Title of Invention

NOVEL ARYLAMIDINE COMPOUND HAVING ANTIFUNGAL AND ANTIPROTOZOAL ACTIVITY

Abstract An arylamidine derivative represented by the general formula (wherein R1 represents optionally protected or substituted amidino; and R2 and R3 are the same or different and each represents hydrogen or halogeno) or a salt of the derivative. The derivative and salt have potent activity against fungi including ones having tolerance to azole type drugs and further have high safety and excellent properties in a repeated dose toxicity test. They are hence useful as an excellent antifungal.
Full Text TECHNICAL FIELD
The present invention relates to a novel
arylamidine derivative or a salt thereof having an
antifungal activity, and an antifungal agent comprising
the same as an active ingredient.
BACKGROUND ART
Serious deep mycosis such as invasive
candidiasis often becomes a fatal disease. Originally,
it has been considered that a principal protective
mechanism on the side of a host organism to fungi such
as Candida would be nonspecific immunization by
neutrophils. When this protective mechanism normally
functions, there is little risk of becoming infected
with fungi. However, in recent years, a risk of
suffering from deep mycosis has been boosted because of
the increased number of patients with underlying
diseases decreasing the immunological function of an
organism, such as malignant tumors and AIDS, frequent
use of anticancer agents or immunosuppressive agents,
heavy use of antibacterial antibiotics or steroid
hormone, long-term use of central venous
hyperalimentation or venous catheterization, and the

like (Non-Patent Document 1).
There are only 6 agents, i.e., amphotericin
B, flucytosine, miconazole, fluconazole, itraconazole,
and micafungin as agents for such deep mycosis.
Amphotericin B has an extremely strong fungicidal
action; however, it has a problem regarding side
effects such as nephrotoxicity, and its clinical use is
therefore limited. Flucytosine is rarely used alone at
present because the agent has problems, e.g.,
development of resistance. Micafungin has a low
activity against the genus Cryptococcus. The other
agents are generically called an azole antifungal
agent, and are most frequently used at present
considering a balance between effectiveness and safety
although their antifungal action tends to be generally
inferior compared to that of amphotericin B (Non-Patent
Document 2).
Currently, fluconazole-resistant Candida
albicans (C. albicans) has been detected with a high
frequency in oropharyngeal candidiasis lesions of AIDS
patients to whom fluconazole has been repeatedly
administered. What is more, most of the resistant
strains show cross resistance to itraconazole and other
azole agents. Further, such resistant strains have
also been reported to be isolated from non-AIDS
patients who developed chronic mucocutaneous
candidiasis or deep candidiasis (Non-Patent Document
3). The problems regarding resistance seriously affect

the management of patients with deep mycosis, the
number of which has been steadily increasing (Non-
Patent Document 3).
[Non-patent document 1]: Rinsho to Biseibutsu
(Clinics and Microorganisms), Vol. 17: pp. 265-266,
1990
[Non-patent document 2]: Rinsho to Biseibutsu
(Clinics and Microorganisms), Vol. 21: pp. 277-283,
1994
[Non-patent document 3]: Rinsho to Biseibutsu
(Clinics and Microorganisms), Vol. 28: pp. 51-58, 2001
DISCLOSURE OF THE INVENTION
PROBLEMS TO BE SOLVED BY THE INVENTION
There is strongly desired an antifungal agent
whose action mechanism differs from those of the
existing agents and which is effective even against
fungi resistant to azole agents while having reduced
side effects. WO03/074476 describes that an
arylamidine derivative has strong antifungal activities
and is useful as an antifungal agent; however, there is
a need for a compound having more reduced side effects
and improved in physical properties such as
hygroscopicity and deliquescence, and, in addition, a
compound also having an excellent effect against
protozoans and the like.
MEANS FOR SOLVING THE PROBLEMS

As a result of intensive studies under such
circumstances, the present inventors have found that a
compound represented by general formula (1):

wherein R1 represents an optionally protected or
substituted amidino group; and R2 and R3 are identical
or different and each represent a hydrogen atom or a
halogen atom; or a salt thereof is effective even
against fungi resistant to azole agents while having
reduced side effects; particularly, a compound of
general formula (1) wherein R1 is an amidino group and
R2 and R3 are each a hydrogen atom exhibits strong
activities against fungi including those resistant to
azole agents while having high safety; and, further, a
trihydrochloride pentahydrate of a compound of general
formula (1) wherein R1 is an amidino group and R2 and R3
are each a hydrogen atom is excellent in chemical
stability while having no deliquescence or
hygroscopicity and suitable as a drug substance, and
additionally has excellent activities against
protozoans, thereby accomplishing the invention.
ADVANTAGES OF THE INVENTION
The compound of the invention is highly
active against fungi including those resistant to azole

agents, has excellent physical properties along with
high safety, and is useful as an antifungal agent. In
addition, the compound has excellent activities against
protozoans and is useful as an antiprotozoan.
BEST MODE FOR CARRYING OUT THE INVENTION
The present invention is described below in
detail.
In the description, unless otherwise noted, a
halogen atom refers to a fluorine atom, a chlorine
atom, a bromine atom, or an iodine atom; an alkyl
group, for example, to a straight-chain or branched C1-12
alkyl group such as methyl, ethyl, propyl, isopropyl,
butyl, sec-butyl, isobutyl, tert-butyl, pentyl,
isopentyl, hexyl, heptyl, and octyl; a lower alkyl
group, for example, to a straight-chain or branched C1-6
alkyl group such as methyl, ethyl, propyl, isopropyl,
butyl, sec-butyl, isobutyl, tert-butyl, pentyl, and
isopentyl; an alkenyl group, for example, to a
straight-chain or branched C2-12 alkenyl group such as
vinyl, allyl, propenyl, isopropenyl, butenyl,
isobutenyl, pentenyl, hexenyl, heptenyl, and octenyl;
an aryl group, for example, to a group such as phenyl
and naphthyl; an aralkyl group, for example, to an ar-
C1-6-alkyl group such as benzyl, diphenylmethyl, trityl,
phenethyl, and naphthylmethyl; an alkoxy group, for
example, to a straight-chain or branched C1-6 alkyloxy
group such as methoxy, ethoxy, propoxy, isopropoxy,

butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy,
and isopentyloxy; an aralkyloxy group, for example, to
an ar-C1-6-alkyloxy group such as benzyloxy,
diphenylmethyloxy, trityloxy, phenethyloxy, and
naphthylmethyloxy; an alkoxyalkyl group, for example,
to a C1-6-alkyloxy-C1-6-alkyl group such as methoxymethyl
and 1-ethoxyethyl; a cycloalkyloxy group, for example,
to a C3-8 cycloalkyloxy group such as cyclopropoxy,
cyclobutoxy, cyclopentyloxy, and cyclohexyloxy; an
aralkyloxyalkyl group, for example, to an ar-C1-6-
alkyloxy-C1-6-alkyl group such as benzyloxymethyl and
phenethyloxymethyl;
an acyl group, for example, to a straight-chain or
branched C2-12 alkanoyl group such as formyl, acetyl,
propionyl, and isovaleryl, an ar-C1-6-alkylcarbonyl
group such as benzylcarbonyl, an aroyl group such as
benzoyl and naphthoyl, a heterocyclic carbonyl group
such as nicotinoyl, thenoyl, pyrrolidinocarbonyl, and
furoyl, a carboxy-C1-6-alkylcarbonyl group such as 3-
carboxypropanoyl and 4-carboxybutanoyl, a C1-6-
alkyloxycarbonyl-C1-6-alkylcarbonyl group such as 3-
(methoxycarbonyl)propanoyl and 4-
(methoxycarbonyl)butanoyl, a succinyl group, a glutaryl
group, a maleoyl group, a phthaloyl group, or a
straight-chain or branched α-aminoalkanoyl group whose
N-terminal is optionally protected, derived from an
amino acid (including, for example, glycine, alanine,
valine, leucine, isoleucine, serine, threonine,

cysteine, methionine, aspartic acid, glutamic acid,
asparagine, glutamine, arginine, lysine, histidine,
hydroxylysine, phenylalanine, tyrosine, tryptophan,
proline, or hydroxyproline);
an alkyloxycarbonyl group, for example, to a straight-
chain or branched C1-12 alkyloxycarbonyl group such as
methoxycarbonyl, ethoxycarbonyl, 1,1-
dimethylpropoxycarbonyl, isopropoxycarbonyl, 2-
ethylhexyloxycarbonyl, tert-butoxycarbonyl, and tert-
pentyloxycarbonyl; a cycloalkyloxycarbonyl group, for
example, to a C3-8 cycloalkyloxycarbonyl group such as
cyclopentyloxycarbonyl and cyclohexyloxycarbonyl; an
aralkyloxycarbonyl group, for example, an ar-C1-6-
alkyloxycarbonyl group such as benzyloxycarbonyl and
phenethyloxycarbonyl; an aryloxycarbonyl group, for
example, to a group such as phenyloxycarbonyl; an
acyloxy group, for example, to a straight-chain or
branched C2-6 alkanoyloxy group such as acetyloxy and
propionyloxy or an aroyloxy group such as benzoyloxy;
an arylthio group, for example, to a group such as
phenylthio; an alkanesulfonyl group, for example, to a
C1-6 alkanesulfonyl group such as methanesulfonyl,
ethanesulfonyl, and propanesulfonyl; an arylsulfonyl
group, for example, to a group such as benzenesulfonyl,
toluenesulfonyl, and naphthalenesulfonyl; an
alkanesulfonyloxy group, for example, to a C1-6
alkanesulfonyloxy group such as methanesulfonyloxy and
ethanesulfonyloxy; an arylsulfonyloxy group, for

example, to a group such as benzenesulfonyloxy and
toluenesulfonyloxy;
an alkylthiocarbonyl group, for example, to a C1-6
alkylthiocarbonyl group such as methylthiocarbonyl and
ethylthiocarbonyl; a cycloalkylidene group, for
example, to a group such as cyclopentylidene and
cyclohexylidene; an aralkylidene group, for example, to
a group such as benzylidene and naphthylmethylene; a
dialkylaminoalkylidene group, for example, to a group
such as N,N-dimethylaminomethylene and N,N-
diethylaminomethylene; a diaralkylphosphoryl group, for
example, to a group such as dibenzylphosphoryl; a
diarylphosphoryl group, for example, to a group such as
diphenylphosphoryl;
an oxygen-containing heterocyclic group, for example,
to a group such as tetrahydrofuryl and
tetrahydropyranyl; an oxygen-containing heterocyclic
alkyl group, for example, to a group such as 5-methyl-
2-oxo-2H-1,3-dioxol-4-ylmethyl; a sulfur-containing
heterocyclic group, for example, to a group such as
tetrahydrothiopyranyl; a heterocyclic oxycarbonyl
group, for example, to a group such as 2-
furfuryloxycarbonyl and 8-quinolyloxycarbonyl; a
nitrogen-containing heterocyclic alkylidene group, for
example, to a group such as 3-hydroxy-4-
pyridylmethylene; and a substituted silyl group, for
example, to a group such as trimethylsilyl,
triethylsilyl, and tributylsilyl.

Each of the above-described groups may be
further substituted with one or more groups selected
from a halogen atom, an optionally protected amino
group, an optionally protected hydroxyl group, a nitro
group, a lower alkyl group, an alkenyl group, an alkoxy
group, an aralkyloxy group, an aryl group, an acyl
group, and an oxo group.
The amino-protecting groups encompass all of
the conventional groups which can be used as protective
groups for an amino group, and include, for example, an
acyl group, an alkyloxycarbonyl group, an
aralkyloxycarbonyl group, an aryloxycarbonyl group, an
aralkyl group, an alkoxyalkyl group, an aralkyloxyalkyl
group, an arylthio group, an alkanesulfonyl group, an
arylsulfonyl group, a dialkylaminoalkylidene group, an
aralkylidene group, a nitrogen-containing heterocyclic
alkylidene group, a cycloalkylidene group, a
diarylphosphoryl group, a diaralkylphosphoryl group, an
oxygen-containing heterocyclic alkyl group, and a
substituted silyl group.
The hydroxyl-protecting groups encompass all
of the conventional groups which can be used as
protective groups for a hydroxyl group, and include,
for example, an acyl group, an alkyloxycarbonyl group,
an aralkyloxycarbonyl group, a heterocyclic oxycarbonyl
group, an alkyl group, an alkenyl group, an aralkyl
group, an oxygen-containing heterocyclic group, a
sulfur-containing heterocyclic group, an alkoxyalkyl

group, an aralkyloxyalkyl group, an alkanesulfonyl
group, an arylsulfonyl group, and a substituted silyl
group.
The amidino-protecting groups encompass all
of the conventional groups which can be used as
protective groups for an amidino group, and include,
for example, an acyl group, an alkyloxycarbonyl group,
an aralkyloxycarbonyl group, an aryloxycarbonyl group,
a cycloalkyloxycarbonyl group, an aralkyl group, an
alkoxyalkyl group, an aralkyloxyalkyl group, an
arylthio group, an alkanesulfonyl group, an
arylsulfonyl group, a cycloalkyloxycarbonyl group, an
alkylthiocarbonyl group, a dialkylaminoalkylidene
group, an aralkylidene group, a nitrogen-containing
heterocyclic alkylidene group, a cycloalkylidene group,
an oxygen-containing heterocyclic alkyl group, and a
substituted silyl group.
The substituents for an amidino group
include, for example, a hydroxyl group optionally
substituted with an acyl group, and optionally
substituted alkoxy and aralkyloxy groups.
The leaving groups include, for example, a
halogen atom, an alkanesulfonyloxy group, an
arylsulfonyloxy group, and an acyloxy group.
The salts of a compound of formula (1)
include, for example, salts with mineral acids such as
hydrochloric acid, hydrobromic acid, phosphoric acid,
and sulfuric acid; salts with organic carboxylic acids

such as formic acid, acetic acid, trichloroacetic acid,
L-lactic acid, L-tartaric acid, citric acid, succinic
acid, maleic acid, fumaric acid, and trifluoroacetic
acid; and salts with sulfonic acids such as
methanesulfonic acid, benzenesulfonic acid, p-
toluenesulfonic acid, mesitylenesulfonic acid, and
naphthalenesulfonic acid.
Preferred salts of a compound of formula (1)
include pharmacologically acceptable salts.
According to the invention, preferred
compounds include the following compounds.
A compound wherein R1 is an amidino group
optionally substituted with a hydroxyl group optionally
substituted with an acyl group is preferable; a
compound wherein R1 is an amidino group optionally
substituted with a hydroxyl group is more preferable;
and a compound wherein R1is an amidino group is still
more preferable.
A compound wherein R2 and R3 are identical or
different and each a hydrogen atom or a fluorine atom
is preferable; and a compound wherein R2 and R3 are each
a hydrogen atom is more preferable.
The compound wherein R1 is an amidino group
and R2 and R3 are each a hydrogen atom is preferably a
salt with hydrochloric acid, phosphoric acid, sulfuric
acid, acetic acid, L-lactic acid, or methanesulfonic
acid, more preferably a salt with hydrochloric acid,
phosphoric acid, or sulfuric acid, still more

preferably a salt with hydrochloric acid.
The hydrochloride of the compound wherein R1
is an amidino group and R2 and R3 are each a hydrogen
atom is preferably a dihydrochloride or
trihydrochloride, more preferably a trihydrochloride.
The trihydrochloride is preferably a
monohydrate or pentahydrate thereof, more preferably a
pentahydrate thereof.
A method for producing the compound of the
invention is then described.
The compound of the invention is produced by
combining methods known per se, and, for example, by
production methods given in the following.

wherein R4 represents a lower alkyl group; and R2 and R3
have the same meanings as described above.
The compound of general formula (1a) can be
produced by reacting the compound of general formula

(2) with the compound of general formula (4) to convert
to the compound of general formula (3), followed by
reacting the compound of general formula (3) with
ammonia or an ammonium salt. This reaction may be
conducted by a method described, for example, in
W096/16947 and J. Org. Chem., 64: 12-13, 1999, or by a
method equivalent thereto.
This series of reactions will be now
described in detail.
(1-1)
The compound of general formula (3) can be
produced by reacting the compound of general formula
(2) with the compound of general formula (4) in the
presence of an acid.
The solvent used in this reaction is not
particularly limited, provided that it does not
adversely affect the reaction, and examples thereof
include alcohols such as methanol, ethanol, 2-propanol,
and 2-methyl-2-propanol; amides such as N,N-
dimethylformamide, N,N-dimethylacetamide, and 1-methyl-
2-pyrrolidone; halogenated hydrocarbons such as
methylene chloride, chloroform, and dichloroethane;
aromatic hydrocarbons such as benzene, toluene, and
xylene; ethers such as dioxane, tetrahydrofuran,
anisole, diethylene glycol dimethyl ether, diethylene
glycol diethyl ether, and ethylene glycol monomethyl
ether; sulfoxides such as dimethylsulfoxide; ketones
such as acetone and 2-butanone; esters such as ethyl

acetate; and carboxylic acids such as acetic acid.
These may be used in the form of a mixture. The
compound of general formula (4) may be also used as a
solvent.
Example of the acid used in the reaction
include hydrogen chloride, hydrogen bromide, perchloric
acid, p-toluenesulfonic acid, and methanesulfonic acid,
and the usage amount thereof is 1- to 200-fold moles,
preferably 5- to 100-fold moles based on the compound
of general formula (2).
In the reaction, the usage amount of the
compound of general formula (4) may be 2- to 1,000-fold
moles based on the compound of general formula (2), and
is preferably used as a solvent.
The reaction may be conducted at -30 to 150°C,
preferably 10 to 50°C for 30 minutes to 24 hours.
(1-2)
The compound of general formula (la) can be
produced by reacting the compound of general formula
(3) with ammonia or an ammonium salt.
The solvent used in this reaction is not
particularly limited, provided that it does not
adversely affect the reaction, and examples thereof
include alcohols such as methanol, ethanol, 2-propanol,
and 2-methyl-2-propanol; amides such as N,N-
dimethylformamide, N,N-dimethylacetamide, and 1-methyl-
2-pyrrolidone; halogenated hydrocarbons such as
methylene chloride, chloroform, and dichloroethane;

aromatic hydrocarbons such as benzene, toluene, and
xylene; ethers such as dioxane, tetrahydrofuran,
anisole, diethylene glycol dimethyl ether, diethylene
glycol diethyl ether, and ethylene glycol monomethyl
ether; nitriles such as acetonitrile; sulfoxides such
as dimethylsulfoxide; heteroaromatics such as pyridine;
and water. These may be used in the form of a mixture.
Examples of the ammonium salt include
ammonium chloride, ammonium bromide, and ammonium
acetate. The usage amount of ammonia or the ammonium
salt may be 3- to 100-fold moles, preferably 3- to 10-
fold moles based on the compound of general formula
(3) .
The reaction may be conducted at 0 to 150°C,
preferably 20 to 120°C for one minute to 24 hours.


wherein R5 represents an optionally substituted acyl,
lower alkyl or aralkyl group; and R2 and R3 have the
same meanings as described above.
The compound of general formula (1b) can be
produced from the compound of general formula (2). The
compound of general formula (1b) can be then alkylated
or acylated to produce the compound of general formula
(1c). The compound of general formula (1c) can be
further reduced to produce the compound of general
formula (1a). In addition, the compound of general
formula (1b) can be reduced to produce the compound of
general formula (1a). These reactions may be conducted
by methods described, for example, in Tetrahedron, 51:
12047-12068, 1995; Synthetic Communication, 26: 4351-
4367, 1996; Synthesis, 16: 2467-2469, 2003;
Heterocycles, 60: 1133-1145, 2003; and Bioorganic and
Medicinal Chemistry Letter, 12: 1203-1208, 2002, or by
methods equivalent thereto.
This series of reactions will be now
described in detail.
(2-1)
The compound of general formula (1b) can be
produced by reacting the compound of general formula
(2) with hydroxylamine or its salt in the presence or
absence of a base.
The solvent used in this reaction is not
particularly limited, provided that it does not
adversely affect the reaction, and examples thereof

include alcohols such as methanol, ethanol, 2-propanol,
and 2-methyl-2-propanol; amides such as N,N-
dimethylformamide, N,N-dimethylacetamide, and 1-methyl-
2-pyrrolidone; halogenated hydrocarbons such as
methylene chloride, chloroform, and dichloroethane;
aromatic hydrocarbons such as benzene, toluene, and
xylene; ethers such as dioxane, tetrahydrofuran,
anisole, diethylene glycol dimethyl ether, diethylene
glycol diethyl ether, and ethylene glycol monomethyl
ether; sulfoxides such as dimethylsulfoxide; ketones
such as acetone and 2-butanone; heteroaromatics such as
pyridine; and water. These may be used in the form of
a mixture.
Examples of the base used in the reaction, if
desired, include metal alkoxides such as sodium
methoxide, sodium ethoxide, potassium tert-butoxide,
and sodium tert-butoxide; inorganic salts such as
sodium hydroxide, potassium hydroxide, sodium
hydrogencarbonate, sodium carbonate, potassium
carbonate, sodium hydride, and potassium hydride; and
organic bases such as triethylamine and pyridine.
The usage amount of the base may be 2- to
100-fold moles, preferably 2- to 20-fold moles based on
the compound of general formula (2).
Examples of the salt of hydroxylamine include
hydrochlorides and sulfates.
The usage amount of hydroxylamine or its salt
may be 2- to 100-fold moles, preferably 2- to 20-fold

moles based on the compound of general formula (2).
The reaction may be conducted at 0 to 150°C,
preferably 50 to 150°C for one minute to 24 hours.
(2-2)
The compound of general formula (1c) can be
produced by reacting the compound of general formula
(lb) with a reactive derivative or an alkylating agent
in the presence or absence of a base.
The solvent used in this reaction is not
particularly limited, provided that it does not
adversely affect the reaction, and examples thereof
include amides such as N,N-dimethylformamide, N,N-
dimethylacetamide, and 1-methyl-2-pyrrolidone;
halogenated hydrocarbons such as methylene chloride,
chloroform, and dichloroethane; aromatic hydrocarbons
such as benzene, toluene, and xylene; ethers such as
dioxane, tetrahydrofuran, anisole, diethylene glycol
dimethyl ether, diethylene glycol diethyl ether, and
ethylene glycol monomethyl ether; nitriles such as
acetonitrile; sulfoxides such as dimethylsulfoxide;
ketones such as acetone and 2-butanone; esters such as
ethyl acetate; carboxylic acids such as acetic acid;
heteroaromatics such as pyridine; and water. These may
be used in the form of a mixture.
Examples of the reactive derivative include
acid anhydrides such as acetylformyloxide, acetic
anhydride, trichloroacetic anhydride, and
trifluoroacetic anhydride; mixed acid anhydrides of

organic carboxylic acids such as acetic acid with
carbonic acid monoalkyl esters such as ethyl
chlorocarbonate and isobutyl chlorocarbonate; mixed
acid anhydrides of organic carboxylic acids such as
acetic acid with organic acids such as pivalic acid;
acid chlorides such as acetyl chloride, trichloroacetyl
chloride, and trifluoroacetyl chloride; acid bromides
such as acetyl bromide; and active esters such as p-
nitrophenyl ester, N-hydroxysuccinimide ester, and N-
hydroxyphthalimide ester. These reactive derivatives
may be used without isolation.
The reactive derivative may be generated in
the system using a coupling agent. Examples of the
coupling agent include carbodiimides such as N,N'-
dicyclohexylcarbodiimide and N-ethyl-N'-(3-
dimethylaminopropyl)carbodiimide; carbonyls such as
carbonyldiimidazole; acid azides such as
diphenylphosphoryl azide; acid cyanides such as
diethylphosphoryl cyanide; 2-ethoxy-l-ethoxycarbonyl-
1,2-dihydroquinoline; O-benzotriazol-1-yl-1,1,3,3-
tetramethyluronium=hexafluorophosphate; and O-(7-
azabenzotriazol-1-yl)-1,1,3,3-
tetramethyluronium=hexafluorophosphate.
Examples of the alkylating agent include
halogenated alkyls such as methyl iodide and ethyl
iodide; halogenated aralkyls such as benzyl chloride
and benzyl bromide; and sulfates such as dimethyl
sulfate.

Examples of the base used in the reaction, if
desired, include metal alkoxides such as sodium
methoxide, sodium ethoxide, potassium tert-butoxide,
and sodium tert-butoxide; inorganic salts such as
sodium hydroxide, potassium hydroxide, sodium
bicarbonate, sodium carbonate, potassium carbonate,
sodium hydride, and potassium hydride; and organic
bases such as triethylamine and pyridine.
The usage amounts of the reactive derivative,
alkylating agent and base may be each 2- to 100-fold
moles, preferably 2- to 10-fold moles based on the
compound of general formula (1b).
The reaction may be conducted at -20 to 100°C,
preferably 0 to 50°C for one minute to 24 hours.
(2-3)
The compound of general formula (1a) can be
produced by subjecting the compound of general formula
(1b) to reduction reaction. In addition, the compound
of general formula (1a) can be produced by subjecting
the compound of general formula (1c) to reduction
reaction.
Examples of the reduction reaction used here
include catalytic hydrogenation reaction employing a
metal catalyst and reduction employing a metal and acid
such as zinc-acetic acid.
When the compound of general formula (1b) or
the compound of general formula (1c) is subjected to
catalytic hydrogenation reaction, the solvent used is

not particularly limited, provided that it does not
adversely affect the reaction, and examples thereof
include alcohols such as methanol, ethanol, 2-propanol,
and 2-methyl-2-propanol; amides such as N,N-
dimethylformamide, N,N-dimethylacetamide, and 1-methyl-
2-pyrrolidone; halogenated hydrocarbons such as
methylene chloride, chloroform, and dichloroethane;
aromatic hydrocarbons such as benzene, toluene, and
xylene; ethers such as dioxane, tetrahydrofuran,
anisole, diethylene glycol dimethyl ether, diethylene
glycol diethyl ether, and ethylene glycol monomethyl
ether; nitriles such as acetonitrile; ketones such as
acetone and 2-butanone; esters such as ethyl acetate;
carboxylic acids such as acetic acid; heteroaromatics
such as pyridine; and water. These may be used in the
form of a mixture.
Examples of the metal catalyst include
palladium catalysts such as palladium-carbon, palladium
oxide, palladium hydroxide, and palladium black, nickel
catalysts such as Raney nickel, and platinum oxide, and
the usage amount thereof may be 0.001- to 1-fold (w/w),
preferably 0.01- to 0.5-fold (w/w) that of the compound
of general formula (1b) or the compound of general
formula (1c).
Examples of reducing agents other than
hydrogen include formic acid; formates such as sodium
formate, ammonium formate, and triethylammonium
formate; cyclohexene; and cyclohexadiene, and the usage

amount thereof may be 2- to 100-fold moles, preferably
2- to 10-fold moles based on the compound of general
formula (1b) or the compound of general formula (1c).
When the compound of general formula (1b) is
subjected to catalytic hydrogenation reaction, the
hydrogen pressure may be normal pressure to 30
atmospheric pressures, preferably 2 to 10 atmospheric
pressures.
When the compound of general formula (lc) is
subjected to catalytic hydrogenation reaction, the
hydrogen pressure may be normal pressure.
The reaction may be conducted at 0 to 200°C,
preferably 0 to 100°C for one minute to 24 hours.

wherein R6 represents an optionally substituted lower
alkyl or aralkyl group; and R2, R3, and R4 have the same
meanings as described above.

The compound of general formula (1d) can be
produced from the compound of general formula (3).
Then, the compound of general formula (1d) can be
reduced to produce the compound of general formula
(la) .
This series of reactions will be now
described in detail.
(3-1)
The compound of general formula (1d) can be
produced by reacting the compound of general formula
(3) with the compound of general formula (5) or a salt
thereof.
Examples of the compound of general formula
(5) include O-methylhydroxylamine and O-
benzylhydroxylamine.
Examples of the salt of the compound of
general formula (5) include hydrochlorides and
sulfates.
This reaction may be conducted according to
production method (1-2).
(3-2)
The compound of general formula (1a) can be
produced by reducing the compound of general formula
(1d). This reaction may be conducted according to
production method (2-3).


wherein R7 represents an optionally substituted lower
alkyl, aralkyl, aryl, alkoxy, cycloalkyloxy or
aralkyloxy group; R2 and R3 have the same meanings as
described above.
The compound of general formula (1e) can be
produced by reacting the compound of general formula
(la) with a reactive derivative in the presence or
absence of a base.
This reaction may be conducted according to
production method (2-2).


wherein R8 represents an amidino-protecting group; R9
represents a hydrogen atom or an optionally substituted
lower alkyl or aralkyl group; L1 represents a leaving
group; and R2 and R3 have the same meanings as described
above.
The compound of general formula (7) can be
produced from the compound of general formula (6). The
compound of general formula (7) can be then reacted
with the compound of general formula (8) to produce the
compound of general formula (1f).
This series of reactions will be now
described in detail.
(5-1)
The compound of general formula (7) can be
produced by converting the hydroxyl group of the
compound of general formula (6) to a leaving group.
When the leaving group is an
alkanesulfonyloxy group or an arylsulfonyloxy group,
the compound of general formula (6) may be reacted in
the presence or absence of a base, for example, with an
alkanesulfonyl chloride such as methanesulfonyl
chloride or an arylsulfonyl chloride such as p-
toluenesulfonic acid chloride.
Examples of the base used in this reaction,
if desired, include metal alkoxides such as sodium
methoxide, sodium ethoxide, potassium tert-butoxide,
and sodium tert-butoxide; inorganic bases such as
sodium hydroxide, potassium hydroxide, sodium

bicarbonate, sodium carbonate, potassium carbonate,
sodium hydride, and potassium hydride; and organic
bases such as triethylamine, N,N-diisopropylethylamine,
and pyridine.
The usage amount of the alkanesulfonyl
chloride or arylsulfonyl chloride may be 1- to 10-fold
moles, preferably 1- to 3-fold moles based on the
compound of general formula (6).
When the leaving group is a halogen atom, the
compound of general formula (6) may be reacted, for
example, with thionyl chloride, thionyl bromide, boron
tribromide, or carbon tetrabromide-triphenylphosphine.
The usage amounts of these reagents may be
each 1- to 10-fold moles, preferably 1- to 3-fold moles
based on the compound of general formula (6).
The solvent used in this reaction is not
particularly limited, provided that it does not
adversely affect the reaction, and examples thereof
include amides such as N,N-dimethylformamide, N,N-
dimethylacetamide, and l-methyl-2-pyrrolidone;
halogenated hydrocarbons such as methylene chloride,
chloroform, and dichloroethane; aromatic hydrocarbons
such as benzene, toluene, and xylene; ethers such as
dioxane, tetrahydrofuran, anisole, diethylene glycol
dimethyl ether, diethylene glycol diethyl ether, and
ethylene glycol monomethyl ether; nitriles such as
acetonitrile; sulfoxides such as dimethylsulfoxide; and
heteroaromatics such as pyridine. These may be used in

the form of a mixture.
(5-2)
The compound of general formula (If) can be
produced by reacting the compound of general formula
(7) with the compound of general formula (8) in the
presence or absence of a base.
The solvent used in this reaction is not
particularly limited, provided that it does not
adversely affect the reaction, and examples thereof
include alcohols such as methanol, ethanol, 2-propanol,
and 2-methyl-2-propanol; amides such as N,N-
dimethylformamide, N,N-dimethylacetamide, and 1-methyl-
2-pyrrolidone; halogenated hydrocarbons such as
methylene chloride, chloroform, and dichloroethane;
aromatic hydrocarbons such as benzene, toluene, and
xylene; ethers such as dioxane, tetrahydrofuran,
anisole, diethylene glycol dimethyl ether, diethylene
glycol diethyl ether, and ethylene glycol monomethyl
ether; nitriles such as acetonitrile; sulfoxides such
as dimethylsulfoxide; ketones such as acetone and 2-
butanone; esters such as ethyl acetate; heteroaromatics
such as pyridine; and water. These may be used in the
form of a mixture.
Examples of the base used in this reaction,
if desired, include metal alkoxides such as sodium
methoxide, sodium ethoxide, potassium tert-butoxide,
and sodium tert-butoxide; inorganic bases such as
sodium hydroxide, potassium hydroxide, sodium

bicarbonate, sodium carbonate, potassium carbonate,
sodium hydride, and potassium hydride; and organic
bases such as triethylamine, N,N-diisopropylethylamine,
and pyridine.
The usage amount of the base may be 1- to 10-
fold moles, preferably 1- to 3-fold moles based on the
compound of general formula (7).
The usage amount of the compound of general
formula (8) used in this reaction may be 1- to 20-fold
moles, preferably 1- to 5-fold moles based on the
compound of general formula (7).
The reaction may be conducted at 0 to 200°C,
preferably 0 to 150°C for one minute to 24 hours.
The removal of the amidino-protecting group
represented by R8 may be carried out by a method
described, for example, in Protective Groups in Organic
Synthesis, 3: 494-653, 1999, or by a method equivalent
thereto.


wherein R10 represents an amidino-protecting group; R11
represents a hydrogen atom or an optionally substituted
lower alkyl or aralkyl group; L2 represents a leaving
group; and R2 and R3 have the same meanings as described
above.
The compound of general formula (10) can be
produced from the compound of general formula (9). The
compound of general formula (10) can be then reacted
with the compound of general formula (11) to produce
the compound of general formula (lg).
This series of reactions will be now
described in detail.
(6-1)
The compound of general formula (10) can be
produced by converting the hydroxyl group of the
compound of general formula (9) to a leaving group.
This reaction may be conducted according to production
method (5-1).
(6-2)
The compound of general formula (lg) can be
produced by reacting the compound of general formula
(10) with the compound of general formula (11) in the
presence or absence of a base. This reaction may be
conducted according to production method (5-2).
The removal of the amidino-protecting group
represented by R10 may be carried out by a method
described, for example, in Protective Groups in Organic
Synthesis, 3: 494-653, 1999, or by a method equivalent

thereto.
Each of the compounds in the above-described
production methods 1 to 6 may be also used in the form
of a salt, and examples of the salt include the same
salts as those described for the compound of general
formula (1) .
Each of the production intermediates obtained
in the above-described production methods (1) to (6)
may be also used without isolation for the subsequent
reaction.
The compounds of general formulas (1a), (1b),
(1c), (1d), (1e), (1f), and (1g) thus obtained or salts
thereof can be derivatized into other compounds of
general formula (1) or salts thereof, for example, by
subjecting to a reaction known per se such as
condensation, addition, oxidation, reduction,
rearrangement, substitution, halogenation, dehydration,
or hydrolysis, or by properly combining these
reactions.
When isomers (e.g., optical isomer,
geometrical isomer, and tautomer) are present in the
compounds in the above-described production methods,
these isomers may be also used, and solvates, hydrates,
and crystals in various forms may be also employed.
In the following, description will be given
of methods for producing compounds of general formulas
(2), (6), (8), (9), and (11) as raw materials in the
production of the compounds of the invention. The

compounds of general formulas (2), (6), (8), (9), and
(11) are produced by combining methods known per se,
and, for example, can be prepared by the following
production methods.

wherein R12 represents an amino-protecting group; L3
represents a leaving group; and R2, R3, and L1 have the
same meanings as described above.
Examples of the compound of general formula
(12) include benzyl=4-(3-bromopropyl)piperidine-1-
carboxylate (J. Med. Chem., 46: 2606-2620, 2003), tert-
butyl=4-(3-bromopropyl)-1-piperidinecarboxylate
(Tetrahedron, 55: 11619-11639, 1999), and 3-[N-[(tert-
butoxy)carbonyl]piperidin-4-yl]propyl iodide (J. Med.
Chem., 37: 2537-2551, 1994). In addition, the
synthesis can be carried out by combining known methods

using tert-butyl=4-(3-hydroxypropyl)-1-
piperidinecarboxylate or the like as a raw material.
Examples of the compound of general formula
(14) include 4-cyanophenol and 4-cyano-3-fluorophenol.
(A-1)
The compound of general formula (13) can be
produced by reacting the compound of general formula
(12) with the compound of general formula (14) in the
presence or absence of a base, followed by
deprotection.
The solvent used in this reaction is not
particularly limited, provided that it does not
adversely affect the reaction, and examples thereof
include alcohols such as methanol, ethanol, 2-propanol,
and 2-methyl-2-propanol; amides such as N,N-
dimethylformamide, N,N-dimethylacetamide, and 1-methyl-
2-pyrrolidone; halogenated hydrocarbons such as
methylene chloride, chloroform, and dichloroethane;
aromatic hydrocarbons such as benzene, toluene, and
xylene; ethers such as dioxane, tetrahydrofuran,
anisole, diethylene glycol dimethyl ether, diethylene
glycol diethyl ether, and ethylene glycol monomethyl
ether; nitriles such as acetonitrile; sulfoxides such
as dimethylsulfoxide; ketones such as acetone and 2-
butanone; esters such as ethyl acetate; heteroaromatics
such as pyridine; and water. These may be used in the
form of a mixture.
Examples of the base used in this reaction,

if desired, include metal alkoxides such as sodium
methoxide, sodium ethoxide, potassium tert-butoxide,
and sodium tert-butoxide; inorganic bases such as
sodium hydroxide, potassium hydroxide, sodium
bicarbonate, sodium carbonate, potassium carbonate,
sodium hydride, and potassium hydride; and organic
bases such as triethylamine, N,N-diisopropylethylamine,
and pyridine.
The usage amount of the base may be 1- to 10-
fold moles, preferably 1- to 3-fold moles based on the
compound of general formula (12).
The usage amount of the compound of general
formula (14) used in this reaction may be 1- to 20-fold
moles, preferably 1- to 5-fold moles based on the
compound of general formula (12).
The reaction may be conducted at 0 to 200°C,
preferably 0 to 150°C for one minute to 24 hours.
The removal of the amino-protecting group
represented by R12 may be carried out by a method
described, for example, in Protective Groups in Organic
Synthesis, 3: 494-653, 1999, or by a method equivalent
thereto.
(A-2)
The compound of general formula (2) can be
produced by reacting the compound of general formula
(13) with the compound of general formula (15). This
reaction may be conducted according to production
method (A-1).


wherein R13 represents a hydroxyl-protecting group; and
R2, R3, L1, and L3 have the same meanings as described
above.
The compound of general formula (17) can be
produced by combining known methods, using tert-
butyl=4-(3-hydroxypropyl)-1-piperidinecarboxylate or
the like as a raw material.
(B-1)
The compound of general formula (16) can be
produced by reacting the compound of general formula
(15) with the compound of general formula (17),
followed by deprotection. This reaction may be
conducted according to production method (A-1).

The removal of the hydroxyl-protecting group
represented by R13 may be carried out by a method
described, for example, in Protective Groups in Organic
Synthesis, 3: 17-245, 1999, or by a method equivalent
thereto.
(B-2)
The compound of general formula (18) can be
produced by converting the hydroxyl group of the
compound of general formula (16) to a leaving group.
This reaction may be conducted according to production
method (5-1).
(B-3)
The compound of general formula (2) can be
produced by reacting the compound of general formula
(18) with the compound of general formula (14). This
reaction may be conducted according to production
method (A-1).


wherein L4 represents a leaving group; and R2 and L3
have the same meanings as described above.
Examples of the compound of general formula
(19) include 4-cyanophenol. Examples of the compound
of general formula (21) include 3-bromo-1-propanol.
(C-1)
The compound of general formula (20) can be
produced by reacting the compound of general formula
(19) with the compound of general formula (21). This
reaction may be conducted according to production
method (A-1).
(C-2)
The compound of general formula (15) can be
produced by converting the hydroxyl group of the
compound of general formula (20) to a leaving group.
This reaction may be conducted according to production
method (5-1) .


wherein R2 and R8 have the same meanings as described
above.
(D-1)
The compound of general formula (22) can be
produced by reacting the compound of general formula
(16) with hydroxylamine or its salt in the presence or
absence of a base. This reaction may be conducted
according to production method (2-1).
(D-2)
The compound of general formula (23) can be
produced by subjecting the compound of general formula
(22) to reduction reaction. This reaction may be
conducted according to production method (2-3).
(D-3)
The compound of general formula (6) can be
produced by protecting the amidino group of the
compound of general formula (23). This reaction may be
carried out by a method described, for example, in
Protective Groups in Organic Synthesis, 3: 494-653,
1999, or by a method equivalent thereto.


wherein R3, R10, and L4 have the same meanings as
described above.
(E-1)
The compound of general formula (24) can be
produced by reacting the compound of general formula
(13) with the compound of general formula (21). This
reaction may be conducted according to production
method (A-l).
(E-2)
The compound of general formula (25) can be
produced by reacting the compound of general formula
(24) with hydroxylamine or its salt in the presence or

absence of a base. This reaction may be conducted
according to production method (2-1).
(E-3)
The compound of general formula (26) can be
produced by subjecting the compound of general formula
(25) to reduction reaction. This reaction may be
conducted according to production method (2-3).
(E-4)
The compound of general formula (9) can be
produced by protecting the amidino group of the
compound of general formula (26). This reaction may be
conducted according to production method (D-3).

wherein R3 and R9 have the same meanings as described
above.
The compound of general formula (8) can be
produced by reacting the compound of general formula
(14) with the compound of general formula (27) or a
salt thereof. This reaction may be conducted according
to production method (2-1).
(Production method G)


wherein R2 and R11 have the same meanings as described
above.
The compound of general formula (11) can be
produced by reacting the compound of general formula
(19) with the compound of general formula (28) or a
salt thereof. This reaction may be conducted according
to production method (2-1).
When used as a medicine, the compound of the
invention may generally be properly mixed with a
pharmaceutical auxiliaries such as an excipient, a
carrier, and a diluent, used for formulation, which can
be orally or parenterally administered in the form of
tablets, capsules, powders, syrups, granules, pills,
suspensions, emulsions, solutions, powder preparations,
suppositories, eye drops, nasal drops, ear drops,
patches, ointments, or injections, according to the
ordinary method. In addition, the administration
method, dosage, and administration frequency can be
properly selected depending on the age, body weight and
symptoms of a patient. Typically, 0.01 to 1,000 mg/kg
thereof may be administered orally or parenterally

(e.g., by injection, drip infusion, or rectal
administration) to an adult once or in several divided
portions in a day.
To establish the usefulness of the compound
of the invention, tests on the antifungal action,
repeated toxicity, Vero cell proliferation inhibiting
activity, stability, and antiprotozoan action thereof
were carried out.
Test Example 1: Antifungal action
The compound of Example 4 was selected as a
compound of the invention. The compound described in
the Example of WO03/074476, having a structure most
analogous to that of the compound of the invention, was
selected as a comparative compound. Their chemical
structures are as follows.

Susceptibility testing of fungi was carried
out using a microbroth dilution method. The medium
used in the susceptibility test was RPMI1640
(RPMI/MOPS) adjusted to pH 7.0 employing 0.165 mol/L
morpholinepropanesulfonic acid (MOPS) and 1.0 mol/L
sodium hydroxide. A test compound was dissolved in

sterile water, which was then subjected to two-fold
serial dilution using 100 (iL of RPMI/MOPS on a 96-well
round bottom plate. Candida albicans TIMM1623 cultured
overnight at 35°C on Sabouraud agar medium was suspended
in a sterilized physiological saline. The number of
cells was counted under a biological microscope; a
suspension of inoculum organism 2x103 cells/mL) was
prepared using RPMI/MOPS, and 100 µL thereof was then
dispensed into each well; and a microplate containing a
predetermined concentration of the test compound, the
medium, and fungal cells was finally prepared. The
plate was cultured at 35°C for 48 hours. After the end
of cultivation, absorbance was determined at 630 nm
using an automatic spectrophotometer. The lowest test
compound concentration at which 50% growth inhibition
was observed as compared to growth control where no
test compound was added was defined as IC50. The
results are shown in Table 1.

The compound of Example 4 had an equivalent
or higher antifungal activity than the comparative
compound.
Test Example 2: Repeated-dose toxicity test
in mice (1)

The compound of Example 4 was selected as a
compound of the invention. Among compounds described
in the Examples of WO03/074476, a compound having a
structure most analogous to that of the compound of the
invention was selected as a comparative compound.
Their chemical structures are as follows.

Six-week old male ICR mice (body weight
range: 30.6 to 34.8 g, the number of animals allocated
to each group: 5) were used to examine the repeated-
dose toxicity. A solution of each test compound was
prepared by dissolution in distilled water. Each
compound was subcutaneously administered at a dose of
6.25 mg/kg once a day for 7 days. A sterilized
physiological saline was administered to a control
group. At the end of the administration period, each
mouse was anesthetized with ether. An injection
syringe containing a heparin solution (Novo-Heparin
Injection 1000, Aventis Pharma K.K.) as an
anticoagulant was used for blood collection from
abdominal portion of the vena cava. A hematological
test was carried out for the following items. Values

in the test compound-treated groups when a value in the
control group is set to 100 are shown in Table 2.
(Hematological test items and determination
methods)
Red blood cell count (RBC): Two-angle laser
flow cytometry
Reticulocyte count: Flow cytometry after RNA
staining

The compound of Example 4 did not decrease
the reticulocyte count and therefore had a higher
1 safety than the comparative compound.
Test Example 3: Repeated-dose toxicity test
in mice (2)
The compound of Example 3 was selected as a
compound of the invention. Among compounds described
in the Examples of WO03/074476, a compound having a
structure most analogous to that of the compound of the
invention was selected as a comparative compound.
Their chemical structures are as follows.


Six-week old male ICR mice (body weight
range: 27.4 to 33.7 g, the number of animals allocated
to each group: 5) were used to examine the repeated-
dose toxicity. A solution of each test compound was
prepared by dissolution in 0.1 mol/L hydrochloric acid.
Each compound was subcutaneously administered at a dose
of 6.25 or 3.13 mg/kg once a day for 14 days. A
sterilized physiological saline was administered to a
control group. At the end of the administration
period, each mouse was anesthetized with ether. An
injection syringe containing a heparin solution (Novo-
Heparin Injection 1000, Aventis Pharma K.K.) as an
anticoagulant was used for blood collection from
abdominal portion of vena cava. A hematological test
was carried out for the following items. Values in the
test compound-treated groups when a value in the
control group is set to 100 are shown in Table 3.
(Hematological test items and determination
methods)
Red blood cell count (RBC): Two-angle laser
flow cytometry
Reticulocyte count: Flow cytometry after RNA


The comparative compound decreased the
reticulocyte count at a dose of 3.13 mg/kg. On the
other hand, the compound of Example 3 did not decrease
the reticulocyte count even at a dose of 6.25 mg/kg,
and therefore had a much higher safety than the
comparative compound.
Test Example 4: Vero cell proliferation
inhibition test
The compound of Example 4 was selected as a
compound of the invention. Among compounds described
in the Examples of WO03/074476, a compound having a
structure most analogous to that of the compound of the
invention was selected as a comparative compound.
Their chemical structures are as follows.


The cytotoxicity of each compound was
evaluated using Vero cells. Each test compound was
dissolved in dimethylsulfoxide (DMSO), which was then
subjected to serial dilution with 10% FBS-containing
E'MEM and added to a 96-well plate. The cells were
suspended in 10% FBS-containing E'MEM, inoculated in an
amount of 3,000 cells/well (96-well plate), and
cultured in a CO2 incubator at 37°C for 3 days. The
degree of growth of Vero cells was evaluated using a
2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-5-
[(phenylamino)carbonyl]-2H-tetrazolium=inner
salt=monosodium salt (XTT) assay. Thus, an XTT
solution containing 1 mg/mL XTT and 25 µmol/L
phenazine=methosulfate (PMS) was added to each well,
which was then incubated in a CO2 incubator for 2 hours,
followed by determining the absorbance of the well at
450 nm (reference: 655 nm) using a micro plate reader.
The absorbance ratio between the control (no added
compound) and each of the wells was calculated, and the
concentration (CC50; µg/mL) of compound inhibiting cell
proliferation by 50% was computed. The results are
shown in Table 4.

The compound of Example 4 had a higher safety
than the comparative compound.

Test Example 5: Hygroscopicity test (1)
The compound of Example 4 was selected as a
compound of the invention. Among compounds described
in the Examples of WO03/074476, a compound having a
structure most analogous to that of the compound of the
invention was selected as a comparative compound.
Their chemical structures are as follows.

The compound of Example 4 and the comparative
compound were stored under conditions of room
temperature and a relative humidity of 75% or 60°C and a
relative humidity of 75% for one week. As a result,
the compound of Example 4 was powder without any change
in the appearance under any of the conditions. On the
other hand, the comparative compound was changed into a
paste form under both of the conditions.
The compound of Example 4 had a higher
stability than the comparative compound.
Test Example 6: Hygroscopicity test (2)
The compound of Example 10 was selected as a
compound of the invention. Among compounds described
in the Examples of WO03/074476, a compound having a
structure most analogous to that of the compound of the

invention was selected as a comparative compound.
Their chemical structures are as follows.

The compound of Example 10 and the
comparative compound were stored under conditions of
room temperature and a relative humidity of 100% for
one week, and weighed. The results are shown in Table
5.

The compound of Example 10 was not
hygroscopic at all, and had a higher stability than the
comparative compound.
Test Example 7: Hygroscopicity test (3)
The compound of Example 10 was selected as a
compound of the invention. Among compounds described
in the Examples of WO03/074476, a compound having a
structure most analogous to that of the compound of the
invention was selected as a comparative compound.
Their chemical structures are as follows.


The compound of Example 10 and the
comparative compound were stored under conditions of
60°C and a relative humidity of 100% for one week. As a
result, the comparative compound deliquesced. On the
other hand, the compound of Example 10 did not
deliquesce and was stable.
Test Example 8: Antiprotozoan action
The antiprotozoan activity of the compound of
Example 3 was determined.
Trichomonas vaginalis CDC337 was cultured in
a Diamond's trypticase-yeast-maltose medium (pH 6.8)
containing 8% fetal bovine serum (FBS). The protozoan
bodies which had been cultured 37°C for 2 days were
centrifuged (1,500 rpm, 10 minutes) and subjected to
medium replacement with a fresh medium before
adjustment to 2x104 bodies/mL, followed by dispensing
them in an amount of 100 µL/well in a micro plate (96-
well, flat bottom). The test compound was dissolved
using 0.1 mol/L hydrochloric acid, which was then
diluted to a predetermined concentration with the
medium, followed by dispensing 100 µL/well thereof into
a micro plate. The minimal test compound concentration

at which the movement of the bodies was not observed
after culture 37°C under anaerobic conditions for 2 days
was defined as MIC.
The MIC of the compound of Example 10 was 16
µg/mL.
Test Example 9: Mouse Candida infection model
test (oral administration)
The compound of Example 47 was selected as a
compound of the invention. The chemical structure
thereof is as follows.

Candida albicans TIMM1623 on a SDA plate,
which had been cultured overnight at 35°C was suspended
in a sterilized physiological saline, which was then
diluted to prepare a suspension of inoculum organism.
To make mice in a transiently compromised state,
cyclophosphamide was intraperitoneally administered at
200 mg/kg 4 days before infection and at 100 mg/kg the
next day after infection. Into the tail vein of mice
was inoculated 0.2 mL of the prepared solution of
inoculum Candida albicans TIMM1623 to induce infection
(about 3xl04 CFU/mouse) . The test compound was
suspended in 0.5% methylcellulose and orally
administered at 1 mg/kg of mouse body weight. The
treatment was started from 2 hours after infection and

carried out for 7 days. The survival number of mice
was observed for 21 days after infection, and recorded.
As a result, all mice died in a group to
w
thich no test compound was administered, but 80% of
ice survived in the group to which the compound of
Example 47 was administered.
The compound of Example 47 also had an
excellent therapeutic effect even when orally
administered.
In the in vitro and in vivo tests, the
compounds of the invention had equivalent or more
excellent antifungal activities than the comparative
compound. In the repeated-dose toxicity tests, the
compounds of the invention did not decrease the
reticulocyte count and had a higher safety than the
comparative compound. In addition, the comparative
compound was difficult in quality control because it
was hygroscopic and deliquescent, but the compounds of
the invention were not hygroscopic, and more excellent
as drug substance than the comparative compound.
Further, the compounds of the invention had an
excellent effect against protozoa.
Examples
The present invention will be now described
in the following Reference Examples and Examples.
However, the invention is not intended to be limited
thereto.
Mixing ratios in eluents are all volume

mixing ratios, and the carrier used in column
chromatography is B.W. Silica Gel or BW-127ZH (Fuji
Silysia Chemical Ltd.) unless otherwise described.
The abbreviations in Examples mean the
following.
Ac: acetyl, Boc: tert-butoxycarbonyl, tBu:
tert-butyl, Et: ethyl, Me: methyl, Ms: methanesulfonyl
DMSO-d6: dimethylsulfoxide-d6
Reference Example 1

To a tetrahydrofuran (110 mL) solution of
10.7 g of tert-butyl=4-(3-hydroxypropyl)-1-
piperidinecarboxylate was added 19.0 g of carbon
tetrabromide under cooling with water, to which 15.0 g
of triphenylphosphine was then added over a period of
13 minutes. This mixture was stirred at room
temperature for 2 hours and 30 minutes and allowed to
stand for 13 hours. To the reaction mixture were added
water, ethyl acetate, and a saturated sodium chloride
aqueous solution. The organic layer was separated,
washed with a saturated sodium chloride aqueous
solution, and dried with anhydrous magnesium sulfate,
followed by distilling off the solvent under reduced
pressure. The resultant residue was purified using
silica gel column chromatography (eluent; hexane:ethyl
acetate = 3:1) to provide 13.2 g of tert-butyl=4-(3-
bromopropyl)-1-piperidinecarboxylate as colorless oily

form.
1H-NMR(CDCl3)δ value: 1. 00-1.20(2H,m),1.20-
1.50(3H,m),1.45(9H,s),1.60-1.70(2H,m),1.80-
1.95(2H,m),2.60-2.75(2H,m),3.40(2H,t,J=6.8Hz),3.90-
4.25(2H,m).
Reference Example 2

To a dimethylsulfoxide (130 mL) solution of
13.2 g of tert-butyl=4-(3-bromopropyl)-1-
piperidinecarboxylate were added 5.13 g of 4-
cyanophenol and 11.9 g of potassium carbonate at room
temperature, which was then stirred at the same
temperature for 2 6 hours. The reaction mixture was
added to a mixture of toluene and water. The organic
layer was separated, washed with a saturated sodium
chloride aqueous solution, and dried with anhydrous
magnesium sulfate, followed by distilling off the
solvent under reduced pressure to provide 14.5 g of
tert-butyl=4-[3-(4-cyanophenoxy)propyl]-1-
piperidinecarboxylate as white solid form.
1H-NMR(CDCl3)δ value: 1.05-1.20(2H,m),1.40-
1.50(3H,m),1.46(9H,s),1.65-1.75(2H,m),1.75-
1.90(2H,m),2.60-2.80(2H,m),3.99(2H,t,J=6.3Hz),4.00-
4.20(2H,m),6.93(2H,d,J=8.7Hz),7.58(2H,d,J=8.7Hz) .
Reference Example 3


To a chloroform (100 mL) solution of 14.0 g
of tert-butyl=4-[3-(4-cyanophenoxy)propyl]-1-
piperidinecarboxylate was dropwise added 40 mL of
trifluoroacetic acid under cooling with water over a
period of 10 minutes. This mixture was stirred at the
same temperature for 20 minutes, and then stirred at
room temperature for 35 minutes. After distilling off
the solvent under reduced pressure, chloroform and
water were added. A sodium hydroxide aqueous solution
was added thereto for adjustment to pH 13.0. The
organic layer was separated, and the aqueous layer was
extracted with chloroform. The organic layer and the
extract were combined, which was then washed with a
sodium hydroxide aqueous solution and dried with
potassium carbonate, followed by distilling off the
solvent under reduced pressure to provide 10.3 g of 4-
[3-(4-piperidinyl)propoxy]benzonitrile as pale yellow
solid form.
1H-NMR(CDCl3).δ value: 1.05-1.20(2H,m),1.35-
1.45(3H,m),1.65-1.90(4H,m),2.50-2.65(2H,m),3.00-
3.15(2H,m),3.99(2H,t, J=6.6Hz),4.78(lH,s),6.93(2H,d,J=9.
0Hz),7.58(2H,d,J=9.0Hz).
Reference Example 4

To an N,N-dimethylformamide (150 mL) solution
of 10.2 g of 4-[3-(4-piperidinyl)propoxy]benzonitrile
were sequentially added 11.2 g of potassium carbonate

and 9.72 g of 4-(3-bromopropoxy)benzonitrile at room
temperature, which was then stirred at the same
temperature for 18 hours. Toluene and water were added
to the reaction mixture. The precipitate was collected
by filtration to provide 13.7 g of 4-(3-{4-[3-(4-
cyanophenoxy)propyl]-1-piperidinyl}propoxy)benzonitrile
as white solid form.
1H-NMR(CDCl3).δ value: 1.20-1.45(5H,m),1.65-
2.05(8H,m),2.40-2.55(2H,m),2.85-
3.00(2H,m),3.99(2H,t,J=6.5Hz),4.06(2H,t,J=6.3Hz),6.93(2
H,d,J=8.8Hz),6.94(2H,d,J=8.8Hz),7.57(2H,d,J=8.8Hz),7.57
(2H,d,J=8.8Hz).
Reference Example 5

A 2-butanone (7.6 niL) solution of 1.12 g of
tert-butyl=4-(3-bromopropyl)-1-piperidinecarboxylate
was added to a 2-butanone (7.0 mL) mixture of 0.50 g of
2-fluoro-4-hydroxybenzonitrile and 0.56 g of potassium
carbonate, which was then heated to reflux for 6 hours
and 30 minutes. After cooling down to room
temperature, the reaction mixture was added to a
mixture of ethyl acetate and water. The organic layer
was separated, washed with water, and dried with
anhydrous magnesium sulfate, followed by distilling off
the solvent under reduced pressure. The resultant
residue was purified using silica gel column
chromatography (eluent; hexane:ethyl acetate =4:1) to

provide 0.72 g of tert-butyl=4-[3-(4-cyano-3-
fluorophenoxy)propyl]-1-piperidinecarboxylate as
colorless oily form.
1H-NMR(CDCl3).δ value: 1.05-1.20(2H,m),1.35-
1.45(3H,m),1.46(9H,s),1.65-1.75(2H,m),1.75-
1.90(2H,m),2.60-2.75(2H,m),3.99(2H,t,J=6.3Hz),4.00-
4.20(2H,m),6.65-6.80(2H,m),7.4 5-7.54(lH,m).

To a methylene chloride (5.5 mL) solution of
0.66 g of tert-butyl=4-[3-(4-cyano-3-
fluorophenoxy)propyl]-1-piperidinecarboxylate was
dropwise added 1.8 mL of trifluoroacetic acid under
cooling with ice over a period of 2 minutes, which was
then stirred at room temperature for 6 hours. The
solvent was distilled off under reduced pressure, and
chloroform and a 1.0 mol/L sodium hydroxide aqueous
solution were added to the resultant residue. The
organic layer was separated and dried with anhydrous
magnesium sulfate, followed by distilling off the
solvent under reduced pressure. The resultant residue
was purified using silica gel column chromatography
(eluent; chloroform:methanol =4:1) to provide 0.28 g
of 2-fluoro-4-[3-(4-piperidinyl)propoxy]benzonitrile as
pale yellow oily form.
1H-NMR(CDCl3).δ value: 1.05-1.20(2H,m),1.30-
1.45(3H,m),1.50-1.75(2H,m),1.75-1.90(2H,m),2.50-

2.65(2H,m),3.00-
3.15(2H,m),3.98(2H,t,J=6.5Hz) , 6.69(1H,dd,J=ll.0,2.3Hz),
6.75(lH,dd,J=8.5,2.3Hz),7.50(1H,dd,J=8.5,8.5Hz).

To an N,N-dimethylformamide solution (2.0 mL)
of 0.10 g of 2-fluoro-4-[3-(4-
piperidinyl)propoxy]benzonitrile were sequentially
added 0.10 g of potassium carbonate and 0.13 g of 4-(3-
bromopropoxy)benzonitrile at room temperature, which
was then stirred at the same temperature for 13 hours.
Ethyl acetate, water, and toluene were added to the
reaction mixture. The organic layer was separated and
dried with anhydrous sodium sulfate, followed by
distilling off the solvent under reduced pressure. The
resultant residue was purified using silica gel column
chromatography (eluent; chloroform:methanol =4:1) to
provide 68 mg of 4-(3-{1-[3-(4-cyanophenoxy)propyl]-4-
piperidinyl}propoxy)-2-fluorobenzonitrile as white
solid form.
1H-NMR(CDCl3).δ value: 1.20-1.45(5H,m),1.65-
2.05(8H,m),2.40-2.55(2H,m),2.85-
3.00(2H,m),3.98(2H,t, J=6.5Hz) , 4.06(2H, t, J=6.3Hz),6.69(1
H,dd,J=11.0,2.4Hz),6.74(1H, dd, J=8.8, 2.4Hz),6.94(2H,d,J=
8.7Hz),7.4 5-7.55(lH,m),7.57(2H,d,J=8.7Hz).
Reference Example 8


As described in Reference Example 7, 0.12 g
of 4-[3-(4-piperidinyl)propoxy]benzonitrile and 0.15 g
of 4-(3-bromopropoxy)-2-fluorobenzonitrile were used to
provide 0.10 g of 4-(3-{4-[3-(4-cyanophenoxy)propyl]-1-
piperidinyl}propoxy)-2-fluorobenzonitrile as white
solid form.
1H-NMR(CDCl3)δ value: 1.20-1.35(3H,m),1.35-
1.4 5(2H,m),1.60-2.05(8H,m),2.4 0-2.50(2H,m),2.85-
3.00(2H,m),3.99(2H,t,J=6.5Hz),4.06(2H,t,J=6.3Hz),6.70-
6.80(2H,m),6.93(2H,d,J=9.0Hz),7.4 5-
7.55(lH,m),7.57(2H,d,J=9.0Hz).

To a dimethylsulfoxide (4.0 mL) solution of
0.26 g of 2-fluoro-4-[3-(4-
piperidinyl)propoxy]benzonitrile and 0.21 g of 4-(3-
chloropropoxy)2-fluorobenzonitrile was added 0.88 mL of
N-ethyldiisopropylamine, which was then stirred at 80
to 90°C for 8 hours and 15 minutes. The reaction
mixture was cooled down to room temperature, to which
water was then added, followed by extraction with ethyl
acetate. The extract was washed twice with water and
dried with anhydrous magnesium sulfate, followed by

distilling off the solvent under reduced pressure. The
resultant residue was purified using silica gel column
chromatography (eluent; chloroform:methanol = 10:1) to
provide 0.25 g of 4-(3-{1-[3-(4-cyano-3-
fluorophenoxy)propyl]-4-piperidinyl}propoxy)-2-
fluorobenzonitrile as brown solid form.
1H-NMR(CDC13) .5 value: 1.20-1.45(5H,m),1.65-
2.05(8H,m),2.4 0-2.50(2H,m),2.85-
3.00(2H,m),3.98(2H,t,J=6.5Hz),4.06(2H,t,J=6.3Hz),6.65-
6.80(4H,m),7.45-7.55(2H,m).

Hydrogen chloride was introduced into an
ethanol (16 mL) suspension of 0.80 g of 4-(3-{4-[3-(4-
cyanophenoxy)propyl]-1-piperidinyl}propoxy)benzonitrile
under cooling with ice, which was then stirred at room
temperature for 15 hours. The solvent was distilled
off under reduced pressure, and the resultant residue
was dissolved in chloroform and added to a mixture of a
saturated sodium bicarbonate aqueous solution and
chloroform. The organic layer was separated and dried
with anhydrous magnesium sulfate, followed by
distilling off the solvent under reduced pressure to
provide 0.77 g of ethyl=4-{3-[4-(3-{4-
[ethoxy(imino)methyl]phenoxy}propyl)-1-

piperidinyl]propoxy}benzimidate as white solid form.
1H-NMR(CDCl3).δ value: 1.20-
1.45(5H,m),1.41(3H,t,J=7.1Hz),1.41(3H,t,J=7.1Hz),1.65-
2.05(8H,m),2.45-2.55(2H,m),2.90-
3.00(2H,m),3.98(2H,t,J=6.5Hz),4.04(2H,t,J=6.3Hz),4.20-
4.40(4H,m),6.89(2H,d,J=8.5Hz),6.90(2H,d,J=8.8Hz),7.60-
7.80(4H,m).

A dichloromethane (21 mL) solution of 3.9 mL
of 2,2,2-trifluoroethanol and 4.3 mL of pyridine was
added dropwise to a dichloromethane (83 mL) solution of
9.0 mL of trifluoromethanesulfonic anhydride under
cooling with ice over a period of 25 minutes. Thereto
was dropwise added a dichloromethane (60 mL) solution
of 8.50 g of N-hydroxyphthalimide and 18.5 mL of N,N-
diisopropylethylamine at the same temperature over a
period of 45 minutes, which was then stirred for 22
hours. To the reaction mixture was added 100 mL of 1
mol/L hydrochloric acid. The organic layer was
separated, washed sequentially with 1 mol/L
hydrochloric acid and a saturated sodium chloride
aqueous solution, and then dried with anhydrous
magnesium sulfate, followed by distilling off the
solvent under reduced pressure. The resultant residue
was purified using silica gel column chromatography

(eluent; hexanerethyl acetate = 3:1) and washed with
hexane to provide 4.43 g of N-(2,2,2-
trifluoroethoxy)phthalimide as white solid form.
1H-NMR(CDCl3).δ value: 4.56(2H,q,J=8.0Hz),7.75-
7.95(4H,m).

To a dichloromethane (45 mL)/methanol (5.9
mL) mixture of 4.30 g of N-(2,2,2-
trifluoroethoxy)phthalimide was dropwise added 0.85 mL
of hydrazine monohydrate at room temperature, which was
then stirred at the same temperature for 3 hours. To
the reaction mixture was added 10 mL of 2.9 mol/L
hydrogen chloride/ethanol, followed by distilling off
the solvent under reduced pressure to provide 2.65 g of
0-(2,2,2-trifluoroethyl)hydroxylamine hydrochloride as
white solid form.
1H-NMR(CDCl3).δ value: 4.60(2H,q,J=9.0Hz),7.80-
7.85(lH,m).

A tetrahydrofuran (10 mL) solution of 1.00 g
of 4-nitrophenyl=chloroformate was added dropwise to a
tetrahydrofuran (5 mL) solution of 0.44 g of pentanol
and 0.76 mL of triethylamine under cooling with ice.

The mixture was stirred at room temperature for 2 hours
and 20 minutes, and ethyl acetate and water were then
added to the reaction mixture. The organic layer was
separated, washed sequentially with a 5% potassium
carbonate aqueous solution and a saturated sodium
chloride aqueous solution, and then dried with
anhydrous magnesium sulfate, followed by distilling off
the solvent under reduced pressure to provide 1.20 g of
4-nitrophenyl=pentyl=carbonate as colorless oily form.
1H-NMR(CDCl3).δ value: 0 . 94 (3H, t, J=7 . 1Hz) , 1. 30-
1.50(4H,m),1.70-
1.85(2H,m),4.29(2H,t, J=6.7Hz),7.39(2H,d,J=9.3Hz),8.28(2
H,d,J=9.3Hz).

As described in Reference Example 13, 1.49 g
of cyclohexyl=4-nitrophenyl=carbonate as white solid
was obtained from 0.50 g of cyclohexanol and 1.00 g of
4-nitrophenyl=chloroformate.
1H-NMR(CDCl3).δ value: 1.20-1.65(6H,m),1.75-
1.85(2H,m),1.95-2.05(2H,m),4.70-
4.80(lH,m),7.39(2H,d, J=9.0Hz) ,8.28(2H, d, J=9.0Hz) .

To an N,N-dimethylformamide (15 mL)
suspension of 9.42 g of potassium=tert-butoxide were

added 10.0 g of 4-cyanophenol and 7.02 mL of 3-chloro-
1-propanol under cooling with water, which was then
stirred at 100°C for one hour. The reaction mixture was
cooled down to room temperature, to which water and
ethyl acetate were then added. The organic layer was
separated, washed sequentially with a 5% potassium
carbonate aqueous solution and a saturated sodium
chloride aqueous solution, and then dried with
anhydrous magnesium sulfate, followed by distilling off
the solvent under reduced pressure. In 100 mL of
dioxane was dissolved 11.9 g of the resultant oily
matter. To this mixture was added 9.28 mL of
triethylamine, to which 5.15 mL of methanesulfonyl
chloride was then added dropwise under cooling with ice
over a period of 8 minutes, followed by stirring at
room temperature for 10 minutes. To the reaction
mixture was dropwise added 100 mL of water, which was
then stirred at room temperature for 45 minutes. The
precipitate was collected by filtration and washed with
water and 2-propanol to provide 12.3 g of 3-(4-
cyanophenoxy)propyl=methanesulfonate as white solid
form.
1H-NMR(CDCl3).δ value:
2.27(2H,tt,J=6.0,6.0Hz),3.02(3H,s),4.15(2H,t,J=6.0Hz),
4.45 (2H,t,J=6.0Hz),6.96(2H,d, J=8.9Hz) , 7.60(2H, d, J=8.9Hz
) -


To an N,N-dimethylformamide (250 mL) solution
of 50.0 g of 3-(4-cyanophenoxy)propyl=methanesulfonate
were added 32.5 g of potassium iodide, 32.9 g of sodium
bicarbonate, and 37.0 g of 3-(4-piperidinyl)-1-propanol
hydrochloride at room temperature, which was then
stirred at 70°C for 6 hours and 50 minutes. The
reaction mixture was cooled down to room temperature,
to which water and toluene were then added, followed by
adjustment to pH 1.0 using hydrochloric acid. The
aqueous layer was separated, adjusted to pH 10.0 using
a 20% sodium hydroxide aqueous solution, and stirred at
room temperature for 15 minutes and then under cooling
with ice for 30 minutes. The precipitate was collected
by filtration and washed with water and toluene to
provide 52.3 g of 4-{3-[4-(3-hydroxypropyl)-1-
piperidinyl]propoxy}benzonitrile as white solid form.
1H-NMR(CDCl3) .δ value: 1. 20-1. 75 (10H,m) , 1. 85-
2.05(4H,m),2.40-2.55(2H,m),2.85-
3.00(2H,m),3.64(2H, t, J=6.6Hz) , 4.06(2H,t,J=6.3Hz),6.94(2
H,d,J=9.0Hz),7.57(2H,d,J=9.0Hz)

To a dimethylsulfoxide (200 mL) solution of
18.7 g of 4-{3-[4-(3-hydroxypropyl)-1-
piperidinyl]propoxy}benzcnitrile was added 8.92 mL of a

50% hydroxylamine aqueous solution at room temperature,
which was then stirred at the same temperature for 5
hours and 30 minutes. Thereto was added 8.92 mL of a
50% hydroxylamine aqueous solution at room temperature,
which was stirred at the same temperature for 15 hours
and 50 minutes. To the reaction mixture were added 2-
propanol and water, which was then stirred at room
temperature for one hour. The precipitate was
collected by filtration and washed with water to
provide 17.5 g of N'-hydroxy-4-{3-[4-(3-hydroxypropyl)-
l-piperidinyl]propoxy}benzamidine as white solid form.
1H-NMR(CDCl3) .δ value: 1.00-1.25(5H,m),1.35-
1.4 5(2H,m),1.55-1.65(2H,m),1.7 5-1.90(4H,m),2.35-
2.45(2H,m),2.80-2.90(2H,m),3.30-
3.40(2H,m),4.00(2H,t, J=6.5Hz),4.34(1H,t,J=5.1Hz),5.60-
5.80(2H,broad),6.90(2H,d,J=8.7Hz),7.58(2H,d,J=8.7Hz),9.
43(lH,s).

To an acetic acid (80 mL) suspension of 10.0
g of N'-hydroxy-4-{3-[4-(3-hydroxypropyl)-1-
piperidinyl]propoxy}benzamidine were added 5.31 mL of
acetic anhydride and 0.50 g of 5% palladium-carbon at
room temperature, which was stirred at room temperature
under hydrogen atmosphere for 12 hours and 50 minutes.
After filtering off insoluble matter, the solvent was

distilled off under reduced pressure. To the resultant
residue was added 6 mol/L hydrochloric acid, followed
by distilling off the solvent under reduced pressure
before purification using silica gel column
chromatography (silica gel: ODS-A from YMC, eluent;
water). The eluate was concentrated to about 100 mL
under reduced pressure and then adjusted to pH 12 using
a 5 mol/L sodium hydroxide aqueous solution. The
precipitate was collected by filtration and washed with
water to provide 8.43 g of 4-{3-[4-(3-hydroxypropyl)-1-
piperidinyl]propoxy}benzamidine as white solid form.
1H-NMR(CDCl3) .δ value: 1. 00-1. 25 (5H,m) , 1. 35-
1.45(2H,m),1.55-1.65(2H,m),1.7 5-1.90(4H,m),2.30-
2.45(2H,m),2.80-2.90(2H,m),3.30-
3.40(2H,m),4.03(2H,t,J=6.3Hz),6.95(2H, d, J=8.8Hz) ,7.72(2
H,d,J=8.8Hz).

To a dioxane (10 mL) suspension of 2.00 g of
4-{3-[4-(3-hydroxypropyl)-1-
piperidinyl]propoxy}benzamidine were sequentially added
1.37 g of di-tert-butyl=dicarbonate, 4 mL of N,N-
dimethylformamide, and 10 mL of a 2.5 mol/L sodium
hydroxide aqueous solution at room temperature, which
was then stirred at the same temperature for 45
minutes. Thereto was added 1.37 g of di-tert-

butyl=dicarbonate at room temperature, which was then
stirred at the same temperature for 2 hours and 45
minutes. Insoluble matter was filtered off, and
chloroform and water were added to the filtrate. The
organic layer was separated, washed with a saturated
sodium chloride aqueous solution, and then dried with
anhydrous magnesium sulfate, followed by distilling off
the solvent under reduced pressure. The resultant
residue was purified using silica gel column
chromatography (eluent; chloroform:methanol =4:1) to
provide 2.35 g of tert-butyl=[1-amino-l-(4-{3-[4-(3-
hydroxypropyl)-1-
piperidinyl]propoxy}phenyl)methylidene]carbamate as
pale red solid form.
1H-NMR(CDCl3) .δ value: 1.15-1.35(5H,m),1.35-
1.75(4H,m),1.55(9H,s),1.85-2.05(4H,m),2.45-
2.55(2H,m),2.85-
3.00(2H,m),3.64(2H,t,J=6.6Hz),4.05(2H,t,J=6.5Hz),6.91(2
H,d, J=8.8Hz),7.83(2H,d,J=8.8Hz) .

To a dimethylsulfoxide (100 mL) solution of
10.0 g of 4-cyanophenol was dropwise added 12.9 mL of a
50% hydroxylamine aqueous solution at room temperature
over a period of 6 minutes, which was then stirred at
the same temperature for 14 hours. Chloroform, a 1
mol/L sodium hydroxide aqueous solution, and water were

added to the reaction mixture. The aqueous layer was
separated and washed sequentially with toluene,
chloroform, and toluene, to which water was then added,
followed by adjustment to pH 7.2 using 6 mol/L
hydrochloric acid. After stirring this mixture at room
temperature for 30 minutes, the precipitate was
collected by filtration and washed with water to
provide 8.88 g of N',4-dihydroxybenzamidine as white
solid form.
1H-NMR(CDCl3) .δ value: 5.50-
5.70(2H,broad),6.73(2H,d,J=8.5Hz),7.47(2H,d,J=8.5Hz),9.
34(lH,s),9.50-9.60(1H,broad).

To an N,N-dimethylformamide (70 mL) solution
of 9.81 g of 4-[3-(4-piperidinyl)propoxy]benzonitrile
were added 8.02 g of potassium carbonate and 2.62 mL of
3-bromo-1-propanol at room temperature, which was then
stirred at the same temperature for 2 hours and 15
minutes. Thereto was added 2.62 mL of 3-bromo-l-
propanol, which was then stirred at the same
temperature for 1 hour and 45 minutes. Chloroform and
water were added to the reaction mixture. The organic
layer was separated, washed sequentially with a 1 mol/L
sodium hydroxide aqueous solution and a saturated
sodium chloride aqueous solution, and then dried with
anhydrous magnesium sulfate, followed by distilling off

the solvent under reduced pressure. The resultant
residue was purified using silica gel column
chromatography (eluent; chloroformrmethanol =4:1) to
provide 4.28 g of 4-{3-[1-(3-hydroxypropyl)-4-
piperidinyl]propoxyJbenzonitrile as white solid form.
1H-NMR(CDCl3) .δ value: 1.15-1.45(5H,m),1.65-
2.00(8H,m),2.55-2.65(2H,m),3.00-
3.10(2H,m),3.81(2H,t,J=5.2Hz),3.98(2H,t,J=6.5Hz),6.92(2
H,d,J=8.5Hz),7.57(2H,d,J=8.5Hz).

To a dimethylsulfoxide (43 mL) suspension of
4.27 g of 4-{3-[1-(3-hydroxypropyl)-4-
piperidinyl]propoxyJbenzonitrile was added 4.32 mL of a
50% hydroxylamine aqueous solution at room temperature,
which was then stirred at 40 to 50°C for 3 hours and 30
minutes. The reaction mixture was cooled down to room
temperature, to which 50 mL of water was then added
dropwise over a period of 10 minutes, followed by
stirring at room temperature for 30 minutes. The
precipitate was collected by filtration and washed with
water to provide 4.59 g of N'-hydroxy-4-{3-[1- (3-
hydroxypropyl)-4-piperidinyl]propoxyJbenzamidine as
white solid form.
1H-NMR(DMSO-d6) .δ value: 1.00-1.40(5H,m),1.50-
1.85(8H,m),2.25-2.35(2H,m),2.75-

2.90(2H,m),3.42(2H,t,J=6.2Hz),3.96(2H,t,J=6.5Hz),4.40-
4.60 (lH,broad),5.60-
5.80(2H,broad) ,6.90(2H,d, J=8.8Hz),7.58(2H,d,J=8.8Hz),9.
43(1H,s).

To an acetic acid (50 mL) suspension of 4.58
g of N'-hydroxy-4-{3-[1-(3-hydroxypropyl)-4-
piperidinyl]propoxy}benzamidine was added 2.59 mL of
acetic anhydride at room temperature, which was then
stirred at the same temperature for one hour. To the
reaction mixture was added 0.50 g of 5% palladium-
carbon, which was then stirred at room temperature
under hydrogen atmosphere for 5 hours and 30 minutes.
Insoluble matter was filtered off, and the solvent was
distilled off under reduced pressure. Water was added
to the resultant residue, which was adjusted to pH 12.5
using a 5 mol/L sodium hydroxide aqueous solution. The
precipitate was collected by filtration and washed with
water to provide 4.82 g of 4-{3-[1-(3-hydroxypropyl)-4-
piperidinyl]propoxy}benzamidine as white solid form.
1H-NMR(DMSO-d5) .δ value: 1.00-1.40(5H,m),1.50-
1.90(8H,m),2.25-2.35(2H,m),2.75-
2.90(2H,m),3.42(2H,t,J=6.2Hz) , 4.01(2H, t, J=6.5Hz) ,7.01(2
H,d, J=8.8Hz) ,7.74(2H, d, J=8.8Hz),8.10-9.20(2H,broad) .
Reference Example 24


To a dioxane (10 mL) suspension of 2.00 g of
4-{3-[1-(3-hydroxypropyl)-4-
piperidinyl]propoxy}benzamidine were added 2.05 g of
di-tert-butyl=dicarbonate, 4 mL of N,N-
dimethylformamide, and 10 mL of a 2.5 mol/L sodium
hydroxide aqueous solution at room temperature, which
was then stirred at the same temperature for 1 hour and
15 minutes. Subsequently, to the reaction mixture was
added 10 mL of a 2.5 mol/L sodium hydroxide aqueous
solution at room temperature, which was then stirred at
the same temperature for 1 hour and 15 minutes.
Chloroform and water were added to the reaction
mixture. The organic layer was separated, and the
aqueous layer was extracted with chloroform. The
organic layer and the extract were combined, which was
then washed with a saturated sodium chloride aqueous
solution and further dried with anhydrous magnesium
sulfate, followed by distilling off the solvent under
reduced pressure. The resultant residue was purified
using silica gel column chromatography (eluent;
chloroform:methanol =4:1) to provide 1.75 g of tert-
butyl=[1-amino-1-(4-{3-[1-(3-hydroxypropyl)-4-
piperidinyl]propoxy}phenyl)methylidene]carbamate as
white solid form.
1H-NMR(CDCl3) .δ value: 1.15-2.00(13H,m),1.55(9H,s),2.55-

2.65(2H,m),3.00-
3.10(2H,m),3.80(2H,t,J=5.2Hz),3.97(2H,t,J=6.5Hz),6.90(2
H,d,J=8.9Hz),7.83(2H,d,J=8.9Hz).

To an N,N-dimethylformamide (10 mL)
suspension of 1.00 g of 4-{3-[1-(3-hydroxypropyl)-4-
piperidinyl]propoxyjbenzamidine was added 1.28 g of
benzyl=4-nitrophenyl=carbonate at room temperature,
which was then stirred at the same temperature for one
hour. To the reaction mixture was added 1 mL of a 5
mol/L sodium hydroxide aqueous solution at room
temperature, which was then stirred at the same
temperature for 15 minutes. Thereto was added 5 mL of
a 5 mol/L sodium hydroxide aqueous solution at room
temperature, which was then stirred at the same
temperature for 5 minutes. Chloroform and a 5%
potassium carbonate aqueous solution were added to the
reaction mixture. The organic layer was separated,
washed sequentially with a 5% potassium carbonate
aqueous solution and a saturated sodium chloride
aqueous solution, and then dried with anhydrous
magnesium sulfate, followed by distilling off the
solvent under reduced pressure. The resultant residue

was purified using silica gel column chromatography
(eluent; chloroform:methanol =4:1) to provide 1.22 g
of benzyl=[1-amino-1-(4-{3-[1-(3-hydroxypropyl)-4-
piperidinyl]propoxy}phenyl)methylidene]carbamate as
pale yellow solid form.
1H-NMR(CDCl3) .δ value: 1.15-1.4 5(5H,m),1.7 5-
2.00(8H,m),2.55-2.65(2H,m),3.00-3.10(2H,m),3.75-
3.85(2H,m),3.98(2H,t,J=6.6Hz),5.21(2H,s),6.91(2H,d,J=8.
9Hz),7.20-7.50(5H,m),7.85(2H,d,J=8.9Hz).

To an N,N-dimethylformamide (20 mL) solution
of 1.00 g of 4-{3-[4-(3-hydroxypropyl)-1-
piperidinyl]propoxy}benzamidine was added 2.14 g of
benzyl=4-nitrophenyl=carbonate at room temperature,
which was then stirred at the same temperature for 18
hours. Chloroform, water, and a 5% potassium carbonate
aqueous solution were added to the reaction mixture.
The organic layer was separated, washed sequentially
with a 5% potassium carbonate aqueous solution and a
saturated sodium chloride aqueous solution, and then
dried with anhydrous magnesium sulfate, followed by
distilling off the solvent under reduced pressure. The
resultant residue was purified using silica gel column

chromatography (eluent; chloroform:methanol =8:1) to
provide 0.93 g of benzyl=[1-amino-1-(4-{3-[4-(3-
hydroxypropyl)-1-
piperidinyl]propoxy}phenyl)methylidene]carbamate as
white solid form.
1H-NMR (CDCl3) .δ value: 1. 20-1. 35 (5H,m) , 1. 50-
1.75(4H,m),1.85-2.05(4H,m),2.45-2.55(2H,m),2.85-
3.00(2H,m),3.62(2H,t,J=6.7Hz) ,4.04(2H, t, J=6.3Hz) ,5.21(2
H,s),6.88-6.93(2H,d,J=8.8Hz),7.25-
7.50(5H,m),7.84(2H,d,J=8.8Hz).

To a dimethylsulfoxide (126 mL) suspension of
12.6 g of 4-(3-{4-[3- (4-cyanophenoxy)propyl]-1-
piperidinyl}propoxy)benzonitrile was added 19.1 mL of a
50% hydroxylamine aqueous solution, which was then
stirred at 50°C for 19 hours. The mixture was cooled
down to room temperature, to which 260 mL of water was
added dropwise over a period of 50 minutes, followed by
stirring at room temperature for 30 minutes and then
under cooling with water for 2 hours. The precipitate
was collected by filtration to provide 15.0 g of 4-{3-
[4-(3-{4-[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}-N'-hydroxybenzamidine as white

solid form.
1H-NMR(DMSO-d6) .δ value: 1.05-1.40(5H,m),1.60-
1.80(4H,m),1.80-1.90(4H,m),2.35-2.45(2H,m),2.80-
2.90(2H,m),3.96(2H,t,J=6.5Hz),4.01(2H,t,J=6.5Hz),5.65-
5.75(4H,m),6.85-6.95(4H,m),7.55-
7.65(4H,m),9.43(lH,s),9.43(lH,s).

(2-1)
To an acetic acid (10 mL) suspension of 1.07
g of 4-{3-[4-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}-N'-hydroxybenzamidine was added
0.64 mL of acetic anhydride at room temperature, which
was then stirred at room temperature for 40 minutes.
To this mixture was added 0.10 g of 5% palladium-
carbon, which was then stirred under hydrogen
atmosphere for 2 hours and 15 minutes. Insoluble
matter was filtered off before adding 4 mL of 6.0 mol/L
hydrochloric acid, and insoluble matter was then
filtered off, followed by distilling off the solvent

under reduced pressure. A 5.0mol/L sodium hydroxide
aqueous solution was added to the resultant residue to
adjust the pH to 12.5, followed by collecting the solid
matter by filtration to provide 0.61 g of 4-{3-[4-(3-
{4-[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxyJbenzamidine as white solid form.
(2-2)
To an acetic acid (150 mL) suspension of 14.9
g of 4-{3-[4-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}-N'-hydroxybenzamidine was added
5.97 mL of acetic anhydride at room temperature, which
was then stirred at room temperature for 1 hour and 20
minutes. To this mixture was added 1.50 g of 5%
palladium-carbon, which was then stirred under hydrogen
atmosphere for 4 hours and 40 minutes. Insoluble
matter was filtered off, and 55 mL of 6.0 mol/L
hydrochloric acid was then added. The solvent was
distilled off under reduced pressure, and ethanol was
added to the resultant residue. The solid matter was
collected by filtration to provide 14.0 g of 4-{3-[4-
(3-{4-[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxyJbenzamidine as white solid form.
1H-NMR(DMSO-d6) .δ value: 1.30-1.45(2H,m),1.45-
1.70(3H,m),1.70-1.90(4H,m),2.15-2.30(2H,m),2.80-
3.00(2H,m),3.10-3.20(2H,m),3.4 5-
3.55(2H,m),4.10(2H,t,J=6.2Hz),4.19(2H,t,J=6.1Hz),7.15(2
H,d,J=8.4Hz),7.16(2H,d, J=8.4Hz),7.84(2H,d,J=8.4Hz),7.86

(2H,d,J=8.4Hz),8.90-9.00(4H,m),9.15-9.30(4H,m),10.60-
10.80(1H,broad).

Hydrogen chloride was introduced into an
ethanol (20 mL) suspension of 1.15 g of 4-(3-{4-[3-(4-
cyanophenoxy)propyl]-1-piperidinyl}propoxy)benzonitrile
under cooling with ice, which was then stirred at room
temperature for 24 hours. The solvent was distilled
off under reduced pressure, and the resultant residue
was dissolved in 20 mL of ethanol. Thereto was added
1.54 g of ammonium acetate, which was then heated to
reflux for 3 hours and 45 minutes. The reaction
mixture was cooled down to room temperature, to which
water was added, followed by distilling off ethanol
under reduced pressure. Chloroform was added to the
resultant residue, to which a 5.0 mol/L sodium
hydroxide aqueous solution was then added to adjust the
pH to 12.5. The precipitate was collected by
filtration to provide 1.13 g of 4-{3-[4-(3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine as white solid form.

1H-NMR(DMSO-d6) .δ value: 1.00-1.40(5H,m),1.60-
1.80(4H,m),1.80-1.95(4H,m),2.35-2.45(2H,m),2.80-
2.90(2H,m),3.98(2H,t,J=6.5Hz),4.03(2H,t,J=6.3Hz),6.30-
7.20(4H,broad),6.85-7.00(4H,m),7.65-7.80(4H,m).

To an ethanol (10 mL) suspension of 0.50 g of
4-{3-[4-(3-{4-[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine was added 1.77 mL of a
2.6 mol/mL hydrogen chloride/ethanol solution at room
temperature, which was then stirred at room temperature
for 4 hours and 15 minutes. The precipitate was
collected by filtration to provide 0.49 g of 4-{3-[4-
(3-{4-[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine hydrochloride as
colorless solid form.
1H-NMR spectral data in DMSO-d6 agreed with
the values of Example 2.


To a dioxane (3.0 mL) suspension of 67 mg of
4-(3-{1-[3-(4-cyanophenoxy)propyl]-4-
piperidinyl}propoxy)-2-fluorobenzonitrile was added 1.0
mL of a 50% hydroxylamine aqueous solution, which was
then heated to reflux for 2 hours. The mixture was
cooled down to room temperature, to which 10 mL of
water was then added dropwise, followed by stirring
under cooling with ice for 30 minutes. The precipitate
was collected by filtration to provide 63 mg of 4-{3-
[1- (3-{4-[amino(hydroxyimino)methyl]phenoxy}propyl)-4-
piperidinyl]propoxy}-2-fluoro-N'-hydroxybenzamidine as
pale yellow solid form.
1H-NMR(DMSO-d6) .δ value: 1.00-1.40(5H,m),1.60-
1.80(4H,m),1.80-1.95(4H,m),2.35-2.4 5(2H,m),2.8 0-
2.90(2H,m),3.98(2H, t, J=6.4Hz) ,4.00(2H,t,J=6.0Hz),5.60-
5.80(4H,m),6.70-6.85(2H,m) ,6.90(2H,d, J=8.8Hz),7.35-
7.45(lH,m),7.58(2H,d,J=8.8Hz),9.43(1H,s),9.50(1H,s).


To an acetic acid (2.0 mL) suspension of 56
mg of 4-{3-[l-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-4-
piperidinyl]propoxy}-2-fluoro-N'-hydroxybenzamidine was
added 0.043 mL of acetic anhydride at room temperature,
which was then stirred at the same temperature for one
hour. To this mixture was added 5.0 mg of 5%
palladium-carbon, which was then stirred under hydrogen
atmosphere for 2 hours. Insoluble matter was filtered
off, and the solvent was distilled off under reduced
pressure. Thereto were added 6.0 mol/L hydrochloric
acid and water, followed by distilling off the solvent
under reduced pressure. The resultant residue was
purified using silica gel column chromatography (silica
gel: ODS-AM120-S50 from YMC, eluent; water). The
resultant residue was dissolved in 5.0 mL of water, to
which a 5.0 mol/L sodium hydroxide aqueous solution was
then added to adjust the pH to 12.2. The solution was
stirred under cooling with ice for 20 minutes, and the
precipitate was collected by filtration to provide 43

mg of 4-{3-[l-(3-{4-
[amino(imino)methyl]phenoxy}propyl)-4-
piperidinyl]propoxy}-2-fluorobenzamidine as white solid
form.
1H-NMR(DMSO-d6) .δ value: 1.05-1.40(5H,m),1.60-
2.05(8H,m),2.30-2.45(2H,m),2.80-
2.90(2H,m),3.98(2H,t,J=6.5Hz),4.02(2H,t,J=6.3Hz),6.20-
6.70(4H,broad),6.75-6.85(2H,m),6.92(2H,d,J=8.4Hz),7.4 5-
7.55(lH,m),7.71(2H,d,J=8.4Hz).

As described in Example 5, 0.10 g of 4-3-{4-
[3-(4-cyanophenoxy)propyl]-1-piperidinyl}propoxy)-2-
fluorobenzonitrile was used to provide 0.11 g of 4-{3-
[4-(3-{4-[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}-2-fluoro-N'-hydroxybenzamidine as
white solid form.
1H-NMR(DMSO-d6) .δ value: 1.00-1.40(5H,m),1.60-
1.75(4H,m),1.75-1.90(4H,m),2.30-2.40(2H,m),2.80-
2.90(2H,m),3.96(2H,t,J=6.5Hz),4.03(2H,t,J=6.3Hz),5.65-
5.80(4H,m),6.75-6.90(2H,m),6.90(2H,d,J=8.9Hz),7.35-
7.45(lH,m),7.58(2H,d,J=8.9Hz),9.43(1H,s),9.50(1H,s).


As described in Example 6, 90 mg of 4-{3-[4-
(3-{4-[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}-2-fluoro-N'-hydroxybenzamidine was
used to provide 34 mg of 4-{3- [4-(3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}-2-fluorobenzamidine as white solid
form.
1H-NMR(DMSO-d6) .δ value: 1.05-1.40(5H,m),1.60-
1.90(8H,m),2.30-2.45(2H,m) ,2.80-
2.90(2H,m),3.98(2H, t, J=6.5Hz) ,4.03(2H,t,J=6.0Hz),6.30-
6.75(4H,broad),6.75-6.85(2H,m),6.93(2H,d,J=8.7Hz),7.45-
7.55(lH,m),7.71(2H,d, J=8.7Hz) .


Hydrogen chloride was introduced into an
ethanol (10 mL) suspension of 0.10 g of 4- (3-{1- [3-(4-
cyano-3-fluorophenoxy)propyl]-4-piperidinyl}propoxy)-2-
fluorobenzonitrile under cooling with ice, which was
then stirred at the same temperature for 1 hour and 10
minutes and at room temperature for 17 hours. The
solvent was distilled off under reduced pressure, and
the resultant residue was suspended in 5.0 mL of
ethanol, to which 44 mg of ammonium acetate was then
added, followed by heating to reflux for 5 hours and 30
minutes. The solvent was distilled off under reduced
pressure, and the resultant residue was dissolved in
8.0 mL of 1.0 mol/L hydrochloric acid, followed by
distilling off the solvent under reduced pressure. The
resultant residue was purified using silica gel column
chromatography (silica gel: ODS-AM120-S50 from YMC,
eluent; water) to provide 46 mg of 4-{3-[1-(3-{4-
[amino(imino)methyl]-3-fluorophenoxy}propyl)-4-
piperidinyl]propoxy}-2-fluorobenzamidine hydrochloride
as white solid form.
1H-NMR (DMSO-d6).δ value: 1.30-1.45(2H,m),1.50-
1.70(3H,m),1.70-1.90(4H,m),2.20-2.30(2H,m),2.80-
2.95(2H,m),3.10-3.20(2H,m),3.4 0-
3.55(2H,m),4.10(2H,t,J=6.0Hz),4.20(2H,t,J=5.7Hz),6.95-
7.05(2H,m),7.05-7.15(2H,m),7.60-7.75(2H,m),9.20-
9.50(8H,m),10.95-11.10(1H, broad).


To a water (2.3 mL) suspension of 1.0 g of 4-
{3-[4-(3-{4-[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine were added 0.72 g of
hydrochloric acid, 6 mL of 2-propanol, and 0.5 mL of
water at room temperature, which was then stirred at
40°C. Thereto was added 9 mL of 2-propanol, which was
then stirred under cooling with ice for one hour. The
precipitated crystals were collected by filtration to
provide 1.4 g of 4-{3-[4-(3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine trihydrochloride
pentahydrate as colorless crystals. These crystals
were used as seed crystals.
Water content: 14.5%
1H-NMR spectral data in DMSO-d6 agreed with
the values of Example 2.
(10-2)
To a water (40 mL)/2-propanol (80 mL)
suspension of 20.0 g of 4-{3-[4-(3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine was added 14.3 g of
hydrochloric acid, which was then stirred at 60°C for 10

minutes. Thereto was added 120 mL of 2-propanol, to
which 100 mg of seed crystals were added, followed by-
stirring at room temperature for 35 minutes and under
cooling with ice for 2 hours. The precipitated
crystals were collected by filtration to provide 28.3 g
of 4—{3—[4—(3—{4—[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine trihydrochloride
pentahydrate as colorless crystals.
Water content: 14.0%
Powder X-ray diffraction peaks (°) : 6.6, 13.2,
16.1, 21.5, 25.5 (20) (shown in Fig. 1)
1H-NMR spectral data in DMSO-d6 agreed with
the values of Example 2.
Example 11
As described in Example 10, 31.1 g of 4-{3-
[4—{3—{4—[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine trimethanesulfonate
dihydrate was obtained from 20.0 g of 4-{3-[4-(3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine and 13.6 g of
methanesulfonic acid.
Water content: 4.6%
1H-NMR(DMSO-d6) .δ value: 1.36-1.79(7H,m),1.86-
1.96(2H,m),2.15-2.25(2H,m),2.34(9H,s),2.85-
2.98(2H,m),3.18-3.26(2H,m),3.50-
3.58(2H,m),4.09(2H,t,J=6.3Hz),4.16(2H,t,J=5.9Hz),7.15(2
H,d,J=8.8Hz) ,7.16(2H,d, J=9.0Hz) , 7.82(2H,d,J=8.5Hz),7.84
(2H,d, J=8.3Hz),8.77-8.87(3H,m),9.10-9.18(3H,m) .

Example 12
As described in Example 10, 21.6 g of 4-{3-
[4-(3-{4-[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine diacetate 1/2-hydrate
was obtained from 20.0 g of 4-{3- [4- (3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine and 8.5 g of acetic
acid.
Water content: 1.4%
1H-NMR(DMSO-d6) .δ value: 1. 10-1.40(5H,m),1.62-
1.80(6H,m),1.75(6H,s),2.4 0-2.47(2H,m) ,2.50-
2.60(2H,m),2.84-2.92(2H,m),4.00-4.15(4H,m),7.10-
7.15(4H,m),7.78(4H, d, J=8.8Hz) .
Example 13
As described in Example 10, 30.3 g of 4-{3-
[4-(3-{4-[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine 3/2-sulfate trihydrate
was obtained from 20.0 g of 4-{3-[4-(3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine and 15.1 g of sulfuric
acid.
Water content: 7.7%
1H-NMR(DMSO-d6) .δ value: 1. 40-1.90(7H,m) , 2.02-
2.10(2H,m) ,2.24-2.33(2H,m),2.94-3.04(2H,m) , 3.30-
3.36(2H,m),3.62-
3.69(2H,m),4.17(2H,t,J=6.3Hz),4.25(2H,t,J=5.7Hz),7.14-
7.16(4H,m),7.78(2H,d,J=9.0Hz),7.78(2H,d, J=9.0Hz) .
Example 14

As described in Example 10, 25.0 g of 4-{3-
[4-(3-{4-[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine triphosphate dihydrate
was obtained from 20.0 g of 4-{3-[4-( 3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine and 16.9 g of
phosphoric acid.
Water content: 4.2%
1H-NMR(DMSO-d6) .δ value: 1.43-1.76(5H,m),1.84-
1.94(2H,m),2.05-2.12(2H,m),2.27-2.33(2H,m),2.96-
3.06(2H,m),3.32-3.38(2H,m),3.64-
3.70(2H,m),4.19(2H,t,J=6.2Hz),4.27(2H,t,J=5.6Hz),7.17(4
H,d,J=8.8Hz),7.80(2H,d,J=9.0Hz),7.80(2H,d, J=9.0Hz) .
Example 15
As described in Example 10, 26.5 g of 4-{3-
[4- (3-{4-[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine di-L-lactate 3/2-
hydrate was obtained from 20.0 g of 4-{3-[4-(3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine and 15.5 g of L-lactic
acid.
Water content: 4.1%
1H-NMR(DMSO-d6) .δ value: 1.33-1.35(6H,m),1.40-
1.70(5H,m),1.84-1.90(2H,m),1.99-2.07(2H,m),2.22-
2.29(2H,m),2.8 6-2.92(2H,m),3.21-3.25(2H,m),3.50-
3.56(2H,m),4.09-
4.14(2H,m),4.19(2H,t,J=6.5Hz),4.26(2H,t,J=5.7Hz),7.15-
7.19(4H,m),7.80(2H,d,J=9.0Hz) ,7.80(2H, d, J=8.3Hz) .


Hydrogen chloride was introduced into an
ethanol (20 mL) suspension of 1.00 g of 4-(3-{4-[3-(4-
cyanophenoxy)propyl]-1-piperidinyl}propoxy)benzonitrile
under cooling with ice, which was then stirred for 18
hours. The solvent was distilled off under reduced
pressure, and the resultant residue was dissolved in 60
mL of ethanol. To this solution was added 2.07 g of 0-
methylhydroxylamine hydrochloride, to which 10.4 mL of
triethylamine was then dropwise added under cooling
with ice over a period of 2 minutes. After stirring at
room temperature for 18 hours, the reaction mixture was
added to a mixture of water and chloroform, which was
then adjusted to pH 10 using a 1.0 mol/L sodium
hydroxide aqueous solution. The organic layer was
separated, washed with water, and then dried with
anhydrous sodium sulfate, followed by distilling off
the solvent under reduced pressure. The resultant
residue was purified using silica gel column
chromatography (eluent; chloroform:methanol = 30:1), to
which ethanol was then added, followed by collecting
the solid by filtration to provide 0.99 g of 4-{3-[4-
(3-{4-[amino(methoxyimino)methyl] phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=O-methyloxime as white

solid form.
1H-NMR(CDCl3) .δ value: 1.20-1.45(5H,m),1.65-
1.84(4H,m),1.88-2.04(4H,m),2.45-2.55(2H,m),2.90-
2.98(2H,m),3.90(6H,s),3.96(2H,t,J=6.6Hz),4.02(2H,t,J=6.
4Hz),4.74(4H,s),6.88(2H,d,J=8.8Hz),6.90(2H,d,J=8.8Hz),7
.55(2H,d,J=8.8Hz),7.55(2H, d, J=8.8Hz) .

To an ethanol (8 mL) suspension of 0.20 g of
ethyl=4-{3-[4-(3-{4-
[ethoxy(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzimidate were sequentially added
0.39 g of O-ethylhydroxylamine hydrochloride and 0.84
mL of triethylamine under cooling with ice, which was
then stirred at room temperature for 3.5 days. The
reaction mixture was added to a mixture of chloroform
and water, which was then adjusted to pH 9.7 using a
20% sodium hydroxide aqueous solution. The organic
layer was separated, washed with water, and then dried
with anhydrous sodium sulfate, followed by distilling
off the solvent under reduced pressure. The resultant
residue was purified using silica gel column
chromatography (eluent; chloroform:ethanol = 20:1) to
provide 0.20 g of 4-{3-[4-(3-{4-

[amino(ethoxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=0-ethyloxime as white
solid form.
1H-NMR(CDCl3) .δ value: 1.20-
1.45 (5H,m),1.32(3H,t,J=7.0Hz),1.32(3H,t,J=7.0Hz),1.65-
1.85(4H,m),1.85-2.05(4H,m) ,2.4 5-2.52(2H,m),2.90-
3.00(2H,m),3.96(2H,t, J=6.6Hz) ,4.02(2H,t,J=6.3Hz),4.14(2
H,q,J=7.0Hz),4.14(2H, q,J=7.0Hz),4.7 4(4H,s),6.8 8(2H,d,J=
8.8Hz),6.8 9(2H,d,J=8.8Hz) ,7.55(2H,d,J=8.8Hz),7.55(2H,d,
J=8.8Hz).

To an ethanol (6 mL) suspension of 0.20 g of
ethyl=4-{3-[4-(3-{4-
[ethoxy(imino)methy1]phenoxy}propyl)-1-
piperidinyl]propoxy}benzimidate were added 0.61 g of 0-
(2,2,2-trifluoroethyl)hydroxylamine hydrochloride and
0.84 mL of triethylamine at room temperature, which was
then stirred at the same temperature for one week.
Chloroform and water were added to the reaction
solution. The organic layer was separated, washed with
a saturated sodium chloride aqueous solution, and then
dried with anhydrous magnesium sulfate, followed by
distilling off the solvent under reduced pressure. The

resultant residue was purified using silica gel column
chromatography (eluent; chloroform:ethanol = 30:1), to
which hexane was then added, followed by collecting the
solid by filtration to provide 0.12 g of 4-{3-[4-(3-{4-
[amino(2,2,2-
trifluoroethoxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=0-(2,2,2-
trifluoroethyl)oxime as white solid form.
1H-NMR(CDCl3) .δ value: 1.20-1.45(5H,m),1.65-
2.05(8H,m),2.45-2.55(2H,m),2.90-
3.00(2H,m),3.96(2H,t,J=6.6Hz) ,4.03(2H,t,J=6.2Hz) ,4.43(2
H,q,J=8.6Hz),4.43(2H,q,J=8.6Hz),4.82(4H,s),6.8 9(2H,d,J=
8.8Hz),6.90(2H,d,J=8.8Hz),7.54(2H,d,J=8.8Hz),7.54(2H,d,
J=8.8Hz).

To an N-methyl-2-pyrrolidone (10 mL) solution
of 0.35 g of propionic acid was added 0.76 g of 1,1'-
carbonyldiimidazole at room temperature, which was then
stirred at the same temperature for one hour. An N-
methyl-2-pyrrolidone (10 mL) solution of 1.00 g of 4-
{3—[4-(3—{4—[amino(hydroxyimino)methyl]phenoxy}propyl)-
l-piperidinyl]propoxy}benzamide=oxime was added to the
mixture at room temperature, which was then stirred for

15 hours. The reaction mixture was added to a mixture
of chloroform and water. The precipitate was collected
by filtration and washed with ethyl acetate to provide
0.58 g of 4-{3-[4-(3-{4-
[amino(propionyloxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=0-(propionyl)oxirne as
white solid form.
1H-NMR(DMSO-d6) .δ value:
1.08(3H,t,J=7.6Hz),1.08(3H,t,J=7.6Hz),1.10-
1.40(5H,m),1.60-1.80(4H,m),1.80-1.95(4H,m),2.35-
2.45(2H,m),2.45(2H,q,J=7.6Hz) ,2.45(2H,q,J=7.6Hz) ,2.80-
2.90(2H,m),3.99(2H,t,J=6.5Hz),4.04(2H,t,J=6.3Hz),6.65(4
H,s),6.95-7.00(4H,m),7.60-7.70(4H,m).

To an N-methyl-2-pyrrolidone (10 mL) solution
of 0.42 g of butyric acid was added 0.76 g of 1,1'-
carbonyldiimidazole at room temperature, which was then
stirred at the same temperature for one hour. An N-
methyl-2-pyrrolidone (10 mL) solution of 1.00 g of 4-
{3-[4-(3-{4-[amino(hydroxyimino)methyl]phenoxy}propyl)-
1-piperidinyl]propoxy}benzamide=oxime was added to the
mixture at room temperature, which was then stirred at
the same temperature for 14 hours. The reaction

mixture was added to a mixture of ethyl acetate and
water. The precipitate was collected by filtration and
washed with ethyl acetate to provide 1.10 g of 4-{3-[4-
(3-{4-[amino(n-butyloxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=0-(butyryl)oxime as white
solid form.
1H-NMR(DMSO-d6) .δ value:
0.92(3H,t,J=7.4Hz),0.92(3H,t,J=7.4Hz),1.05-
1.40(5H,m),1.55-1.90(12H,m),2.35-
2.45(2H,m),2.42(2H,t,J=7.3Hz),2.42(2H,t,J=7.3Hz),2.80-
2.90(2H,m),3.99(2H,t,J=6.5Hz),4.04(2H,t,J=6.5Hz),6.64(4
H,s),6.95-7.00(4H,m),7.63-7.66(4H,m).

To an N-methyl-2-pyrrolidone (10 mL) solution
of 0.47 g of pivalic acid was added 0.76 g of 1,1'-
carbonyldiimidazole at room temperature, which was then
stirred at the same temperature for 1 hour and 20
minutes. An N-methyl-2-pyrrolidone (10 mL) solution of
1.00 g of 4-{3-[4-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=oxime was added to the
mixture at room temperature, which was then stirred at
the same temperature for 2 hours. The reaction mixture

was added a mixture of ethyl acetate and water. The
precipitate was collected by filtration and washed with
ethyl acetate to provide 0.81 g of 4-{3-[4-(3-{4-
[amino(2,2-
dimethylpropionyloxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=0-(2,2-
dimethylpropionyl)oxime as white solid form.
1H-NMR(CDCl3) .δ value: 1.20-
1.45(5H,m),1.34(9H,s),1.34(9H,s),1.65-1.85(4H,m),1.85-
2.05(4H,m),2.45-2.55(2H,m),2.90-
3.00(2H,m),3.97(2H,t,J=6.5Hz) ,4.04(2H, t, J=6.5Hz) ,4.92(4
H,s),6.90(2H,d,J=8.8Hz),6.91(2H,d,J=8.8Hz),7.64(2H,d,J=
8.8Hz),7.64(2H,d,J=8.8Hz).

To an N-methyl-2-pyrrolidone (5 mL) solution
of 0.42 g of methyl=hydrogen=succinate was added 0.52 g
of 1,1'-carbonyldiimidazole at room temperature, which
was then stirred at the same temperature for 2 hours.
An N-methyl-2-pyrrolidone (10 mL) solution of 0.50 g of
4-{3-[4-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=oxime was added to the
mixture at room temperature, which was then stirred at

the same temperature for 3 hours. The reaction mixture
was added to a mixture of ethyl acetate and water.
Insoluble matter was filtered off, followed by
adjustment to pH 7.0 using a 5 mol/L sodium hydroxide
aqueous solution before adding a saturated sodium
chloride aqueous solution. The organic layer was
separated and dried with anhydrous magnesium sulfate,
followed by distilling off the solvent under reduced
pressure. Ethyl acetate and water were added to the
resultant residue. The organic layer was separated,
washed with water, and then dried with anhydrous
magnesium sulfate, followed by distilling off the
solvent under reduced pressure. The resultant residue
was suspended in diisopropyl ether, and the solid
matter was collected by filtration and washed with
ethyl acetate and diisopropyl ether to provide 0.24 g
of 4-(3-{4-[3-(4-{amino[(4-methoxy-4-
oxobutylyl)oxyimino]methyl}phenoxy)propyl]-1-
piperidinyl}propoxy)benzamide=0-[(4-methoxy-4-
oxo)butylyl]oxime as white solid form.
1H-NMR(CDCl3) .δ value: 1.20-1.45(5H,m),1.65-
2.05(8H,m),2.45-2.55(2H,m),2.70-2.80(4H,m),2.80-
2.90(4H,m),2.90-
3.00 (2H,m),3.71(3H,s),3.71(3H,s) ,3.97(2H, t,J=6.6Hz) ,4.0
4(2H,t,J=6.3Hz),5.00-
5.20(4H,broad) ,6.90(2H,d,J=8.7Hz),6.91(2H,d,J=8.7Hz),7.
63(2H,d,J=8.7Hz),7.63(2H,d,J=8.7Hz).


To an N-methyl-2-pyrrolidone (4 mL) solution
of 0.23 g of N-(tert-butoxycarbonyl)-L-valine was added
0.18 g of 1.1'-carbonyldiimidazole at room temperature,
which was then stirred at the same temperature for 3
hours. An N-methyl-2-pyrrolidone (6 mL) solution of
0.25 g of 4-{3-[4-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=oxime was added to the
mixture at room temperature, which was then stirred at
the same temperature for 18 hours. The reaction
mixture was added to a mixture of ethyl acetate and
water. The organic layer was separated, washed with a
saturated sodium chloride aqueous solution, and then
dried with anhydrous magnesium sulfate, followed by
distilling off the solvent under reduced pressure. The
resultant residue was suspended in diisopropyl ether,
and the solid matter was collected by filtration and
purified using silica gel column chromatography
(eluent; chloroform:methanol = 20:1) to provide 0.27 g
of 4-(3-{4-[3-(4-{amino[(2S)-2-(tert-
butoxycarbonyl) amino-3-
methylbutylyloxyimino]methyl}phenoxy)propyl]-1-

piperidinyl}propoxy)benzamide=0-[(2S)-2(tert-
butoxycarbonyl)amino-3-methylbutylyl]oxime as white
solid form.
1H-NMR(DMSO-d6) .δ value:
0.90(6H,d, J=6.8Hz),0.90(6H,d,J=6.8Hz),1.05-
1.45(5H,m),1.41(9H,s),1.41(9H,s),1.60-1.95(8H,m),2.00-
2.10(2H,m),2.35-2.45(2H,m),2.80-2.90(2H,m),3.95-
4.10(6H,m),6.65-6.85(4H,broad),6.97-7.00(4H,m),7.30-
7.40(2H,broad),7.60-7.7 0(4H,m).

To an ethanol (2 mL) solution of 0.10 g of 4-
(3—{4—[3—(4-{amino[(2S)-2-(tert-butoxycarbonyl)amino-3-
methylbutylyloxyimino]methyl}phenoxy)propyl]-1-
piperidinyl}propoxy)benzamide=0-[(2S)-2-(tert-
butoxycarbonyl) amino-3-methylbutylyl] oxime was added 5
mL of 2.9 mol/L hydrogen chloride/ethanol at room
temperature, which was then stirred at the same
temperature for 18 hours and 30 minutes. Thereto was
added 1 mL of 2.9 mol/L hydrogen chloride/ethanol,
which was then stirred for 30 minutes. The solvent was
distilled off under reduced pressure, and the resultant
residue was suspended in diisopropyl ether, followed by

was added to a mixture of ethyl acetate and water. The
organic layer was separated, washed with a saturated
sodium chloride aqueous solution, and then dried with
anhydrous magnesium sulfate, followed by distilling off
the solvent under reduced pressure. The resultant
residue was purified using silica gel column
chromatography (eluent; chloroform:methanol = 10:1) to
provide 0.14 g of 4-(3-{4-[3-(4-{amino[(2S)-2-(tert-
butoxycarbonyl)amino-4-
methylpentanoyloxyimino]methyl}phenoxy)propyl]-1-
piperidinyl}propoxy)benzamide=0-[(2S)-2(tert-
butoxycarbonyl)amino-4-methylpentanoyl]oxime as white
solid form.
1H-NMR(CDCl3) .δ value:
0.97 (3H,d,J=5.9Hz) ,0.97 (3H, d, J=5 . 9Hz) , 0 . 99 (3H, d, J=6.1Hz
),0.99(3H,d,J=6.lHz),1.25-
1.90(15H,m),1.45(9H,s),1.45(9H,s) , 1.95-2.15(4H,m),2.55-
2.65(2H,m),3.00-
3.10(2H,m),3.97(2H,t,J=6.3Hz) ,4.04(2H,t,J=6.3Hz),4.4 0-
4.50(2H,m),4.95-5.05(2H,m),5.15-5.35(4H,broad),6.88-
6.91(4H,m),7.63(4H,d,J=8.4Hz).


To a chloroform (5.0 mL) solution of 0.10 g
of 4-(3-{4-[3-(4-{amino[(2S)-2-(tert-
butoxycarbonyl)amino-4-
methylpentanoyloxyimino]methylJphenoxy)propyl]-1-
piperidinyl}propoxy)benzamide=O-[(2S)-2-(tert-
butoxycarbonyl)amino-4-methylpentanoyl]oxime was added
20 µL of trifluoroacetic acid under cooling with ice,
which was then stirred at room temperature for 2 hours
and 10 minutes. To the reaction mixture was added 0.1
mL of trifluoroacetic acid , which was then stirred for
21 hours. Thereto was further added 1 mL of
trifluoroacetic acid, which was then stirred for 3
hours. The solvent was distilled off under reduced
pressure, and 2.9 mol/L hydrogen chloride/ethanol was
added, followed by distilling off the solvent under
reduced pressure. The resultant solid was suspended in
diisopropyl ether, followed by collecting solid matter
by filtration before washing with diisopropyl ether to
provide 0.04 g of 4-{3-[4-(3-{4-[amino((2S)-2-amino-4-
methylpentanoyloxy)imino]methyl}phenoxy)propyl]-1-
piperidinyl]propoxy}benzamide=0-((2S)-2-amino-4-
methylpentanoyl)oxime hydrochloride as white solid
form.
1H-NMR(DMSO-d6) .δ value:
0.90(3H,d, J=6.6Hz) ,0.90(3H, d, J=6.6Hz),0.91(3H,d,J=6.3Hz
),0.91(3H,d,J=6.3Hz),1.30-1.45(2H,m),1.50-
1.95(15H,m),2.20-2.30(2H,m),2.80-2.95(2H,m),3.10-
3.20(2H,m),3.75-4.30(6H,m),7.10-7.20(4H,m),7.65-

7.80(4H,m),8 . 25-8.40(2H,broad),8.4 0-
8.60(4H, broad) , 11.05-11.20(2H,broad) .

To an N-methyl-2-pyrrolidone (10 niL) solution
of 0.65 g of N-(tert-butoxycarbonyl)-L-isoleucine 1/2-
hydrate was added 1.14 g of 1,1'-carbonyldiimidazole at
room temperature, which was then stirred at the same
temperature for 1 hour and 30 minutes. An N-methyl-2-
pyrrolidone (15 mL) solution of 1.00 g of 4-{3-[4-(3-
{4- [amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=oxime was added to the
mixture at room temperature, which was then stirred at
the same temperature for 2 days. The reaction mixture
was added to a mixture of ethyl acetate and water, and
insoluble matter was filtered off. The organic layer
was separated, washed with water, and then dried with
anhydrous magnesium sulfate, followed by distilling off
the solvent under reduced pressure. The resultant
residue was purified using silica gel column
chromatography (eluent; chloroformrmethanol = 30:1),
and the resultant solid was suspended in diisopropyl
ether, followed by collecting the solid matter by
filtration before washing with diisopropyl ether to

provide 0.38 g of 4-(3-{4-[3-(4-{amino[(2S,3S)-2-(tert-
butoxycarbonyl)amino-3-
methylpentanoyloxyimino]methyl}phenoxy)propyl]-1-
piperidinyl}propoxy)benzamide=0-[(2S,3S)-2-(tert-
butoxycarbonyl)amino-3-methylpentanoyl]oxime as white
solid form.
1H-NMR(CDCl3) .δ value:
0.95 (3H,t, J=7.4Hz) , 0 . 95 (3H, t, J=7 . 4Hz) ,1.02 (3H, d, J=6 . 8Hz
) ,1.02(3H,d,J=6.8Hz),1.15-
1.50(7H,m),1.45(9H,s),1.45(9H,s),1.50-2.20(12H,m),2.60-
2.70(2H,m),3.05-
3.20(2H,m),3.97(2H,t,J=6.5Hz),4.04(2H,t,J=6.2Hz),4.25-
4.40(2H,m),5.10-5.25(4H,m),6.8 5-6.95(4H,m),7.60-
7.65(4H,m) .

To an ethanol (5 mL) solution of 0.20 g of 4-
(3-{4-[3-(4-{amino[(2S,3S)-2-(tert-
butoxycarbonyl)amino-3-
methylpentanoyloxyimino]methylJphenoxy)propyl]-1-
piperidinyl}propoxy)benzamide=0-[(2S,3S)-2(tert-
butoxycarbonyl)amino-3-methylpentanoyl]oxime was added
2 mL of 2.9 mol/L hydrogen chloride/ethanol at room

temperature, which was then stirred at the same
temperature for 5 hours. Thereto was added 2 mL of 2.9
mol/L hydrogen chloride/ethanol, which was then stirred
for 26 hours. Thereto was further added 1 mL of 2.9
mol/L hydrogen chloride/ethanol, which was then stirred
for 3 days. The solvent was distilled off under
reduced pressure, and the resultant solid was suspended
in diisopropyl ether, followed by collecting the solid
matter by filtration before washing with diisopropyl
ether to provide 0.13 g of 4-{3-[4-(3-{4-
[amino((2S,3S)-2-amino-3-
methylpentanoyloxyimino)methyl]phenoxy}propyl)-1-
piperidinyljpropoxy}benzamide=0-((2S,3S)-2-amino-3-
methylpentanoyl)oxime hydrochloride as white solid
form.
1H-NMR(DMSO-d6) .δ value: 0.85-
0.95(6H,m),0.99(3H,d,J=6.8Hz),0.99(2H,d,J=6.8Hz),1.2 0-
2.10(17H,m),2.15-2.30(2H,m),2.80-2.95(2H,m),3.10-
3.20(2H,m),3.95-4.30(6H,m),6.95-7.05(4H,m),7.05-
7.20(4H,m),7.65-7.75(4H,m),8.50-8.60(2H,broad),8.75-
8.90(6H,broad).

To an N,N-dimethylformamide (10 mL)

suspension of 1.00 g of 4-{3-[4- (3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine was added 4-
nitrophenyl=acetate at room temperature, which was then
stirred at the same temperature for 1 hour and 15
minutes. Chloroform and a 5% potassium carbonate
aqueous solution were added to the reaction mixture,
and insoluble matter was filtered off. The organic
layer was separated, washed sequentially with a 5%
potassium carbonate aqueous solution and a saturated
sodium chloride aqueous solution, and then dried with
anhydrous magnesium sulfate, followed by distilling off
the solvent under reduced pressure. Ethyl acetate and
water were added to the resultant residue, which was
then adjusted to pH 3.0 using hydrochloric acid. The
aqueous layer was separated and washed with ethyl
acetate, which was then adjusted to pH 12.0 using a
sodium hydroxide aqueous solution. The precipitate was
collected by filtration to provide 0.80 g of N'-acetyl-
4-{3-[4-(3-{4-
[(acetylimino)(amino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine as white solid form.
1H-NMR (DMSO-d6) .δ value: 1.00-1.40(5H,m),1.60-
1.90(8H,m),2.09(3H,s),2.09(3H,s),2.35-2.45(2H,m),2.80-
2.85(2H,m),4.02(2H,t,J=6.3Hz) ,4.07(2H,t,J=6.3Hz) , 6.90-
7.10(4H,m),7.95-8.05(4H,m).


As described in Example 29, 0.62 g of 4-{3-
[4-(3-{4-[amino(hexanoylimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}-N'-(hexanoyl)benzamidine as white
solid form was obtained from 0.50 g of 4-{3-[4-(3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine and 0.68 g of 4-
nitropnenyl=hexanoate.
1H-NMR(CDCl3) .δ value:
0.91(3H,t,J=6.7Hz),0.91(3H,t,J=6.7Hz),1.20-
1.45(13H,m),1.65-2.05(12H,m),2.4 5-
2.55(2H,m),2.54(2H, t, J=7 . 5Hz) ,2.54(2H,t,J=7.5Hz),2.90-
3.00(2H,m),4.00(2H,t, J=6.5Hz),4.06(2H,t,J=6.3Hz),6.93(2
H,d,J=8.7Hz),6.94(2H,d,J=8.7Hz),7.84(2H,d,J=8.7Hz),7.84
(2H,d,J=8.7Hz).

To an N,N-dimethylformamide (10 mL)
suspension of 0.50 g of 4-{3-[4- (3-{4-

[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine was added 0.78 g of
benzyl=4-nitrophenyl=carbonate at room temperature,
which was then stirred at the same temperature for 50
minutes. Chloroform and water were added to the
reaction mixture. The organic layer was separated,
washed sequentially with a 5% potassium carbonate
aqueous solution and a saturated sodium chloride
aqueous solution, and then dried with anhydrous
magnesium sulfate, followed by distilling off the
solvent under reduced pressure. The resultant residue
was purified using silica gel column chromatography
(eluent; chloroformrmethanol =4:1) to provide 0.67 g
of 4-{3-[4-(3-{4-
[amino(benzyloxycarbonylimino)methyl]phenoxyJpropyl)-1-
piperidinyl]propoxy}-N'-(benzyloxycarbonyl)benzamidine
as white solid form.
1H-NMR(CDCl3) .δ value: 1.20-1.4 5(5H,m),1.65-
2.05(8H,m),2.45-2.55(2H,m),2.85-
3.00(2H,m),3.99(2H,t,J=6.6Hz),4.06(2H,t,J=6.3Hz),5.21(2
H,s),5.21(2H,s),6.91(2H,d,J=8.7Hz),6.93(2H,d,J=8.7Hz),7
.20-7.50(10H,m),7.85(2H,d,J=8.7Hz),7.85(2H, d, J=8.7Hz) .


To an N,N-dimethylformamide (10 mL)
suspension of 0.50 g of 4-{3-[4-(3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine was added 0.60 g of
ethyl=4-nitrophenyl=carbonate at room temperature,
which was then stirred at the same temperature for 2
hours and 30 minutes. Chloroform and water were added
to the reaction mixture. The organic layer was
separated, washed sequentially with water, a 5%
potassium carbonate aqueous solution, and a saturated
sodium chloride aqueous solution, and then dried with
anhydrous magnesium sulfate, followed by distilling off
the solvent under reduced pressure. The resultant
residue was purified using silica gel column
chromatography (eluent; chloroform:ethanol =4:1) to
provide 0.58 g of 4-{3-[4-(3-{4-
[amino(ethoxycarbonylimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}-N'-(ethoxycarbonyl)benzamidine as
white solid form.
1H-NMR(CDCl3) .δ value: 1.20-
1.45(5H,m),1.36(3H,t,J=7.1Hz),1.36(3H,t,J=7.1Hz),1.65-
2.05(8H,m),2.45-2.55(2H,m),2.90-
3.00(2H,m),4.00(2H,t,J=6.5Hz),4.06(2H,t,J=6.3Hz),4.22(2
H,q,J=7.1Hz),4.22(2H,q,J=7.1Hz),6.92(2H,d,J=8.8Hz),6.93
(2H,d,J=8.8Hz),7.8 6(2H,d,J=8.8Hz),7.86(2H,d,J=8.8Hz).


As described in Example 31, 0.23 g of 4-{3-
[4-(3-{4-
[amino(pentyloxycarbonylimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}-N'-(pentyloxycarbonyl)benzamidine
as white solid form was obtained from 0.50 g of 4-{3-
[4-(3-{4-[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine and 1.20 g of 4-
nitrophenyl=pentyl=carbonate.
1H-NMR(CDCl3) .δ value:
0.91(3H,t,J=7.1Hz),0.91(3H,t,J=7.1Hz),1.20-
2.05(25H,m),2.45-2.55(2H,m),2.90-
3.00(2H,m),3.99(2H,t,J=6.6Hz),4.06(2H,t, J=6.6Hz) ,4.15(2
H,t,J=6.8Hz),4.15(2H,t,J=6.8Hz),6.92(2H,d,J=8.6Hz),6.93
(2H,d,J=8.6Hz),7.85(2H,d,J=8.6Hz),7.8 5(2H,d,J= 8.6Hz).

As described in Example 31, 0.33 g of 4-{3-
[4-{3-{4-

[amino(cyclohexyloxycarbonylimino)methyl]phenoxy}propyl
)-1-piperidinyl]propoxy}-N'-
(cyclohexyloxycarbonyl)benzamidine as white solid form
was obtained from 0.50 g of 4-{3-[4-(3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine and 0.76 g of
cyclohexyl=4-nitrophenyl=carbonate.
1H-NMR(CDCl3) .δ value: 1.15-2.10(33H,m),2.45-
2.55(2H,m),2.90-
3.00(2H,m),3.99(2H,t,J=6.5Hz),4.06(2H,t,J=6.3Hz),4.60-
4.75(2H,m),6.91(2H,d,J=8.8Hz),6.92(2H,d,J=8.8Hz),7.85(2
H,d, J=8.8Hz),7.85(2H,d,J=8.8Hz) .

As described in Example 31, 0.72 g of 4-{3-
[4-(3-{4-[amino(2,2,2-
trichloroethoxycarbonylimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}-N'-(2,2,2-
trichloroethoxycarbonyl)benzamidine as white solid form
was obtained from 0.50 g of 4-{3-[4-(3-{4-
[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine and 0.90 g of 4-
nitrophenyl=2,2,2-trichloroethyl=carbonate.
1H-NMR(CDCl3) .δ value: 1.20-1.45(5H,m),1.65-

2.05(8H,m),2.45-2.55(2H,m),2.90-
3.00(2H,m),4.01(2H,t,J=6.5Hz) ,4.08(2H,t,J=6.3Hz) ,4.87(2
H,s),4.87(2H,s),6.94(2H,d,J=8.8Hz),6.95(2H,d,J=8.8Hz),7
.89(2H,d,J=8.8Hz),7.90(2H, d, J=8.8Hz) .

To a chloroform (20 mL) solution of 2.17 g of
tert-butyl=[1-amino-1-(4-{3-[4-(3-hydroxypropyl)-1-
piperidinyl]propoxy}phenyl)methylidene]carbamate was
added 0.86 mL of triethylamine at room temperature, to
which 0.48 mL of methanesulfonyl chloride was added
dropwise under cooling with ice, followed by stirring
at room temperature for one hour. To this mixture were
added 0.36 mL of triethylamine and 0.20 mL of
methanesulfonyl chloride under cooling with ice, which
was then stirred at room temperature for 50 minutes.
Water was added to the reaction mixture. The organic
layer was separated, washed with a saturated sodium
chloride aqueous solution, and then dried with
anhydrous magnesium sulfate, followed by distilling off
the solvent under reduced pressure. The resultant
residue was dissolved in 20 mL of dimethylsulfoxide, to
which 1.43 g of potassium carbonate and 0.79 g of N',4-
dihydroxybenzamidine were added, followed by stirring

at 70°C for one hour. The reaction mixture was cooled
down to room temperature, to which water, chloroform,
and a sodium hydroxide aqueous solution were then
added. The organic layer was separated, washed
sequentially with a sodium hydroxide aqueous solution
and a saturated sodium chloride aqueous solution, and
then dried with anhydrous magnesium sulfate, followed
by distilling off the solvent under reduced pressure.
The resultant residue was purified using silica gel
column chromatography (eluent; chloroform:methanol =
4:1) and suspended in a mixture of chloroform and a
sodium hydroxide aqueous solution, followed by
collecting the solid matter by filtration to provide
0.38 g of tert-butyl=[l-amino-l-(4-{3-[4-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}phenyl)methylidene]carbamate as
white solid form.
1H-NMR(DMSO-d6) .δ value: 1.05-
1.40(5H,m),1.44(9H,s),1.60-1.80(4H,m),1.80-
1.95(4H,m),2.35-2.45(2H,m),2.80-
2.90(2H,m),3.96(2H,t,J=6.6Hz),4.06(2H,t,J=6.3Hz),5.70(2
H,s),6.90(2H,d,J=8.8Hz),6.98(2H,d,J=8.8Hz),7.58(2H,d,J=
8.8Hz),7.94(2H,d,J=8.8Hz),8.70-
9.30(2H,broad),9.43(lH,s).


To an ethanol (5 mL) suspension of 0.30 g of
tert-butyl=[1-amino-1-(4-{3-[4-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}phenyl)methylidene]carbamate was
added 5 mL of hydrochloric acid at room temperature,
which was then stirred at the same temperature for 15
hours. The solvent was distilled off under reduced
pressure, and water was subsequently added to the
resultant residue, which was then adjusted to pH 12.5
using a 1 mol/L sodium hydroxide aqueous solution. The
precipitate was collected by filtration and washed with
water to provide 0.22 g of 4-{3-[4-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine as white solid form.
1H-NMR(DMSO-d6) .δ value: 1.05-1.40(5H,m),1.60-
1.90(8H,m),2.35-2.45(2H,m),2.80-
2.90(2H,m),3.96(2H,t,J=6.3Hz) , 4.02(2H,t,J=6.2Hz),5.70(2
H,s),6.85-
6.95(4H,m),7.58(2H,d,J=8.8Hz),7.71(2H,d,J=8.5Hz).


To a tetrahydrofuran (20 mL) suspension of
1.59 g of tert-butyl=[1-amino-1-(4-{3-[1-{3-
hydroxypropyl)-4-
piperidinyl]propoxy}phenyl)methylidene]carbamate were
added 10 mL of methylene chloride and 0.63 mL of
triethylamine at room temperature. To this mixture was
dropwise added 0.35 mL of methanesulfonyl chloride
under cooling with ice, which was then stirred at room
temperature for 20 minutes. Chloroform and water were
added to the reaction mixture. The organic layer was
separated, washed with a saturated sodium chloride
aqueous solution, and then dried with anhydrous
magnesium sulfate, followed by distilling off the
solvent under reduced pressure. The resultant residue
was dissolved in 20 mL of N,N-dimethylformamide, to
which 1.05 g of potassium carbonate and 0.58 g of N',4-
dihydroxybenzamidine were then added, followed by
stirring at 60 to 70°C for one hour. The reaction
mixture was cooled down to room temperature, to which
chloroform and water were then added, followed by
collecting the precipitate by filtration to provide
2.23 g of tert-butyl=[1-amino-1-(4-{3-[l-(3-{4-

[amino(hydroxyimino)methyl]phenoxy}propyl)-4-
piperidinyl]propoxy}phenyl)methylidene]carbamate as
white solid form.
1H-NMR(DMSO-d6) .δ value: 1.00-
1.50(5H,m),1.44(9H,s),1.60-1.90(8H,m),2.35-
2.55(2H,m),2.80-2.90(2H,m),3.95-
4.05(4H,m),5.70(2H,s),6.90(2H,d,J=8.8Hz),6.98(2H,d,J=8.
8Hz),7.58(2H,d,J=8.8Hz),7.93(2H,d,J=8.8Hz),8.60-
9.4 0(2H,broad),9.43(lH,s).

To an ethanol (5 mL) suspension of 0.50 g of
tert-butyl=[1-amino-1-(4-{3-[1-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-4-
piperidinyl]propoxy}phenyl)methylidene]carbamate was
added 1 mL of hydrochloric acid at room temperature,
which was then stirred at the same temperature for 15
hours and 30 minutes. To the reaction mixture was
added 4 mL of hydrochloric acid at room temperature,
which was then stirred at the same temperature for 2
hours and 30 minutes. The solvent was distilled off
under reduced pressure, and water was then added,
followed by adjustment to pH 12.5 using a 5 mol/L
sodium hydroxide aqueous solution. The precipitate was

collected by filtration and washed with water. The
resultant solid was dissolved in 1 mol/L hydrochloric
acid, followed by distilling off the solvent under
reduced pressure before purification using silica gel
column chromatography (silica gel: ODS-A from YMC,
eluent; water:ethanol = 95:5). The eluate was
concentrated to about 20 mL, which was then adjusted to
pH 12.5 using a 5 mol/L sodium hydroxide aqueous
solution. The precipitate was collected by filtration
and washed with water to provide 0.17 g of 4-{3-[4-(3-
{4-[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}-N'-hydroxybenzamidine as white
solid form.
1H-NMR(DMSO-d6) .δ value: 1.05-1.40(5H,m),1.60-
1.85(8H,m),2.35-2.45(2H,m),2.80-2.90(2H,m),3.95-
4.05(4H,m),5.70(2H,s),6.90(2H,d,J=8.7Hz),6.91(2H,d,J=8.
7Hz),7.58(2H,d,J=8.7Hz) , 7.71(2H,d,J=8.7Hz) .

To a tetrahydrofuran (10 mL) solution of 1.06
g of benzyl=[1-amino-1-(4-{3-[1-(3-hydroxypropyl)-4-
piperidinyl]propoxy}phenyl)methylidene]carbamate was
added 0.39 mL of triethylamine under cooling with ice.
Thereto was dropwise added 0.22 mL of methanesulfonyl

chloride, which was then stirred at the same
temperature for 15 minutes. Chloroform and water were
added to the reaction mixture. The organic layer was
separated, washed with a saturated sodium chloride
aqueous solution, and then dried with anhydrous
magnesium sulfate, followed by distilling off the
solvent under reduced pressure. The resultant residue
was dissolved in 20 mL of N,N-dimethylformamide, to
which 0.36 g of N',4-dihydroxybenzamidine and 0.65 g of
potassium carbonate were then added, followed by
stirring at 60°C for 40 minutes. The reaction mixture
was cooled down to room temperature, to which ethyl
acetate and a 5% potassium carbonate aqueous solution
were then added. The precipitate was collected by
filtration and washed sequentially with water and ethyl
acetate to provide 0.55 g of benzyl=[1-amino-1-(4-{3-
[1-(3-{4-[amino(hydroxyimino)methyl]phenoxy}propyl)-4-
piperidinyl]propoxy}phenyl)methylidene]carbamate as
white solid form.
1H-NMR(DMSO-d6) .δ value: 1.05-1.40(5H,m),1.60-
1.90(8H,m),2.30-2.4 5(2H,m),2.80-2.90(2H,m),3.95-
4.05(4H,m),5.09(2H,s),5.70(2H,s),6.90(2H,d,J=8.9Hz),7.0
0(2H,d,J=8.9Hz),7.25-
4.45(5H,m),7.58(2H,d,J=8.9Hz),7.97(2H,d, J=8.9Hz),8.90-
9.60(3H,broad).


To an N,N-dimethylformamide (5 mL) solution
of 89 mg of N-(tert-butoxycarbonyl)-L-valine was added
66 mg of 1,1'-carbonyldiimidazole at room temperature,
which was then stirred at the same temperature for 2
hours and 30 minutes. To this mixture was added 0.20 g
of benzyl=[1-amino-1-(4-{3-[1-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-4-
piperidinyl]propoxy}phenyl)methylidene]carbamate at
room temperature, which was then stirred at the same
temperature for 3 days. Ethyl acetate and water were
added to the reaction mixture. The organic layer was
separated, washed with a saturated sodium chloride
aqueous solution, and then dried with anhydrous
magnesium sulfate, followed by distilling off the
solvent under reduced pressure. The resultant residue
was purified using silica gel column chromatography
(eluent; chloroform:methanol =4:1) to provide 0.34 g
of 4-{3-[4-(3-{4-
[amino(benzyloxycarbonylimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=0-[(2S)-2-(tert-
butoxycarbonyl) amino-3-methylbutylyl] oxime as white

solid form.
1H-NMR(CDCl3) .δ value:
1.01(3H,d,J=6.8Hz),1.05(3H,d, J=6.6Hz),1.25-
1.50(5H,m),1.45(9H,s),1.65-1.85(4H,m),1.95-
2.05(4H,m),2.10-2.25(lH,m) , 2.50-2.60(2H,m),2.95-
3.05(2H,m),3.99(2H,t,J=6.5Hz) ,4.04(2H,t,J=6.3Hz),4.25-
4.35(lH,m),5.05-5.20(3H,m) , 5.21(2H,s),6.85-
6.95(4H,m),7.25-7.50(5H,m) ,7.60-7.65(2H,m),7.80-
7.90(2H,m).

To a chloroform (10 mL) solution of 0.10 g of
4-{3-[4-(3-{4-
[amino(benzyloxycarbonylimino)methyljphenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=0-[(2S)-2-(tert-
butoxycarbonyl)amino-3-methylbutylyl]oxime was added 2
mL of trifluoroacetic acid at room temperature, which
was then stirred at the same temperature for one hour.
The solvent was distilled off under reduced pressure,
and 2.9 mol/L hydrogen chloride/ethanol was then added,
followed by distilling off the solvent under reduced
pressure. The resultant solid was suspended in
diisopropyl ether, and solid matter was collected by

filtration and washed with diisopropyl ether to provide
0.04 g of 4-{3-[4-(3-{4-
[amino(benzyloxycarbonylimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=0-((2S)-2-amino-3-
methylbutylyl)oxime hydrochloride as white solid form.
1H-NMR(DMSO-d6) .δ value:
1.01(3H,d,J=6.8Hz),1.03(3H,d,J=6.8Hz),1.30-
1.95(9H,m),2.10-2.35(3H,m),2.80-3.00(2H,m),3.10-
3.20(2H,m),3.4 5-3.55(2H,m),3.90-4.00(lH,m),4.05-
4.20(4H,m),5.36(2H,s),7.02(2H,d,J=9.0Hz),7.05-
7.15(2H,broad),7.15(2H,d,J=8.8Hz),7.35-
7.55(5H,m),7.69(2H,d,J=8.8Hz),7.81(2H,d,J=9.0Hz),8.60-
8.75(3H,broad).

To a tetrahydrofuran (20 mL) solution of 0.93
g of benzyl=[1-amino-l-(4-{3-[4-(3-hydroxypropyl)-1-
piperidinyljpropoxy}phenyl)methylidene]carbamate was
added 0.37 mL of triethylamine at room temperature, to
which 0.19 mL of methanesulfonyl chloride was then
added under cooling with ice, followed by stirring at
room temperature for 30 minutes. Ethyl acetate, water,
and a saturated sodium chloride aqueous solution were
added to the reaction mixture. The organic layer was

separated and dried with anhydrous magnesium sulfate,
followed by distilling off the solvent under reduced
pressure. The resultant oily matter was dissolved in
20 mL of N,N-dimethylformamide, to which 0.25 g of
N',4-dihydroxybenzamidine and 0.45 g of potassium
carbonate were then added, followed by stirring at 70°C
for 4 hours. Thereto was added 0.12 g of N',4-
dihydroxybenzamidine, which was then stirred at 70°C for
one hour. The reaction mixture was cooled down to room
temperature, and then added to a mixture of ethyl
acetate and water. The organic layer was separated,
washed with water, and then dried with anhydrous
magnesium sulfate, followed by distilling off the
solvent under reduced pressure. The resultant residue
was suspended in diisopropyl ether, and the solid
matter was collected by filtration and purified using
silica gel column chromatography (eluent;
chloroforrn:methanol = 5:1) to provide 0.22 g of
benzyl=[1-amino-1-(4-{3-[4-(3-{4-
[ amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}phenyl)methylidene]carbamate as
white solid form.
1H-NMR(CCDCl3) .δ value: 1.20-1.45(5H,m),1.65-
2.05(8H,m),2.45-2.55(2H,m),2.90-
3.00(2H,m),3.96(2H,t,J=6.5Hz),4.0 6(2H,t,J=6.3Hz),4.79(2
H,s),5.21(2H,s),6.89(2H,d,J=8.5Hz),6.93(2H,d,J=8.8Hz),7
.25-7.60(5H,m),7.55(2H,d,J=8.5Hz) ,7.85(2H,d, J=8.8Hz) .
Example 44


To an N-methyl-2-pyrrolidone (10 mL) solution
of 44 mg of N-(tert-butoxycarbonyl)-L-valine was added
33 mg of 1,1'-carbonyldiimidazole at room temperature,
which was then stirred at the same temperature for 3
hours. Thereto was added 0.10 g of benzyl=[1-amino-l-
(4-{3-[4-(3-{4-
[amino(hydroxyimino)methyl]phenoxyJpropyl)-1-
piperidinyl]propoxy}phenyl)methylidene]carbamate at
room temperature, which was then stirred at the same
temperature for 17 hours. The reaction mixture was
added to a mixture of ethyl acetate and water. The
organic layer was separated, washed with water, and
then dried with anhydrous magnesium sulfate, followed
by distilling off the solvent under reduced pressure.
The resultant residue was purified using silica gel
column chromatography (eluent; chloroform:methanol =
30:1) to provide 0.11 g of 4-(3-{4-[3-(4-{amino[(2S)-2-
(tert-butoxycarbonyl)amino-3-
methylbutylyloxyimino]methyl}phenoxy)propyl]-1-
piperidinyl}propoxy)-N'-benzyloxycarbonylbenzamidine as
white solid form.
1H-NMR(CDCl3) .δ value:

1.01(3H,d,J=6.8Hz),1.05(3H,d,J=6.8Hz),1.25-
1.45(5H,m),1.45(9H,s),1.60-2.25(9H,m),2.50-
2.60(2H,m),2.95-
3.05(2H,m),3.97(2H,t,J=6.5Hz),4.06(2H,t,J=6.3Hz),4.25-
4.35(lH,m),5.05-5.20(2H,m),5.21(2H,s),6.85-
6.95(4H,m),7.25-7.40(3H,m),7.40-7.50(2H,m),7.60-
7.70(2H,m),7.8 0-7.90(2H,m).

To a chloroform (1.0 mL) solution of 0.02 g
of 4-(3-{4-[3-(4-{amino[(2S)-2-(tert-
butoxycarbonyl)amino-3-
methylbutylyloxyimino]methyl}phenoxy)propyl]-1-
piperidinyl}propoxy)-N'-benzyloxycarbonylbenzamidine
was added 0.5 mL of trifluoroacetic acid at room
temperature, which was then stirred at the same
temperature for 2 hours. The solvent was distilled off
under reduced pressure, and 2.9 mol/L hydrogen
chloride/ethanol was then added to the resultant
residue, followed by distilling off the solvent under
reduced pressure. The resultant residue was suspended
in diisopropyl ether, and the solid matter was
collected by filtration and washed with diisopropyl

ether to provide 14 mg of 4-{3-[4-(3-{4-[amino((2S)-2-
amino-3-methylbutylyloxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}-N'-benzyloxycarbonylbenzamidine
hydrochloride as pale brown solid form.
1H-NMR(DMSO-d6) .δ value:
1.01(3H,d,J=6.8Hz),1.02(3H,d,J=6.8Hz),1.30-
1.65(5H,m),1.70-1.95(4H,m),2.15-2.35(3H,m) ,2.80-
2.95(2H,m),3.10-3.25(2H,m),3.4 5-3.55(2H,m),3.90-
4.25(5H,m),5.36(2H,s),7.00(2H,d,J=8.8Hz),7.15(2H,d,J=9.
0Hz),7.38-7.50(5H,m),7.65-
7.75(2H,m),7.83(2H,d, J=8.8Hz),8.55-8.65(2H,broad) .

To an acetic acid (15 mL) suspension of 1.00
g of 4-{3-[4-(3-{4-
[ amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=oxime was added 0.4 8 mL
of acetic anhydride at room temperature, which was then
stirred at the same temperature for one hour. The
solvent was distilled off under reduced pressure, and
the resultant residue was subsequently added to a
mixture of water and chloroform, which was then
adjusted- to pH 7.5 using a saturated sodium bicarbonate
aqueous solution. The precipitate was collected by

filtration and washed with water and chloroform to
provide 1.07 g of 4-{3-[4-(3-{4-
[amino(acetyloxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=0-acetyloxime as white
solid form.
1H-NMR(DMSO-d6) .δ value: 1.05-1.40(5H,m),1.60-
1.80(4H,m),1.80-1.95(4H,m),2.12(3H,s),2.12(3H,s),2.35-
2.45(2H,m),2.80-
2.90(2H,m),3.99(2H,t,J=6.5Hz),4.03(2H,t,J=6.3Hz),6.60-
6.80 (4H,broad),6.95-
7.00(4H,m),7.64(2H,d,J=7.6Hz) ,7.64(2H, d,J=7.6Hz) .

To a 2-propanol (20 mL) suspension of 4.00 g
of 4-{3-[4-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=oxime were added 4 mL of
water and 2.37 mL of hydrochloric acid at room
temperature, which was then heated to reflux for 3
minutes. The mixture was cooled down to room
temperature, to which 10 mL of 2-propanol was then
added under cooling with ice, followed by stirring at
the same temperature for one hour. The precipitate was
collected by filtration and washed with a 90% (v/v) 2-

propanol aqueous solution to provide 3.94 g of 4—{3—[4—
(3-{4-[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=oxime hydrochloride.
1H-NMR(DMSO-d6) .δ value: 1. 30-1. 65 (5H,m) , 1. 70-
1.95(4H,m),2.15-2.30(2H,m),2.8 0-3.00(2H,m),3.10-
3.20(2H,m),3.45-
3.55(2H,m),4.08(2H, t, J=6.3Hz) ,4.17(2H,t,J=6.1Hz),7.05-
7.20(4H,m),7.70-7.80(4H,m),8.50-9.50(4H,broad),10.70-
10.90(1H,broad),11.00-11.20(2H, broad) .

To an N-methyl-2-pyrrolidone (10 mL) solution
of 0.56 g of N-(tert-butoxycarbonyl)-L-phenylalanine
was added 0.35 g of 1,1'-carbonyldiimidazole at room
temperature, which was then stirred at the same
temperature for 2 hours. Thereto was added 0.50 g of
4-{3-[4-(3-{4-
[amino(hydroxyimino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamide=oxime at room
temperature, which was then stirred at the same
temperature for 3 hours and 40 minutes. The reaction
mixture was added to a mixture of ethyl acetate and
water, and the precipitate was collected by filtration
to provide 0.82 g of 4-(3-{4-[3-(4-{amino[(2S)-2-(tert-

butoxycarbonyl)amino-3-
phenylpropionyloxyimino]methyl}phenoxy)propyl]-1-
piperidinyl}propoxy)-N'-[(2S)-2-(tert-
butoxycarbonyl)amino-3-phenylpropionyloxy]benzamidine
as white solid form.
1H-NMR(CDCl3) .δ value: 1.24-
1.28(3H,m),1.43(9H,s),1.43(9H,s),1.65-1.85(4H,m),1.90-
2.10(5H,m),2.45-2.50(2H,m),2.90-2.95(2H,m),3.05-
3.40(5H,m),3.90-4.05(4H,m),4.75-4.85(4H,m),5.15-
5.20(2H,m),6.8 5-6.90(4H,m),7.20-7.35(10H,m),7.55-
7.60(4H,m):

To an ethanol (10 mL) suspension of 0.50 g of
4-(3-{4-[3-(4-{amino[(2S)-2-(tert-butoxycarbonyl)amino-
3-phenylpropionyloxyimino]methyl}phenoxy)propyl]-1-
piperidinyl}propoxy)-N'-[(2S)-2-(tert-
butoxycarbonyl) amino-3-phenylpropionyloxy]benzamidine
was added 5 mL of 2.9 mol/L hydrogen chloride/ethanol
under cooling with ice, which was then stirred at the
same temperature for 20 minutes, followed by allowing
to stand at room temperature for 26 hours. The solvent

was distilled off under reduced pressure, and the
resultant residue was suspended in chloroform, followed
by collecting solid matter by filtration to provide
0.50 g of 4-{3-[4-(3-{4-[amino((2S)-2-amino-3-
phenylpropionyloxyimino) methyl ] phenoxy} propyl) -1-
piperidinyl]propoxy}-N'-((2S)-2-amino-3-
phenylpropionyloxy)benzamidine hydrochloride as
slightly pale yellow solid form.
1H-NMR(DMSO-d6) .δ value: 1.35-1.40(2H,m),1.55-
1.70(3H,m),1.70-1.95(5H,m),2.20-2.30(2H,m),2.85-
2.95(2H,m),3.10-3.35(4H,m),3.45-3.50(2H,m),4.05-
4.15(4H,m),4.15-4.25(3H,m),7.10-7.20(4H,m),7.25-
7.40(10H,m),7.70-7.80(4H,m).
Formulation Example 1
In water for injection were dissolved 1.25 g
of the compound obtained in Example 2 and 5.0 g of D-
mannitol so as to provide a total amount of 100 ml.
The solution was filtered through a 0.22-|am membrane
filter, and 10 mL of the resultant drug solution was
packed in an ampule and sealed, followed by steam
sterilization to provide an injection.
Formulation Example 2
In water for injection were dissolved 1.02 g
of the compound obtained in Example 10 and 31.5 g of
sodium chloride so as to provide a total amount of 3.5
L. The solution was filtered through a 0.22-µm
membrane filter, and 10 mL of the resultant drug
solution was packed in an ampule and sealed, followed

by steam sterilization to provide an injection.
Formulation Example 3
There were mixed 500 mg of the compound
obtained in Example 1, 350 mg of lactose, 250 mg of
corn starch, and 400 mg of crystalline cellulose (trade
name: Ceolus .PH101; Asahi Kasei Chemicals Corporation) ,
to which 0.6 mL of a 5% hydroxypropylcellulose aqueous
solution and water were then added before kneading.
The resultant mixture was dried at 60°C, followed by
adding 100 mg of crospovidone (trade name: Kollidon CL:
BASF), 100 mg of light anhydrous silicic acid, and 20
mg of magnesium stearate before mixing. Into the form
of a round tablet 8 mm in diameter was tableted 175 mg
of the mixture to provide a tablet.
Formulation Example 4
There were mixed 500 mg of the compound
obtained in Example 1, 200 mg of lactose, and 530 mg of
corn starch, to which 0.6 mL of a 5%
hydroxypropylcellulose aqueous solution and water were
then added before kneading. The resultant mixture was
dried at 60°C, followed by adding 70 mg of crosspovidone
(trade name: Kollidon CL: BASF), 180 mg of crystalline
cellulose (trade name: Ceolus PH302; Asahi Kasei
Chemicals Corporation), and 20 mg of magnesium stearate
before mixing. In a No. 3 gelatine capsule was packed
150 mg of the mixture to provide a capsule.
INDUSTRIAL APPLICABILITY

The compound of the present invention is
useful as an excellent antifungal agent because it is
highly active against fungi including those resistant
to azole agents and has excellent physical properties
as well as being highly safe in the repeated-dose
toxicity tests, and also useful as an antiprotozoan
because it has an excellent antiprotozoan activity.
BRIEF DESCRIPTION OF THE DRAWING
Fig. 1 is a powder X-ray diffraction pattern
of 4-{3-[4-(3-{4-[amino(imino)methyl]phenoxy}propyl)-1-
piperidinyl]propoxy}benzamidine trihydrochloride
pentahydrate.

WE CLAIM:
1. 4-{3-[4-(3-{4-[amino(imino)methyl]phenoxy}-propyl)-1-
piperidinyl]propoxy}benzamidine trihydrochloride pentahydrate.


An arylamidine derivative represented by the
general formula (wherein R1 represents optionally
protected or substituted amidino; and R2 and R3 are the
same or different and each represents hydrogen or
halogeno) or a salt of the derivative. The derivative
and salt have potent activity against fungi including
ones having tolerance to azole type drugs and further
have high safety and excellent properties in a repeated
dose toxicity test. They are hence useful as an
excellent antifungal.

Documents:

03827-kolnp-2006-abstract.pdf

03827-kolnp-2006-claims.pdf

03827-kolnp-2006-correspondence others-1.1.pdf

03827-kolnp-2006-correspondence others.pdf

03827-kolnp-2006-correspondence-1.2.pdf

03827-kolnp-2006-correspondence-1.3.pdf

03827-kolnp-2006-correspondence-1.4.pdf

03827-kolnp-2006-description(complete).pdf

03827-kolnp-2006-drawings.pdf

03827-kolnp-2006-form-1.pdf

03827-kolnp-2006-form-18.pdf

03827-kolnp-2006-form-2.pdf

03827-kolnp-2006-form-26.pdf

03827-kolnp-2006-form-3.pdf

03827-kolnp-2006-form-5.pdf

03827-kolnp-2006-international search authority report-1.1.pdf

03827-kolnp-2006-international search authority report.pdf

03827-kolnp-2006-pct others.pdf

03827-kolnp-2006-pct request others.pdf

03827-kolnp-2006-priority document.pdf

3827-KOLNP-2006-ABSTRACT 1.1.pdf

3827-kolnp-2006-abstract-1.2.pdf

3827-kolnp-2006-amanded claims.pdf

3827-kolnp-2006-cancelled pages-1.1.pdf

3827-KOLNP-2006-CANCELLED PAGES.pdf

3827-KOLNP-2006-CLAIMS 1.1.pdf

3827-kolnp-2006-correspondence.pdf

3827-KOLNP-2006-DESCRIPTION (COMPLETE) 1.1.pdf

3827-kolnp-2006-description (complete)-1.2.pdf

3827-kolnp-2006-examination report.pdf

3827-kolnp-2006-form 1-1.3.pdf

3827-KOLNP-2006-FORM 1.1.2.pdf

3827-kolnp-2006-form 18.pdf

3827-kolnp-2006-form 2-1.2.pdf

3827-KOLNP-2006-FORM 2.1.1.pdf

3827-kolnp-2006-form 26.pdf

3827-KOLNP-2006-FORM 3.1.1.pdf

3827-kolnp-2006-form 3.pdf

3827-kolnp-2006-form 5.pdf

3827-kolnp-2006-granted-abstract.pdf

3827-kolnp-2006-granted-claims.pdf

3827-kolnp-2006-granted-description (complete).pdf

3827-kolnp-2006-granted-form 1.pdf

3827-kolnp-2006-granted-form 2.pdf

3827-kolnp-2006-granted-specification.pdf

3827-KOLNP-2006-OTHERS.pdf

3827-kolnp-2006-petition under rule 137.pdf

3827-kolnp-2006-reply to examination report-1.1.pdf

3827-KOLNP-2006-REPLY TO EXAMINATION REPORT.pdf

3827-kolnp-2006-reply to examination report1.2.pdf

3827-kolnp-2006-translated copy of priority document.pdf


Patent Number 249525
Indian Patent Application Number 3827/KOLNP/2006
PG Journal Number 43/2011
Publication Date 28-Oct-2011
Grant Date 25-Oct-2011
Date of Filing 19-Dec-2006
Name of Patentee M/S TOYAMA CHEMICAL CO., LTD.
Applicant Address 2-5-3-CHOME, NISHISHINJUKU, SHINJUKU-KU, TOKYO
Inventors:
# Inventor's Name Inventor's Address
1 KAZUYA HAYASHI 2764-4, SUMIYOSHI, UOZU-SHI, TOYAMA
2 SAYURI UEHARA 682-22, HORIKAWAKOIZUMICHO, TOYAMA-SHI TOYAMA
3 TEIICHI MORITA 477-1, AKADA, TOYAMA-SHI, TOYAMA
4 KAZUTO KUNITANI 2731-1,SANGA, KOSUGI-MACHI, IMIZU-GUN, TOYAMA
PCT International Classification Number C07D 211/22
PCT International Application Number PCT/JP2005/011809
PCT International Filing date 2005-06-28
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 2004-193386 2004-06-30 Japan