Note: Descriptions are shown in the official language in which they were submitted.
~300145
DESCRIPTION
TAURINE-TYPE COMPOUN3S
Technical Field
This invention relates to a novel taurine-type
compound.
Prlor Art
Since the lumen diameter of blood vessels is
regarded as one of the factors which have great influence
on blood flow, vasodilators are used. However, the lumen
diameter of blood vessels with arteriosclerosis, etc. is
less responsive to drugs, and smooth muscle does not exist
in microvessels. Further, from the standpoint of
rheology, unless a pressure gradient is varied, blood flow
is not sufficiently improved even if lumen diameter of
vessels is changed. Therefore, for the improvement of
microcirculation under ischemic condition, treatment with
vasodilators alone is not expected to produce sufficent
effect. Rather, it is very important to positively
improve hematological properties from the standpoint of
promoting the deformability of erythrocytes and of
controlling platelet functions. Thus, therapies for
thrombosis or microcirculation disorder from this
standpoint have been attaching attention in recent years,
and the investigation of the conventional agents for
i30C)~45
-- 2 --
cardiovascular system from hemorheological standpoint has
revealed that pentoxifylline, trapidil and dilazep are
useful as drugs for improving hematological properties as
well.
However, few agent have been developed so far
which put stress on the promotion of deformability of
erythrocytes and the control of platelet function that
have significant hemorheological influence on the flow
within microvessels. Therefore, agents that display
excellent effects thereon have been demanded.
Disclosure of the Invention
The main object of the present invention is to
satisfy the above demand.
That is, the invention provides agents which can
accelerate deformability of erythrocytes and control
platelet function to thereby improve microcirculation.
We conducted extensive reserch on compounds
having said effects, and found that certain taurine-type
compounds can fulfil the above object, and accomplished
the present invention.
The present invention provides a taurine-type
compound represented by the formula
R-N ~ H (I)
( C~2 ) nS3
i3~81~5
3 --
wherein R is a phenyl group optionally having
substituent(s) selected from lower alkyl, halogen-
substituted lower alkyl, lower alkoxy, lower alkanoyl and
halogen atom, and n is 2 or 3.
The taurine-type compounds of the present
invention are novel compounds undisclosed in any
literatures. They not only have extremely strong effect
of improving erythrocyte deformability when compared with
conventional agents, but also possess inhibitory effect on
platelet aggregation, and thus exert remarkable effect of
improving hematological properties and are useful as
medicaments for mammals including humans. That is to say,
the compounds (I) of the present invention have an
excellent effect of promoting erythrocyte deformability
and effect of inhibiting platelet aggregation, and improve
blood fluidity in microcirculatory regions, i.e.,
microcirculation. Therefore, the compounds (I) of the
present invention are useful as agents for preventing and
treating arteriosclerosis, cerebral infarction, myocardial
infarction, peripheral thrombosis and obstruction, etc.
Thus the present invention also provides an
agent for the improvement of microcirculation which
contains an effective amount of the compound of the above
formula (I) and a pharmaceutically acceptable carrier or
excipient.
13()0~4S
4 --
Further the present invention provides a method
of improving microcirculation in a patient in need of
amelioration of microcirculation comprising administering
an effective amount of the compound of the formula (I) to
said patient.
- Throughout the specification and particularly in
the definition of the formula (I), lower alkyl and alkyl
moiety of halogen-substituted lower alkyl, lower alkoxy
and lower alkanoyl are intended to mean straight or
branched saturated hydrocarbon chains having 1 to 6 carbon
atoms or alicyclic saturated hydrocarbon groups having 3
to 6 carbon atoms. Examples thereof are methyl, ethyl,
propyl, cyclopropyl, isopropyl, butyl, isobutyl, sec-
butyl, tert-butyl, cyclobutyl, pentyl, isopentyl,
neopentyl, tert-pentyl, cyclopentyl, hexyl, isohexyl,
cyclohexyl, etc. Examples of halogen atoms are fluorine,
chlorine, bromine and the like. When R in the formula (I)
is a phenyl group having substituents, the number of the
substituents is preferably 1 to 3.
Among the compounds of the formula (I),
preferable are those wherein R is a phenyl group
optionally having one or two substituents selected from
Cl-C4 alkyl, halogen-substituted Cl-C4 alkyl, Cl-C4
alkoxy, Cl-C4 alkanoyl and halogen atom. More preferable
are the compounds of the formula (I) wherein R is a phenyl
i300145
group optionally having one or two substituents selected
from Cl-C4 alkyl, Cl-C2 alkoxy, chlorine atom and fluorine
atom, and n is 3.
The compounds (I) of the present invention can
be prepared by various processes. The typical processes
are illustrated below.
Process ~1)
R - N N - H (~)
~ H 2 ) n ~ 0 ( m )
R--N N (I)
\J (C H2 ) n S 03
In the above reaction scheme, R and n are as
defined above.
The starting material, i.e., a phenylpiperazine
derivative (II) is prepared according to the method
described in the following literatures:
i) C. B. Pollard et al., J. Am. Chem. Soc., 56, 2199
(1934)
ii) the same author et al, the same literature as above,
76, 1853 (1954)
1300~45
iii) the same author et al, J. Org. Chem., 23, 1333
(1958)
The compound of the present invention is
prepared by adding a cyclic sulfonic acid ester (III) in
an equimolar or slightly excess amount, relative to the
phenylpiperazine derivative (II) prepared by the method
described in the above literatures, to a solution of the
phenylpiperazine derivative (II) in a lower alcohol,
preferably methanol, ethanol, propanol and isopropanol,
di lower alkyl ketone, preferably acetone, methyl ethyl
ketone and diethyl ketone or a mixture of these solvents
at a temperature between 0C and room temperature, and
then effecting the reaction by stirring the mixture at a
temperature between room temperature and reflux
temperature for several hours to several days. The
compound of the present invention thus obtained is
isolated by filtering a precipitate formed spontaneously
or by filtering a precipitate formed either upon
concentrating the reaction mixture by evaporating the
solvent under reduced pressure or upon adding a solvent in
which the reaction product is sparingly soluble. The
precipitate is purified by recrystallization from water,
methanol, ethanol, propanol, isopropanol, acetone, methyl
ethyl ketone, diethyl ketone or a mixture of these
solvents.
i3~5
-- 7 --
Process _(2)
A
R--N 1~--H (II)
~ X-- (CH2 ) n S 03 M (IV)
Ao/
R--N N\ e (I)
\ J (C H2 ) n S 03
In the above reaction scheme, X is a halogen
atom, M is an alkali metal or alkaline earth metal,
preferably sodium or patassium, and R and n are as defined
above.
The compound of the present invention is
prepared by reacting the phenylpiperazine derivative (II~
with equimolar or slightly excess amount of the sulfonic
acid derivative (IV) in water or in a solvent which is a
mixture of water and the solvent exemplified in connection
with Process (1) in arbitrary proportions for several
hours to several days at a temperature between room
temperature and reflux temperature of the solvent used.
The compound of the present invention thus obtained is
isolated and purified in the same manner as described in
Process (1).
130014~i
The compound of the present invention may be
administered orally in such dosage forms as tablets,
pills, capsules, granules, powders, liquids, etc, or
pareterally in such dosage forms as injections including
intravenous, intramuscular and like injections,
suppositories, etc.
Each of these dosage forms can be prepared by
formulation methods which are conventional in the art. In
preparing oral solid pharmaceutical compositions, vehicles
and, if required, bindersr disintegrants, lubricants,
coloring agents, sweetening agents, flavors, etc. may be
added to the active principle of the invention, and then
tablets, coated tablets, granules, powders, capsules, etc.
may be prepared by a conventional method. In preparing
injections, to the active component of the present
invention may be added pH adjusting agent, buffer,
stabilizers, isotonic agents, local anesthetics, etc., and
then subcutaneous, intramuscular, intravenous and like
injections may be prepared by a conventional method. In
preparing suppositories, to the active component of the
present invention may be added bases and, if required,
surfactants, etc., and then suppositories can be prepared
according to a conventional method.
In preparing tablets, capsules, granules and
powders, useful vehicles include lactose, sucrose, starch,
~300145
talc, magnesium stearate, calcium stearate, crystalline
cellulose, methyl cellulose, carboxymethyl cellulose,
glycerin, sodium alginate, arabic gum, etc; useful binders
include polyvinyl alcohol, polyvinyl ether, ethyl
cellulose, arabic gum, shellac, sucrose, etc; useful
lubricants include magnesium stearate, talc, etc.;
coloring agents and disintegrants can be those commonly
used in this field. Tablets may be coated by a
conventional method.
Bases to be used in preparing suppositories
include oleaginous bases such as cacao butter,
polyethyleneglycol, lanolin, fatty acid triglycerides,
Witepsol (registered trademark, product of Dynamite
Nobel).
In the pharmaceutical compositions of the
invention, the content of the compound of the present
invention may vary according to dosage form, solubility of
the compound, chemical properties, administration route,
dosage plan, etc. Generally, the content is preferably
about 10-15 w/w% in oral compositions (tablets, capsules,
etc.), about 0.1-1 w/v% in injections, and about 1-5 w/w%
in suppositories.
The dose of the compound of the formula (I) of
the present invention is appropriately determined case by -
case depending on symptoms, age and sex of subjects,
~300145
-- 10 --
etc. When administered orally, a daily dose of about 1 to
about 300 mg is generally given to human adults in 2-4
divided doses. When administered in the form of
injection, for example, intravenous injection, a dose of 2
ml (1-10 mg), which may be diluted with saline or glucose
solution for injection if re~uired, is generally injected
gradually to human adults over 5 minutes or more once a
day. In the case of suppositories, a daily dose of 1-300
mg is generally inserted intrarectally to human adults
once or twice a day at intervals of 6-12 hours.
The present invention will be described in
greater detail with reference to the following examples
and pharmacological tests.
Example 1
4-(3-Sulfopropyl)-l-(p-tolyl)plperazine, inner salt
(Compound 1)
A 1.48 g quantity (8.40 mmols) of l-(p-
tolyl)piperazine is dissolved in 30 ml of ethanol and the
solution is ice-cooled. A solution of 1.22 g (10 mmols)
of 1,3-propanesultone in 10 ml of acetone is slowly added
dropwise with stirring. The mixture is stirred with ice-
cooling for one hour and subsequently stirred at room
temperature overnight to effect reaction, and then 50 ml
of acetone is added thereto. The crystals precipitated
are collected by filtration and recrystallized from water-
1300145
methanol-acetone.
Yield: 1.3 9 (52%)
M.p.: 268-270 C (decomp.)
TLC: Rf=0.33 (silica gel; chloroform/methanol
=3/1 (v/v))
IR spectrum (KBr tablet):
1160 cm~l (vSO (asymmetric))
1030 cm~ SO2(symmetric
FAB mass spectrum (m/e)
299 (M+1)
H-NMR spectrum (DMSO-d6, TMS as an internal
standard, ~)
2.01 (2H, quintet, J=6.8 Hz, -CH2-)
2.21 (3H, singlet, -CH3)
2.5-4.0 (12H, multiplet, -CH2-)
6.9 (2H, doublet, J=8.7 Hz, ~-H)
7.1 ~2H, doublet, J=8.7 Hz, ~-H)
9.9 (lH, broad singlet,~N~H)
Elemental analysis (for C14H22N2S13' MW= 298-40)
Calculated value (%)
H, 7.43; C, 56.35; N, 9.39
Found value (%)
H, 7.74; C, 56.12; N, 9.27
Example 2
4-(3-Sulfopropyl)-l-phenylpiperazine, inner salt ~Compound
1300i4S
- 12 -
Using 3.25 g (20 mmols) of l-phenylpiperazine
and 2.44 g (20 mmols) of 1,3-propanesultone, the reaction
is conducted in the same manner as in Example 1, and the
resulting crude crystals are recrystallized from water.
Yield: 1.4 g (25 ~)
M.p.: decomposition at 260 C or higher
TLC: Rf=0.23 (silica gel; chloroform/methanol
=3/1 (v/v))
IR spectrum I KBr tablet3:
1155 cm~l (vsO2 (asymmetric))
1030 cm~l (vsO2 (symmetric))
FAB mass spectrum (m/e~: 285 (M+l)
H-NMR spectrum (DMSO-d6, TMS as an internal
standard, ~)
2.00 (2H, quintet, J=6.8 Hz, -CH2-)
2.64 (2H, triplet, J=6.8 Hz, -CH2-)
3.0-3.5 (lOH, multiplet, -CH2-)
6.5-7.0 (3H, multiplet, ~-H)
7.19 (2H, multiplet, ~-H)
Elemental analysis (for C13H20N2S13' MW
Calculated value (~)
H, 7.09; C, 54.91; N, 9.85
Found value (~)
H, 7.38; C, 55.19; N, 10.06
~30C~1~5
- 13 -
Example 3
4-(2-Sulfoethyl)-l-phenylpiperazine, inner salt (Compound
3)
A 3.25 g quantity (20 mmols) of l-phenyl-
piperazine is dissolved in 10 ml of water, and a solution
of 4.3 g (20 mmols) of sodium 2-bromoethanesulfonate in 10
ml of water is added dropwise thereto at room temperature
with stirring. The mixture is subjected to reaction at
room temperature for 1 hour and then at 70 C overnight,
and cooled. The crystals precipitated are collected by
filtration and recrystallized from water-acetone.
Yield: 1.8 g (33%)
M.p.: decomposition at 250C or higher
TLC: Rf=0.12 (silica gel; chloroform/methanol=
3/1 (v/v))
IR spectrum (KBr tablet):
1170 cm~l (vsO2 (asymmetric))
1050 cm~l (vsO2 (symmetric))
FAB mass spectrum (m/e): 271 (M+l)
H~NMR spectrum (DMSO-d6, TMS as an internal
standard, ~)
2.8-3.5 (12H, multiplet, -CH2-)
6.7-7.0 (3H, multiplet, ~-H)
7.26 (2H, multiplet, ~-H)
Elemental analysis (for C12H18N2S13~ MW 270-35)
~3~A~
- 14 -
Calculated value (%)
H, 6.71; C, 53.31; N, 10.36
Found value (%)
~, 7.06; C, 52.98; N, 10.20
Example 4
4-(3-Sulfopropyl)-l-(p-acetylphenyl)piperazine, inner salt
(Compound 4)
A 4.1 g quantity (20 mmols) of l-(p-
acetylphenyl)piperazine is dissolved in 40 ml of acetone,
and a solution of 2.44 g (20 mmols) of 1,3-propanesultone
in 10 ml of acetone is added dropwise thereto at room
temperature with stirring. After reaction at room
temperature with stirring overnight, the yellow crystals
precipitated are collected by filtration and then
recrystallized from water-acetone.
Yield: 1.7 g (26%)
M.p.: decomposition at 250 C or higher
TLC: Rf=0.10 (silica gel; chloroform/methanol
=3/1 (v/v))
IR spectrum (KBr tablet):
1650 cm~l (vC=O)
1150 cm~l (vsO2 (asymmetric))
1030 cm~l (vsO2 (symmetric))
FAB mass spectrum (m/e)
327 (M+l)
1300~45
H-NMR spectrum (DMSO-d6, TMS as an internal
standard, ~)
1.98 (2H, quintet, J=6.8 Hz, -CH2-)
2.47 (3H, singlet, CH3CO-)
2.64 (2H, triplet, J=6.8 Hz, -CH2-)
2.8-3.5 (lOH, multiplet, -CH2-)
7.04 (2H, doublet, J=8.7 Hz, ~-H)
7.82 (2H, doublet, J=8.7 Hz, ~-H)
Elemental analysis (for C15H22N2SlO4, MW
Calculated value (%)
H, 6.79; C, 55.20; N, 8.58
Found value (%)
H, 7.02; C, 54.95; N, 8.66
Example 5
4-(3-Sulfopropyl)-l-(p-fluorophenyl)piperazine, inner salt
(Compound 5)
The reaction is conducted in the same manner as
in Example 4 except that 5.2 g (29 mmols) of l-(p-
fluorophenyl)piperazine and 3.5 g (29 mmols) of 1,3-
propanesultone are used. The resulting product is
recrystallized from ethanol.
Yield: 2.5 g (29 %)
M.p.: 265-268 C (decomp.)
TLC: Rf=0.18 (silica gel; chloroform/methanol
=3/1 (v/v))
~300~A5
- 16 -
IR spectrum ~Br tablet):
1165 cm~l (vsO2 (asymmetric))
1025 cm~l (vsO2 (symmetric))
FAB mass spectrum (m/e): 303 (M+l)
H-NNR spectrum (DMSO-d6, TMS as an internal
standard, ~)
2.04 (2H, quintet, J=6.8 Hz, -CH2-)
2.69 (2H, triplet, J=6.8 Hz, -CH2-)
2.8-4.0 (lOH, multiplet, -CH2-)
7.06 (4H, multiplet, ~-H)
9.9 (lH, broad sinylet, ~N~H)
Elemental analysis (for C13HlgN2SlO3Fl, MW
Calculated value (%)
H, 6.33; C, 51.64; N, 9.26
Found value (%)
H, 6.58; C, 51.77; N, 9.25
Example 6
4-(3-Sulfopropyl)-l-(o-methoxyphenyl)piperazine, inner
salt (Compound 6)
The reaction is conducted in the same manner as
in Example 4 except that 5.0 g (26 mmols) of l-(o-
methoxyphenyl)piperazine and 3.2 g (26 mmols) of 1,3-
propanesultone are used. The resulting product is
recrystallized from methanol.
Yield: 6.5 g (80%)
13001~S
M.p.: 271-273 C (decomp.)
TLC: Rf=0.38 (silica gel; chloroform/methanol=
3/1 (v/v))
IR spectrum (KBr tablet):
1160 cm~l (vs02 (asymmetric))
1035 cm~l (vs02 (symmetric))
FAB mass spectrum (m/e): 315 (M+l)
H-NMR spectrum (DMSO-d6, TMS as an internal
standard, O)
2.04 (2H, quintet, J=6.8 Hz, -CH2-)
2.68 (2H, triplet, J=6.8 Hz, -CH2-)
2.5-4.0 (lOH, multiplet, -CH2-)
3.79 (3H, singlet, -OCH3)
6.7-7.3 (4H, multiplet, ~-H)
9.86 (lH, broad singlet, /N~ H)
Elemental analysis (for C14H22N2S104, MW= 314.40)
Calculated value (%)
H, 7.05; C, 53.48; N, 8.91
Found value (%)
H, 7.34; C, 53.22; N, 8.83
Example 7
4-(3-Sulfopropyl)-l-(o-trifluoromethylphenyl)piperazine,
inner salt (Compound 7)
The reaction is conducted in the same manner as
in Example 4 except that 5.0 g (22 mmols) of l-(o-
1~00145
- 18 -
trifluoromethylphenyl)piperazine and 2.7 g (22 mmols) of
1,3-propanesultone are used. The resulting product is
recrystallized from methanol-water.
Yield: 5.3 g (69%)
M.p.: 292-294 C (decomp.)
TLC: Rf=0.15 (silica gel; chloroform/methanol
=3/1 (v/v))
IR spectrum (KBr tablet):
1145 cm~l tVso2 (asymmetric))
1030 cm~l (~SO2 (symmetric))
FAB mass spectrum (m/e)
353 (M+l)
H-NMR spectrum (DMSO-d6, TMS as an internal
standard, ~)
2.03 (2H, quintet, J=6.8 Hz, -CH2-)
2.68 (2H, triplet, J=6.8 Hz, -CH2-)
2.7-4.2 (lOH, multiplet, -CH2-)
7.1-7.6 (4H, multiplet, ~-H)
Elemental analysis (for C14H19N2SlO3F3, MW= 352.37)
Calculated value (%)
H, 5.43; C, 47.72; N, 7.95
Found value (%)
H, 5.62; C, 47.53; N, 7.97
Example 8
4-(3-Sulfopropyl)-l-(m-chlorophenyl)piperazine, inner salt
~301~
-- 19 --
(Compound 8)
The reaction is conducted in the same manner as
in Example 4 except that 4.22 g (21.5 mmols) of l-(m-
chlorophenyl)piperazine and 2.62 g (21.5 mmols) of 1,3-
propanesultone are used. The resulting product is
recrystallized from water-acetone.
Yield: 4.4 g (64 %)
M.p.: decomposition at 245 C or higher
TLC: Rf=0.17 (silica gel; chloroform/methanol
=3/1 (v/v))
IR spectrum (KBr tablet):
1150 cm 1 (~S02 (asymmetric))
1035 cm~l (vs02 (sym~etric))
FAB mass spectrum (m/e): 319 (M+l)
H-NMR spectrum (DMSO-d6, TMS as an internal
standard, ~)
2.03 (2H, quintet, J=6.8 Hz, -CH2-)
2.67 (2H, triplet, J=6.8 Hz, -CH2-)
2.5-4.0 (lOH, multiplet, -CH2-)
6.8-7.3 (4H, multiplet, ~-H)
9.86 (lH, broad singlet, ~N~H)
Elemental analysis (for C13H19N2S13Cll' MW
Calculated value (%)
H, 6.01; C, 48.98; N, 8.79
Found value (~)
~30~1~5
- 20 -
H, 6.15; C, 49.30; N, 8.66
Example 9
4-(3-Sulfopropyl)-l-(p-methoxyphenyl)piperazine, inner
salt (Compound 9)
The reaction is conducted in the same manner as
in Example 4 except that 4.2 g (22 mmols) of l-(p-
methoxyphenyl)piperazine and 3.2 g (26 mmols) of 1,3-
propanesultone are used. The resulting product is
recrystallized from water-acetone.
Yield: 4.2g (61~)
M.p.: 285-287 C (decomp.)
TLC: Rf=0.43 (silica gel; chloroform/methanol/water=
65/25/4 (v/v/v))
IR spectrum (KBr tablet):
1160 cm~l (vsO2 (asymmetric))
1030 cm~l (~SO2 (symmetric))
FAB mass spectrum (m/e): 315 (M+l)
H-NMR spectrum (DMSO-d6, TMS as an internal
standard, ~)
2.05 (2H, quintet, J=6.8 Hz, -CH2-)
2.68 (2H, triplet, J=6.8 Hz, -CH2-)
2.8-3.6 (lOH, multiplet, -CH2-)
3.7 (3H, singlet, -CH3)
6.84 (2H, doublet, J=8.9 Hz, ~-H)
6.97 (2H, doublet, J=8.9 Hz, ~-H)
~1300~4~i
9.9 (lH, broad singlet, / ~ )
Elemental analysis (for C14H22N2S14' MW 314- 0)
Calculated value (%)
H, 7.05; C, 53.48; N, 8.91
Found value (%)
H, 7.32; C, 53.36; N, 8.87
Example 10
4-(3-Sulfopropyl)-1-(3,4-dimethoxyphenyl)piperazine, inner
salt (Compound 10)
The reaction is conducted in the same manner as
in Example 4 except that 6.4 g (29 mmols) of 1-(3,4-
dimethoxyphenyl)piperazine and 4.3 g (35 mmols) of 1,3-
propanesultone are used. The resulting product is
recrystallized from water-acetone.
Yield: 9.2 9 (92%)
M.p.: 275-276 C
TLC: Rf=0.46 (silica gel; chloroform/methanol/water=
65~25/4 (v/v/v))
IR spectrum (KBr tablet):
1160 cm~l (~SO2 (asymmetric))
1030 cm~l (~SO2 (symmetric))
FAB mass spectrum (m/e)
345 (M+l)
H-NMR spectrum (DMSO-d6, TMS as an internal
standard, ~)
~3001~5
- 22 -
2.05 (2H, quintet, J=6.8 Hz, -CH2-),
2.70 (2H, triplet, J=6.8 Hz, -CH2-)
2.8-3.6 (lOH, multiplet, -CH2-)
3.68 (3H, singlet, -CX3)
3.75 (3H, singlet, -CH3)
6.4-6.9 (3H, multiplet, ~-H)
9.9 (lH, broad singlet, ~N~H)
Elemental analysiS (C15H24N2S1O5' MW
Calculated value (%)
H, 7.02; C, 52.31; N, 8.13
Found value (%)
H, 7.22; C, 52.16; N, 8.11
Example 11
4-(3-Sulfopropyl)-l-(p-tert-butylphenyl)piperazine, inner
salt (Compound 11)
The reaction is conducted in the same manner as
in Example 4 except that 2.2 g (10 mmols) of l-(p-tert-
butylphenyl)piperazine and 1.8 g (15 mmols) of 1,3-
propanesultone are used. The resulting product is
recrystallized from water-methanol-acetone.
Yield: 2.2 g (65 ~)
M.p.: decomposition at 285 C or higher
TLC: Rf=0.5 (silica gel; chloroform/methanol
=2/1 (v/v))
IR spectrum (RBr tablet):
~300145
1160 cm~l (vsO2 (asymmetric))
1035 cm~l (vsO2 (symmetric))
FAB mass spectrum (m/e): 341 (M+l)
H-NMR spectrum (DMSO-d6, TMS as an internal
standard, ~)
1.24 (9H, singlet, CH3)
2.0 (2H, quintet, J=6.8 Hz, -CH2-)
2.66 (2H, triplet, J=6.8 Hz, -CH2-)
2.5-4.0 (lOH, multiplet, -CH2-)
6.9 (2H, doublet, J=8.9 Hz, ~-H)
7.3 (2H, doublet, J=8.9 Hz, ~-H)
9.9 (lH, broad singlet, / ~ )
Elemental analysis (for C17H28N2S13/ MW 340.48)
Calculated value (%)
H, 8.29; C, 59.97; N, 8.23
Found value (%)
H~ 8.53; C, 59.87; N, 8.14
Example 12
4-(3-Sulfopropyl)-l-(o-tolyl)piperazine, inner salt
(Compound 12)
The reaction is conducted in the same manner as
in Example 4 except that 4.1 g (23 mmols) of l-(o-
tolyl)piperazine and 3.4 g (28 mmols) of 1,3-
propanesultone are used. The resulting product is
recrystallized from water.
~30~145
~ 24 -
Yield: 4.49 (63%)
M.p.: 289-291 C (decomp.)
TLC: Rf=0.57 (silica gel; chloroform/methanol/water=
65/25/~ (v/v/v))
IR spectrum (KBr tablet):
SO2 Y ) )
1030 cm~~ SO (symmetric))
FAB mass spectrum (m/e): 299 (M+l)
H-NMR spectrum (DMSO-d~, TMS as an internal
standard, ~)
2.05 (2H, quintet, J=6.8 Hz, -CH2-)
2.26 (3H, singlet/ -CH3)
2.70 (2H, triplet, J=6.8 Hz, -CH2-)
2.9-3.7 (lOH, multiplet, -CH2-)
6.94-7.23 (5H, multiplet, ~-H)
10.0 (lH, broad singlet, ~N~ )
Elemental analysis (for C14H22N2S13' MW= 298-40)
Calculated value (~)
H, 7.43; C, 56.35; N, 9.39
Found value (%)
H, 7.66; C, 56.21; N, 9.29
Example 13
4-(3-Sulfopropyl)-l-(o-chlorophenyl)piperazine, inner salt
(Compound 13)
The reaction is conducted in the same manner as
~30~5
in Example 4 except that 4.2 g (21.4 mmols) of 1-(o-
chlorophenyl)piperazine and 3.2 g (26.2 mmols) of 1,3-
propanesultone are used. The resulting product is
recrystallized from water.
Yield: 4.4 g (64 %)
M.p.: decomposition at 290 C or higher
TLC: Rf=0.55 (silica gel; chloroform/methanol/water=
=65/25/4 (v/v/v))
IR spectrum (KBr tablet):
1155 cm~l (vsO2 (asymmetric))
1030 cm~l (vsO2 (symmetric))
FAB mass spectrum (m/e): 319 (M+l)
H-NMR spectrum (DMSO-d6, TMS as an internal
standard, ~)
2.04 (2H, quintet, J=6.8 Hz, -CH2-)
2.70 (2H, triplet, J=6.8 Hz, -CH2-)
2.8-3.8 (lOH, broad multiplet, -CH2-)
7.04-7.51 (4H, multiplet, ~-H)
10.05 (lH, broad singlet, ~N~H)
Elemental analysis (for Cl3Hl9N2slo3cllr MW 3
Calculated value (%)
H, 6.01; C, 48.98; N, 8.79
Found value (%)
H, 6.30; C, 48.83; N, 8.77
Pharmacological test
13~01~5
- 26 -
The pharmacological tests with respect to the
compounds (I) according to the present invention are
described below.
(A) Effect on acceleration of erythrocyte deformability
Experiment 1: filter filtration method
A dispersion of erythrocytes used was prepared
by cetrifuging heparinized venous blood of Japanese albino
rabbit (1100 r.p.m. x 7 minutes, at 4 C), repeating the
steps of washing the precipitate with phosphate buffered
saline (140.5 mM NaCl, 8 mM Na2HPO4, 2 mM KH2PO4, pH 7.4)
having a temperature of 4 C and centrifuging the
precipitate (2800 r.p.m. x 10 minutes, at 4C) for several
times, and dispersing the centrifugate in a phosphate
buffered saline having a temperature of 4C to give a
dispertion of 5%-Hct Ihematocrit).
Subsequently NaCl was added to the dispersion to
adjust it to a high osmotic pressure oE 400 mOsm/kg.
Then, time (FR) required for 0.5 ml of the dispersion of
erythrocytes to pass through a filter at 37C under the
influence of gravity was determined with or without
addition of drug. The filter employed was that for
determination of erythrocyte deformability (5 ~m of pore
diameter, 13 mm~, product of Nomura Micro Science). The
variation rate(%) calculated from the following equation
was used as an index of improvement of deformability.
~300~S
- 27 -
FR(drug) - FR(control)
Variation rate (%) =
FR(control) - FR(blank)
wherein FR(drug) stands for FR value determined with
addition of drug, FR(control) stands for FR value
determined without addtion of drug, and FR(blank) stands
for FR value of phosphate buffered saline.
The results indicate that the variation rate was
-11.9% in the case of pentoxifylline (Trental~, product of
Hoechst Janan), while it was -24% in the case of 4-(3-
sulfopropyl)-l-(p-tolyl)piperazine (inner salt) of the
present invention (Compound 1). Therefore it is seen that
the compound of the present invention remarkably improves
the erythrocyte deformability.
Experiment 2: viscosity determination method
Blood viscosity was determined at 37C at the
shear rate of 150, 75, 37.5 and 18.75 sec l using ELD
rotational viscosimeter (product of Tokyo Keiki, 0.8
cone).
After heparinized venous blood of Japanese
albino rabbit was preserved at 4C for 24 hours, the blood
was tested with or without addtion of drug (20 ~/ml).
The results indicated that Compound 1 of the
present invention exerted stronger vicosity reducing
effect than the controls, i.e., dilazep (Comelian~,
product of Kowa) and pentoxifylline (Trental~, product of
~300145
- 28 -
Hoechst Japan). The results are shown in Table 1.
Table 1
. . . _
DrugViscosity of preserved blood (cp)
~20 ~g/ml)150sec~1 75sec~l 37.5sec~l 18.75sec~
Compound 1 3.55 4.26 5.34 6.72
Dilazep 4.02 5.46 7.20 8.88
Pentoxifylline 4.15 5.64 8.70 13.4
Blank 4.26 5.94 9.00 16.8
.
Experiment 3: morphological observation
A dispersion of erythrocytes of Japanese albino
rabbit (46~-Hct (hematocrit), solvent: phosphate buffered
saline) was prepared in the same manner as in Experiment
1, and a drug (20 ~g/ml) was added thereto. The mixture
was then diluted 25-fold with phosphate buffered saline
havlng a high osmotic pressure of 400 mOsm/kg, and
observed by an optical microscope.
According to the results, when Compound 1 or 4-
(3-sulfopropyl)-1-(p-fluorophenyl)piperazine, inner salt
(Compound 5) or 4-(3-sulfopropyl)-1-(o-methoxyphenyl)-
piperazine, inner salt (Compound 6) of the present
invention was added, each of these drugs exerted the
effect of normalizing the shape of erythrocytes which had
been made abnormal due to the high osmotic pressure. The
effect was remarkably stronger than that of pentoxifylline
(Trental~, product of Hoechst Japan) serving as control,
-and equal or superior to that of dilazep (Comelian~,
~ ~3 [)O~L~5
- 29 -
product of Kowa) serving as control.
While pentoxifylline, trapidil and dilazep are
recognized as typical agents for promoting erythrocyte
deformability, the above test demonstrated that this
erythrocyte deformability promoting effect of the
compounds of the present invention are stronger than that
of dilazep which was reported to exhibit 10 times the
effect of trapidil and 100 times the effect of
pentoxifylline (Susumu Yamamoto et al., Japanese
Pharmacology & Therapeutics, 11(10), 4273 (1983)).
(B) Inhibitory effect of platelet aggregation
Platelet rich plasma (PRP) and platelet poor
plasma (PPP) used were prepared from citrlc acid-added
arterial blood of Japanese albino rabbit. The platelet
aggregation test was conducted according to the me~hod
described in literature ~G.V.R. Born, Nature, 194, 927
tl962)). The inhibitory effect of the drug on platelet
aggregation induced independently by collagen (final
concentration: 5 ~g/ml) and by ADP (adenosine diphosphate
final concentration: 10 ~M) were determined using
aggregometer.
The results indicated that, at a concentration
of 100 ~g/ml, the aggregation inhibitory effects of
Compounds 1, 5 and 6 of the present invention were 1.1-2.3
times (collagen-induced) and 1.7-3.9 times (ADP-induced)
1~00145
- 30 -
as strong as dilazep (Comelian~, product of Kowa).
(C) Effect on blood viscositv on disturbing blood flow
The common carotid arteries and jugular veins of
Wister-rats 300-370 g in body weight were exposed under
anesthesia with pentobarbital. Blood sample was obtained
as heparinized blood (1) from one of the veins. Then both
of arteries were ligated. Sixty minutes after the
ligation, the other vein was ligated, and, from the end of
the vein, blood returning from the head was similarly
collected as heparinized blood (2).
The percent increase in the viscosity of the
heparinized blood (2) was determined based on the
viscosity of the heparinized blood (1). The blood
viscosity was determined immediately after blood
collection using the same rotational viscosimeter as used
in Experiment 2 of the above item (A) Effect on
acceleration of erythrocyte deformability. The test drugs
were dissolved in physiological saline and injected via
tail vein 15 minutes before the ligation of the vein.
The results obtained at shear rates of 37.5, 75
and 150 sec 1 were shown in Table 2.
In the control group which was given only
physiological saline, marked increase in viscosity was
observed at all shear rates of 18.75, 37.5, 75, 150 and
375 sec 1. In contrast, as evident from the results shown
~3~4~
- 31 -
in Table 2, the compound of the present invention such as
4-~3-sulfopropyl)-1-(p-tert-butylphenyl)piperazine, inner
salt (Compound 11) displayed siginificant inhibitory
effect on the increase in blood viscosity, which proved to
be stronger than that of known agents for improving
microcirculation, i.e. pentoxifylline and trapidil.
On the other hand, diltiazem which is a
vasodilator was not effective in this test at a dose ~0.5
mg/kg) at which this drug produce a vasodilating action.
Table 2
Percent increase in blood viscosity
Test drug Shear rate (sec 1)
(dosage) 37.5 75 150
Control 29.6+1.8 22.3+0.915.4+0.5
Compound 1115.8+3.2 # 13.0+2.9 6.8+2.3**
(1 mg/kg)
Compound 1117.8+1.7 # 14.5+1.510.9+1.2
Pentoxifylline27.8+2.7 20.5+2.113.5+1.8
Trapidil 26.0+1.7 20.7+0.314.lil.7
Diltiazem 27.5+0.6 22.6+1.014.9+2.3
(Note) Means + S.D.
* p<0.05 (in comparision with control)
** p<0.01 (in comparision with control)
**# p<0.05 (in comparision with pentoxifilline)
~300~S
(D) Effect on blood pressure in anesthetized rat
Blood pressure was determined by cannulation in
the carotid artery of pentobarbital-anesthetized rat and
connecting the cannula to a blood pressure transducer.
When 0.5 mg/kg of diltiazem was administered,
the effect on blood pressure was observed immediately
after the administration and the blood pressure was
recovered after about 20 minutes later. When 1 mg/kg or
10 mg/kg of pentoxifylline was administered, the transient
dose-dependent effect on blood pressure was observed
immediately after the administration.
On the other hand, it was observed that, when 1
mg/kg or 10 mg/kg of the compound of the present
invention, e.g., Compound 11 was administered, the
compound had substantially no influence on blood pressure,
demonstrating its hemorheological effect selectively.
The following preparation examples illustrate
pharmaceutical compositions containing the compound of the
present invention and preparing methods thereof.
Preparation Example 1 (Preparation of tablets)
Comound 11 50 g
Lactose 200 g
Corn starch 80 g
Hydrolyzed starch 20 g
~30014S
- 33 -
Calcium stearate lO g
360 g
Compound 11, lactose, corn starch and hydrolyzed
starch were mixed, and granulation was conducted after
adding water thereto, giving active paste. After drying
overnight at 45C, the granules were sieved, and thereto
was added calcium stearate, followed by compression,
giving tablets 360 mg in weight and 10 mm in diameter.
Preparation Example 2 tPreparaiton of tablets)
Compound 11 25.0 g
Lactose 115.0 g
Corn starch 50.0 g
Gelatinized corn starch 8.0 g
Calcium stearate 2.0 g
200.0 9
Compound ll, lactose, corn starch and
gelatinized corn starch were mixed. After crushing, the
mixture was made pasty by adding water. The paste was
dried overnight at 45C, mixed with calcium stearate and
shaped by compression into tablets 200 mg in weight and 8
mm in diameter.
Preparation Example 3 (Preparation of capsules)
Compound ll 25.0 g
Lactose 150.0 g
Corn starch 40.0 g
13Q014S
- 34 -
Talc5.0 9
220.0 g
Compound 11, lactose and corn starch were mixed
and crushed. After mixing with talc, the mixture was
packed into hard gelatin capsules.
Preparation Example 4 (Preparation of injections)
A 50 g quantity of Compound 11 and 400 g of
glucose were successively dissolved in 8000 ml of
distilled water for injection with stirring. Distilled
water for injection was further added thereto to adjust
the total amount to 10000 ml. The mixture was sterile-
filtered, and placed into 2 ml-colorless amples. AftPr
passing nitrogen gas therethrough, the amples were sealed.
Preparation Example 5 (Preparation of suppositories)
Compound 11 50 mg
Witepsol S55 2 g
(trademark, product of
Dynamite Nobel, mixture
of mono, di and triglycerides
of saturated fatty acids
ranging from lauric acid
to stearic acid)
Witepsol S55 was heated at 120C for 30 minutes
and cooled at room temperature to a temperature below
50C, and mixed sufficiently with Compound 11 with
i3~3114S
- 35 -
stirrin~. The mixture was charged into a mold at about
38C. The suppository was prepared by cooling after
solidification upon coolin~.
Preparation Examples 6 to 20
Following the procedure of preparation Examples
1-5 and using Compounds 1, 2 and 6 in place of Compound
11, each of the preparations were formulated with the same
composition.
