. 2
B/2), the piperazine ring was introduced according to a standard
method. Reduction of ketones 9a,b,c,d then yielded the alcohols
(.+-.)-24a,b,c,d which were transformed into the corresponding
derivatives (.+-.)-25a,b,c,d by means of PBr.sub.3. By treating
(.+-.)-25a,b,c,d with N-methylpiperazine, the end product
(.+-.)-3a,b,c,d was obtained. The thiazepine (.+-.)-3a,b was
resolved into the enantiomorphs (+)-3b and (-)-3b by means of HPLC,
using a Chiralpak AD amylose column, or equivanent means.
[0055] 1.2 Materials and Methods
[0056] The melting points were determined using an Electrothermal
8103 device and were not corrected. The IR spectra were recorded
with Perkin-Elmer 398 and FT 1600 spectrophotometers. The
.sup.1H-NMR spectra were recorded with a Bruker 200 MHz
spectrometer and a Varian 500 MHz spectrometer with TMS as internal
standard; the chemical shift values (.delta.) are given in ppm and
the coupling constants (J) in Hertz. All reactions were carried out
in an argon atmosphere. Progress of the reactions was monitored by
TLC on silica gel plates (Riedel-de-Haen, Art. 37341). Merck silica
gel (Kieselgel 60) was used for the chromatography columns (70-230
mesh) and for the flash chromatography columns (230-400 mesh).
Exacts were dried on MgSO.sub.4 and the solvents removed at reduced
pressure. The HPLC separation was carried out using a Chiralpak AD
Amylose column (097-702-40808) (length.times.diameter=250
mm.times.10 mm). The elemental analyses were carried out on a
Perkin-Elmer 240C elemental analyzer, and the results are within
0.4% of the theoretical value, unless otherwise specified. The
yields refer to purified, non-optimised products.
[0057] 1.3 Preparation of the Compounds
[0058] 2-(phenylthio)pyrrole (12)
[0059] To a solution of phenyl magnesium bromide (prepared from
bromobenzene (0.75 ml) and magnesium chips (0.19 g, 7.8 mmol) in
anhydrous THF (20 ml), cooled to 0.degree. C., was slowly added a
solution of 2-thiocyanopyrrole 11 (0.5 g, 4.0 mmol) in anhydrous
THF (20 ml). After stirring at 0.degree. C. for 30 min, the mixture
was poured into crushed ice and extracted with ethyl acetate. The
organic phase was washed with 20% NH.sub.4Cl, anhydride and
evaporated. The residue was purified by chromatography (35% hexane
in chloroform) to give 0.6 g (93% yield) of 12 as colourless
prisms: melting point 86-87.degree. C. (hexane); IR (CHCl.sub.3)
3420 cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta.8.20 (br s, 1H),
7.25-6.85 (m, 5H), 6.92 (m, 1H), 6.55 (m, 1H), 6.31 (m, 1H). Anal.
(C.sub.9H.sub.9NS): C, H, N.
[0060] Ethyl ester of 2-(phenylthio)pyrrole-1-acetic acid (13)
[0061] To a mixture of 18-Crown-6 (20 mg, 0.074 mmol) and potassium
terbutoxide (0.166 mg, 1.48 mmol) in anhydrous THF (5 ml) was added
a solution of 12 (0.2 g, 1.14 mmol) in ahydrous THF (5 ml) under
nitrogen. After 2 hours at ambient temperature, a solution of ethyl
bromoacetate (0.254 ml, 2.28 mmol) in anhydrous THY (1 ml) was
added dropwise. After stirring for 30 min at ambient temperature
and the addition of 5 ml of water, the solvent was removed at
reduced pressure and the residue extracted with EtOAc. The organic
layers were washed with a saturated solution of NaCl, anhydrified
and evaporated. The residue was chromatographed (30% hexane in
chloroform) to give 0.28 g (96% yield) of 13 as colourless prisms:
melting point 101-102.degree. C. (cyclohexane); IR (CHCl.sub.3)
1760 cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta.7.25-6.90 (m, 5H);
6.61 (m, 1H), 6.31 (m, 1H), 4.68 (s, 2H), 4.05 (q, 2H, J=7.0 Hz),
1.14 (t, 3H, J=7.1 Hz). Anal. (C.sub.14H.sub.15NO.sub.2S): C, H,
N.
[0062] Methyl ester of 2-(phenylthio)pyrrole-1-selenoacetic acid
(15)
[0063] A solution of dimethylaluminium methylselenolate (2.2 mmol)
prepared by heating a solution of trimethylaluminium in toluene
with selenium in powder form for 2 hours at reflux temperature
under argon) in anhydrous toluene (1.1 ml) was added dropwise to a
solution of 13 (0.57 g, 2.2 mmol) in anhydrous dichloromethane (5
ml) cooled to 0.degree. C., under nitrogen. The mixture was
agitated at 0.degree. C. for 45 min, heated to ambient temperature
and stirred for another 45 min. Water (2 ml) was added and the
mixture was extracted with EtOAc. The organic layers were washed
with a saturated NaCl solution, anhydrified and evaporated. The
crude product was purified by distillation (85.degree. C./0.1 mm
Hg) to give 0.6 g (97% yield) of 15 as a colourless oil: IR (t.q.)
1720 cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta.7.24-6.95 (m, 6H);
6.69 (m, 1H), 6.37 (m, 1H), 4.69 (s, 2H), 2.11 (s, 3H) m/z 311 (40,
M.sup.+), 188, 155, 109, 91 (100). Anal. (C.sub.13H.sub.13SeNOS):
C, H, N.
[0064] 1-[2-(methylthio)phenyl]ethanone (18)
[0065] To a suspension of lithium hydride (0.57 g, 6.7 mmol) in
anhydrous 1,2-dimethoxyethane (5 ml), stirred vigorously, was added
dropwise a solution of acid 17 (1.0 g, 5.9 mmol) in anhydrous
1,2-dimethoxyethane. The suspension was refluxed for 2,5 hours,
cooled to -10.degree. C. and added with methyl-lithium (4.2 ml, 6.7
mmol, 1,6 M) in the space of 30 min. The reaction mixture was
stirred for 2 hours at ambient temperature. HCl 1N was added to the
mixture, which was extracted with ethyl ether. The organic layers
were washed with a saturated NaCl solution, anhydrified and
concentrated. Chromatography of the crude product (5% benzene in
dichloromethane) gave 0.78 g (79% yield) of 18 as colourless
prisms, the spectroscopic data for which were identical to those
reported in the literature
[0066] 1-[5-chloro-2-(methylthio)phenyl]ethanone (20b)
[0067] A mixture of 19b (1.0 g, 6.3 mmol), anhydrous aluminium
chloride (1.88 g, 13.5 mmol) and carbon sulphide (20 ml) was heated
to reflux under argon and acetic anhydride (0.46 ml, 6.3 mmol) was
added dropwise. After reflux for 4 h, the solution was poured onto
crushed ice and 20 ml of HCl 6N were added. The mixture was
extracted with EtOAc and the organic layers were washed with a
saturated NaCl solution, anhydrified and concentrated. The oily
residue was chromatographed (30% hexane in chloroform) to give 0.5
g (40% yield) of 20b as a waxy solid: IR (t.q.) 1670 cm.sup.-1;
.sup.1H NMR (CDCl.sub.3).delta.7.83 (d, 1H, J=2.1 Hz); 7.44 (dd,
1H), 7.29 (d, 1H, J=8.1 Hz), 2.63 (s, 3H), 2.41 (s, 3H). Anal.
(C.sub.9H.sub.9CIOS): C, H
[0068] 2-Bromo-1-[2-(methylthio)phenyl]ethanone (21a).
[0069] The title compound was obtained starting from 18 and
following the procedure as described below to obtain 21c. 21a was
obtained as colorless prism (69% yield):mp 81-82.degree.
C.(hexanes); IR (nujol) 1690 cm.sup.-1, .sup.1H NMR
(CDCl.sub.3).delta.8.00-7.80 (m,2H), 7.35 (m,2H), 5.53 (s,2H), 2.27
(s,3H) Anal. (C.sub.9H.sub.9BrOS)C,H,
[0070] 2-Bromo-1-[5-chloro2(methylthio)phenyl]ethanone (21b).
[0071] The title compound was obtained starting from 20b and
following the procedure as described below to obtain 21c. 21b was
obtained as colorless prism (620% yield); mp 97-98.degree.
C.(hexanes); IR (CHCl.sub.3) 1685 cm.sup.-1; .sup.1H NMR
(CDCCl.sub.3) .delta.7.98-7.73 (m,3H), 5.57 (s,2H), 2.31 (s,3H).
Anal.(C.sub.9H.sub.8BrClOS) C,H.
[0072] 1-[2-(methylthio)phenyl]-2-pyrrol-1-yl)ethanone (22a)
[0073] To a solution of 21a (2.1 g, 8.6 mmol) in 20 ml of anhydrous
DMF, was added hexamethylene tetramine in portions. The solid
formed was filtered, washed with chloroform and dried.
[0074] The hexamethylenetetrammonium sat thus obtained was added to
a concentrated HCl solution (3 ml) in 8 ml of ethanol.
[0075] The mixture was stirred for 96 hours in the dark at ambient
temperature. The white solid (NH.sub.4Cl) was filtered d the
solution vacuum-concentrated.
[0076] The residue was crystallised by ethanol, and
2-amino-1-[2-(methyl-thio)phenyl]ethanone hydrochloride was
obtained with a yield of 78%; .sup.1H NMR (DMSO-d.sub.6)
.delta.8.43 (br s, 2H); 8.11-7.31 (m, 5H), 4.59 (d, 2H, J=3.2 Hz),
2.46 (s, 3). Anal. (C.sub.9H.sub.12CINOS): C, H, N.
[0077] The 2-amino-1-[2-(methylthio)phenyl]ethaone hydrochloride
was dissolved in an aqueous solution of sodium acetate, glacial
acetic acid and 2.5 dimethoxy tetrahydrofran. After stirring for 15
min at 100.degree. C., the mixture was cooled and extracted with
ethyl acetate. The organic layer was washed with NaHCO.sub.3, dried
and evaporated. The crude product was chromatographed (CHCl.sub.3)
to give 22a with a yield of 50%: melting point 113-114.degree. C.
(hexane); IR (nujol) 1690 cm.sup.-1; .sup.1H NMR (CDCl.sub.3)
.delta.7.74-7.23 (m, 4H), 6.66 (m, 2H), 6.21 (m, 2H), 5.25 (d, 2H,
J=3.4 Hz), 2.44 (s, 3H). Anal. (C.sub.13H.sub.13NOS): C, H, N.
[0078] 1-[5-chloro-2-(methylthio)phenyl]-2-pyrrol-1-yl)ethanone
(22b)
[0079] Starting from 21b (5.58 g, 20.0 mmol),
2-amino-1-[5-chloro-2-(methy- l-thio)phenyl]ethanone hydrochloride
was obtained using the procedure described in the previous example:
yield 75%; .sup.1H NMR PMSO-d.sub.6) .delta.8.41 (br s, 2H);
8.10-7.28 (m, 3H), 4.48 (d, 2H, J=3.2 Hz), 2.42 (s, 3H). Anal.
(C.sub.9H.sub.11CI.sub.2NOS): C, H, N.
[0080] Starting from
2-amino-1-[5-chloro-2-(methylthio)phenyl]ethanone hydro-chloride,
the titre compound was obtained as colourless prisms using the
procedure described to obtain 22a: 51% yield; melting point
124-125.degree. C. (hexane); IR (nujol) 1720 cm.sup.-1; .sup.1H NMR
(CDCl.sub.3) .delta.7.62 (d, 1H, J=2.3 Hz); 7.45 (dd, 1H, J=8.2,
2.3 Hz), 7.29 (d, 1H, J=8.2 Hz), 6.65 (m, 2H), 6.22 (m, 2H), 5.21
(s, 2H), 2.43 (s, 3H); .sup.13C NMR (CDCl.sub.3) .delta.194.2,
140.6, 134.5, 132.4, 130.1, 128.9, 127.7, 121.8, 109.2, 56.6, 16.6.
Anal. (C.sub.13H12CINOS): C, H. N.
[0081] 1-[2-(methylsulphinyl)phenyl]-2-pyrrol-1-yl)ethanone
(23a)
[0082] To a suspension of sodium periodate (0.55 g, 2.6 mmol) in
methanol (7 ml) and water (1.4 ml) was added a solution of 22a (0.6
g 2.6 mmol) in methanol (2 ml). After stirring for 24 hours at
ambient temperature the sodium iodate was removed by filtration and
the filtrate was evaporated. The residue was chromatographed (5%
EtOAc in dichloromethane) to give 0.59 g of 23a (92% yield) as
colourless prisms: melting point 174-175.degree. C. (ethanol); IR
(nujol) 1710, 1090 cm.sup.-1; .sup.1H NMR (CDCl.sub.3)
.delta.8.42-7.64 (m, 4H), 6.66 (m, 2H), 6.27 (m, 2H), 5.44 (0.5
ABq, 1H, J=18.0 Hz), 5.27 (0.5 ABq, 1H, J=18.0 Hz), 2.80 (s, 3H).
Anal. (C.sub.13H.sub.13NO.sub.2S): C, H, N.
[0083] 1-[5-chloro-2-(methylsulphinyl)phenyl]-2-pyrrol-1-yl)
ethanone (23b)
[0084] The titre compound was prepared starting from 22b (1.1 g,
4.45 mmol) using the procedure described above for 23a; colourless
prisms (89% yield): melting point 218-219.degree. C. (ethanol); IR
(nujol) 1710, 1080 cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta.8.37
(d, 1H, J=8.0 Hz); 7.85 (m, 2H), 6.66 (m, 2H), 6.28 (m, 2H), 5.40
(0.5 Abq, 1H, J=17.7 Hz), 5.25 (0.5 Aq, 1H, J=17.8 Hz), 2.79 (s,
3H); .sup.13C NMR (CDCl.sub.3) .delta.193.4, 149.6, 136.9, 134.7,
132.5, 128.8, 127.0, 121.8, 109.8, 55.7, 44.3. Anal.
(C.sub.13H.sub.12CINO.sub.2S): C, H, N.
[0085] 9,10-dihydropyrrolo[2,1-b][1,3]benzothiazepine-9-one
(9a)
[0086] Method A: to a highly reactive solution of the crystalline
complex of triflate copper(I) and benzene (0.81 g, 1.6 mmol) in
anhydrous benzene (20 ml), cooled to 0.degree. C., was added a
solution of selenoester 15 (0.5 g. 1,6 mmol) in anhydrous benzene
(11 ml) and the mixture was left to stir at ambient temperature for
16 hours. Ethyl ether (10 ml) was added, the organic phase was
washed with ammonia 6N, anhydrified and concentrated. The crude
product was chromatographed (5% hexane im dichloromethane) to give
51 mg of 9a (12% yield) as colourless prisms. Ketone 16 (55% yield)
was also recovered as a dense oil. Compound 9a: melting point
94-95.degree. C. (hexane); IR (CDCl.sub.3) 1690 cm.sup.-1; .sup.1H
NMR (CDCl.sub.3) 3 8.14-7.30 (m, 4H); 6.88 (m, 1H), 6.42 (m, 1H),
6.12 (mn, 1H), 5.15 (s, 2H); 13C NM (CDCl.sub.3) .delta.190.9,
136.1, 133.3, 132.3, 130.8, 127.6, 123.9, 120.2, 114.4, 109.2,
57.6. MS m/z 265 (10, M.sup.+), 215 (100), 187, 154,115, 97. Anal.
(C.sub.12H.sub.9NOS): C, H, N.
[0087] Compound 16: IR (t.q.) 1670 cm.sup.-1; .sup.1H NMR
(CCDl.sub.3) .delta.7.22-6.81 (m, 6H); 6.70 (m, 1H), 6.42 (m, 1H),
4.61 (s, 2H), 2.20 (s, 3H). MS m/z 231 (100, M.sup.+). Anal.
(C.sub.13H.sub.13NOS): C, H, N.
[0088] Method B: trifluoroacetic anhydride (1.0 ml, 7.4 mmol) was
added dropwise to just distilled N,N-dimethylformamide (8 ml)
cooled to 0.degree. C. After stirring for 20 min at 0.degree. C., a
solution of 23a (1.0 g, 4.0 mmol) was added in Just distilled
N,N-dimethyl formamide (24 ml) and after 15 min at 0.degree. C. and
1 hour at ambient temperature, the pH of the dark red solution was
brought to 7 with NaOH 1N and the mixture was stirred for another
30 min. Extraction with dichloromethane, anhydrification of the
extracts and evaporation of the solvent gave an oily residue which
was chromatographed (30% hexane in chloroform). Compound 9a was
obtained with a 45% yield.
[0089]
7-chloro-9,10-dihydropyrrolo[2,1-b][1,3]benzothiazepine-9-one
(9b)
[0090] The titre compound was obtained with a yield of 42%, as
colourless prisms, starting from 23b and adopting the procedure as
described for 9a (Method B): melting point 106-107.degree. C.
(hexane); IR (CDCl.sub.3) 1690 cm.sup.-1; .sup.1H NMR (CDCl.sub.3)
.delta.8.14 (d, 1H, J=2.2 Hz); 7.49 (d, 1H, J=8.1 Hz), 7.38 (dd,
1H, J=8.0, 2.3 Hz), 6.88 (in, 1H), 6.43 (m, 1H), 6.12 (m, 1H), 5.14
(s, 2H); .sup.13C NMR (CDCl.sub.3) .delta.189.7, 138.3, 133.3,
137.3, 134.1, 133.1, 132.2, 131.9, 124.1, 119.5, 114.7, 109.5,
57.3. MS m/z 250 (20, M.sup.+), 249 (100), 221, 216, 188, 158, 110.
Anal. (C.sub.12H.sub.8CINOS): C, H, N.
[0091]
(.+-.)-9,10-dihydro-9-hydroxpyrrolo[2,1-b][1,3]benzothiazepine
(24a)
[0092] To a solution of 9a (61 mg, 0.23 mol) in arhydrous methane
(1 ml), cooled to 0.degree. C. and under nitrogen, were added
aliquots of sodium borohydride (80 mg, 0.23 mmol). After stirring
for 1 hour at 0.degree. C., the reaction was stopped with water (1
ml) and the mixture extracted with EtOAc. The organic layers were
washed with brine, anhydrified and concentrated. The residue was
chromatographed (15% EtOAc in dichloromethane) to give 24a (64 mg,
92% yield) as colourless prisms: melting point 101-102.degree. C.
(ethanol); IR (nujol) 3300 cm.sup.-1; .sup.1H NMR (CDCl.sub.3)
.delta.7.47-7.12 (m, 4H), 6.86 (m, 1H), 6.33 (m, 1H), 6.02 (m, 1H),
5.08 (m, 1H), 4.89 (dd, 1H, J=13.9, 1.7 Hz), 4.28 (dd, 1H, J=13.9,
6.0 Hz), 2.05 (d, 1H, J=9.6 Hz). Anal. (C.sub.12H.sub.11NOS): C, H,
N.
[0093]
(.+-.)-7-chloro-9,10-dihydro-9-hydroxpyrrolo[2,1-b][1,3]benzothiaze-
pine (24b)
[0094] The titre compound was obtained starting from 9b (112 mg,
0.45 mmol) using the procedure described above: 88% yield; melting
point 118-119.degree. C. (ethanol); IR (CHCl.sub.3) 3300 cm.sup.-1;
.sup.1H NMR (CDCl.sub.3) .delta.7.48 (d, 1H, J=2.1 Hz); 7.32 (d,
1H, J=8.0 Hz), 7.15 (dd, 1H, J=8.1, 2.1 Hz), 6.88 (m, 1H), 6.34 (m,
1H), 6.11 (m, 1H), 5.02 (m, 1H), 4.85 (dd, 1H, J=13.9, 1.9 Hz),
4.29 (dd, 1H, J=13.9, 6.6 Hz), 2.10 (dd, 1H, J=9.6 Hz). Anal
(C.sub.12H.sub.10CINOS): C, H, N.
[0095]
(.+-.)-9-bromo-9,10-dihydropyrrolo[2,1-b][1,3]benzothiazepine
(255a)
[0096] To a solution of 24a (0.26 g, 1.0 mmol) in anhydrous ethyl
ether (4 ml) was added dropwise a solution of PBr3 (0.13 g, 0.5
mmol) in anhydrous ethyl ether (1 ml) and the reaction mixture was
kept at reflux temperature for 2 hours under nitrogen. Anhydrous
ethanol was added (0.2 ml) and the resulting solution was heated to
reflux temperature for another hour. Five ml of an aqueous solution
of 5% Na.sub.2CO.sub.3 were then added, the organic phase was
separated, anhydrified and evaporated. The crude product was
chromatographed (hexane and chloroform 1:1) to give 25a (0.2 g, 64%
yield) as colourless prisms: melting point 115-116.degree. C.
(cyclohexene); .sup.1H NMR (CDCl.sub.3) .delta.7.46-7.09 (m, 4H);
6.93 (m, 1H), 6.39 (m, 1H), 6.12 (m, 1H), 5.75 (dd, 1H, J=6.9, 2.6
Hz), 5.07 (dd, 1H, J=14.7, 2.6 Hz), 4.61 (dd, 1H, J=14.7, 6.9 Hz).
Anal. (C.sub.12H.sub.10BrNOS): C, H, N.
[0097]
(.+-.)-9-bromo-7-chloro9,10-dihydropyrrolo[2,1-b][1,3]benzothiazepi-
ne (25b)
[0098] The titre compound was obtained starting from 24b (0.31 g,
1.6 mmol) using the procedure described above: 51% yield; melting
point 106-107.degree. C. (cyclohexane); .sup.1H NMR (CDCl.sub.3) 5
7.45 (d, 1H, J=2.1 Hz); 7.27 (d, 1H, J=8.6 Hz), 7.11 (dd, 1H,
J=8.6, 2.1 Hz), 6.92 (m, 1H), 6.39 (m, 1H), 6.12 (m, 1H), 5.63 (dd,
1H, J=7.0, 2.3 Hz), 5.06 (dd, 1H, J=14.4, 2.3 Hz), 4.61 (dd, 1H,
J=14.0, 7.0 Hz). .sup.13C NMR (CDCl.sub.3) .delta.139.9, 134.1,
133.2, 131.7, 131.5, 128.9, 125.6, 119.6, 114.6, 108.1, 51.2, 51.0.
Anal. (C.sub.12H.sub.9BrCINOS): C, H, N.
EXAMPLE 1
[0099]
(.+-.)-9-(4-methylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-b][1,3]en-
zo-thiazepine (3a)
[0100] A mixture of 25a (0.65 g, 2.0 mmol) and N-methylpiperazine
(1.1 ml, 10.0 mmol) was heated to 130.degree. C. for 2 hours under
argon, cooled, poured onto crushed ice and extracted with ethyl
ether. The organic extracts collected were washed with brine,
anhydrified. and concentrated. The residue was chromatographed
(EtOAc) to give 0.45 g (75% yield) of 3a as colourless prisms:
melting point 206-207.degree. C. (hexane); .sup.1H NMR (CDCl.sub.3)
.delta.7.49-7.09 (m, 4H); 6.87 (m, 1H), 6.29 (m, 1H), 4.68 (dd, 1H,
J=14.4, 8.6 Hz), 4.51 (dd 1H, J=14.4, 3.7 Hz), 2.56-2.34 (m, 8H),
2.23 (s, 3H); .sup.13C NMR (CDCl.sub.3) .delta.138.1, 134.6, 132.9,
130.4, 127.3, 126.9, 123.9, 121.7, 113.3, 107.7, 66.1, 55.9, 48.8,
46.6. 46.1. MS m/z 299 (100, M.sup.+), 219, 200, 167, 149, 113.
Anal. (C.sub.17H.sub.21N.sub.3S) C, H, N.
EXAMPLE 2
[0101]
(.+-.)-7-chloro9-(4-methylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-b-
][1,3]benzothiazepine (3b) (ST1455)
[0102] The titre compound was obtained starting from 25b (0.3 g,
0.95 mmol) using the procedure described above. 3b was obtained as
colourless prisms (68% yield): melting point 210-211.degree. C.
(hexane); .sup.1H NMR (CDCl.sub.3) .delta.7.51 (d, 1H, J=2.4 Hz);
7.30 (d, 1H, J=8.5 Hz), 7.06 (dd, 1H, J=8.5, 2.4 Hz), 6.86 (m, 1H),
6.29 (m, 1H), 6.05 (m, 1H), 4.71 (dd, 1H, J=14.0, 8.6 Hz), 4.45
(dd, 1H, J=14.0, 3.4 Hz), 3.95 (dd, 1H, J=8.6, 3.4 Hz), 2.65-2.25
(m, 8H), 1.42 (s, 3H); .sup.13C NMR (CDCl.sub.3) .delta.140.1,
133.2, 133.0, 132.4, 131.6, 127.3, 123.9, 121.1, 113.6, 107.9,
65.9, 55.8, 55.7, 47.7, 45.9, 44.9, 26.8. MS m/z 333 (10, M.sup.+),
250, 233 (100), 201, 166, 139. Anal. (C.sub.17H.sub.20CIN.sub.3S):
C, H, N. The dihydrochloride salt (named hereinafter ST1468) was
obtained by dissolving an analytical sample in HCl 1N in methanol.
The solvent was evaporated and the residue recrystallized (methanol
and ethyl ether 1:1). Anal. (C.sub.17H.sub.22CI.sub.3N.sub.3S), C,
H, N.
EXAMPLE 3
[0103] Semipreparatory Chiral Separation of (.+-.)-3b
[0104] First of all, the hydrochloride salt of (.+-.)-3b was
purified on a short column filled with silica gel, using
dichloromethane and methanol (9:1) as the eluent. The purified
solvent was converted to the free base. Evaporation of the solvent
gave an oily residue which was dissolved in isopropanol and the
solution was diluted with hexane until the 95:5 ratio was obtained.
For the separation of the enantiomers, a 10-15 mg/ml concentration
of the racemic mixture was made. A mixture of hexane (plus 0.1%
triethylamide) and isopropanol was used as the mobile phase.
[0105] A gradient-type mixer maintained the ratio between the
solvents hexane and isopropanol at 95:5. Injection amounts were 100
.mu.l per injection. The enantiomers were collected using a
fraction collector. Only fractions with a signal above 10% (10 mV)
of the total scale were collected. The amounts with signals below
10% were collected separately and used for a second purification.
The purity of both enantiomers was determined by weighing the trace
peaks separately.
[0106] (+)-3b: .sup.1H NMR (500 Mhz, CDCl.sub.3) .delta.7.52 (d,
1H, J=2.4 Hz); 7.32 (d, 1H, J=8.3 Hz), 7.09 (dd, 1H, J=2.4, 8.3
Hz), 6.88 (m, 1H), 6.30 (n, 1H), 6.07 (m, 1H), 4.71 (dd, 1H, J=8.8,
14.2 Hz), 4.50 (dd, 1H, J=3.9, 14.7 Hz), 3.97 (dd, 1H, J=3.4, 8.8
Hz), 2.55 (m, 4H), 2.40 (m, 4H), 2.27 (s, 3H); .sup.13C NMR (500
MHz, CDCl.sub.3) .delta.139.9, 133.0, 132.9, 132.3, 131.6, 127.3,
123.8, 121.0, 113.5, 107.9, 88.2, 65.8, 55.6, 48.6, 45.9, 45.8;
purity (ee) 94.6%; [.alpha.].sub.D=+46.0.de- gree. (c 0.48, MeOH).
The respective dihydrochloride, obtained as in the case of compound
(.+-.)3b, was named ST1469.
[0107] (-)-3b: .sup.1H NMR (500 Mhz, CDCl.sub.3) .delta.7.53 (d,
1H, J=2.3 Hz); 7.32 (d, 1H, J=8.3 Hz), 7.09 (dd, 1H, J=6.8 Hz),
6.88 (m, 1H), 6.30 (m, 1H), 6.07 (m, 1H), 4.71 (dd, 1H, J=9.3, 14.5
Hz), 4.48 (dd, 1H, J=3.4, 14.2 Hz), 3.98 (dd, 1H, J=2.9, 8.8 Hz),
2.49 (m, 8H), 2.27 (s, 3H); .sup.13C NMR (500 MHz, CDCl.sub.3)
.delta.140.0, 133.0, 132.9, 132.3, 131.6, 127.3, 123.8, 121.0,
113.5, 107.9, 88.2, 65.8, 55.6, 48.6, 45.9, 45.8; purity (ee) 98%;
[.alpha.].sub.D=-47.90 (c 0.54, MeOH). The respective
dihydrochloride, obtained as in the case of compound (.+-.)3b, was
named ST1470.
EXAMPLE 4
[0108]
(.+-.)-7-Fluoro-9-(4-methylpiperazin-1-yl)-9,10-dihydropyrrolo
[2,1b][1,3]benzothiazepine (3c) (ST1456)
[0109] The synthesis of the (3c) was been performed as described in
Scheme 2B/1 and 2B/2 where c=F.
[0110] 1-[5-Fluoro-2-(methylthio)phenyl]ethanone (20c).
[0111] A mixture of 4-fluorothioanisole (19c) (4 g, 28.13 mmol),
anhydrous aluminium chloride (8.40 g, 63.02 mmol) and carbon
disulphide (89 ml) was heated at reflux under argon atmosphere, and
acetic anhydride (2.65 ml, 28.07 mmol) was added dropwise in 2 h.
After reflexing for 24 h, the solution was poured into crushed ice,
water (62.48 ml) and concentrated hydrochloric acid (2.68 ml). The
organic phase was separated and water exacted with dichloromethane
(3.times.30 ml), the organic layers were washed with brine, dried
and concentrated. The oily residue was chromatographed (50%
petroleum ether 40-60.degree. in dichioromethane) to afford 20c
(2.98 g) as a colourless crystalline solid, mp 82.0-84.3.degree. C.
(58% yield). .sup.1NMR (CDCl.sub.3) .delta.7.49-7.43 (dd, 1H,
J=9.23, 2.44 Hz); 7.29-7.11 (m, 2H); 2.57 (s, 3H); 2.39 (s,
3H).
[0112] 2-Bromo-1-[5-fluoro-2-(methylthio)phenyl]ethanone (21
c).
[0113] To a sting solution of 20c (2.07 g, 11.34 mmol), carbon
tetrachloride (62 ml) and glacial acetic acid (2.07 ml) was added
at room temperature a solution of bromine (.mu.L 546.6, 10.66 mmol)
in carbon tetrachloride (34 mmol). The first drop was added and
after 20 minutes the solution was added dropwise in 4 hours. After
stirring for 16 hours the solvent was distilled and to the residue
was added water and solid sodium bicarbonate (to pH 7), the organic
phase was separated and water extracted with dichioromethane
(3.times.30 mn), the organic layers were dried and evaporated. The
crude product was chromatographed (70% petroleum ether
40-60.degree. in dichloromethane) to give 1.90 g of 21c as a
yellowish solid (64% yield). .sup.1NMR (CDCl.sub.3)
.delta.7.47-7.42 (dd, 1H, J=8.85, 2.84 Hz); 7.38-7.18 (m, 2H); 4.42
(s, 2H); 2.43 (s, 3H).
[0114] 1-[5-Fluoro-2-(methylthio)phenyl]-2-(pyrrol-1-yl)ethanone
(22c).
[0115] To a stirring solution of hexamethylenetetramnine (3.18 g,
22.70 nmol) in chloroform (29.6 ml) at room temperature was added
dropwise in 5 minutes a solution of 21c (1.90 g, 7.20 mmol) in
chloroform (16 ml). As soon as the solid formed the solution was
rapidly filtered and the desired product was collected as a yellow
amorphous solid that was washed with chloroform, dried and used for
the following reaction; (99% yield).
[0116] A suspension of
1-[5-fluoro-2-(methylthio)phenyl]ethanon-2-hexamini- um bromide
(1.62 g, 4.02 mmol) in methanol (13.3 ml) was warmed to 0.degree.
C. and was added of concentrated hydrochloric acid (1.86 ml). The
mixture was stirred for 96 hours in the dark at room temperature.
The white solid (ammonium chloride) was removed by filtration and
the obtained solution was evaporated. The residue was
recrystallized from ethanol to give
2-amino-1-[5-fluoro-2-(methylthio)phenyl]ethanone hydrochloride as
a yellow solid, that was used in the next step without further
purification. (98% yield).
[0117] To a solution of
2-amino-1-[5-fluoro-2-(methylthio)phenyl]ethanone hydrochloride
(4.54 g, 19.27 mmol) in water (29 ml), heated at 90.degree. C.,
were added trihydrated sodium acetate (2.62 g, 19.27 mmol), glacial
acetic acid (17 ml) and 2,5-dimethoxytetrahydrofuran (2.40 ml,
18.50 mmol). After 20 seconds at 90-100.degree. C. the mixture was
cooled and extracted with ethyl acetate. The organic layers were
washed with a 20% solution of sodium bicarbonate and brine, dried
ad evaporated. The residue was chromatographed (50% petroleum ether
40-60.degree. in dichloromethane) to afford 2.07 g of 22c as white
crystals mp 133.2-134.0.degree. C. (50% yield). .sup.1NMR
(CDCl.sub.3) .delta.7.39-7.15 (m, 3H); 6.66-6.65 (m, 2H); 6.22-6.20
(m, 2H); 5.20 (s, 2H); 2.42 (s, 3H). MS m/z 252 (M.sup.++H), 234,
202 (100), 183, 169, 154, 141, 126, 109, 80.
[0118] 1-[5-Fluoro-2-(methysulfinyl)phenyl]-2-(pyrrol-1-yl)ethanone
(23c).
[0119] To a stirred cooled solution of
1-[5-Fluoro-2-(methylthiophenyl]-2-- (pyrrol-1-yl)ethanone (22c)
(1.76 g, 7.06 mmol) in dichloromethane (12 ml) was added dropwise
in 30 minutes a solution of m-chloroperbezoic acid (71.5% grade,
1.70 g, 7.06 mmol) in dichioromethane (10 ml). After stirring for
45 minutes at 0.degree. C., the mixture was treated with a 5%
solution of sodium carbonate in water (41 ml) and was stirred for
15 minutes at room temperature. The organic phase was separated and
water was exacted with dichloromethane (3.times.10 ml). The organic
layers were dried and evaporated, the residue was chromatographed
(10% dichloromethane in ethyl acetate) to afford 1.02 g of 23c as
white crystals that darkened rapidly (64% yield). .sup.1NMR
(CDCl.sub.3 ) .delta.8.42-8.35 (m, 1H); 7.61-7.51 (m, 2H);
6.68-6.62 (m, 2H); 6.26-6.24 (m, 2H); 5.41-5.17 (q, 2H, J=31.32,
17.87 Hz); 2.77 (s, 3H).
[0120] 7-Fluoro-9,10-dihydropyrrolo[2,1-b][1,3]benzotiazepin-9-one
(9c).
[0121] Trifluoroacetic anhydride (1.02 ml) was added dropwise under
argon atmosphere to a freshly distilled N,N-dimethylformamide (8
ml) cooled to 0.degree. C. After s ting for 20 minutes at 0.degree.
C. a solution of 23c (109 g, 4.12 mmol) in N,N-dimethylformamide
(29 ml) was added. After 15 minutes at room temperature water (41
ml) was added to the dark yellow solution and pH was adjusted to 7
with sodium acetate, the mixture obtained was stirred at room
temperature for 1 night. Extraction with dichloromethane, drying of
the eracts, and evaporation of the solvent gave an oily residue
which was chromatographed (30% petroleum ether 40-60.degree. in
dichloromethane). The compound 9c was crystallised from n-hexane as
yellowish crystals mp 133.8-134.2.degree. C. (20% yield). .sup.1H
NMR (CDCl.sub.3) .delta.7.82-7.76 (m, 1H);7.55-7.49 (m, 1H);
7.18-7.09 (m, 1H); 6.88-6.87 (m, 1H); 6.42-6.40 (m, 1H); 6.12-6.09
(m, 1H); 5.14 (s, 2H). MS m/z 233 (100) (M+), 205, 200, 172,
126.
[0122]
(.+-.)-7Fluoro-9,10-dihydro-9-hydroxpyrrolo[2,1-b][1,3]benzothiazep-
ine (24c).
[0123] To a solution of 9c (0.037 g 0.16 mmol) in dry
tetrahydrofuran (0.5 ml) and dry methanol (0.7 ml), cooled to
0.degree.0 C. under argon atmosphere, was added in portions sodium
borohydride (0.09 g, 0.16 mmol). After stirring for 1 hour at
0.degree. C. the reaction was quenched with a saturated solution of
ammonium chloride (1 ml), the solvent was removed and the mixture
was extracted with ethyl acetate (3.times.2 ml). The organic layers
were dried and evaporated, the crude product was chromatographed
(30% petroleum ether 40-60.degree. in dichloromethane) to give 24c
0.036 g (96% yield). 1H NMR (CDCl.sub.3) .delta.7.40-7.33 (m, 1H);
7.24-7.18 (m, 1H); 6.93-6.84 (m, 2H); 6.33-6.31 (m, 1H); 6.11-6.08
(m, 1H); 5.12-5.04 (m, 1H); 4.91-4.83 (dd, 1H, J=14.22, 2.25 Hz);
4.34-4.24 (dd, 1H, J=14.02, 6.51 Hz); 2.09-2.04 (d, 1H, J=9.83
Hz).
[0124]
(.+-.)-9-Bromo-7-fluoro-9,10-dihydropyrrolo[2,1-b][1,3]benzothiazep-
ine (25c).
[0125] To a solution of 24c (0.17 g 0.71 mmol) in dry ethyl ether
(3 ml) was added dropwise a solution of phosphorus tribromide
(.mu.L 33.5, 0.36 mmol) in dry ethyl ether (0.7 ml); the reaction
mixture was refluxed for 2 hours under argon atmosphere. After
cooling to room temperature dry ethanol (.mu.L 143) was added and
the resulting solution was heated at reflux for 1 hour. Then 4 ml
of aqueous sodium carbonate was added; the organic phase was
separated, dried and evaporated. The crude product was
chromatographed (50% petroleum ether 40-60.degree. in
dichloromethane) to give 0.103 g of pure 25c (48% yield). .sup.1H
NMR (CDCl.sub.3) .delta.7.35-7.16 (m, 2H); 6.92-6.82 (m, 2H);
6.39-6.37 (m, 1H); 6.13-6.09 (m, 1H); 5.69-5.64 (m, 1H); 5.10-5.01
(dd, 1H, J=14.57, 2.65 Hz); 4.70-4.59 (dd, 1H, J=14.88, 7.05
Hz)
EXAMPLE 5
[0126]
(.+-.)-7-Fluoro-9-(4methylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1b]-
[1,3]benzothiazepine(3c) ST 1456.
[0127] A mixture of 25c (0.05 g, 0.18 mmol) and N-methylpiperazine
(1 ml) was heated at 140.degree. C. for 17 hours under argon
atmosphere. The reaction mixture was then cooled, diluted with
ethyl acetate (30 ml) and washed with brine. The organic layers
were dried, evaporated and the oily residue was chromatographed
(10% trimethylamine in ethyl acetate) to afford 0.037 g of 3c as a
colourless solid mp 213-214.degree. C. (63% yield). .sup.1NMR
(CDCl.sub.3) .delta.7.37-7.26 (m, 2H); 6.85-6.76 (m, 2H); 6.29-6.27
(m, 1H); 6.06-6.03 (m, 1H); 4.78-4.67 (m, 1H); 4.49-4.40 (dd, 1H,
J=14.19, 3.48 Hz); 3.99-3.93 (dd, 1H, J=8.97, 3.39 Hz); 2.64-2.37
(m, 8H); 2.25 (s, 3H). MS m/z 318 (100) (M.sup.++H), 277, 218,
185.
EXAMPLE 6
[0128]
(.+-.)-7-Fluoro-9-(4ethylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-b]-
[1,3]benzothiazepine(4c) ST1457.
[0129] The desired product 4c was obtained starting from 25c (0.053
g, 0.178 mmol), using 4-methylpiperazine (1 ml). The colourless
liquid 4c was obtained with 73% yield. 1H NMR (CDCl.sub.3)
.delta.7.37-7.26 (m, 2H); .delta.6.86-6.75 (m, 2H); 6.29-6.26 (m,
1H); 6.05-6.02 (m, 1H); 4.80-4.68 (mn, 1H); 4.48-4.39 (dd, 1H,
J=13.95, 3.78 Hz); 3.97-3.91 (dd, 1H, J=9.23) 3.65 Hz); 2.60-2.32
(m, 10H); 1.08-1.00 (t, 3H, J=7.33 Hz). MS m/z 332 (100)
(M.sup.++H), 277, 218, 185, 115.
EXAMPLE 6
[0130]
(.+-.)-7-Fluoro-9-[4-(2'-hydroxyethyl)piperazin-1-yl]-9,10-dihydrop-
yrrolo[2,1b][1,3]benzothiazepiue (5c) ST 1458.
[0131] A solution of 25c (0.065 g, 0.218 mmol) hydroxyethylpiperine
(.mu.L 59, 0.218 mmol) and 2-buthanone (2 ml) was refluxed for 21
hours. The reaction ixte was then evaporated and to the residue was
added water and was extracted with ethyl acetate, combined extracts
were dried and evaporated. The crude product was chromatographed to
give 5c as a colourless amorphous solid (69% yield). .sup.1H NMR
(CDCl.sub.3) .delta.7.37-7.24 (m, 2H); 6.86-6.77 (m, 2H); 6.29-6.27
(m, 1H); 6.06-6.03 (m, 1H); 4.77-4.65 (m, 1H); 4.48-4.40 (dd, 1H,
J=14.15, 3.46 Hz); 3.98-3.92 (dd, 1H, J=8.93, 3.50 Hz); 3.59-3.54
(t, 2H, J=5.37 Hz); 2.85 (bs, 1H); 2.57-2.47 (m, 10H). MS m/z 348
(100) (M.sup.30+H) 288, 218, 185.
EXAMPLE 7
[0132]
(.+-.)-7-Bromo-9-(4-methylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-b-
][1,3]benzothiazepine (3d).
[0133] The synthesis of the (3d) was been performed as described in
Scheme C and 2B/2 where d=Br.
[0134] Bis-(2-hydroxycarbonyl-4-bromo)phenyldisulphide (26).
[0135] To a cooled (0-5.degree. C.) sing solution of
2-amino-5-bromobenzoic acid (1 g, 4.63 mmol), sodium hydroxide
(0.185 g, 4.63 mmol), water (7.71 ml) and sodium nitrite (0.32 g,
4.63 mmol) was slowly added a solution of concentrated hydrochloric
acid (1.44 ml) in water (2.5 ml), the mixture was stirred at
0-5.degree. C. for 1 h, than was neutralised with potassium
carbonate and potassium acetate. The cold diazonium salt solution
was run into a vigorously stirred solution of potassium ethyl
xanthate (2.23 g, 13.89 mmol) and water (7.7 ml) previously heated
at 75-80.degree. C. and was maintaining this temperature during
addition and for her 1 h. The reaction mixture was cooled to room
temperature and stirred for 1 h. Than hydrogen peroxide (3.22 ml)
was added and the solution was stirred for 1 night at room
temperature. The xue was filtered and the solution was acidified
(on an ice bath) and filtered again. The product that was collected
as a yellow amorphous solid was dissolved with aqueous sodium
hydroxide and reprecipitated with hydrochloric acid to afford pure
26 (1.02 g) (95% yield). The compound was used in the next step
without further purification
[0136] 1-[5-Bromo-(2-methylthio)phenyl]hydroxycarbonyl (27).
[0137] To a solution of (26) (1 g, 2.15 mmol) in 85% ethanol (17.2
ml) and sodium hydroxide, sodium borohydride (0.163 g) was added in
portions. The resulting solution was stirred 30 minutes at room
temperature and for additional 3 hours at reflux. Then ice was
added and the mixture was stirred for 15 minutes at room
temperature, a solution of sodium hydroxide (0.302 g, 7.55 mmol) in
water (1.9 ml) and dimethyl sulphide (376 .mu.L, 3.97 mmol) were
added and the reaction mixture was stirred 2.5 hours at reflux.
After cooling 1 drop of ammonium hydroxide 30% was added (to
destroy the excess of sodium hydroxide), hydrochloric acid was
added (pH 3). The solid obtained was collected by filtration. The
crude product was chromatographed (4% formic acid, 20% ethyl
acetate in toluene) to afford 1.017 g of 27 as a yellow solid (96%
yield). 1H NMR (DMSO-d.sub.6) .delta.7.95-7.94 (d, 1H, J=2.69 Hz);
7.72-7.66 (dd, 1H, J=8.80, 1.95 Hz); 7.29-7.25 (d, 1H, J=8.77 Hz);
2.37 (s, 3H).
[0138] 1-[5-Bromo-(2-methylthio)phenyl]ethanone (28).
[0139] A stirred solution of (27) (0.1 g, 0.40 mmol) in dry
tetrahydrofran (3.03 ml) was cooled to 0.degree. C. (ice bath) and
treated with methyllithium (1.4 M solution in ether, 1.156 ml. 1.62
mmol). After 2 hours at 0.degree. C. under sing,
tirethylchorosilane (1.03 ml, 8.09 mmol) was rapidly added while
stirring continued, the ice bath was removed and the reaction
mixture was allowed to came to room temperature, then 1 N
hydrochloric acid (3.05 ml) was added and the resulting two phase
system was stirred at room temperature for 30 minutes. The organic
phase was separated and water was extracted with ether (3.times.5
ml), the combined extracts were dried and evaporated. The crude
product was chromatographed (30% petroleum ether 40-60.degree. in
dichloromethane) to afford 0.064 g of 28 as a yellowish solid mp
71.5-73.0.degree. C. (64% yield). .sup.1H NMR (CDCl.sub.3)
.delta.7.89-7.88 (d, 1H, J=1.91 Hz); 7.57-7.51 (dd, 1H, J=8.45,
2.48 Hz); 7.18-7.14 (d, 1H, J=8.48 Hz); 2.58 (s, 3H); 2.39 (s,
3H).
[0140] 2-Bromo-1-[5-bromo-2-(metylthio)phenyl]ethanone (21 d).
[0141] Starting from 28 (0.60 g, 2.43 mmol), the desired product
21d was obtained following the procedure described for 21c. The
crude product was chromatographed (50% petroleum ether
40-60.degree. in dichloromethane) to give the pure product 0.57 g.
(72% yield). .sup.1H NMR (CDCl.sub.3) .delta.7.87-7.58 (d, 1H,
J=1.60 Hz);7.48-7.42 (dd, 1H, J=8.05, 2.08); 7.12-7.16(d, 1H,
J=8.06 Hz); 4.43 (s, 2H); 2.43 (s, 3H)
[0142] 1-[5-Bromo-2-(methylthio)phenyl]-2-(pyrrol-1-yl) ethaone
(22d).
[0143] The desired product 22d was obtained as white crystals,
following the procedure described for 22c; mp 138.0-139.2.degree.
C. (32% yield). .sup.1H NMR (CDCB) .delta.7.76-7.55 (d, 1H, J=1.92
Hz);7.60-754 (dd, 1H, J=8.43, 2.16 Hz);7.23-7.19 (d, 1H, J=8.82
Hz); 6.65-6.63 (m, 2H); 6.22-6.20 (m, 2H); 5.20 (s, 2H); 2.41 (s,
3H).
[0144] 1-[5-Bromo-2-(methylsulfylphenyl]-2-(pyrrol-1-yl)ethanone
(23d).
[0145] Starting from 22d (0.27 g, 0.86 mmol), the desired product
was obtained following the above-described procedure, as white
crystals mp 138.0-139.2.degree. C. (75% yield) .sup.1H NMR
(CDCl.sub.3) .delta.8.30-8.25 (m, 1H); 8.0-7.96 (m, 2H); 6.64-6.62
(m, 2H); 6.27-6.25 (ml, 2H); 5.43-5.18 (q, 2H, J=32.28, 17.92 Hz);
2.77 (s, 3H).
[0146] 7-Bromo-9,10-dihydropyrrolo[2,1-b][1,3]benzothiazepine-9-one
(9d)
[0147] The reaction to obtain 9d was carried out, accordingly the
procedure described for 9c, using trifluoroacetic acid as solvent
(0.63 ml) and adding solid 23d (0.20 g, .062 mmol) to the cold
(0.degree. C.) solution of trifluoroacetic acid and trifluoroacetic
anhydride. The desired product 9d was obtained as yellowish
crystals (64% yield). .sup.1H NMR (CDCl.sub.3) .delta.8.22-8.21 (d,
1H, J=1.99 Hz);7.56-7.51 (dd, 1H, J=8.28, 2.24 Hz); 7.42-7.38 (d,
1H, J=8.42 Hz); 6.87-6.86 (m, 1H); 6.43-6.41 (m, 1H); 6.12-6.09 (m,
1H); 5.13 (s, 2H). MS m/z 293 (100) (M.sup.+), 265, 261, 232, 214,
186, 154, 115.
[0148]
(.+-.)-7-Bromo-9,10-dihydro-9-hydroxpyrrolo[2,1-b][1,3]benzothiazep-
ine (24d).
[0149] Starting from 9d (0.12 g, 0.39 mmol), the desired product
was obtained accordingly the procedure described for 24c (65%
yield). .sup.1NMR (CDCl.sub.3) .delta.7.63-7.62 (d, 1H, J=1.70 Hz);
7.32-7.21 (m, 2H); 6.88-6.87 (m, 2H); 6.34-6.32 (m, 1H); 6.12-6.09
(m, 1H); 5.02 (s, 1H; 4.90-4.82 (dd, 1H, J=13.86, 1.83 Hz);
4.36-4.25 (dd, 1H, J=14.18, 6.33 Hz); 1.99(s, 1H)
[0150]
(.+-.)-7,9-dibromo-9,10-dihydropyrrolo[2,1-b][1,3]benzothiazepine
(25d).
[0151] Starting from 24d (0.07 g, 0.25 mmol) the title compound was
obtained following the above-described procedure (36% yield). 1H
NMR (CDCl.sub.3) .delta.7.59 (i, 1H); 7.28-7.16 (m, 2H); 6.92-6.91
(m, 1H); 6.39-6.37 (m, 1H); 6.13-6.10 (m, 1H); 5.65-5.60 (dd, 1H,
J=6.98, 2.57 Hz); 5.09-5.00 (dd, 1H, J=14.65, 2.35 Hz); 4.66-4.55
(dd, 1H, J=14.68, 7.03 Hz).
[0152]
(.+-.)-7-Bromo-9-(4methylpiperazin-1-yl)-9,10-dihydropyrrolo[2,1-b]-
[1,3]benzothiazepine (3d).
[0153] Starting from 25d (0.033 g, 0.092 mmol) the title compound
was obtained following the procedure described for 3c (58% yield).
.sup.1H NMR (CDCl.sub.3) .delta.7.64 (s, 1H); 7.35-7.21 (m, 2H);
6.86-6.84 (m, 1H); 6.29-6.26 (m, 1H); 6.06-6.02 (m, 1H); 4.73-4.61
(m, 1H); 4.49-4.40 (dd, 1H, J=14.30, 8.57 Hz); 3.98-3.92 (dd, 1H,
J=8.64, 3.53 H/z); 2.57-2.37 (m, 11H).
[0154] 2. Pharmacology
EXPERMENTAL PROCEDURES
In Vitro Binding Assay
[0155] D.sub.1,D.sub.2,D.sub.3 and 5-HT.sub.2a affinity.
[0156] Male CRL:CD(SD)BR-COBS rats (Charles River, Italy) were
killed by decapitation (procedures involving animals and their care
were conducted in conformity with the institutional guidelines that
are in compliance with national (D.L. n. 116, G.U., suppl. 40, Feb.
18, 1992) and international laws and policies (EEC Council
Directive 86/609, OJ L 358, 1, Dec. 12, 1987; Guide for the Care
and Use of Laboratory Animals,U.S. National Research Council,
1996); their brains were rapidly dissected into the various areas
(striatum for DA-.sub.1 and DA-.sub.2 receptors, olfactory tubercle
for DA-.sub.3 receptors and cortex for 5-HT.sub.2A receptors) and
stored at -80.degree. C. until the day of assay. Tissues were
homogemnsed in about 50 volumes of Tris HCl, 50 mM, pH 7.4 (for
DA-.sub.1, DA-.sub.2 and 5-HT.sub.2A receptors) or Hepes Na, 50 mM,
pH 7.5 (for DA-.sub.3 receptors), using an Ultra Turrax TP-1810
(2.times.20 sec.), and centriuged at 50000 g for 10 min. The
pellets were resuspended in fresh buffer, incubated at 37.degree.
C. for 10 min and centrifuged as before. The pellets were then
washed once by resuspension in fresh buffer and centrifuged again.
The pellets obtained were resuspended in the appropriate incubation
buffer (Tris HCl, 50 mM, pH 7.1, containing 10 .mu.M pargyline,
0.1% ascorbic acid, 120 mM NaCl, 5 mM KCl, 2 mM CaCl.sub.2, 1 mM
MgCI.sub.2 for DA-.sub.1 and DA-.sub.2 receptors; Hepes Na, 50 mM,
pH 7.5, containing 1 mM EDTA, 0.005% ascorbic acid, 0.1% albumin,
200 nM eliprodil for DA-.sub.3 receptors) just before the binding
assay.
[0157] [.sup.3H]SCH 23390 (specific activity 70.3 Ci/mmol, NEN)
binding to DA-.sub.1 receptors was assayed in a final incubation
volume of 0.5 ml, consisting of 0.25 ml of membrane suspension (2
mg tissue/sample)) 0.25 ml of [.sup.3H]ligand (0.4 nM) and 10 .mu.l
of displacing agent or solvent. Non-specific binding was obtained
in the presence of 10 .mu.M (-)-cis-flupentixol.
[0158] [.sup.3H]Spiperone (specific activity 16.5 Ci/mmol, NEN)
binding to DA-.sub.2 receptors was assayed in a final incubation
volumne of 1 ml, consisting of 0.5 ral of membrane suspension (1 mg
tissue/samnple), 0.5 ml of [.sup.3H]ligand (0.2 nM) and 20 .mu.l of
displacing agent or solvent. Non-specific binding was obtained in
the presence of 100 M (-)sulpiride.
[0159] [.sup.3H]-7-OH-DPAT (specific activity 159 Ci/mmol,
Amersham) binding to DA-.sub.3 receptors was assayed in a final
incubation volume of 1 ml, consisting of 0.5 ml of membranes
suspension (10 mg tissue/sample), 0.5 ml of [.sup.3H]ligand and 20
.mu.l of displacing agent or solvent. Non-specific binding was
obtained in te presence of 1 .mu.M dopamine.
[0160] [.sup.3H]Ketanserin (specific actvity 63.3 Ci/mol, NEN)
binding to 5-HT.sub.2A receptors was assayed in a final incubation
volume of 1 ml consisting of 0.5 ml of minebrane suspension (5
mg,tissue/sample), 0.5 ml of [.sup.3H]ligand (0.7 nM) and 20 .mu.l
of displacing agent or solvent. Non-specific binding was obtained
in the presence of 1 .mu.M methysergide. Incubations (15 min at
37.degree. C. for DA-.sub.2 and 5-HT.sub.2A receptors; 15 min at
25.degree. C. for DA-.sub.1 receptors; 60 min at 25.degree. C. for
DA-.sub.3 receptors) were stopped by rapid filtration under vacuum
through GF/B (for DA-.sub.1, DA-.sub.2 and 5-HT.sub.2A receptors)
or GF/C (for DA-.sub.3 receptors) filters which were then washed
with 12 ml of ice-cold buffer) using a Brandel M-48R. The
radioactivity trapped on the filters was counted in 4 ml of Ultima
Gold MV (Packard) in a LKB 1214 rack beta liquid scintillation
spectrometer with a counting efficiency of 60%.
[0161] H.sub.1 affinity
[0162] Whole cortexes from imale Fischer rats (300-350 g) were
homogenised with a Polytron in nine volumes (w/v) of 50 mzM
Na.sup.+-K.sup.+ phosphate buffer (pH 7.5). The homogenate was
centrifuged at 16500.times. g for 10 min and the particulate
fraction was resuspended in the original volume of buffer. In a
typical experiment, aliquots of the homaogenate (0.3 - 0.4 mg
prot.) were incubated at 25.degree. C. in 0.50 ml of the same
buffer containing [.sup.3H] pyisamiine (2 nM) and the displacing
drugs.
[0163] After 30 min of incubation, 4 ml of ice-cold buffer was
added and the bond and free [.sup.3H] pyrilaminie were separated by
filtration under vacuum through glass fiber filters (Whatman GF/B).
The filters were washed three times with 4 ml of buffer, dried,
placed in 15 ml of Optifluor, and counted by liquid scintillation
spectrometry after a 12 h extraction period.
[0164] All assays were done in triplicate and specific binding was
defined as the total amount of [.sup.3H] pyrilamine bound minus
that bound in the presence of 10.sup.-4M of pyluaxnine. Protein
concentration was determined by the Bradford method.
[0165] [.sup.3H] pyrilamine, (20 ci/nmol) was obtained from New
England Nuclear Corp.
[0166] Muscarinic Affinity
[0167] Male Fischer rats (300-350 g) were killed by decapitation
and cortexes were rapidly removed and homogenised using a Polytron
in 20 vol (w/v) of ice-cold 50 mM PBS buffer (pH 7.4). Homogenates
were centrifuged at 20000.times. g for 15 min. The precipitated
material was resuspended in assay buffer and was used for binding
assay.
[0168] Triplicate incubation tubes contained [.sup.3H] QNB (0.16
nM), various concentrations of drug and an aliquot of freshly
resuspended tissue (.about.0.4 mg prot) in a fal volume of 2 ml.
Tubes were incubated at 37.degree. C. for 60 min and the incubation
was terminated by rapid filtration under vacuum through GF/B glass
fiber filters. The filters were rinsed three times with ice-cold
buffer using a Brandel filtration apparatus (Gaithersburg, Md.,
USA) and were placed in vials containing 15 ml of Optifluor .RTM.,
cooled overnight, and counted in a liquid scintillation. Specific
binding was defined as the excess over blanks containing 1 .mu.M
Atropine.
[0169] [.sup.3H] QNB 42 ci/nmole was obtained from New England
Nuclear Corp.
[0170] Calculations
[0171] Drugs were tested in triplicate at different concentrations,
from 10.sup.-5 to 10.sup.-10 M. IC.sub.50's, the concentration of
drug that caused 50% inhibition of [.sup.3H]Ligand binding, was
obtained using "Allfit" program running on an IBM personal
computer.
IN VIVO TEST
[0172] Antagonism of Apomorphine-induced climbing in mouse
[0173] Groups of ten mice (CD1 male) were dosed with test compounds
by the subcutaneous route 30 jutes before apomorphine and placed
individually into cylindical wire mesh cages (height 14 cm,
diameter 12 cm, mesh size 2 mm). Climbing behaviour was assessed at
5 min intervals for 30 min, is starting 5 min after apomorphine
(1.3 mg/Kg, s.c.) (Greg C. Figdon et al. Neuropsychopharmacology
Vol. 15,.pp231-242 (1996)).
[0174] Antaionism of 5-MeO-DMT-induced Head Twitches in mouse
[0175] Groups of ten mice of the CD1 strain (male) were utilised to
evaluate head twitches induced by 5 methoxy-N,N-dimethyl-tryptamine
(5-MeO-DMT) at a subcutaneous dosage of 10 mg/Kg,
[0176] The evaluations were begun 6 minutes from the 5-MeO-DMT and
lasted 15 minutes (counting the numxiber of head-twitches produced
by the animal). The substances were administered subcutaneously 30
minutes before the 5-MeO-DMT (Greg C. Rigdon et al.
Neuropsychopharmacology Vol. 15,.pp231-242 (1996)).
[0177] Extrapyramidal Symptom
[0178] The test was performed on male Wistar rats (7-8 animals per
group); the catalepsy evaluation was carried out by means of a
metallic bar 0.6 cm in diameter positioned 10 cm from the
workplace. The substances under study were administered
subcutaneously 30 minutes before the first evaluation. The
subsequent observations were recorded at 60, 90, 120, 1807 240, 300
minutes from administration.
[0179] The test consisted in positioning the animal with its
forepaws on the bar and training how long the animal remained
hanging onto the bar employing an end-point of 60 seconds (N. A.
Moore et al. Journal of Pharmacology and Experimental Therapeutics
Vol. 262 pp 545-551 (1992).
RESULTS AND DISCUSSION
[0180] D.sub.1, D.sub.2,D.sub.3 and 5HT.sub.2a affinity
[0181] Table 1 reports the averages and standard errors in the
affinity values expressed as Ki (nM) reported by each product under
study with regard to the SHT.sub.2, D.sub.2, D.sub.1 and D.sub.3
receptors.
[0182] These values were compared with the ones relative to
reference compounds which are structurally similar to the compounds
of the present invention (RS-Octoclothepin, R-(-) Octoclothepin;
S-(+)-Octoclothepin) or belonging to the pharmacological class of
the atypical antipsychotic (Olanzapine and Clozapine).
1TABLE 1 Compounds 5HT.sub.2A D.sub.1 D.sub.2 D.sub.3 Clozapine
11.00 .+-. 1.00 353.00 .+-. 35 250.00 .+-. 57 312.80 .+-. 65.07
Olanzapine 12.00 .+-. 1.00 85.00 .+-. 3.5 69.00 .+-. 17 25.80 .+-.
7.75 RS-Octoclothepin 0.22 .+-. 0.02 2.28 .+-. 0.15 0.36 .+-. 0.07
2.38 .+-. 0.37 R-(-)-Octoclothepin 0.16 .+-. 0.01 2.02 .+-. 0.17
3.64 .+-. 0.46 20.90 .+-. 4.35 S-(+)-Octoclothepin 0.14 .+-. 0.01
1.97 .+-. 0.53 0.40 .+-. 0.04 0.75 .+-. 0.12 (.+-.)-3a 7.85 .+-.
2.2 160 .+-. 77 70 .+-. 17 57 .+-. 6.2 ST1455 1.14 .+-. 0.12 27.00
.+-. 10 3.80 .+-. 0.5 ST1460 1.48 .+-. 0.20 16.40 .+-. 1.0 49.60
.+-. 6.0 ST1461 1.72 .+-. 0.25 22.00 .+-. 1.3 2.06 .+-. 0.2 ST1456
5.1 .+-. 0.40 78.00 .+-. 8.76 37.50 .+-. 3.01 12.20 .+-. 3.65
Haloperidol 164.10 .+-. 23.6 318.30 .+-. 59.2 4.81 .+-. 1.0 15.4
.+-. 3.23 Methysergide 5.3 .+-. 0.8 (-)-cis-Flupentixol 37.8 .+-.
1.7 Sulpiride 240 .+-. 58 Dopamine 11 .+-. 3.4
[0183] The binding evaluations relative to the compound (.+-.)-3a,
to the chloro derivative ST1455 and to the fluoro derivative
ST1456, show that the substitution in position 7 with a halogen is
an important condition for improving the affinities of the formula
(1) compounds towards the 5HT.sub.2A D.sub.2 and D.sub.3
receptors.
[0184] Relative to receptor 5-HT.sub.2A the 7-chloro derivatives,
raceme (ST 1455) and single isomers (form (+) ST1460, form (-) ST
1461) demonstrate an affinity that improves, by virtue of
substitution with halogen, and proves moderately lower than the one
shown by the structurally analogous reference compounds
(RS-Octoclothepin, R-(-)-Octoclothepin, S-(+)-Octoclothepin) and
greater to that shown by the atypical neuroleptics Clozapine and
Olanzapine.
[0185] The enantiomers of the ST 1455 chloro-derivative, like those
of RS-Octoclothepin, do not present significant differences of
affinity towards the 5HT.sub.2a receptor, but reveal a marked
stereoselectivity of action regarding the capacity of interaction
towards the doparinergic D.sub.2 receptor.
[0186] While it is observed that R-(-)-Octoclothepin shows
approximately 10 times less affinity with regard to the D.sub.2
receptor with respect to isoform(+), the preferred compound ST
1460, shows approximately 25 times less affinity than the (-)
isomer, ST 1461.
[0187] It is therefore interesting to note that the preferred
product ST 1460 presents a lower activity on the D.sub.2 receptors
(involved in the extraphyramidal effects), as compared to what its
closest structural analogue R-(-)-Octoclothepin shows, together
with an improved affinity for the 5HT.sub.2 and D.sub.1 receptors
(involved in the neuroleptic action), as compared to what the
reference atypical antipsychotic Olansapinie demonstrates.
[0188] In the case of the preferred product ST 1460, it is
therefore possible to obtain therapeutic effects associated with a
control of extrapyramidal symptoms using lower doses with respect
to those necessary for Clozapine or Olanzapine
[0189] Concerning the racemic 7-fluoro derivative, ST 1456, for
which has been shown an interaction capacity towards the D.sub.2
receptor even below that shown by the best Octoclothepin enantiomer
and towards that shown by Olanzapine, it is believed that it could
possess a stereoselectivity of interaction towards the very same
receptor, analogously to the racemic chloro-derivative.
[0190] Table 2 shows the inhibition constants (pKi) of the formula
(I) compounds and of the reference compounds towards the D.sub.1,
D.sub.2 D.sub.3 and 5HT.sub.2 receptors, and the following ratios
of relative affinity D.sub.1/D.sub.2 and 5HT.sub.2/D.sub.2 . This
last value, if above 1.12, is considered a valid indication for
describing the "atypical" profile of an antipsychotic (Meltzer et
al. J. Pharmacol Exp. Ther 251 (1) pp 238-245 1989).
[0191] Also reported is the LogY parameter which, considering the
relative affinities towards the 5HT.sub.2, D.sub.2, and D.sub.1
receptors of each product identifies and distinguishes a classic
antipsychotic (Log Y>6.48) from an atypical one (Log Y<6.48)
(Meltzer et al J. Pharmacol Exp. Ther 251 (1) pp 238-245 1989).
2TABLE 2 D.sub.1 D.sub.2 D.sub.1/ Log Compound pKi pKi
5HT.sub.2ApKi D.sub.2 5HT.sub.2/D.sub.2 Y Clozapine 6.45 6.60 7.96
0.98 1.21 3.95 Olanzapine 7.07 7.16 7.92 0.99 1.11 5.43
RS-Octoclothepin 8.64 9.44 9.66 0.92 1.02 8.02 R-(-)-Octoclothepin
8.69 8.44 9.80 1.03 1.16 5.37 S-(+)-Octoclothepin 8.71 9.40 9.85
0.93 1.05 7.66 (.+-.)-3a 6.8 7.15 8.1 0.95 1.13 4.99 ST1455 7.57
8.42 8.94 0.90 1.06 6.56 ST1460 7.79 7.30 8.83 1.07 1.21 4.67
ST1461 7.66 8.69 8.76 0.88 1.01 7.40 ST1456 7.11 7.43 8.29 0.96
1.12 5.39 Haloperidol 6.50 8.32 6.78 0.78 0.82 9.14
[0192] Observation of these results suggests that the preferred
compound ST 1460 is superior to its closest structural analogue
(-)-Octoclothepin (respectively 1.21 and 1.16) and different from
its owning racemic form ST1455 and from the isoform(-) ST 1461, for
which the above-mentioned parameters describe a profile of classic
antipsychotic.
[0193] Concerning a comparison with compounds having a known
atypical antipsychotic activity, the relative affinity ratio
5-HT.sub.2/D.sub.2 and the LogY value of the preferred compound ST
1460 describe a atypical profile comparable to that of clozapine
and superior to olanzapine.
[0194] The raceme fluoro derivative ST 1456 is, unlike the racemic
chloro derivative ST1455, an atypical antipsychotic comparable to
the reference compound Olanzapine.
[0195] Another interesting aspect of some formula (I) compounds
that emerges from an examination of the parameters represented in
Table 3 is the high value of the D.sub.3/D.sub.1 receptonal
affinity ratio they showed.
3 TABLE 3 Compound D.sub.1 pKi D.sub.3 pKi D.sub.3/D.sub.1
Clozapine 6.45 6.50 1.01 Olanzapine 7.07 7.59 1.07 RS-Octoclothepin
8.64 8.62 1.00 R-(-)-Octoclothepin 8.69 7.68 0.88
S-(+)-Octoclothepin 8.71 9.12 1.05 (.+-.)-3a 6.8 7.24 1.07 ST1456
7.11 7.91 1.11 Haloperidol 6.50 7.81 1.20
[0196] Said value is, for everyone, comparable to that determined
for the reference compound Olanzapine.
[0197] Moreover, the D.sub.3/D.sub.1 value reported by formula (I)
compounds clearly distinguishes them from the partially atypical
neuroleptic R-(-)-Octoclothepin which shows an 0.88 activity
ratio.
[0198] The relatively greater D.sub.3/D.sub.1 ratio renders formula
(I) products useful in the treatment of the negative symptoms of
schizophrenia which involve the emotional and cognitive sphere,
such as for example dementia, with respect to Octoclothepin, which
with its more active D.sub.1 receptor is oriented towards the
control of symptoms linked to muscle tone.
[0199] Affinity relative to the H1 receptors of histamine and
muescarinics.
[0200] Table 4 reports the average and standard deviations of three
determinations which describe the interaction capacity (Ki, nM) of
each formula (I) and reference compound towards the H.sub.1
receptor of Histamine and towards the muscarinic receptors.
4 TABLE 4 Muscarinic H.sub.1 Receptor Receptors Compound Ki (nM)
E.S. Ki (nM) E.S. Clozapine 14.00 0.00 54.50 0.00 Olanzapine 0.35
0.20 22.10 13.12 RS-Octoclothepin 1.00 0.08 434.10 31.70
R-(-)-Octoclothepin 2.32 0.06 154.60 12.70 S-(+)-Octoclothepin 0.62
0.07 748.60 87.30 ST1455 9.46 1.98 418.20 47.65 ST1460 21.33 12.00
286.40 13.20 ST1461 1.40 0.06 514.50 154.10 ST1456 7.33 1.36
2224.00 105.60 Haloperidol 384.00 0.00 Pyrilamine 12.20 0.09
Atropine 2.59 0.01
[0201] A study of the mentioned parameters shows that the
interaction capacity of the preferred compound ST 1460 towards the
H.sub.1 receptor and towards the muscairtic receptors is less
marked than that shown by its direct structural analog, the
partially atypical neuroleptic R-(-)-Octoclothepin, and by the
reference compounds of the pharmacological class of the atypical
antipsychotic, Clozapine and Olanzapine.
[0202] These results render the compound ST 1460 particularly
useful in the treatment of schizophrenia and distinguish it from
the atypical reference compounds which, by virtue of their greater
interaction capacity towards the above-mentioned receptors,
associate antipsychotic efficacy with the appearance of the
following side effects: dryness of the throat and the respiratory
tract, constipation and weight gain.
5TABLE 5 Dosage that determines approximately 50% climbing
inhibition induced by apomorphine in the mouse Molecule DOSE
(MG/KG) DOSE (.mu.MOLES/KG) Olanzapine 0.12 0.38 Octoclothepin RS
0.02 0.043 Octoclothepin R 0.013 0.026 Octoclothepin S 0.041 0.083
ST 1468 0.052 0.13 ST 1469 0.10 0.24 ST 1470 0.025 0.06
[0203]
6TABLE 6 Dosage that determines 50% of the Head-twitching
inhibition induced by 5-MeO-DMT in the mouse Molecule DOSE (MG/KG)
DOSE (.mu.MOLES/KG) Olanzapine 0.18 0.58 Octoclothepin RS 0.064
0.14 Octoclothepin R 0.089 0.18 Octoclothepin S 0.079 0.16 ST 1468
0.19 0.47 ST 1469 0.196 0.49 ST 1470 0.10 0.24
[0204]
7TABLE 7 Dosage that determines 100% of catalepsy in rats TIME IN
MINUTES OF INSURGENCE OF CATALEPSY IN DOSE DOSE 100% OF THE
MOLECULE (MG/KG) (.mu.MOLES/KG) ANIMALS RS-Octoclothepin 0.6 1.3
120 R-(-)-Octoclothepin 14.85 30 120 S-(+)-Octoclothepin 0.3 0.55
300 ST 1468 1.22 3 180 ST 1469 >12.2 >30 ST 1470 4.06 10
90
[0205] Table 5 shows the calculated dosage of each single compound
that determines a 50% inhibition to climbing behavior induced by
apomorphine in the mouse, an indication of a dopaminergic
activity.
[0206] From the results a reduction of the arty bond which leads to
an increase in dosage by the compound under study (ST 1469) can be
observed both in vivo as in vitro, in that 0.1 mg/Kg (0.24
.mu.moles/Kg) are necessary in order to have an antagonism to the
effect produced by stimulation of the dopaminergic system, of
approximately 50% with respect to R-(-)-Octoclothepin whose dosage
is much lower at 0.013 mg/Kg (0.026 .mu.moles/Kg), an indication of
R-(-)-Octoclothepin's greater affinity bond.
[0207] A lesser affinity to the system classifies it as an Atypical
Antipsychotic and makes it comparable to the class of drugs
represented by olanzapine.
[0208] Stimulation of the serotoniergic system (table 6) by means
of a non-selective agonist of serotomin receptors (5-MeO-DMT)
places the compound (ST 1469) in the in vivo studies comparable to
Olanzapine (0.19 mg/Kg as opposed to 0.18 mg/Kg) and with less
affinity to the serotoninergic system with respect to
R-(-)-Octoclothepin (0.089 mg/Kg).
[0209] The onset of the extrapyramidal syndrome (catalepsy), a
negative effect of the typical neuroleptics, (see table 7) which
effect should be lacking in those that are atypical, favours ST
1469 in that at a dosage corresponding to 30 .mu.moles/Kg
R-(-)-Octoclothepin causes catalepsy in all animals after 120
minutes of treatment while the compound under study does niot
present catalepsy throughout the entire observation period.
[0210] From the in vivo results produced it can be concluded that
ST 1469, as characterized by its demonstrated activity on the
neurotransmitting systems examined and no insurgence of catalepsy,
can be placed in the atypical neuroleptic class of drugs.
* * * * *