N-(12-Nitroxydodecyl)-6-(4-ethyl or isopropyl-1-piperazinyl)pyridine-3-carboxamide or physiologically acceptable salts thereof. The said compounds have excellent inhibiting activity of cerebral edema, especially ischemic cerebral edema, and inhibiting activity of delayed neuronal death (an inhibiting activity of Ca-influx in neuronal cells). Cerebral edema is a pathologic condition accompanying cerebrovascular disorders, especially the acute stage cerebrovascular disorders and then the compounds are useful as an inhibiting agent for cerebral edema or a therapeutic agent for cerebrovascular disorders. Moreover, because the compounds do hardly show a behavior suppressing action, which is considered to be side effect in treating cerebrovascular disorders at the acute stage, they are an excellent therapeutic agent for, in particular, the acute stage cerebrovascular disorders. Moreover, the compounds show a cerebral protective activity (an anti-anoxic activity), an activity of increasing cerebral blood flow, and an activity of inhibiting lipid peroxidation, and these activities may lead to the increased utility as a therapeutic agent for cerebrovascular disorders.

TECHNICAL FIELD 
This invention relates to a pyridinecarboxamide derivative. More 
particularly, this invention relates to an 
N-(12-nitroxydodecyl)-6-piperazinylpyridine-3-carboxamide derivative, a 
process for the preparation thereof and a pharmaceutical preparation which 
comprises as an active ingredient said derivative. Moreover, this 
invention relates to a therapeutic agent for cerebrovascular disorders or 
cerebral edema which comprises as an active ingredient said pyridine 
carboxamide derivative. Moreover, this invention relates to a method for 
the treatment of cerebrovascular disorders or cerebral edema which 
comprises administering said pyridinecarboxamide derivative. Moreover, 
this invention relates to an intermediate for the synthesis of said 
pyridinecarboxamide derivative. 
BACKGROUND ART 
Neuronal cells are weak to ischemia and may easily be damaged, but there is 
a recoverable area around the ischemic neuronal cells, which is referred 
to as the "Penumbra" (Astrup, J., Siesjo, B., Symon, L.; Stroke, 
12:723-725 1981). In the therapy of cerebrovascular disorders at the acute 
stage, it is important to protect the neuronal cells in the penumbra area 
from cell damage and maintain cerebral functions. 
It has been known that cerebrovascular disorders caused by ischemia may 
accompany cerebral edema with an unusually increased moisture content in 
the brain in the ischemic center and penumbra area (Kenji Inamura and 
Akiro Terashi: Brain Nerv. 44(9): 779-785, 1992). Cerebral edema may be 
also caused by cerebral tumor, encephalitis, heat stroke, cerebral trauma 
by a traffic accident. The edema may increase the cerebral capacity, which 
results in the increase in cerebral pressure, because the brain is closed 
within the hard skull. A precipitous increase in cerebral pressure may 
cause cerebral hernia, which makes patients fall in the dangerous state of 
their life. 
Cerebral edema may accompany sodium and calcium influx into neuronal cells, 
which are found at a higher concentration extracellularly as compared with 
the intracellular one (Takao Asano, Hiroo Johshita, Osamu Gotoh and 
others: Cerebral Surgery 13: 1147-1159, 1985), and it is believed that 
calcium influx may activate calcium-dependent enzymes (proteases, 
phospholipases or the like), which results in the damage of cytoskeleton 
or cell membrane. 
Activation of phospholipase A2, a phospholipase, may release arachidonic 
acid from the phospholipid in cell membrane. Accumulation of the 
arachidonic acid may inhibit respiration of mitochondria to decrease ATP. 
Moreover, it is believed that peroxidation of lipids by the free radicals 
produced during the metabolism of arachidonic acid may cause disorders of 
cell membrane or increased permeability of the membrane to provoke the 
progress of the edema. 
In addition to such acute disorders of neuronal cells, the phenomenon 
referred to as the delayed neuronal death has been found out (Kirino T., 
Brain Res., 239: 57-69, 1982). This means the phenomenon that the neuronal 
cells after a short period of ischemia fall off after several days to 
several weeks. It has now been elucidated that delayed cellular death such 
as delayed neuronal death is related with a calcium concentration in 
neuronal cells (Ogura, A., Miyamoto, M., Kudo, Y., Exptl. Brain Res., 
73:447-458, 1988). Such being the case, it is the important object in the 
treatment of cerebrovascular disorders at the acute stage to inhibit 
cerebral edema which would greatly influence upon the prognosis for life 
of patients and also could be the cause of acute and delayed neuronal 
death. 
Presently there has been mainly applied an osmotherapy for the treatment of 
cerebral edema. In this method, a liquid of hyperosmorality is injected 
into blood, whereby an osmotic pressure in blood is raised and moisture is 
withdrawn from edema tissues. However, satisfactory effects have not been 
attained as yet and there has been desired a novel anti-cerebral edema 
agent other than the osmotherapy. 
On the other hand, our copending JP-A-5-32630 discloses that 
pyridinecarboxamide derivatives having a methylene chain of 9-13 carbon 
atoms and bonded to the amido nitrogen have an activity of increasing 
cerebral blood flow. Moreover, it was reported by Sakurai et al. that the 
compound of Example 10 of said JP-A-5-32630, namely, 
N-(11-nitroxy-1-undecanyl)-6-(4-methyl-1-piperazinyl)nicotinamide could 
show a cerebral protective effect on the hypoxia and anoxia models 
(Sakurai Einosuke, Jpn. J. Pharmacol., Vol. 61, No. suppl. 1, PAGE 289p 
1993). 
As the compounds having a cerebral protective action (an anti-anoxia 
action) would be expected to show an inhibiting action on cerebral edema, 
it has been attempted to review and pick up those compounds having an 
anti-anoxia action. However, the compounds disclosed in Example 10 of 
JP-A-5-32630 have been regarded as undesirable for the therapy of 
cerebrovascular disorders at the acute stage, because they were observed 
to possess a behavior suppressing activity that Nizofenone and others 
possess as a side effect. 
DISCLOSURE OF INVENTION 
This invention relates to a pyridinecarboxamide derivative represented by 
the formula (1) 
##STR1## 
(wherein R represents an ethyl group or an isopropyl group) or a 
physiologically acceptable salt thereof. 
Moreover, this invention relates to a compound represented by the formula 
(2) 
##STR2## 
(wherein R is as defined above and X represents a hydroxyl group, a 
mesyloxy group, a tosyloxy group, a bromine atom or an iodine atom). 
Moreover, this invention relates to a 6-piperazinylpyridine-3-carboxylic 
acid represented by the formula (3) 
##STR3## 
(wherein R is as defined above) or a metal salt or acid addition salt 
thereof. 
Moreover, this invention relates to a process for the production of the 
pyridinecarboxamide derivative represented by the formula (1) which 
comprises reacting the compound represented by the abovementioned formula 
(2) with a nitrating agent. 
Moreover, this invention relates to a process for the production of the 
pyridinecarboxamide derivative represented by the formula (1) which 
comprises reacting an alkali metal salt, a halide or an anhydride of the 
6-piperazinylpyridine-3-carboxylic acid represented by the formula (3) 
with a 12-aminoalkyl nitrate represented by the formula (4) 
EQU H.sub.2 N(CH.sub.2).sub.12 ONO.sub.2 (4) 
or an acid addition salt thereof. 
Moreover, this invention relates to a pharmaceutical composition for 
treating cerebrovascular disorders, especially cerebrovascular disorders 
at the acute stage, which comprises the pyridinecarboxamide derivative 
represented by the formula (1) or a physiologically acceptable salt 
thereof and a pharmaceutically acceptable excipient. 
Moreover, this invention relates to the therapeutic agent for 
cerebrovascular disorders at the acute stage which is used for treating 
cerebrovascular disorders caused by cerebral infarction or subarachnoid 
hemorrhage. 
Moreover, this invention relates to the therapeutic agent for cerebral 
edema which comprises the pyridinecarboxamide derivative represented by 
the formula (1) or a physiologically acceptable salt thereof and a 
pharmaceutically acceptable excipient. 
Moreover, this invention relates to a method for the treatment of 
cerebrovascular disorders, especially cerebrovascular disorders at the 
acute stage, which comprises administering the pyridinecarboxamide 
derivative represented by the formula (1) or a physiologically acceptable 
salt thereof to patients suffering from cerebrovascular disorders, 
especially cerebrovascular disorders at the acute stage. 
Moreover, this invention relates to a method for the treatment of cerebral 
edema which comprises administering the pyridinecarboxamide derivative 
represented by the formula (1) or a physiologically acceptable salt 
thereof to patients suffering from cerebral edema. 
The pyridinecarboxamide derivatives (1) of this invention have excellent 
inhibiting activity of cerebral edema and inhibiting activity of delayed 
neuronal death (inhibiting activity of Ca-influx in neuronal cells), and 
further a cerebral protective action (an anti-anoxia action), and an 
inhibiting action on peroxidation of lipids, while they have no behavior 
suppressing activity which is believed to be side effects so that they are 
highly useful as a therapeutic agent for cerebrovascular disorders. The 
pathologic type of cerebrovascular disorders to which the therapeutic 
agent for cerebrovascular disorders of the present invention may be 
applied includes cerebral hemorrhage, brain infarction (cerebral 
thrombosis, cerebral infarction), transient ischemic attack, subarachnoid 
hemorrhage and others. The cerebrovascular disorders at the acute stage as 
stated herein is meant to indicate the cerebrovascular disorders at the 
period of time of less than one month after the onset of cerebrovascular 
disorders. 
Almost all cerebrovascular disorders at the acute stage are accompanied 
with cerebral edema which may accelerate microcirculation disorders around 
lesions to make cerebral disorders far worse. 
Delayed neuronal death is meant to indicate the neuronal death wherein 
neuronal cells such as hippocampus CA fall off several days after a severe 
transient global cerebral ischemia due to temporal cardiac arrest and 
others. The mechanism of this action is believed to be an increase in 
glutamic acid and subsequent increase in intracellular calcium, and thus 
the pyridinecarboxamide derivatives of this invention, which can inhibit 
the delayed neuronal death, are useful as a therapeutic agent for 
cerebrovascular disorders. 
Moreover, energy deficiency caused by ischemia or release of 
neurotransmitters such as glutamine and the like may cause influx of 
calcium ions into cells and generation of free radicals. Excessive 
production of free radicals may accelerate the formation of lipoperoxide 
and may cause irreversible disorders of cell membrane and rise in 
permeability of membrane, which leads to cerebral edema and neuronal 
death. Accordingly, the pyridinecarboxamide derivatives of the invention 
which inhibit the influx of calcium ions and the formation of lipoperoxide 
are useful as a therapeutic agent for cerebrovascular disorders at the 
acute stage. 
The compounds (2) and compounds (3) of this invention are useful as 
intermediates for the synthesis of the pyridinecarboxamide derivatives 
(1). 
The pyridinecarboxamide derivatives represented by the formula (1) can be 
produced by reacting the compounds of the formula (2) with a nitrating 
agent such as nitric acid, fuming nitric acid, tetrabutylammonium nitrate, 
strongly basic ion exchange resins (e.g., Amberlyst) of a nitrate form, 
silver nitrate or potassium nitrate in the presence or absence of a 
solvent at -40.degree. C. to 120.degree. C., preferably -40.degree. C. to 
room temperature. As the solvent, there may be mentioned dichloromethane, 
chloroform, carbon tetrachloride, diethyl ether, tetrahydrofuran, toluene, 
acetonitrile, acetic anhydride, sulfuric acid, etc. and a mixed solvent 
thereof. 
Moreover, the pyridinecarboxamide derivatives represented by the formula 
(1) may be also produced by condensing 12-aminododecyl nitrate or a salt 
thereof with a salt of the compound of the formula (3) or a reactive 
intermediate thereof. 
Specific examples of salts of 12-aminododecyl nitrate may include nitrate, 
hydrochloride or the like. 
Specific examples of the salts or reactive intermediates of the compounds 
of the formula (3) may include sodium 
6-(4-ethyl-1-piperazinyl)pyridine-3-carboxylate, potassium 
6-(4-ethyl-1-piperazinyl)pyridine-3-carboxylate, 
6-(4-ethyl-1-piperazinyl)pyridine-3-carbonyl chloride, sodium 
6-(4-isopropyl-1-piperazinyl)-pyridine-3-carboxylate, potassium 
6-(4-isopropyl-1-piperazinyl)pyridine-3-carboxylate, 
6-(4-isopropyl-1-piperazinyl)pyridine-3-carbonyl chloride and the like. 
The compounds (1) can be preferably produced by reacting a salt of the 
compound (3) with 0.5 to 4 equivalents of 12-aminododecyl nitrate or a 
salt thereof in a solvent in the presence of 0.5 to 4 equivalents of a 
condensing agent at -40.degree. C. to 40.degree. C., preferably 0.degree. 
C. to room temperature. There may be incorporated in situ 0.5 to 4 
equivalents of an additive and a base. As the solvent, there may be 
mentioned dichloromethane, chloroform, carbon tetrachloride, 
tetrahydrofuran, diethyl ether, dioxane, benzene, toluene, xylene, 
acetonitrile, dimethylformamide, dimethyl sulfoxide and the like and a 
mixed solvent thereof. As the condensing agent, there may be mentioned 
carbodiimides such as dicyclohexylcarbodiimide, 
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and the like, 
azides such as diphenylphosphoryl azide and the like, carbonyldiimidazole, 
diethyl pyrocarbonate and the like. As the additive, there may be 
mentioned N-hydroxysucccinimide, 1-hydroxybenzotriazole and the like and, 
as the base, there may be mentioned organic bases such as triethylamine, 
diisopropylethylamine, pyridine, and the like. 
12-Aminododecyl nitrate (4) or a salt thereof can be synthesized from 
12-aminododecanol or an active intermediate thereof such as 
12-bromododecylamine, 12-iodododecylamine, 12-aminododecyl 
methanesulfonate, 12-aminododecyl p-toluenesulfonate and the like, in 
similar conditions to those for the compound (1). 
The compounds of the formula (2) wherein X is a bromine atom or an iodine 
atom can be synthesized by reacting the compounds wherein X is a hydroxyl 
group with hydrobromic acid or hydriodic acid. 
The compounds of the formula (2) wherein X is an iodine atom can be also 
synthesized by reacting the compounds wherein X is a mesyloxy group or a 
tosyloxy group with sodium iodide or potassium iodide in acetone. 
The compounds of the formula (2) wherein X is a mesyloxy group or a 
tosyloxy group can be synthesized by reacting the compounds wherein X is a 
hydroxyl group with mesyl chloride or tosyl chloride in the presence of a 
base. 
The compounds of the formula (2) can be synthesized by reacting a compound 
of the formula (3) or a reactive intermediate thereof such as a sodium 
salt or potassium salt or acid halide thereof with a compound of the 
formula (5) 
EQU H.sub.2 N(CH.sub.2).sub.12 X (5) 
(wherein X is a bromine atom, an iodine atom, a mesyloxy group, a tosyloxy 
group or a hydroxyl group) in a solvent in the presence of 0.5 to 4 
equivalents of a condensing agent at a temperature from -40.degree. C. to 
40.degree. C. There may be incorporated in situ 0.5 to 4 equivalents of an 
additive and a base. As the solvent, there may be mentioned 
dichloromethane, chloroform, carbon tetrachloride, tetrahydrofuran, 
diethyl ether, dioxane, benzene, toluene, xylene, acetonitrile, 
dimethylformamide, dimethyl sulfoxide, and the like and a mixed solvent 
thereof. As the condensing agent, there may be mentioned carbodiimides 
such as dicyclohexylcarbodiimide, 
1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride, and the 
like, azides such as diphenylphosphoryl azide, and the like, 
carbonyldiimidazoles, diethyl pyrocarbonate and the like. As the additive, 
there may be mentioned N-hydroxysucccinimide, 1-hydroxybenzotriazole and 
the like and, as the base, there may be mentioned organic bases such as 
triethylamine, diisopropylethylamine, pyridine and the like. 
The compounds of the formula (2) wherein X is a hydroxyl group can be 
produced by reacting a compound of the formula (6) 
##STR4## 
(wherein Y is a halogen atom such as chlorine, bromine, iodine or the 
like) with 1 to 100 equivalents of a compound of the formula (7) 
##STR5## 
(wherein R is as defined above) in the presence or absence of a solvent at 
a temperature from room temperature to a reflux temperature or in a sealed 
tube. There may be also incorporated 0.05 to 10 equivalents of sodium 
iodide or potassium iodide in situ, or 0.5 to 10 equivalents of a base may 
be incorporated. As the solvent, there may be mentioned dichloromethane, 
chloroform, carbon tetrachloride, tetrahydrofuran, diethyl ether, dioxane, 
benzene, toluene, xylene, acetonitrile, dimethylformamide, dimethyl 
sulfoxide, methanol, ethanol, and the like and a mixed solvent thereof. As 
the base, there may be mentioned inorganic bases such as sodium hydrogen 
carbonate, sodium carbonate, potassium carbonate, cesium carbonate, and 
the like or organic bases such as triethylamine, diethylamine, 
diisopropylamine, diisopropylethylamine, pyridine, and the like 
The compounds of the formula (2) can be also prepared by reacting a 
compound of the formula (6) with 1 to 100 equivalents of piperazine in the 
presence or absence of a solvent at a temperature from room temperature to 
a reflux temperature or in a sealed tube to form the compound of the 
formula (8) 
##STR6## 
and then reacting the compound of the formula (8) thus obtained with 1 to 
10 equivalents of a compound of the formula RZ [wherein R is as defined 
above and Z represents a halogen atom such as chlorine, bromine, iodine 
and the like or a leaving group such as a sulfonate (e.g., a 
methanesulfonyloxy group, a toluenesulfonyloxy group) in the presence or 
absence of a solvent at a temperature from room temperature to a reflux 
temperature or in a sealed tube. There may be also incorporated 0.05 to 4 
equivalents of sodium iodide or potassium iodide in situ, or 0.5 to 10 
equivalents of a base may be incorporated. As the solvent, there may be 
mentioned dichloromethane, chloroform, carbon tetrachloride, 
tetrahydrofuran, diethyl ether, dioxane, benzene, toluene, xylene, 
acetonitrile, dimethylformamide, dimethyl sulfoxide, methanol, ethanol, 
and the like and a mixed solvent thereof. As the base, there may be 
mentioned inorganic bases such as sodium hydrogen carbonate, sodium 
carbonate, potassium carbonate, cesium carbonate, and the like or organic 
bases such as triethylamine, diethylamine, diisopropylamine, 
diisopropylethylamine, pyridine, and the like. 
The compounds of the formula (6) can be prepared by reacting a compound of 
the formula (9) 
##STR7## 
(wherein Y is as defined above) with 0.5 to 4 equivalents of the compound 
of the formula (10) 
EQU H.sub.2 N(CH.sub.2).sub.12 OH (10) 
in a solvent in the presence of 0.5 to 4 equivalents of a condensing agent 
at a temperature from -40.degree. C. to 40.degree. C. There may be 
incorporated in situ 0.5 to 4 equivalents of an additive and a base. As 
the solvent, there may be mentioned dichloromethane, chloroform, carbon 
tetrachloride, tetrahydrofuran, diethyl ether, dioxane, benzene, toluene, 
xylene, acetonitrile, dimethylformamide, dimethyl sulfoxide, and the like 
and a mixed solvent thereof. As the condensing agent, there may be 
mentioned carbodiimides such as dicyclohexylcarbodiimide, 
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and the like, 
azides such as diphenylphosphoryl azide, and the like, 
carbonyldiimidazole, diethyl pyrocarbonate and the like. As the additive, 
there may be mentioned N-hydroxysucccinimide, 1-hydroxybenzotriazole and 
the like and, as the base, there may be mentioned organic bases such as 
triethylamine, diisopropylethylamine, pyridine and the like. 
The compounds of the formula (3) can be also prepared by reacting a 
compound of the formula (11) 
##STR8## 
(wherein Y is as defined above and W represents a halogen atom such as 
chlorine, bromine, iodine, and the like) with 0.5 to 4 equivalents of the 
compound of the formula (10) in a solvent at a temperature from 
-40.degree. C. to 40.degree. C. There may be incorporated in situ 0.5 to 4 
equivalents of a base. As the solvent, there may be mentioned 
dichloromethane, chloroform, carbon tetrachloride, tetrahydrofuran, ether, 
dioxane, benzene, acetonitrile, dimethylformamide, dimethyl sulfoxide, and 
the like and a mixed solvent thereof. As the base, there may be mentioned 
inorganic bases such as sodium hydrogen carbonate, sodium carbonate, 
potassium carbonate, cesium carbonate and the like or organic bases such 
as triethylamine, diethylamine, diisopropylamine, diisopropylethylamine, 
pyridine, and the like. 
The compound of the formula (10) is synthesized from the compound of the 
formula (13) 
##STR9## 
which is obtained from the compound of the formula (12) 
EQU HO(CH.sub.2).sub.12 OH (12). 
The compound of the formula (12) is allowed to react with 0.2 to 2 
equivalents of an azodicarboxylic acid esters, phosphines and phthalimide 
in a solvent at a temperature from -10.degree. C. to 40.degree. C. As the 
solvent, there may be mentioned dichloromethane, chloroform, carbon 
tetrachloride, tetrahydrofuran, ether, dioxane, benzene, acetonitrile, 
dimethylformamide, dimethyl sulfoxide and the like and a mixed solvent 
thereof. As the azodicarboxylic acid esters, there may be mentioned 
azodicarboxylic acid diethyl ester, azodicarboxylic acid diisopropyl ester 
and the like. As the phosphine, there may be mentioned triphenylphosphine, 
tributylphosphine and the like. 
The compound of the formula (13) can be also prepared by reacting the 
compound of the formula (12) with hydrobromic acid to form 
12-bromododecanol followed by reacting with potassium phthalimide. 
The compound of the formula (13) thus obtained may be allowed to react with 
an acid, a base or hydrazine in a solvent at a temperature of 0.degree. C. 
to a reflux temperature to prepare the compound of the formula (10). As 
the solvent, there may be mentioned dichloromethane, chloroform, carbon 
tetrachloride, tetrahydrofuran, ether, dioxane, benzene, acetonitrile, 
dimethylformamide, dimethyl sulfoxide, methanol, ethanol, acetic acid, 
water and the like and a mixed solvent thereof. As the acid, there may be 
mentioned hydrochloric acid, sulfuric acid, acetic acid, and the like and, 
as the base, sodium hydroxide, potassium hydroxide, and the like. 
The compound of the formula (10) can be also synthesized by reducing the 
compound of the formula (14) 
EQU H.sub.2 N(CH.sub.2).sub.11 COOH (14) 
or a derivative thereof such as an ester or Boc derivative thereof with 
LiAlH.sub.4, BH.sub.3 or the like. 
The compound of the formula (15) obtained by converting to the Boc 
derivative an d su bsequent BH.sub.3 reduction 
EQU BocNH(CH.sub.2).sub.12 OH (15) 
may be also reacted with iodine, mesyl chloride or tosyl chloride and, if 
necessary, in the presence of a base or triphenylphosphine followed by 
deprotection of the Boc with an acid, thereby synthesizing the compound 
wherein X is an iodine atom, a mesyloxy group or a tosyloxy group. 
Specific examples of the pyridinecarboxamide derivatives (1) of the present 
invention are given below: 
N-(12-nitroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide, 
N-(12-nitroxydodecyl)-6-(4-isopropyl-1-piperazinyl)pyridine-3-carboxamide. 
Specific examples of the compounds of the formula (2) are given below: 
N-(12-hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide, 
N-(12-hydroxydodecyl)-6-(4-isopropyl-1-piperazinyl)pyridine-3-carboxamide, 
N-(12-iodododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide, 
N-(12-iodododecyl)-6-(4-isopropyl-1-piperazinyl)pyridine-3-carboxamide, 
N-(12-bromododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide, 
N-(12-bromododecyl)-6-(4-isopropyl-1-piperazinyl)pyridine-3-carboxamide, 
N-(12-mesyloxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide, 
N-(12-mesyloxydodecyl)-6-(4-isopropyl-1-piperazinyl)-pyridine-3-carboxamide 
N-(12-tosyloxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide, 
N-(12-tosyloxydodecyl)-6-(4-isopropyl-1-piperazinyl)-pyridine-3-carboxamide 
. 
Specific examples of the compounds of the formula (7) are given below: 
1-Ethylpiperazine, 
1-isopropylpiperazine. 
Specific examples of the compounds of the formula (9) are given below: 
6-Chloronicotinic acid, 
6-bromonicotinic acid. 
Specific examples of the compounds of the formula (5) are given below: 
12-Aminododecanol, 
12-bromododecylamine, 
12-iodododecylamine, 
12-aminododecyl methanesulfonate, 
12-aminododecyl p-toluenesulfonate. 
Specific examples of the compounds of the formula (11) are given below: 
6-Chloronicotinoyl chloride, 
6-bromonicotinoyl chloride. 
As the physiologically acceptable salts of the pyridinecarboxamide 
derivatives (1) of the present invention, there may be mentioned, for 
example, hydrochloride, sulfate, nitrate, hydrobromide, phosphate, 
maleate, fumarate, tartarate, malate, succinate, malonate, propionate, 
methanesulfonate, benzenesulfonate, p-toluenesulfonate, formate, acetate, 
trifluoroacetate and the like, and those compounds containing plural 
acidic functional groups such as carboxyl may be isolated in the form of 
an inorganic salt with a paired ion such as sodium, potassium, lithium, 
calcium, magnesium or the like. 
A part of the pyridinecarboxamide derivatives represented by the formula 
(1) of this invention is metabolized and converted in vivo to a novel 
pyridine derivative which is effective in the treatment of cerebrovascular 
disorders. Main metabolic sites are listed below. 
1) N-Oxidation at the 4-position of the piperazine ring and at the 
1-position of the pyridine ring. 
2) Hydroxylation of the piperazine ring, the pyridine ring and 
(CH.sub.2).sub.12 and ring cleavage of the piperazine ring incidental to 
the hydroxylation. 
3) Hydroxylation and dealkylation of the alkyl group at the 4-position of 
the piperazine ring. 
4) Hydrolysis of the nitric ester and the pyridine-3-carboxamide. 
Main metabolites are recited below: 
N-(12-nitroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
N-oxide, 
N-(12-nitroxydodecyl)-6-(4-isopropyl-1-piperazinyl)-pyridine-3-carboxamide 
N-oxide. 
The compounds (1) or pharmaceutically acceptable salts thereof according to 
the invention may be formulated to dosage forms such as tablets, granules, 
fine granules, powders, capsules, syrups, elixirs, suspensions, emulsions, 
injections and the like by incorporating suitable excipients, auxiliary 
agents, lubricants, antiseptics, disintegrating agents, buffer agents, 
binding agents, wetting agents, emulsifiers, coloring agents, corringents 
or flavors, and they may be administered orally or parenterally, 
preferably via intravenous injection or intravenous instillation. 
In preparing a pharmaceutical preparation as drugs for internal use, the 
conventionally applicable auxiliaries such as lactose, sucrose, sorbitol, 
mannitol, potato starch, corn starch, cellulose derivatives, gelatin and 
the like are suitable as a carrier and lubricants such as magnesium 
stearate, Carbowax, polyethylene glycol and the like may be further added. 
The active compound in admixture with the above may be formed into 
granules, tablets, capsules and the like according to a conventional 
method. 
In preparing a pharmaceutical preparation in the form of an aqueous 
preparation, the active ingredient may be dissolved in distilled water for 
injection and, as required, antioxidants, stabilizers, solubilizing 
agents, water-soluble surfactants, nonaqueous solvents, buffer agents, pH 
adjusters, preservatives, isotonic agents or soothing agents may be added 
and the resultant aqueous solution may be filtered, filled and sealed in a 
conventional manner and then sterilized by means of autoclaved 
sterilization or hot air sterilization to prepare injections. 
In preparing a pharmaceutical preparation in the form of an emulsifiable 
injection, the sterilized active ingredient may be dissolved in a 
nonaqueous solvent and, if necessary, distilled water for injection, 
antioxidants, stabilizers, solubilizing agents, water-soluble surfactants, 
buffer agents, pH adjusters, preservatives, isotonic agents or soothing 
agents may be added and then the resultant emulsion may be filtered, 
filled and sealed in a conventional manner to prepare injections. 
A dose of the compound or pharmaceutically acceptable salt thereof 
according to this invention may be selected depending upon the body 
weight, age, sex, lapsed time after onset, classification of diseases and 
others, and it is 1-1000 mg per day. 
BEST MODE FOR CARRYING OUT THE INVENTION

This invention will be more specifically illustrated by way of the 
following Preparation Examples, Pharmacological Effect and Formulation 
Examples, but this invention is not intended to be limited thereto. 
EXAMPLE 1 
N-(12-Nitroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR10## 
To 30 ml of fuming nitric acid cooled to -30.degree. C. was gradually added 
7.9 g of 
N-(12-hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide over 
30 minutes and the mixture was stirred at -30 to -20.degree. C. for 40 
minutes. The reaction solution was poured into ice, neutralized with 
sodium hydrogen carbonate and extracted with chloroform. The extract was 
dried over anhydrous sodium sulfate and distilled under reduced pressure. 
The residue thus obtained was chromatographed over a silica gel column and 
recystallized from methanol-water to afford the title compound as a 
crystal. 
mp 71-72.degree. C. 
.sup.1 H NMR (CDC1.sub.3).delta. 1.13 (t, J=7.3 Hz, 3H), 1.27-1.42 (m, 
16H), 1.59 (quint, J=6.8 Hz, 2H), 1.71 (quint, J=6.8 Hz, 2H), 2.46 (q, 
J=7.3 Hz, 2H), 2.54 (t, J=4.8 Hz, 4H), 3.42 (q, J=6.8 Hz, 2H), 3.66 (t, 
J=4.8 Hz, 4H), 4.44 (t, J=6.8 Hz, 2H), 5.94 (brs, 1H), 6.62 (d, J=9.2 Hz, 
1H), 7.89 (dd, J=2.4, 9.2 Hz, 1H), 8.68 (d, J=2.4 Hz, 1H) 
EXAMPLE 2 
N-(12-Nitroxydodecyl)-6-(4-isopropyl-1-piperazinyl)pyridine-3-carboxamide 
##STR11## 
Synthesis was carried out in the same manner as in Example 1 using as a 
starting material 
N-(12-hydroxy-dodecyl)-6-(4-isopropyl-1-piperazinyl)pyridine-3-carboxamide 
. The title compound was obtained as a colorless crystal. 
mp 56-58.degree. C. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.08 (d, J=6.3 Hz, 6H), 1.26-1.36 (m, 
16H), 1.57-1.60 (m, 2H), 1.67-1.73 (m, 2H), 1.61 (t, J=4.8 Hz, 4H), 2.73 
(sept, J=6.3 Hz, 1H), 3.41 (q, J=6.8 Hz, 2H), 3.65 (t, J=4.8 Hz, 4H), 4.44 
(t, J=6.8 Hz, 2H), 5.96 (brs, 1H), 6.61 (d, J=9.2 Hz, 1H),7.89 (dd, J=2.4, 
8.7 Hz, 1H), 8.53 (d, J=2.4 Hz, 1H) 
EXAMPLE 3 
N-(12-Nitroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR12## 
To a solution of 5.7 g of 12-aminododecyl nitrate hydrochloride in 80 ml of 
methylene chloride were added successively under ice-cooling 5.21 g of 
sodium 6-(4-ethylpiperazinyl)pyridine-3-carboxylate, 3.91 g of 
1-hydroxybenzotriazole (HOBt) and 2.76 g of 
1-ethyl-3-(3-dimethylaminopropyl )carbodiimide hydrochloride (WSCI) and 
then the mixture was stirred at room temperature overnight. The reaction 
solution was diluted with chloroform and washed successively with water, a 
saturated aqueous solution of sodium hydrogen carbonate and a saturated 
aqueous solution of sodium chloride, dried over anhydrous sodium sulfate 
and then distilled under reduced pressure. The residue thus obtained was 
chromatographed over a silica gel column to afford 8.56 g of the title 
compound as a crystal. 
EXAMPLE 4 
N-(12-Nitroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR13## 
Preparation of Amberlyst A-26 (manufactrured by Rohm & Haas Co.) of a 
nitrate form 
50 g of Amberlyst A-26 (Cl form) was washed successively with 300 ml each 
of methanol, water and a 2.5N aqueous solution of sodium hydroxide and 350 
ml of ion exchanged water and then converted to the nitrate form with 300 
ml of 1N-nitric acid. After the conversion, it was washed with ion 
exchanged water until it became neutral and then replaced with 200 ml of 
ethanol and 100 ml of acetone. The Amberlyst A-26 (a nitrate form) thus 
contained was dried at 50.degree. C. under reduced pressure for 2 hours. 
To a solution of 1.0 g of 
N-(12-mesyloxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide in 
toluene was added 4.0 g of Amberlyst A-26 (a nitrate form) and the 25 
mixture was heated under reflux for 3 hours. The ion exchange resin was 
filtered off and the filtrate was distilled off. The residue thus obtained 
was chromatographed over a silica gel column to afford 0.85 g of the title 
compound. 
Similarly, the title compound was obtained using as a starting material 
N-(12-tosyloxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide, 
N-(12-iodododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide or 
N-(12-bromododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide. 
EXAMPLE 5 
N-(12-Nitroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR14## 
To a solution of 76 mg of 
N-(12-mesyloxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide in 
toluene was added 70 mg of tetrabutylammonium nitrate and the mixture was 
refluxed for 3 hours. To the reaction solution were added water and 
chloroform and the mixture was washed with a saturated aqueous solution of 
sodium chloride. The organic layer was dried over anhydrous magnesium 
sulfate and distilled off, and the residue thus obtained was 
chromatographed over a silica gel column to afford 63.1 mg of the title 
compound. 
Similarly, the title compound was obtained using as a starting material 
N-(12-tosyloxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide, 
N-(12-iodododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide or 
N-(12-bromododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide. 
EXAMPLE 6 
N-(12-Nitroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR15## 
To a solution of 1.30 g of 
N-(12-iodododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide in 
toluene-acetonitrile was added 1.46 g of silver nitrate and the mixture 
was stirred at 35.degree. C. for 3.5 hours. After the reaction solution 
was filtered, water and chloroform were added and the mixture was washed 
with a saturated aqueous solution of sodium chloride. The organic layer 
was dried over anhydrous magnesium sulfate and distilled off, and the 
residue thus obtained was chromatographed over a silica gel column to 
afford 1.11 g of the title compound. 
Similarly, the title compound was obtained using as a starting material 
N-(12-bromododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide. 
EXAMPLE 7 
N-(12-Iodododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR16## 
To a solution of 0.26 g of potassium iodide in 2 ml of phosphoric acid was 
added 0.10 g of 
N-(12-hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide and 
the mixture was stirred at 110.degree. C. for 3 hours. The reaction 
solution was diluted with water, and a saturated aqueous solution of 
sodium hydrogen carbonate and an aqueous solution of sodium thiosulfate 
were added. The mixture was extracted with chloroform. The organic layer 
was washed with water and then dried over anhydrous sodium sulfate. The 
solvent was distilled off under reduced pressure to afford 0.11 g of the 
title compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.13 (t, J=7.2 Hz, 3H), 1.27-1.38 (m, 
16H), 1.56-1.60 (m, 2H), 1.82 (quint, J=7.1 Hz, 2H), 2.47 (q, J=7.2 Hz, 
2H), 2.54 (t, J=5.1 Hz, 4H), 3.19 (t, J=7.1 Hz, 4H), 3.42 (q, J=6.7 Hz, 
2H), 3.67(t, J=5.1 Hz, 4H), 5.90 (bs, 1H), 6.62 (d, J=8.8 Hz, 1H), 7.90 
(dd, J=2.4 Hz, 8.8 Hz, 1H), 8.53 (d,J=2.4 Hz, 1H) 
EXAMPLE 8 
N-(12-Iodododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR17## 
A solution of 2.09 g of 
N-(12-hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide in 
57% hydriodic acid was stirred at 120.degree. C. for 30 minutes. The 
reaction solution was diluted with water and extracted with chloroform. 
The chloroform layer was washed with a saturated aqueous solution of 
sodium hydrogen carbonate and a saturated aqueous solution of sodium 
chloride, then dried over anhydrous magnesium sulfate. The solvent was 
distilled off under reduced pressure, and the residue thus obtained was 
chromatographed over a silica gel column to afford 1.94 g of the title 
compound. 
EXAMPLE 9 
N-(12-Bromododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR18## 
To a solution of 419 mg of 
N-(12-hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide in 
20 ml of toluene was added 0.60 ml of a 47% hydrobromic acid and the 
mixture was heated under reflux for one hour. The reaction solution was 
washed successively with water, a saturated aqueous solution of sodium 
hydrogen carbonate and a saturated aqueous solution of sodium chloride, 
dried over anhydrous magnesium sulfate. The solvent was distilled off 
under reduced pressure, and the residue thus obtained was chromatographed 
over a silica gel column to afford 400 mg of the title compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.13 (t, J=7.2 Hz, 3H), 1.27-1.44 (m, 
16H), 1.56-1.63 (m, 2H), 1.85 (quint, J=7.2 Hz, 2H), 2.47 (q, J=7.2 Hz, 
2H), 2.54 (t, J=5.1 Hz, 4H), 3.39-3.45 (m, 4H), 3.67 (t, J=5.1 Hz, 4H), 
5.90 (bs, 1H), 6.62 (d, J=8.8 Hz, 1H), 7.90 (dd, J=2.4 Hz, 8.8 Hz, 1H), 
8.53 (d, J=2.4 Hz, 1H) 
EXAMPLE 10 
N-(12-Iodododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR19## 
To 170 mg of 12-iodododecylamine hydroiodide were added 120 mg of sodium 
6-(4-ethyl-1-piperazinyl)-pyridine-3-carboxylate, 10 ml of methylene 
chloride, 95 mg of 1,3-dicyclohexylcarbodiimide (DCC) and 12 mg of 
1-hydroxybenzotriazole (HOBt) and the mixture was stirred at room 
temperature for 3 hours. Insolubles were filtered off and the filtrate was 
chromatographed over a silica gel column to afford 169.5 mg of the title 
compound. 
EXAMPLE 11 
N-(12-Iodododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR20## 
To a suspension of 0.20 g of sodium 
6-(4-ethyl-1-piperazinyl)pyridine-3-carboxylate in 10 ml of methylene 
chloride was added 0.2 ml of oxalyl chloride and the mixture was heated 
under reflux for one hour. The reaction solution was distilled off under 
reduced pressure and 10 ml of methylene chloride was added to the residue. 
0.3 ml of triethylamine and 0.17 g of 12-iodododecylamine hydrochloride 
were further added and the mixture was stirred overnight. The reaction 
solution was diluted with chloroform and washed successively with a 1N 
aqueous solution of sodium hydroxide, water and a saturated aqueous 
solution of sodium chloride, dried over anhydrous sodium sulfate. The 
solvent was distilled off, and the residue thus obtained was 
chromatographed over a silica gel column to afford 0.21 g of the title 
compound. 
EXAMPLE 12 
N-(12-Iodododecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR21## 
To a solution of 72 mg of 
N-(12-mesyloxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide in 
acetone was added 130 mg of sodium iodide and the mixture was heated under 
reflux for 10 hours. To the reaction solution were added water and 
chloroform and extracted with chloroform. The chloroform layer was washed 
with a saturated aqueous solution of sodium chloride and then dried over 
anhydrous magnesium sulfate. The solvent was then distilled off under 
reduced pressure. The residue thus obtained was chromatographed over a 
silica gel column to afford 69.0 mg of the title compound. 
Similarly, the title compound was obtained using as a starting material 
N-(12-tosyloxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide. 
EXAMPLE 13 
N-(12-Mesyloxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR22## 
To a solution of 3.77 g of 12-aminododecyl methanesulfonate hydrochloride 
in 100 ml of methylene chloride were added successively under ice-cooling 
3.60 g of sodium 6-(4-ethyl-1-piperazinyl)pyridine-3-carboxylate, 2.89 g 
of 1-hydroxybenzotriazole (HOBt), 1.99 g of 
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (WSCI) and 
then the mixture was stirred overnight. The reaction solution was diluted 
with chloroform and washed successively with water, a saturated aqueous 
solution of sodium hydrogen carbonate and a saturated aqueous solution of 
sodium chloride, dried over anhydrous sodium sulfate and then distilled 
under reduced pressure. The residue thus obtained was chromatographed over 
a silica gel column to afford 4.96 g of the title compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.13 (t, J=7.3 Hz, 3H), 1.27-1.33 (m, 
16H), 1.57 -1.61 (m, 2H), 1.75 (quint, J=7.2 Hz, 2H), 2.47 (q, J=7.3 Hz, 
2H), 2.54 (t, J=5.1 Hz, 4H), 3.00 (s, 3H), 3.42 (q, J=6.6 Hz, 2H), 3.67 
(t, J=5.1 Hz, 4H), 4.22 (t, J=6.6 Hz, 2H), 5.92 (bs, 1H), 6.62 (d, J=8.8 
Hz, 1H), 7.90 (dd, J=2.4 Hz, 8.8 Hz, 1H), 8.53 (d, J=2.4 Hz, 1H) 
EXAMPLE 14 
N-(12-Tosyloxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR23## 
To a suspension of 210 mg of 
N-(12-hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide in 
10 ml of methylene chloride were added 0.03 ml of pyridine, 115 mg of 
tosyl chloride and a catalytic amount of 4-dimethylaminopyridine and the 
mixture was stirred at room temperature for one hour. The reaction 
solution was washed with water and a saturated aqueous solution of sodium 
chloride and then dried over anhydrous magnesium sulfate. The solvent was 
distilled off under reduced pressure and the residue thus obtained was 
chromatographed over a silica gel column to afford 219 mg of the title 
compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.14 (t, J=7.2 Hz, 3H), 1.21-1.26 (m, 
16H), 1.57 -1.65 (m, 4H), 2.45 (s, 3H), 2.46-2.50 (m, 16H), 2.55 (t, J=5.2 
Hz, 4H), 3.42 (q, J=6.6 Hz, 2H), 3.67 (t, J=5.2 Hz, 4H), 4.02 (t, J=6.3 
Hz, 2H), 5.94 (bs, 1H), 6.62 (d, J=8.8 Hz, 1H), 7.34 (d, J=8.3 Hz, 2H), 
7.78 (d, J=8.3 Hz, 2H), 7.89 (dd, J=2.4 Hz, 8.8 Hz, 1H), 8.53 (d, J=2.4 
Hz, 1H) 
EXAMPLE 15 
N-(12-Mesyloxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR24## 
To a solution of 1.26 g of 
N-(12-hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide in 
20 ml of chloroform were added 4.2 ml of triethylamine, 5 ml of pyridine 
and 1.0 ml of mesyl chloride and the mixture was stirred at room 
temperature for 30 minutes. The reaction solution was washed successively 
with water, 3N-hydrochloric acid and a saturated aqueous solution of 
sodium chloride and then dried over anhydrous magnesium sulfate. The 
solvent was distilled off under reduced pressure and the residue thus 
obtained was chromatographed over a silica gel column to afford 1.11 g of 
the title compound. 
EXAMPLE 16 
12-Aminododecyl Nitrate Hydrochloride 
EQU H.sub.2 N(CH.sub.2).sub.12 ONO.sub.2.HCl 
To 12 ml of fuming nitric acid was added at -20.degree. C. 3.0 g of 
12-aminododecanol and the mixture was stirred for 30 minutes. The reaction 
solution was poured into ice-water and extracted with chloroform. The 
extract was washed successively with a saturated aqueous solution of 
sodium hydrogen carbonate and a saturated aqueous solution of sodium 
chloride, dried over anhydrous sodium sulfate. Then 3.7 ml of a hydrogen 
chloride-ethyl acetate solution was added under ice-cooling and distilled 
under reduced pressure. The residue thus obtained was recrystallized from 
chloroform-hexane to afford 3.03 g of the title compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.20-1.42 (m, 16H), 1.68-1.80 (m, 4H), 
2.94-3.01 (m, 2H), 4.44 (t, J=6.8 Hz, 2H), 8.22-8.33 (brs, 3H) 
PREATION EXAMPLE 1 
N-(12-Hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR25## 
To 0.6 g of N-(12-hydroxydodecyl)-6-chloropyridine-3-carboxamide was added 
5 ml of 1-ethylpiperazine and the mixture was stirred at 160.degree. C. 
for 30 minutes. The reaction solution was diluted with chloroform, washed 
with water, dried over anhydrous sodium sulfate and distilled off under 
reduced pressure. The residue was chromatographed over a silica gel column 
to afford the title compound. 
mp 108.5-110.degree. C. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.13 (t, J=7.3 Hz, 3H), 1.27-1.32 (m, 
16H), 1.52-1.63 (m, 4H), 2.47 (q, J=7.3 Hz, 2H), 2.54 (t, J=4.8 Hz, 4H), 
3.42 (q, J=6.8 Hz, 2H), 3.62-3.67 (m, 6H), 5.93 (brs, 1H), 6.62 (d, J=8.7 
Hz, 1H), 7.90 (dd, J=2.4, 8.7 Hz, 1H), 8.53 (d, J=2.4 Hz, 1H) 
PREATION EXAMPLE 2 
N-(12-Hydroxydodecyl)-6-(4-isopropyl-1-piperazinyl)pyridine-3-carboxamide 
##STR26## 
To 2.78 g of N-(12-hydroxydodecyl)-6-chloropyridine-3-carboxamide was added 
4.15 g of 1-isopropylpiperazine and the mixture was stirred at 160.degree. 
C. for 30 minutes. The reaction solution was diluted with chloroform, 
washed with water, dried over anhydrous sodium sulfate and distilled off 
under reduced pressure. The residue was chromatographed over a silica gel 
column to afford 2.2 g of the title compound as a crystal. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.08 (d, J=6.8 Hz, 6H), 1.27-1.32 (m, 
16H), 1.52-1.65 (m, 4H), 2.61 (t, J=4.8 Hz, 4H), 2.73 (7th, J=6.8 Hz, 1H), 
3.42 (q, J=6.8 Hz, 2H), 3.62-3.66 (m, 6H), 5.92 (brs, 1H), 6.61 (d, J=8.7 
Hz, 1H), 7.89 (dd, J=2.4, 8.7 Hz, 1H), 8.52 (d, J=2.4 Hz, 1H) 
PREATION EXAMPLE 3 
N-(12-Hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR27## 
PREATION EXAMPLE 3-1 
6-(4-Ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR28## 
A mixture of 0.5 g of 6-chloronicotinamide and 3 ml of 1-ethylpiperazine 
was stirred at 110.degree. C. for 2 hours. The reaction solution was 
diluted with ethyl acetate, washed with water and then dried over 
anhydrous sodium sulfate. The solvent was distilled off under reduced 
pressure to afford 0.10 g of the title compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.13 (t, J=7.3 Hz, 3H), 2.46 (q, J=7.3 Hz, 
2H), 2.54 (t, J=5.3 Hz, 4H), 3.69 (t, J=5.3 Hz, 4H), 5.60-5.80 (brs, 2H), 
6.63 (d, J=8.7 Hz, 1H), 7.93 (dd, J=2.4, 8.7 Hz, 1H), 8.58 (d, J=2.4 Hz, 
1H) 
PREATION EXAMPLE 3-2 
N-(12-Hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
To a suspension of 0.055 g of 
6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide, 0.011 g of 
tetrabutylammonium hydrogen sulfate, 0.11 g of sodium hydroxide and 0.05 g 
of potassium carbonate in 3 ml of toluene was added dropwise at 70.degree. 
C. a solution of 12-bromododecanol in 2 ml of toluene and the mixture was 
stirred at 70.degree. C. for 13 hours. The reaction solution was diluted 
with chloroform, washed with water and a saturated aqueous solution of 
sodium chloride, dried over anhydrous sodium sulfate. The solvent was 
distilled off under reduced pressure. The residue thus obtained was 
chromatographed over a silica gel column to afford 0.025 g of the title 
compound. 
PREATION EXAMPLE 4 
N-(12-Hydroxydodecyl)-6-(4-isopropyl-1-piperazinyl)pyridine-3-carboxamide 
PREATION EXAMPLE 4-1 
N-(12-Hydroxydodecyl)-6-(1-piperazinyl)pyridine-3-carboxamide 
##STR29## 
To a suspension of 2.38 g of 
N-(12-hydroxydodecyl)-6-chloropyridine-3-carboxamide in 50 ml of toluene 
was added 3.0 g of piperadine and the mixture was heated under reflux for 
10 hours and then the reaction solution was distilled under reduced 
pressure. The residue thus obtained was chromatographed over a silica gel 
column to afford the title compound as a pale yellow crystal. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.27-1.33 (m, 16H), 1.52-1.61 (m, 4H), 
2.97 (t, J=4.8 Hz, 4H), 3.42 (q, J=6.8 Hz, 2H), 3.59-3.65 (m, 6H), 5.92 
(brs,1H), 6.61 (d, J=8.7 Hz, 1H), 7.90 (dd, J=2.7, 8.7 Hz, 1H), 8.53 (d, 
J=2.4 Hz, 1H) 
PREATION EXAMPLE 4-2 
N-(12-Hydroxydodecyl)-6-(4-isopropyl-1-piperazinyl)pyridine-3-carboxamide 
A solution of 0.7 g of 
N-(12-hydroxydodecyl)-6-(1-piperazinyl)pyridine-3-carboxamide, 0.24 g of 
isopropyl bromide and 0.7 g of potassium carbonate in 20 ml of 
dimethylformamide was stirred at 100.degree. C. for 3 hours. The reaction 
solution was diluted with chloroform, washed with water and a saturated 
aqueous solution of sodium chloride, dried over anhydrous sodium sulfate. 
The solvent was distilled off under reduced pressure. The residue thus 
obtained was chromatographed over a silica gel column to afford the title 
compound as a crystal. 
PREATION EXAMPLE 5 
N-(12-Hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR30## 
To 4.07 g of potassium 6-(4-ethyl-1-piperazinyl)-pyridine-3-carboxylate and 
3.00 g of 12-aminododecanol was added 50 ml of chloroform and then 2.01 g 
of 1-hydroxybenzotriazole (HOBt) and 3.07 g of 
1,3-dicyclohexylcarbodiimide (DCC) were added thereto while stirring at 
room temperature and then the mixture was stirred at room temperature for 
17 hours. A further 50 ml of chloroform was added, the mixture was stirred 
and then insolubles were filtered off. After the residue on a filter paper 
was washed with chloroform, the washing and the chloroform solution were 
combined and washed with a 10% aqueous solution of sodium hydroxide and 
dried over anhydrous magnesium sulfate. After the chloroform solution was 
concentrated, diisopropyl ether was added and the crystal thus separated 
out was recovered by filtration and dried under reduced pressure to afford 
5.10 g of the title compound as a crystal. 
PREATION EXAMPLE 6 
N-(12-Hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR31## 
To 4.07 g of potassium 6-(4-ethyl-1-piperazinyl)-pyridine-3-carboxylate and 
3.00 g of 12-aminododecanol was added 50 ml of methylene chloride and then 
2.86 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride 
(WSCI) and 2.01 g of 1-hydroxybenzotriazole (HOBt) were added while 
stirring at room temperature. The mixture was stirred at room temperature 
for 15 hours. To the reaction solution were added water and diisopropyl 
ether and subsequent filtration and drying under reduced pressure afforded 
5.67 g of the title compound as a crystal. 
PREATION EXAMPLE 7 
N-(12-Hydroxydodecyl)-6-(4-ethyl-1-piperazinyl)pyridine-3-carboxamide 
##STR32## 
To a suspension of 0.22 g of sodium 
6-(4-ethyl-1-piperazinyl)pyridine-3-carboxylate in 10 ml of methylene 
chloride were added 0.2 ml of oxalyl chloride and the mixture was heated 
under reflux for one hour. The reaction solution was distilled off under 
reduced pressure, to the residue were added 10 ml of methylene chloride, 
0.3 ml of triethylamine and 0.15 g of 12-aminododecanol and the mixture 
was stirred overnight. The reaction solution was diluted with chloroform, 
washed successively with a 1N aqueous solution of sodium hydroxide, water 
and a saturated aqueous solution of sodium chloride and then dried over 
anhydrous sodium sulfate. The solvent was distilled off and the residue 
thus obtained was chromatographed over a silica gel column to afford 0.21 
g of the title compound. 
PREATION EXAMPLE 8 
12-Iodododecylamine hydroiodide 
EQU H.sub.2 N(CH.sub.2).sub.12 I.HI 
An aqueous solution of 100 mg of 12-aminododecanol in 57% hydriodic acid 
was heated under reflux for one hour. To the reaction solution was added 
chloroform and the mixture was washed successively with water, a 10% 
aqueous solution of sodium thiosulfate and a saturated aqueous solution of 
sodium chloride and then dried over anhydrous magnesium sulfate. The 
solvent was distilled off and the residue thus obtained was 
chromatographed over a silica gel column to afford 171 mg of the title 
compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.27-1.39 (m, 16H), 1.65 (bs, 2H), 
1.73-1.86 (m, 4H), 2.98 (t, J=7.8 Hz, 2H), 3.19 (t, J=7.1 Hz, 2H), 8.21 
(bs, 1H), 6.62 (d, J=8.8 Hz, 1H) 
PREATION EXAMPLE 9 
12-Aminododecanol 
EQU H.sub.2 N(CH.sub.2).sub.12 OH 
In 600 ml of methanol was dissolved while warming 48.0 g of 
N-(12-hydroxydodecyl)phthalimide, 50 ml of hydrazine monohydrate was added 
at room temperature with stirring and the mixture was stirred overnight. 
The crystal thus separated out was filtered off and the solvent was 
distilled off. The residue was extracted with hot chloroform and 
insolubles were filtered off. The filtrate was distilled, the residue was 
extracted with hot chloroform (100 ml.times.3) and then the solvent was 
distilled off. The residue was dissolved in methanol and recrystallization 
from methanol-ethyl acetate gave 21.6 g of the title compound as a 
crystal. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.22-1.47 (m, 16H), 1.43 (quint, J=7 Hz, 
2H), 1.56 (quint, J=7 Hz, 2H), 2.68 (q, J=7 Hz, 2H), 3.64 (t, J=7 Hz, 2H) 
PREATION EXAMPLE 10 
N-(12-Hydroxydodecyl)phthalimide 
##STR33## 
To a solution of 45 g of 12-bromododecanol in 500 ml of dimethylformamide 
was added 75 g of potassium phthalimide and the mixture was stirred at 
100.degree. C. for 3 hours. After completion of the reaction, the solvent 
was distilled off and the residue was dispersed with water. The crude 
crystal thus separated out was recovered by filtration and recrystallized 
from methanol to afford 54 g of the title compound as a crystal. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.19-1.41 (m, 16H), 1.56 (quint, J=7 Hz, 
2H), 1.67 (quint, J=7 Hz, 2H), 3.64 (q, J=6 Hz, 2H), 3.68 (t, J=7 Hz, 2H), 
7.71 (dd, J=3 Hz, 5 Hz, 2H), 7.84 (dd, J=3 Hz, 5 Hz, 2H) 
PREATION EXAMPLE 11 
12-Bromododecanol 
EQU Br(CH.sub.2).sub.12 OH 
To 30.25 g of 1,12-dodecanediol were added 350 ml of toluene and 52 ml of 
48% hydrobromic acid and the mixture was heated under reflux under 
azeotropic condition for 2 hours. The reaction solution was cooled and the 
starting material precipitated out was filtered off. The filtrate was 
distilled and recrystallized from hexane to afford 30.2 g of the title 
compound as a crystal. 
PREATION EXAMPLE 12 
12-Aminododecanol hydrochloride 
EQU H.sub.2 N(CH.sub.2).sub.12 OH.HCl 
To a suspension of 5.0 g of tert-butyl N-(12-hydroxydodecyl)carbamate in 25 
ml of methanol was added 3 ml of hydrochloric acid and the mixture was 
heated under reflux for one hour. The reaction solution was distilled off 
under reduced pressure, ethyl acetate was added to the residue, the 
crystal thus precipitated out was recovered by filtration and dried under 
reduced pressure to afford 3.23 g of the title compound. 
.sup.1 H NMR (CD.sub.3 OD).delta. 1.28-1.40 (m, 16H), 1.51 (quint, J=6.8 
Hz, 2H), 1.63 (quint, J=7.3 Hz, 2H), 2.89 (t, J=7.8 Hz, 2H), 3.52 (t, 
J=6.8 Hz, 2H) 
PREATION EXAMPLE 13 
12-Aminododecyl methanesulfonate hydrochloride 
EQU H.sub.2 N(CH.sub.2).sub.12 OMs.HCl 
To a solution of 8.06 g of tert-butyl N-(12-mesyloxydodecyl)carbamate in 60 
ml of ethyl acetate was added 10 ml of hydrochloric acid under ice-cooling 
and the mixture was stirred at room temperature for one hour. The reaction 
solution was cooled with ice, hexane was added, the crystal thus 
precipitated out was recovered by filtration and dried under reduced 
pressure to afford the title compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.23-1.42 (m, 16H), 1.71-1.83 (m, 4H), 
2.94-3.01 (m, 2H), 3.00 (s, 3H), 4.22 (t, J=6.3 Hz, 2H), 8.19-8.27 (brs, 
3H) 
PREATION EXAMPLE 14 
tert-Butyl N-(12-mesyloxydodecyl)carbamate 
BocNH(CH.sub.2).sub.12 OMs 
To a solution of 8.96 g of tert-butyl N-(12-hydroxydodecyl)carbamate in 60 
ml of tetrahydrofuran were successively added dropwise under ice-cooling 
2.1 ml of mesyl chloride and 3.8 ml of triethylamine and the mixture was 
stirred for one hour. To the reaction solution was added ethyl acetate and 
the mixture was washed successively with water, a saturated aqueous 
solution of sodium hydrogen carbonate, water and a saturated aqueous 
solution of sodium chloride, dried over anhydrous sodium sulfate and the 
solvent was distilled off under reduced pressure to afford 8.06 g of the 
title compound. 
.sup.2 H NMR (CDCl.sub.3).delta. 1.23-1.47 (m, 18H), 1.44 (s, 9H), 
1.71-1.78 (m, 2H), 3.00 (s, 3H), 3.07-3.12 (m, 2H), 4.22 (t, J=6.3 Hz, 
2H), 4.45-4.51 (m, 1H) 
PREATION EXAMPLE 15 
12-Iodododecylamine hydrochloride 
EQU H.sub.2 N(CH.sub.2).sub.12 I.HCl 
To a solution of 0.33 g of tert-butyl N-(12-iodododecyl)carbamate in 25 ml 
of ethyl acetate was added 2.6 ml of hydrochloric acid and the mixture was 
stirred for 0.5 hour. The reaction solution was distilled off under 
reduced pressure. The residue was dissolved in chloroform, washed with 
water and then dried over anhydrous sodium sulfate. The solvent was 
distilled off under reduced pressure. Crystallization from ethyl acetate 
and hexane afforded 0.17 g of the title compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.20-1.43 (m, 16H), 1.70-1.85 (m, 4H), 
2.92-3.02 (m, 2H), 3.19 (t, J=7.3 Hz, 2H), 8.13-8.35 (m, 3H) 
PREATION EXAMPLE 16 
tert-Butyl N-(12-iodododecyl)carbamate 
EQU BocNH(CH.sub.2).sub.12 I 
To a suspension of 0.5 g of tert-butyl N-(12-hydroxydodecyl)carbamate in 15 
ml of toluene were added 0.28 g of imidazole, 1.09 g of triphenylphosphine 
and 0.84 g of iodine and the mixture was stirred for 0.5 hour. Then, 5 ml 
of tetrahydrofuran was further added and the mixture was stirred for 2 
hours. To the reaction solution was added ethyl acetate and the mixture 
was washed successively with a 10% aqueous solution of sodium thiosulfate 
and water and dried over anhydrous sodium sulfate and the solvent was 
distilled off under reduced pressure. The residue thus obtained was 
chromatographed over a silica gel column to afford 0.42 g of the title 
compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.23-1.47 (m, 18H), 1.44 (s, 9H), 1.81 
(quint, J=7.3 Hz, 2H), 2.99-3.12 (m, 2H), 3.18 (t, J=7.3 Hz, 2H), 
4.45-4.53 (m,1H) 
PREATION EXAMPLE 17 
tert-Butyl N-(12-hydroxydodecyl)carbamate 
EQU BocNH(CH.sub.2).sub.12 OH 
To a suspension of 1.2 g of sodium borohydride in 10 ml of tetrahydrofuran 
(THF) were added under ice-cooling a solution of 3.6 g of iodine in 20 ml 
of THF and the mixture was stirred for one hour. Then, a solution of 7.5 g 
of 12-(tert-butoxycarboxamido)dodecanoic acid in 25 ml of THF was added 
dropwise and the mixture was stirred at room temperature for one hour. To 
the reaction solution were added successively water and an aqueous 
solution of citric acid and extracted with ethyl acetate. The organic 
layer was washed successively with an aqueous solution of citric acid, a 
1N-sodium hydroxide solution, water and a saturated aqueous solution of 
sodium chloride and dried over anhydrous sodium sulfate. The solvent was 
distilled off under reduced pressure. The residue thus obtained was 
recrystallized from ethyl acetate-hexane to afford 4.7 g of the title 
compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.23-1.47 (m, 18H), 1.44 (s, 9H), 1.56 
(quint, J=7.3 Hz, 2H), 3.07-3.12 (m, 2H), 3.64 (t, J=6.8 Hz, 2H), 
4.45-4.53 (m, 1H) 
PREATION EXAMPLE 18 
12-(tert-Butoxycarboxamido)dodecanoic acid 
EQU BocNH(CH.sub.2).sub.11 COOH 
To 2.15 g of 12-aminododecanoic acid were added 20 ml of dioxane and 10 ml 
of water and then 2.39 g of di-tert-butyl dicarbonate and 10 ml of an 
aqueous 1N-sodium hydroxide solution were added under ice-cooling. The 
mixture was stirred at room temperature overnight. The reaction solution 
was distilled off under reduced pressure, the residue was made acidic by 
the addition of a 10% aqueous solution of citric acid and extracted with 
ethyl acetate. The organic layer was washed successively with water and a 
saturated aqueous solution of sodium chloride and dried over anhydrous 
sodium sulfate. The solvent was distilled off under reduced pressure to 
afford 3.09 g of the title compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.22-1.49 (m, 16H), 1.44 (s, 9H), 
1.59-1.64 (m, 2H), 2.34 (t, J=7.3 Hz, 2H), 3.05-3.12 (m, 2H), 4.49-4.54 
(m, 1H) 
PREATION EXAMPLE 19 
Sodium 6-(4-ethyl-1-piperazinyl)pyridine-3-carboxylate 
##STR34## 
A solution of 3.1 g of sodium hydroxide in 15 ml of water was added to 75 
ml of methanol. 15 g of methyl 
6-(4-ethyl-1-piperazinyl)pyridine-3-carboxylate was added and the mixture 
was heated under reflux for 1.5 hours. The solvent was distilled off under 
reduced pressure, to the resulting residue was added hot acetone and the 
crystal thus precipitated out was recovered by filtration. It was again 
washed with hot acetone to afford 17.2 g of the title compound as a 
crystal. 
PREATION EXAMPLE 20 
Potassium 6-(4-ethyl-1-piperazinyl)pyridine-3-carboxylate 
##STR35## 
To a solution of 69.81 g of ethyl 
6-(4-ethyl-1-piperazinyl)pyridine-3-carboxylate in 280 ml of methanol were 
added 37 g of potassium hydroxide and a small volume of water and the 
mixture was heated under reflux for one hour. To the reaction solution 
were added methanol and isopropyl alcohol, the crystal thus precipitated 
out was recovered by filtration and then dried under reduced pressure to 
afford 70.65 g of the title compound as a colorless crystal. 
.sup.1 H NMR (CD.sub.3 OD).delta. 1.14 (t, J=7.2 Hz, 3H), 2.48 (q, J=7.2 
Hz, 2H), 2.58 (t, J=5.1 Hz, 4H), 3.61 (t, J=5.1 Hz, 4H), 6.75 (d, J=8.8 
Hz, 1H), 8.05 (dd, J=2.4 Hz, 8.8 Hz, 1H), 8.69 (d, J=2.4 Hz, 1H) 
PREATION EXAMPLE 21 
Methyl 6-(4-ethyl-1-piperazinyl)pyridine-3-carboxylate 
##STR36## 
To 100 g of 1-ethylpiperazine warmed to 80.degree. C. was added at 
80-100.degree. C. with stirring 50 g of methyl 
6-chloropyridine-3-carboxylate. To the reaction solution was added 0.5 1 
of water and the mixture was stirred. The crystal thus precipitated out 
was recovered by filtration, dried under reduced pressure to afford 68.8 g 
of the title compound as a crystal. 
PREATION EXAMPLE 22 
Ethyl 6-(4-ethyl-1-piperazinyl)pyridine-3-carboxylate 
##STR37## 
To 55.69 g of ethyl 6-chloropyridine-3-carboxylate was added 115 ml of 
1-ethylpiperazine and the mixture was heated with stirring (55-137.degree. 
C.) for 5 minutes. After completion of the reaction, the 1-ethylpiperazine 
was distilled off under reduced pressure, the residue was crystallized 
from water-metanol. The crystal was washed with water and then dried under 
reduced pressure to afford 71.89 g of the title compound as a colorless 
crystal. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.13(t, J=7.2 Hz, 3H), 1.36 (t, J=7.1 Hz, 
3H), 2.47 (q, J=7.2 Hz, 2H), 2.54 (t, J=5.1 Hz, 4H), 3.71 (t, J=5.1 Hz, 
4H), 4.33 (q, J=7.1 Hz, 2H), 6.58 (d, J=8.8 Hz, 1H), 8.02 (dd, J=2.4 Hz, 
8.8 Hz, 1H), 8.80 (d, J=2.4 Hz, 1H) 
PREATION EXAMPLE 23 
1-Isopropylpiperazine 
##STR38## 
PREATION EXAMPLE 23-1 
1-Acetyl-4-isopropylpiperazine 
##STR39## 
To 12.95 g of piperazine, 0.75 g of sodium iodide and 3.46 g of potassium 
carbonate were added successively 25 ml of methanol and 6.15 g of 
2-bromopropane and the mixture was stirred at 60.degree. C. for 4 hours. 
After the addition of chloroform and washing with a saturated aqueous 
solution of sodium chloride, the mixture was dried over anhydrous 
magnesium sulfate. The solvent was distilled off under reduced pressure, 
12 ml of acetic anhydride was slowly added and the mixture was stirred at 
room temperature for 2.5 hours. The reaction solution was poured into ice 
and chloroform was added and the mixture was neutralized with sodium 
carbonate. After extracting with chloroform, the extract was dried over 
anhydrous magnesium sulfate. The solvent was distilled off under reduced 
pressure and the residue thus obtained was chromatographed over a silica 
gel column. The solvent was distilled off and the residue was allowed to 
stand overnight. The diacetylpiperazine thus precipitated out was filtered 
off to afford 7.26 g of the title compound as an oily substance. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.04 (d, J=6 Hz, 6H), 2.08 (s, 3H), 2.47 
(t, J=5 Hz,2H), 2.51 (t, J=5 Hz, 2H), 2.71 (sept, J=6 Hz, 1H), 3.46 (t, 
J=5 Hz, 2H), 3.62 (t, J=5 Hz, 2H) 
PREATION EXAMPLE 23-2 
1-Isopropylpiperazine 
To 7.26 g of 1-acetyl-4-isopropylpiperazine were added 100 ml of methanol 
and 10 g of potassium hydroxide and the mixture was heated under reflux 
for 17 hours. After the solvent was distilled off, water and chloroform 
were added. The mixture was extracted with chloroform and dried over 
anhydrous magnesium sulfate. The solvent was distilled off under reduced 
pressure to afford 5.78 g of the title compound as an oily substance. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.05 (d, J=6 Hz, 6H), 2.49 (bt, J=5 Hz, 
4H), 2.75 (sept, J=6 Hz, 1H), 2.90 (t, J=5 Hz, 4H) 
PREATION EXAMPLE 24 
1-Isopropylpiperazine 
##STR40## 
1-Formyl-4-isopropylpiperazine 
##STR41## 
To a solution of 1.02 g of formylpiperazine in 90 ml of acetone was added 
0.1 g of 5% palladium carbon and the mixture was stirred under hydrogen 
atmosphere for 60 hours. The catalyst was filtered off and distilled off 
under reduced pressure to afford 1.31 g of the title compound. 
.sup.1 H NMR (CDCl.sub.3).delta. 1.04 (d, J=6.3 Hz, 6H), 2.48 (t, J=4.8 Hz, 
2H), 2.51 (t, J=4.8 Hz, 2H), 2.74 (sept, J=6.3 Hz, 1H), 3.37 (t, J=4.8 
Hz,2H), 3.55 (t, J=4.8 Hz, 2H), 8.01 (brs, 1H) 
1-Isopropylpiperazine 
To a solution of 0.29 g of 1-formyl-4-isopropylpiperazine in 10 ml of 
methanol were added 0.5 ml of hydrochloric acid and 1 ml of water and the 
mixture was heated under reflux for 2 hours. The reaction solution was 
distilled off under reduced pressure, a iN aqueous solution of sodium 
hydroxide was added and the mixture was extracted with chloroform and 
dried over anhydrous magnesium sulfate. The solvent was distilled off 
under reduced pressure to afford 0.1 g of the title compound. 
Pharmacological test results of the pyridinecarboxamide derivatives (1) of 
the present invention will be shown below. 
For comparison, the compound of Example 10 and the compound of Example 2 of 
JP-A-5-32630 were used, namely, 
N-(ll-nitroxy-1-undecanyl)-6-(4-methyl-1-piperazinyl)-nicotinamide 
(hereinafter referred to as "Comparative Compound 1"), and 
N-(12-nitroxy-1-dodecanyl)-6-(4-methyl-1-piperazinyl)nicotinamide 
(hereinafter referred to as "Comparative Compound 2"). 
The compounds of the present invention will be shown below in terms of the 
corresponding Example numbers. As an example, "Compound 1" is meant to 
indicate the compound obtained by the present Example 1. 
1. Behavior Suppressing Action 
General behavior of mice caused by the pyridinecarboxamide derivatives (I) 
of the present invention was studied. 
Using ddy-strain mice of 7 weeks old, test substance was administered at 
100 .mu.l/10 sec to the tail vein and behavior of the animal was observed 
for one hour. After the administration, sedative animal was evaluated as 
sedated, while animal with behavioral suppression was evaluated as 
suppressed. 
TABLE 1 
______________________________________ 
R n Behavioral suppression (10 mg/kg) 
______________________________________ 
Compound 1 
Et 12 Not shown 
Compound 2 
i-Pr 12 Not shown 
Comparative 
Me 11 Sedative action 
Compound 1 
______________________________________ 
Behavioral suppression was not observed with Compound 1 and Compound 2. 
However, it has become apparent that Comparative Compound 1 showed the 
behavioral suppression and therefore is not desirable as a therapeutic 
agent for cerebrovascular disorders. 
2. Cerebral Protective Action 
Cerebral protective action of the pyridinecarboxamide derivatives of this 
invention were studied (an anti-anoxia action) using a hypoxic model of 
mice. 
Using ddY-strain male mice of 6 weeks old, the test substance of Compound 1 
or Compound 2 was intravenously administered to the tail vein at the dose 
of 1.0 mg/kg. After 30 minutes from the administration, animal were 
decapitated to measure a gasping duration. This measurement was made by 
two persons who had not been informed of the test substances and the 
measured values were averaged to obtain the data. Also, Comparative 
Compound 1 was administered in the same manner as described above. 
It is believed that gasping after decapitation is controlled by the 
respiratory center and, if the nerve functions are maintained by the 
respiratory center, the gasping duration would be prolonged. Moreover, it 
is suggested that the ischemia with decapitation is related to the 
decrease in the intracerebral glucose which is believed to be essential as 
a nutrition component in the brain. In view of the foregoing, the 
pyridinecarboxamide derivatives of this invention can prolong the gasping 
duration and then are useful as a cerebral protective agent. 
The action of each substance will be shown in Table 2, with the gasping 
duration of Comparative Compound 1 being defined as 1. 
TABLE 2 
______________________________________ 
R n Gasping duration 
______________________________________ 
Compound 1 
Et 12 1.4 
Compound 2 
i-Pr 12 1.5 
Comparative 
Me 11 1 
Compound 1 
______________________________________ 
The Compound 1 and Compound 2 had 1.4 and 1.5 times higher cerebral 
protective action than Comparative Compound 1, respectively. 3. Inhibiting 
Action on Calcium-Influx 
From SD-strain male rats, the brain was excised under light ether 
anesthesia. The hippocampus slice with a thickness of 400 mm was prepared 
by means of a microslicer and stored in artificial cerebrospinal fluid. 
The slice was immersed in fura-2/Am 10 mM for one hour to load the 
pigment. The fluorescent images obtained with the excited wavelengths of 
380 nm and 340 nm were taken and analyzed by means of Argus-50 System. The 
ischemic state was produced by perfusing a solution containing no glucose 
and saturated with a N.sub.2 /CO.sub.2 gas from 3 minutes to 10 minutes 
after initiation of the measurement. The drug was dissolved in a perfusion 
solution and applied from 20 minutes prior to the ischemia to 10 minutes 
after the ischemia. 
The ratios of the data obtained at 340 nm/380 nm were calculated and an 
average from 2 minutes through 16 minutes after completion of the ischemia 
was determined. 
The perfusate containing no drug was used and similar experiment was 
performed to prepare a control. The change rates are as shown below in 
terms of the control value being defined as 100%. 
______________________________________ 
Compound 1 95.2% 
Comparative Compound 2 
105.6% 
______________________________________ 
Compound 1 controlled the Ca-influx in cerebral neuronal cells. No 
considerably great difference in the change rates was observed between the 
group given Compound 1 and the group given Comparative Compound 2, but 
Compound 1 showed an earlier reversion to the baseline during the recovery 
stage (approximately 10 minutes after the ischemia). 
4. Anti-cerebral Edema Action (Obstructive Cerebral Ischemia Model Using 
Polyvinyl Acetate) 
Inhibiting action on cerebral ischemia was studied in an obstructive 
cerebral ischemia model using polyvinyl acetate. 
The obstructive model was prepared as described below. (Hiroyoshi Nishi et 
al., Stroke 1989, 20:1236-1240) 
Wistar-strain male rats weighing 200-350 g were used and fixed at the 
dorsal position under anesthesia of ether and then the left common carotid 
artery, internal carotid artery, external carotid artery were isolated. 
The external carotid artery was ligated, the pterygopalatine artery was 
fastened with clamp and a cannula was inserted from the external carotid 
artery toward the branch between the internal carotid artery and the 
common carotid artery. 5 .mu.l of a 3% polyvinyl acetate/52% ethanol 
solution in water was injected. After 30 seconds, the clamp was removed 
from the artery and then the wound sutured. After 24 hours, animal was 
decapitated and the cerebrum was separated, within 120 seconds from which 
wet weights of the left cerebrum and the right cerebrum were measured. The 
left and right cerebra were dried in an oven at 105.degree. C. for 24 
hours and then the dry weights were measured. A cerebral water content was 
calculated according to the following equation: 
##EQU1## 
Compound 1, Compound 2 or Comparative Compound 2 was intravenously injected 
to the tail vein at the doses of 1.0 mg/kg and 3.0 mg/kg before 5 minutes 
from the administration of polyvinyl acetate. 
Inhibitory rates of cerebral edema in the cerebral edema model using 
polyvinyl acetate are shown in Table 3. 
TABLE 3 
______________________________________ 
R n 1 mg/kg 3 mg/kg 
______________________________________ 
Compound 1 Et 12 29.0% 61.9% 
Compound 2 i-Pr 12 59.1% 
Comparative Me 12 21.5% 11.3% 
Compound 2 
______________________________________ 
Compounds 1 and 2 inhibited cerebral edema by around 60%, whereas 
Comparative Compound 2 inhibited only by around 20%. 
5. Anti-Cerebral Edema Action (Ischemic Reperfusion Model Using SHR-SP) 
The inhibitory action on cerebral edema of the pyridinecarboxamide 
derivatives of this invention were studied using the SHR-SP models 
(Spontaneous hypertensive rats which are vulnerable to cerebral 
hemorrhage). 
Male SHR-SP of 15-17 weeks old were measured for the blood pressure one day 
prior to the test and classified into groups. Both common carotid arteries 
were separated under pentobarbital anesthesia at 35 mg/kg and common 
carotid arteries were fastened by clamp to cause ischemia. After 2 hours, 
clamp was removed from the arteries to reperfuse. After a further 2 hours, 
the animal was decapitated and cerebrum was excised under ether 
anesthesia. The cerebrum was dried in an overn at 105.degree. C. for 24 
hours and the cerebral water content (%) was calculated. 
##EQU2## 
Compound 1, Compound 2 and Comparative Compound 2 were intravenously 
administered twice, that is, immediately after the ischemia and 
immediately before the reperfusion, at the dose of 1 mg/kg, respectively. 
Inhibitory rates of cerebral ischemia by Compound 1, Compound 2 and 
Comparative Compound 2 are shown in Table 3-2. 
TABLE 3-2 
______________________________________ 
R n 
______________________________________ 
Compound 1 Et 12 63% 
Compound 2 i-Pr 12 71% 
Comparative 
Me 12 -90% 
Compound 2 
______________________________________ 
Compound 1 and Compound 2 showed a potent anti-cerebral edema activity, 
whereas Comparative Compound 2 made cerebral edema worse. 
6. Inhibitory Rate of Delayed Neuronal Death 
MON/Jms/Gbs-strain male jirds (weighing 60-80 g) were fixed at the dorsal 
position under 1.5-2.0% halothane anesthesia and the common carotid artery 
was isolated. Ischemia was caused by fastening the common carotid artery 
with clamp for 3 minutes and then reperfused. After 7 days, the brain was 
excised under ether anesthesia and fixed with 10% formalin for 2 days and 
then tissue slices of the hippocampus were prepared. The hippocampus CA1 
cells were stained using HE staining and the survival rate of the CA1 
cells was evaluated. 
Compound 1 was administered to the carotid artery at the dose of 0.5 mg/kg 
immediately after the ischemia. Using as a control the case wherein the 
drug was not administered, inhibitory rate was calculated according to the 
following equation: 
##EQU3## 
TABLE 4 
______________________________________ 
R n Inhibitory rate 
______________________________________ 
Compound 1 
Et 12 20% 
______________________________________ 
7. Inhibitory Action on Lipid Peroxidation 
Inhibitory action on lipid peroxidation was studied using rat brain 
homogenate. 
Wistar-strain male rats (weighing 230-300 g) were decapitated and the 
cerebrum was quickly excised. A 4 times volume of a solution of 50 mM 
phosphoric acid and 0.142 mM NaCl (pH 7.4) was added and the mixture was 
homogenized and then centrifuged at 3000 rpm for 10 minutes. The 
supernatant thus obtained was prepared so as to give a protein level of 2 
mg/ml. 
Determination of lipid peroxide was carried out using TBA method 
(thiobarbituric acid). To the brain homogenate was added Compound 2 so as 
to give a final concentration of 10.sup.-4 M, thereby initiating the 
reaction. After incubation at 37.degree. C. for 15 minutes, the reaction 
was stopped in ice by adding a 35% perchloric acid solution. The reaction 
mixture was centrifuged at 4.degree. C. and 1000 rpm for 5 minutes and to 
the supernatant thus obtained was added a 0.5% TBA solution. The mixture 
was boiled at 100.degree. C. for 15 minutes. After cooling, absorbance was 
determined at 532 nm. 1,1,3,3-Tetraethoxypropane as a standard was 
subjected to the reaction in the same manner as above and an amount of 
lipid peroxide, that is, an amount of the malondialdehyde (MDA) produced 
was determined from the absorbances. 
Inhibitory rate of lipid peroxidation was calculated according to the 
following equation: 
______________________________________ 
Inhibitory rate of lipid peroxidation = 
[1 - {MDA nmol (Compound 2; 
incubated for 15 min.) - MDA 
nmol (Compound 2; incubated for 
0 min.)}/{MDA nmol (solvent; 
incubated for 15 min.) - MDA 
nmol (solvent; incubated for 0 
min.)}] .times. 100(%) 
______________________________________ 
TABLE 5 
______________________________________ 
Inhibitory rate of lipid 
R n peroxidation (%) 
______________________________________ 
Compound 2 
i-Pr 12 71 
______________________________________ 
Table 5 demonstrates that the pyridinecarboxamide derivatives of this 
invention have an inhibitory action on lipid peroxidation.