?2764126 Summary - Canadian Patents Database (2023)

Note: Descriptions are shown in the official language in which they were submitted.

CA 02764126 2011-11-30

DIPHENYL SULFIDE DERIVATIVES AND MEDICINES CONTAINING
SAID: AS ACTIVE INGREDIENT

TECHNICAL FIELD
[0001]

The present invention relates to a novel diphenyl sulfide
derivative that is effective as a medicine, or a pharmaceutically
acceptable salt or hydrate thereof, and a sphingosine-1 -phosphate
3 (S1P3) receptor-antagonist and a medicine containing the same as
an active ingredient.

BACKGROUND ART
[0002]

Sphingosine-l-phosphate (SIP) was considered to be merely an
intermediary metabolite in sphingosine metabolism. However, it has
been reported that SIP has a cell growth promoting action and a control
action of a cell motility function, and it is now clear that SIP
is a new lipid mediator that exhibits various physiological actions,
such as an apoptosis action, a cell morphology regulation action,
and vasoconstriction (Non-Patent Literatures 1 and 2).

[0003]

This SIP combines two actions, an action as an intracellular
second messenger and an action as an intercellular mediator. Studies
into SIP' s action as an intercellular mediator are especially active.
It has been reported that information is transmitted via a plurality
of G protein-coupled receptors present on the cell membrane
surface(Endothelial Differentiation Gene, EDG)(Non-Patent
l

CA 02764126 2011-11-30

Literatures 1 and 3). Currently, five sub-types of SiP receptors
are known, including Edg-1, Edg-3, Edg-5, Edg-6, and Edg-8 which
are called as S1P1, S1 P'3' S1P2, S1P4, and S1P5, respectively.
[0004]

From various studies into these SiP receptors, it has been
reported that so-called Sip receptor regulator, which exhibits an
agonistic or antagonistic action against this receptor, is effective
against a wide range of diseases. Patent Literature 2 and Non-Patent
Literatures 4 to 7 report that the S1P3 antagonist is effective as
a therapeutic or preventive medicine for respiratory tract
contraction, bronchial asthma, chronic obstructive pulmonary
disease (COPD), pulmonary emphysema, tracheal stenosis, diffuse
panbronchiolitis, bronchitis resulting from infection, connective
tissue disease, or transplantation, diffuse pulmonary
hamartoangiomyomatosis, adult respiratory distress syndrome (ARDS),
interstitial pneumonitis, lung cancer, pneumonia hypersensitivity,
idiopathic interstitial pneumonia, fibrosis of the lung, sepsis,
or cytokine storm caused by an influenza virus or RS virus infection.
[0005]

Further, Patent Literatures 3 to 6 show that the S1P3 antagonist
is also effective against arterial sclerosis, blood vessel intimal
hypertrophy, solid tumors, diabetic retinopathy, rheumatoid
arthritis, cardiacarrest,ischemia-reperfusion disorders, cerebral
blood vessel spasms after subarachnoid bleeding, angina pectoris
or myocardial infarction caused by coronary vessel spasms,
2

CA 02764126 2011-11-30

glomerulonephritis, thrombosis, lung disease caused by pulmonary
edema such as ARDS, cardiac arrhythmia, eye disease, eye hypertension,
glaucoma, glaucomatous retinopathy, optic neuropathy, macula-lutea
degeneration and the like.

[00061
Further, although currently there are recombinants form of human
activated protein C (rhAPC) inmedicines thatare effective as sepsis
therapeutic medicines, rhAPC may also cause hemorrhaging as a side
effect. Therefore, there is a need to develop a novel sepsis
therapeutic or preventive medicine that does not exhibit such side
effects. Non-Patent Literatures 5 and 7 report that the S1P3
receptor contributes to multiple organ failure caused by sepsis
based on analysis using S1P3 knockout mice, thereby suggesting that
the S1P3 antagonist may be effective as a sepsis therapeutic or
preventive medicine. In addition, it has been reported that the
S1P1 antagonist increases vascular wall permeability, and causes
pulmonary edema (Non-Patent Literature 8). Therefore, in order
for a novel sepsis therapeutic or preventive medicine to have a
high level of safety, that therapeutic or preventive medicine should
have a weak S1P1 antagonistic action, preferably exhibit an S1P1
agonistic action, and more preferably not exhibit an action against
the S1P1 receptor.

[0007]

Known S1P receptor regulators include, for example, the
compounds represented by the following general formula (A) described
3

CA 02764126 2011-11-30
in Patent Literature 1,

[0008]
[Formula 1]

I`' x' N112
PC1(t7Rs)2 (A)
{-{2)n +Y

[0009]
(In the formula (A) , R1 represents a hydrogen atom, a halogen atom,
a halogenated or unhalogenated lower alkyl group having 1. to 4 carbon
atoms, a hydroxy group, a phenyl group, an aralkyl group, a lower
alkoxy group having 1 to 4 carbon atoms, a trifluoromethyloxy group,
an optionally substituted aralkyloxy group, an optionally
substituted phenoxy group, a cyclohexylmethyloxy group, an
optionally substituted aralkyloxy group, a pyridylmethyloxy group,
a cinnamyloxy group, a naphthylmethyloxy group, a phenoxymethylgroup,
a hydroxymethyl group, a hydroxyethyl group, a lower alkylthio group
having 1 to 4 carbon atoms, a lower alkylsulfinyl group having 1
to 4 carbon atoms, a lower alkylsulfonyl group having 1 to 4 carbon
atoms, a benzylthio group, an acetyl group, a nitro group, or a cyano
group; R2 represents a hydrogen atom, a halogen atom, a halogenated
or unhalogenated lower alkyl group having 1 to 4 carbon atoms, a
lower alkoxy group having 1 to 4 carbon atoms, an aralkyl group,
or an aralkyloxy group; R3 represents a hydrogen atom, a halogen
atom, a trifluoromethyl group, a lower alkyl group having 1 to 4

CA 02764126 2011-11-30

carbon atoms, a lower alkoxy group having 1 to 4 carbon atoms, a
hydroxy group,a benzyloxy group, a phenyl group, a lower alkoxymethyl
group having 1 to 4 carbon atoms or a lower alkylthio group having
1 to 4 carbon atoms; R4 represents a hydrogen atom, a halogen atom,
a trifluoromethyl group, a lower alkyl group having 1 to 4 carbon
atoms, a lower alkoxymethyl group having 1 to 4 carbon atoms, a lower
alkylthiomethyl. group having 1 to 4 carbon atoms, a hydroxymethyl
group, a phenyl group, or an aralkyl group; R5 represents a hydrogen
atom or a lower alkyl group having 1 to 4 carbon atoms; X represents
0, S, SO, or SO2; and Y represents -CH2O-, -CH2-, -CH=CH-, -CF=CF-,
-CH2CH2-, -CH2CFH-, -CH2CF2-, or -CH(OH)CF2-.).

[0010]

However, Patent Literature 1 does not include
2-aminophosphoric acid monoester derivatives or 3-aminophosphonic
acid derivatives having a diphenyl sulfide skeleton in which a
hydroxyl group is substituted for a phenyl group. Further, the fact
that 2-aminophosphoric acid monoester derivatives or
3-aminophosphonic acid derivatives having such a structure exhibit
an excellent S1P3 receptor-antagonistic action is also not known.
[0011.]

Other examples of known SiP receptor regulators include the
compounds represented by the following general formula (B) in Patent
Literature 6,

CA 02764126 2011-11-30
[0012]

[Formula 21

1 %Q X R2 NH;
'" ^ OPO(OH)2 (B)
H2 ~ 1
R3
[0013]

(In the formula (B), R1 represents a chlorine atom, a linear alkyl
group having 1 to 3 carbon atoms, or a trifluoromethyl group; R2
represents a fluorine atom or a chlorine atom; R3 represents a linear
alkyl group having 1 to 3 carbon atoms; X represents an oxygen atom
or a sulfur atom; and n denotes an integer of 2 or 3.).

[0014]

Further, among the compounds representedby the general formula
(B), it has been reported that the optically active compounds
represented by the general formula (Ba), have a weak S1P3 agonistic
action and an excellent agonistic action against S1P1 and/or S1P4.
[0015]

[Formula 3]

CI NH2

CH2 ,OPO(OH)2 (Ba)
R3

[0016]
(in the formula (Ba),R1, R3, and X are as defined above.)
6

CA 02764126 2011-11-30
(0017]

However, the compounds having an inverse asymmetric center
to the optically active compounds represented by the general formula
(Ba), are not known. Further, the fact that such optically active
compounds exhibit an excellent S1P3 receptor-antagonistic action
is also not known.

(0018]
Patent Literature 1 W004074297 pamphlet
Patent Literature 2 W003020313 pamphlet

Patent Literature 3 Japanese Patent Application Laid-Open No.
2005-247691

Patent Literature 4 W007043568 pamphlet
Patent Literature 5 W006063033 pamphlet
Patent Literature 6 W008018427 pamphlet

Non-Patent Literature 1 Y. Takuma et al., Mol. Cell. Endocrinol.,
177, 3(2001).

Non-Patent Literature 2 Y. Igarashi, Ann, N.Y. Acad. Sci., 845,
19(1998).

Non-Patent Literature 3 H. Okazaki et al., Biochem. Biophs. Res.
Commun., 190, 1104(1993).

Non-Patent Literature 4 Y. Gon et.al., Proc Natl Acad Sci U S A.
102(26),9270(2005).

Non-Patent Literature 5 F. Nissen et al., Nature, 452, 654(2008)
Non-Patent Literature 6 D. Christina et al.,
Am.J.Pathol.,170(1),281(2007)

CA 02764126 2011-11-30

Non-Patent Literature? F. Nissen etal., Blood, 113(12), 2859(2009)
Non-Patent Literature 8 M.G.Sanna et al., Nature Chemical
biology, 2,434(2006)

SUMMARY OF THE INVENTION

PROBLEMS TO BE SOLVED BY THE INVENTION
[0019]

It is an object of the present invention to provide a diphenyl
sulfide derivative having an excellent S1P3 antagonistic activity.
MEANS FOR SOLVING THE PROBLEMS

[0020]

As a result of intensive studies into the S1P3 antagonist,
the present inventors discovered that a novel diphenyl sulfide
derivative has an excellent S1P3 antagonistic action, thereby
completing the present invention.

[0021]

Specifically, a first aspect of the invention relates to a
diphenyl sulfide derivative, or a pharmaceutically acceptable salt
or hydrate thereof, represented by the following general formula
(1),

[0022]
[Formula 4]

CA 02764126 2011-11-30
R',,~ S ti 2 R2

.' cH '' X-'**0 .c3H
NH2 H
[0023]

(In the formula (1) , Rl represents a hydrogen atom, a halogen atom,
a trif luoromethyl group, an optionally substituted alkyl group having
1to6carbon atoms, an optionally substituted cycloalkyl group having
3 to 6 carbon atoms, an optionally substituted aryl group having
6 to 10 carbon atoms, an optionally substituted alkoxy group having
1 to 4 carbon atoms, an optionally substituted benzyloxy group, an
optionally substituted acyl group having 1 to 4 carbon atoms, a cyano
group, or a carboxyl group; R2 represents an optionally substituted
alkyl group having 1 to 6 carbon atoms or an optionally substituted
alkenyl group having 2 to 6 carbon atoms; X represents a methylene
group which maybe substituted with 1 or 2 fluorine atoms or represents
an oxygen atom; Y represents a hydrogen atom or an optionally
substituted alkyl group having 1 to 6 carbon atoms; and Z represents
a halogen atom).

[0024]

Further, a second aspect of the invention relates to the
diphenyl sulfide derivative according to the first aspect of the
invention, or a pharmaceutically acceptable salt or hydrate thereof,
wherein the compound represented by the general formula (1) is
9

CA 02764126 2011-11-30
represented by the general formula (la),
[0025]

[Formula 5]

R CS !
2 O
R
CX.PL OH {1 )
NHz OH
[0026]

(In the formula (la), R1, R2, X, and Y are as defined above),
[0027]

In addition, a third aspect of the invention relates to the
diphenyl sulfide derivative according to the first aspect of the
invention, or a pharmaceutically acceptable salt or hydrate thereof,
wherein the compound represented by the general formula (1) is
represented by the general formula (lb),

[0028]
[Formula 6]

Rz O
R S*** 01
OH Xar- .OH (1 b)
NH2 OH
[0029]

(In the formula (lb), Xa represents an oxygen atom or -CH2-, and
R1, R2, and Y are as defined above).

CA 02764126 2011-11-30
[0030]

Still further, a fourth aspect of the invention relates to
the diphenyl sulfide derivative according to the first aspect of
the invention, or a pharmaceutically acceptable salt or hydrate
thereof, wherein the compound represented by the general formula
(1) is represented by the general formula (lc),

[0031]
[Formula 71

2 O
H''' O_P.-OH (1c)
NHS OH
[0032]

(In the formula (lc), R1 and R2 are as defined above).
[0033]

in addition, a fifth aspect of the invention relates to the
diphenyl sulfide derivative according to the fourth aspect of the
invention, or a pharmaceutically acceptable salt or hydrate thereof,
wherein R' represents a trifluoromethyl group, an alkyl group having
1 to 6 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms,
an aryl group having 6 to 10 carbon atoms, or a benzyloxy group.

Still further, a sixth aspect of the invention relates to the
diphenyl sulfide derivative according to the first aspect of the
invention, or a pharmaceutically acceptable salt or hydrate thereof,
11

CA 02764126 2011-11-30

wherein the compound represented by the general formula (1) is
(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-trifluoromethylphenylthi
o)phenyl]-2-propylbutylphosphoric acid monoester,
(R)-2-allyl-2-amino-4-[2-chloro-4-(2-hydroxy-5-tri.flu.oromethylp
henylthio)phenyl]butylphosphoric acid monoester,
(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-trifluoromethylphenylthi
o)phenyl]-2-methylbutylphosphoric acid monoester,
(S)-2-amino-4-[2-chloro-4-(2-hydr.oxy-5-isopropylphenylthio)phen
y1]-2-methylbutylphosphoric acid monoester,
(S)-2-amino-4-[2-chloro-4-(5-cyclopropyl-2-hydroxyphenylthio)ph
enyl]-2-methylbutylphosphoric acid monoester,
(S)-2-amino-4-[4-(5-benzyloxy-2-hydroxyphenylthio)-2-chlorophen
yl]-2-methylbutylphosphoric acid monoester,
(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-propylphenylthio)phenyl]
2-propylbutylphosphoric acid monoester,

(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-isopropylphenylthio)phen
yl]-2-propylbutylphosphoric acid monoester,
(S)-2-amino-4-[2-chloro-4-(5-cyclopropyl-2-hydroxyphenylthio)ph
enyl]-2-propylbutylphosphoric acid monoester,
(S)-2-amino-4-[4-(5-t-butyl-2-hydroxyphenylthio)-2-chlorophenyl
1-2-propylbutylphosphoric acid monoester,
(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-biphenylthio)phenyl]-2-p
.ropylbutylphosphoric acid monoester, or
(S)-2-amino-4-[4-(5-benzyloxy-2-hydroxyphenylthio)-2-chlorophen
yl]-2--propylbutylphosphoric acid monoester.

CA 02764126 2011-11-30

In addition, a seventh aspect of the invention relates to the
diphenyl sulfide derivative according to the first aspect of the
invention, or a pharmaceutically acceptable salt or hydrate thereof,
wherein the compound represented by the general formula (1) is
(-)-2-amino-4-[2-chloro-4-(2-hydroxy-5-trifluoromethylphenylthi
o)phenyl]-2-propylbutylphosphoric acid monoester,
(-)-2--a3.lyl-2-amino-4-[2-chloro-4-(2-hydroxy-5-trifluoromethylp
henylthio)phenyl]butylphosphoric acid monoester,
(-)-2-amino-4-[2-chloro-4-(2-hydroxy-5-trifluoromethylphenylthi
o)phenyl]-2-methylbutylphosphoric acid monoester,
(-)-2-amino-4-[2-chloro-4-(2-hydroxy-5-isopropylphenylthio)phen
yl]-2-methylbutylphosphoric acid monoester,
(-)-2-amino-4-[2-chloro-4-(5-cyclopropyl-2-hydroxyphenylthio)ph
enyl]-2-methylbutylphosphoric acid monoester,
(-)-2-amino-4-[4-(5-benzyloxy-2-hydroxyphenylthio)-2-chlorophen
yl)-2-methylbutylphosphoric acid monoester,
(-)-2-amino-4-[2-chloro-4-(2-hydroxy-5-propylphenylthio)phenyl]
-2-propylbutylphosphoric acid monoester,
(-)-2-amino-4-[2-chloro-4-(2-hydroxy-5-isopropylphenylthio)phen
yl]-2-propylbutylphosphoric acid monoester,
(-)-2-amino-4-[2-chloro-4-(5-cyclopropyl-2-hydroxyphenylthio)ph
enyl]-2-propylbutylphosphoric acid monoester,
(-)-2-amino-4-[4-(5-t-butyl-2-hydroxyphenylthio)-2-chlorophenyl
1-2-propylbutylphosphoric acid monoester,
(-)-2-amino-4-[2-chloro-4-(2-hydroxy-5-biphenylthio)phenyl]-2-
13

CA 02764126 2011-11-30

propylbutylphosphoric acid monoester, or
(-)-2-amino-4-[4-(5-benzyloxy-2-hydroxyphenylthio)-2-chlorophen
yl)-2-propylbutylphosphoric acid monoester.

Still further, an eighth aspect of the invention relates to
a medicine that is based on a sphingosine-1-phosphate 3 (S1P3)
receptor-antagonistic action, which comprises as an active
ingredient the diphenyl sulfide derivative according to any one of
the first to seventh aspects of the invention, or a pharmaceutically
acceptable salt or hydrate thereof.

In addition, a ninth aspect of the invention relates to the
medicine according to the eighth aspect of the invention, wherein
the medicine is a therapeutic or preventive medicine for respiratory
tract contraction, bronchial asthma, chronic obstructive pulmonary
disease (COPD), pulmonary emphysema, tracheal stenosis, diffuse
panbronchiolitis, bronchitis resulting from infection, connective
tissue disease, or transplantation, diffuse pulmonary
hamartoangiomyomatosis, adult respiratory distress syndrome (ARDS),
interstitial pneumonitis, lung cancer, pneumonia hypersensitivity,
idiopathic interstitial pneumonia, fibrosis of the lung, sepsis,
or cytokine storm caused by an influenza virus or RS virus infection.

Still further, a tenth aspect of the invention relates to the
medicine according to the eighth aspect of the invention, wherein
the medicine is a therapeutic medicine for arterial sclerosis, blood
vessel intimal hypertrophy, solid tumors, diabetic retinopathy,
rheumatoid arthritis, cardiac arrest, ischemia-reperfusion
14

CA 02764126 2011-11-30

disorders, cerebral blood vessel spasms af ter subarachnoidbleeding,
angina pectoris or myocardial infarction caused by coronary vessel
spasms, glomerulonephritis, thrombosis, lung disease caused by
pulmonary edema, cardiac arrhythmia, eye disease, eye hypertension,
glaucoma, glaucomatous retinopathy, optic neuropathy, or
macula-lutea degeneration.

In addition, an eleventh aspect of the invention relates to
the medicine according to the eighth aspect of the invention, wherein
the medicine is a therapeutic or preventive medicine for sepsis.

Further, a twelfth aspect of the invention relates to a
pharmaceutical composition comprising the diphenyl sulfide
derivative according to any one of the first to seventh aspects of
the invention, or a pharmaceutically acceptable salt or hydrate
thereof, and a pharmaceutically acceptable carrier.

In addition, a thirteenth aspect of the invention relates to
a use of the diphenyl sulfide derivative according to any one of
the first to seventh aspects of the invention, or a pharmaceutically
acceptable salt or hydrate thereof, in manufacture of a medicine
based on an S1P3 receptor-antagonistic action.

Further, a fourteenth aspect of the invention relates to the
use according to the thirteenth aspect of the invention, wherein
the medicine based on an S1P3 receptor-antagonistic action is a
therapeutic or preventive medicine for respiratory tract contraction,
bronchial asthma, chronic obstructive pulmonary disease (COPD),
pulmonary emphysema, tracheal stenosis, diffuse panbronchiolitis,

CA 02764126 2011-11-30

bronchitis resulting from infection, connective tissue disease, or
transplantation, diffuse pulmonary hamartoangiomyomatosis, adult
respiratory distress syndrome (ARDS), interstitial pneumonitis,
lung cancer, pneumonia hypersensitivity, idiopathic interstitial
pneumonia, fibrosis of the lung, sepsis, or cytokine storm caused
by an influenza virus or RS virus infection.

In addition, a fifteenth aspect of the invention relates to
the use according to the thirteenth aspect of the invention, wherein
the medicine based on an S1P3 receptor-antagonistic action is a
therapeutic medicine for arterial sclerosis, blood vessel intimal
hypertrophy, solid tumors, diabetic retinopathy, rheumatoid
arthritis, cardiac arrest, ischemia-reperfusion disorders, cerebral
blood vessel spasms after subarachnoid bleeding, angina pectoris
or myocardial infarction caused by coronary vessel spasms,
glomerulonephritis, thrombosis, lung disease caused by pulmonary
edema, cardiac arrhythmia, eye disease, eye hypertension, glaucoma,
glaucomatous retinopathy, optic neuropathy, or macula-lutea
degeneration.

Further, a sixteenth aspect of the invention relates to the
use according to the thirteenth aspect of the invention, wherein
the medicine based on an S1P3 receptor-antagonistic action is a
therapeutic or preventive medicine for sepsis.

In addition, a seventeenth aspect of the invention relates
to the diphenyl sulfide derivative according to any one of the first
to seventh aspects of the invention, or a pharmaceutically acceptable
16

CA 02764126 2011-11-30

salt or hydrate thereof, for use in inducing an. S1P3
receptor-antagonistic action.

Further, an eighteenth aspect of the invention relates to the
diphenyl sulfide derivative according to any one of the first to
seventh aspects of the invention, or a pharmaceutically acceptable
salt or hydrate thereof, for use in treatment or prevention of
respiratory tract contraction, bronchial asthma, chronic
obstructive pulmonary disease (COPD),pulmonary emphysema, tracheal
stenosis, diffuse panbronchiolitis, bronchitis resulting from
infection, connective tissue disease, or transplantation, diffuse
pulmonary hamartoangiomyomatosis, adult respiratory distress
syndrome (ARDS), interstitial pneumonitis, lung cancer, pneumonia
hypersensitivity, idiopathic interstitial pneumonia, fibrosis of
the lung, sepsis, or cytokine storm caused by an influenza virus
or RS virus infection.

In addition, a nineteenth aspect of the invention relates to
the diphenyl sulfide derivative according to any one of the first
to seventh aspects of the invention, or a pharmaceutically acceptable
salt or hydrate thereof, for use in treatment of arterial sclerosis,
blood vesselintimalhypertrophy,solid tumors, diabetic retinopathy,
rheumatoid arthritis, cardiac arrest, ischemia-reperfusion
disorders, cerebral blood vessel spasms after subarachnoid bleeding,
angina pectoris or myocardial infarction caused by coronary vessel
spasms, glomerulonephritis, thrombosis, lung disease caused by
pulmonary edema, cardiac arrhythmia, eye disease, eye hypertension,
17

CA 02764126 2011-11-30

glaucoma, glaucomatous retinopathy, optic neuropathy, or
macula-lutea degeneration.

Further, a twentieth aspect of the invention relates to the
diphenyl sulfide derivative according to any one of the first to
seventh aspects of the invention, or a pharmaceutically acceptable
salt or hydrate thereof, for use in treatment or prevention of sepsis.

In addition, a twenty-first aspect of the invention relates
to the diphenyl sulfide derivative according to any one of the first
to seventh aspects of the invention, or a pharmaceutically acceptable
salt or hydrate thereof, for inducing an S1P3 receptor-antagonistic
action.

Further, a twenty-second aspect of the invention relates to
amethod for inducing an S1P3 receptor-antagonistic action in a target,
comprising administrating effective amount of the diphenyl sulfide
derivative according to any one of the first to seventh aspects of
the invention, or a pharmaceutically acceptable salt or hydrate
thereof to a target requiring the induction of an S1P3
receptor-antagonistic action.

ADVANTAGEOUS EFFECTS OF THE INVENTION
[0034]

According to the present invention, a diphenyl sulfide
derivative having an excellent S1P3 antagonistic action can be
provided. The inventive compound is effective as a preventive or
a therapy f or sepsis, respiratory tract contraction, bronchial asthma,
chronic obstructive pulmonary disease (COPD), pulmonary emphysema,
18

CA 02764126 2011-11-30

tracheal stenosis, diffuse panbronchiolitis, bronchitis resulting
from infection, connective tissue disease, or transplantation,
diffuse pulmonary hamartoangiomyomatosis, adult respiratory
distress syndrome (ARDS), interstitial pneumonitis, lung cancer,
pneumonia hypersensitivity, idiopathic interstitial pneumonia,
fibrosis of the lung, or cytokine storm (hyperproduction) caused
by an influenza virus or RS virus infection, arterial sclerosis,
blood vesselintimalhypertrophy,solidtumors,diabetic retinopathy,
rheumatoid arthritis, cardiac arrest, ischemia-reperfusion
disorders, cerebral blood vessel spasms after subarachnoid bleeding,
angina pectoris or myocardial infarction caused by coronary vessel
spasms, glomerulonephritis, thrombosis, lung disease caused by
pulmonary edema such as ARDS, cardiac arrhythmia, eye disease, eye
hypertension, glaucoma, glaucomatous retinopathy, optic neuropathy,
and macula-lutea degeneration.

MODE FOR CARRYING OUT THE INVENTION
[00351

The "halogen atom" used in the present invention is a fluorine
atom, a chlorine atom, a bromine atom, or an iodine atom. Examples
of the "alkyl group having 1 to 6 carbon atoms" include a linear
hydrocarbon group having 1 to 6 carbon atoms, such as a methyl group,
an ethyl group, an n-propyl group, an n-butyl group, an n-pentyl
group, or an n-hexyl group, and a branched hydrocarbon group having
1 to 6 carbon atoms, such as an i-propyl group or a t-butyl group.
Examples of the "cycloalkyl group having 3 to 6 carbon atoms" include
19

CA 02764126 2011-11-30

a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, and
a cyclohexyl group. Examples of the "aryl group having 6 to 10 carbon
atoms" include a phenyl group and a naphthyl group. Examples of
the "alkoxy group having i to 4 carbon atoms" include a methoxy group,
an ethoxy group, an n-propoxy group, an n-butoxy group, an i-propoxy
group, and a t-butoxy group. Examples of the "alkenyl group having
2 to 6 carbon atoms" include an allyl group. Examples of the "acyl
group having 1 to 4 carbon atoms" include an acetyl group. Examples
of the "methylene group which may be substituted with 1 or 2 fluorine
atoms" include - CH2-, -CHF-, and -CF2-.

(0036]

Further, the "alkyl group having 1 to 6 carbon atoms," the
"cycloalkyl group having 3 to 6 carbon atoms, " the "aryl group having
6 to 10 carbon atoms, " the "alkoxy group having 1 to 4 carbon atoms, "
the "benzyloxy group, " the "acyl group having 1 to 4 carbon atoms, "
and the "alkenyl group having 2 to 6 carbon atoms " mayhave a subs ti tuent.
Examples of this "substituent" include a halogen atom, a
trifluoromethyl group, an alkyl group having 1 to 6 carbon atoms,
a cycloalkyl group having 3 to 6 carbon atoms, an aryl group having
6 to 10 carbon atoms, an alkoxy group having 1 to 4 carbon atoms,
a benzyloxy group, an acyl group having 1 to 4 carbon atoms, a cyano
group, an alkenyl group having 2 to 6 carbon atoms, a hydroxyl group,
a nitro group, and an amino group.

(0037]

In view of the object of the present invention, which is to

CA 02764126 2011-11-30

obtain an excellent S1P3 antagonistic action, it is preferred that
R1 be a trifluoromethyl group, an alkyl group having 1 to 6 carbon
atoms, a cycloalkyl group having 3 to 6 carbon atoms, an aryl group
having 6 to 10 carbon atoms, or a benzyloxy group. More preferably,
R' is a trifluoromethyl group, a benzyloxy group, an n-propyl group,
an i-propyl group, a t-butyl group, a cyclopropyl group, or a phenyl
group. Still more preferably, R' is a trifluoromethyl group, a
t-butyl group, or a phenyl group. Further, it is preferred that
R2 be an alkyl group having 1 to 6 carbon atoms or an alkenyl group
having 2 to 6 carbon atoms. More preferably, R2 is a methyl group,
an n-propyl group, or an allyl group. To achieve a high level of
safety, it is still more preferred that R2 be an n-propyl group.
In addition, it is preferred that R2 have the steric configuration
illustrated in the general formula (la). Moreover, it is preferred
that X be a methylene group or an oxygen atom, and more preferably
an oxygen atom. Still further, it is preferred that z be a chlorine
atom.

[0038]

Examples of a pharmaceutically acceptable salt in the present
invention include an acid addition salt, such as a hydrochloride
salt, a hydrobromide salt, an acetate salt, a trifluoroacetate salt,
a methanesulfonate salt, a citrate salt, or a tartrate salt, and
an alkaline addition salt, such as a sodium salt, a potassium salt,
a calcium salt, a magnesium salt, or an aluminum salt.

[0039]

CA 02764126 2011-11-30

According to the present invention, among the compounds
represented by the general formula (1), a compound in which X is
an oxygen atom and Y is a hydrogen atom, specifically, a compound
represented by the general formula(id) (excluding compounds in which
R1 is a carboxyl group), can be prepared based on the following
synthesis pathway A, for example.

[0040]
[Formula 8]

R ~ z R2
(1d)
C}H se OPO(OH)2
NH2
[0041]

(In the formula (ld) , R1 represents a hydrogen atom, a halogen atom,
a trifluoromethyl group, an optionally substituted alkyl group having
1to6carbon atoms, an optionally substituted cycloalkyl group having
3 to 6 carbon atoms, an optionally substituted aryl group having
6 to 10 carbon atoms, an optionally substituted alkoxy group having
1 to 4 carbon atoms, an optionally substituted benzyloxy group, an
optionally substituted acyl group having 1 to 4 carbon atoms, a cyano
group, or a carboxyl group; Rz represents an optionally substituted
alkyl group having 1 to 6 carbon atoms or an optionally substituted
alkenyl group having 2 to 6 carbon atoms; and z represents a halogen
atom.)

[0042]

CA 02764126 2011-11-30
<Synthesis pathway A>

[0043]
[Formula 9]

z
R3 R3 A 'A r. Z R3
! R2 "`N I P.,
N
R 1 A- f R r A-2 (6) oRs r
(2) (4)
R1 SH
A-3 A "'" Z A-4 A .~õ z '' ORS (9)
(7) N8R4 HR4 " A-5
(~) N.
R, A
R2 R# S R2
(10) NHR4 (12) NHR4
RI S Z
A-~,,,. I P I~ z
off O(OH)2
(1d) NH2
[0044]

In the synthesis pathway A, an optically active compound
represented by the general formula (4),

[0045]
[Formula 10]

CA 02764126 2011-11-30
R3
R2
N
N (4) .
QR`

[00461
(In the formula (4), R3 represents an optionally substituted alkyl
group having 1 to 6 carbon atoms; and R2 is as defined above.),
[0047]

can be prepared by reacting an optically active compound represented
by the general formula (2),

[00481
[Formula 11]
N
N=. y~ (2)
OR3

[0049]
(In the formula (2), R3 is as defined above.),
[0050]

with a compound represented by the general formula (3),
24

CA 02764126 2011-11-30
[0051]

[Formula 12]

R2A8 (3)
[0052]

(In the formula (3), Aa represents a typical leaving group such as
a halogen atom, a methanesulfonyloxy group, a
para-toluenesulfonyloxy group, or a trifluoromethanesulfonyloxy
group; and R2 is as defined above.) , in the presence of a base (Step
A-1).

[0053]

Specifically, first, ina reaction solvent suchas 1, 4-dioxane,
tetrahydrofuran, or diethyl ether, the compound represented by the
general formula (2) is treated at -78 C using abase. Then, a compound
represented by the general formula (3) is reacted at -78 C on the
obtained anion of the compound represented by the general formula
(2). Next, the temperature is gradually increased to normal
temperature to obtain a compound represented by the general formula
(4). Examples of the base that can be used in this reaction include
n-butyllithium and lithium diisopropylamide, and n-butyllithium is
preferred.

In the present specification, the term "normal temperature"
means 15 to 25 C as defined in the Japanese Pharmacopoeia.

CA 02764126 2011-11-30

In the synthesis pathway A, an optically active compound
represented by the general formula (6),

[0054]
[Formula 13]

N
I
OR 3

[0055]
(In the formula (6), Ab represents a typical leaving group such as
a halogen atom, a methanesulfonyloxy group, a
para-toluenesulfonyloxy group, or a trifluoromethanesulfonyloxy
group; and R2, R3, and Z are as defined above.),

[0056]
can be prepared by reacting the optically active compound represented
by the general formula (4) with a compound represented by the general
formula (5),

[0057]
[Formula 14]

CA 02764126 2011-11-30
[0058]

(In the formula (5), Ac represents a typical leaving group such as
a halogen atom, a methanesulfonyloxy group, a
para-toluenesulfonyloxy group, or a trifluoromethanesulfonyloxy
group; and Ab and Z are as defined above.), in the presence of a
base (Step A-2).

[00593

Specifically, first, in a reaction solvent such as 1, 4 -dioxane,
tetrahydrofuran, or diethyl ether, the compound represented by the
general formula (4) is treated at -78 C using a base. Then, the
compound represented by the general formula (5) is reacted at -78 C
on the obtained anion of the compound represented by the general
formula (4). Next, the temperature is gradually increased to normal
temperature to obtain the compound represented by the general formula
(6) . Examples of the base that can be used in this reaction include
n-butyllithium and lithium diisopropylamide, and n-butyllithium is
preferred.

In the synthesis pathway A, a compound represented by the
general formula (7), can be prepared by subjecting the compound
represented by the general formula (6) to acid hydrolysis, and then
protecting the amino group with a typical protecting reagent.
[0060]

[Formula 15]

CA 02764126 2011-11-30
R2
C02R3 (7)
HR'H

[0061]
(In the formula (7), R4 represents a general protecting group for
amino group; and Ab, R2, R3, and Z are as defined above.),

[0062]

The R4 in the formula is not especially limited as long as
it protects the amino group. For example, an acyl group, such as
an acetyl group, or a carbamate, such as t-butoxycarbonyl or
benzyloxycarbonyl, can be used (Step A-3).

Specifically, first, in an inorganic or organic acid, or in
a mixed solvent of an inorganic or organic acid and an organic solvent,
the compound represented by the general formula (6) is subjected
to acid hydrolysis at normal temperature. Here, as the inorganic
acid, hydrochloric acid, hydrobromic acid or the like can be used.
As the organic acid, trifluoromethanesulfonic acid or the like can
be used. Further, as the organic solvent, methanol, ethanol,
tetrahydrofuran, 1, 4-dioxane, ethyl acetate or the like can be used.
Among these, it is preferred to carry out the acid hydrolysis using
a hydrochloric acid in 1,4-dioxane.

Next, after neutralization with abase to obtain an amino ester,
28

CA 02764126 2011-11-30

this amino ester and an acyl chloride or an acid anhydride are reacted
at 0 C to normal temperature in a solvent to obtain the compound
represented by the general formula (7). Examples of the solvent
that can be used in this step include ethyl acetate, tetrahydrofuran,
N, N-dimethylf ormamide, 1, 4-dioxane, methylene chloride, chloroform,
methanol, ethanol, and acetonitrile. As the acyl chloride, acetyl
chloride, benzyloxycarbonyl chloride or the like can be used. As
the acid anhydride, acetic anhydride, di-t-butyldicarbonate or the
like can be used. Among these, it is preferred to carry out the
reaction using di-t-butyldicarbonate.

In the synthesis pathway A, a compound represented by the
general formula (8) can be prepared by reducing the compound
represented by the general formula (7) (Step A-4).

[0063]
[Formula 16]

t3H (8)
tNHRW

[0064]
(In the formula (8), Ab, R2, R , and z are as defined above.)
[0065]

CA 02764126 2011-11-30

For example, in a reaction solvent such as tetrahydrofuran,
1, 4-dioxane, ethanol, methanol or the like, the compound represented
by the general formula (7) is prepared by reduction using a reductant
at 0 C to the ref lux temperature, andpreferably at normal temperature.
Examples of the reductant that can be used include borane, alkyl
borane derivatives such as 9-borabicyclo[3.3.1]nonane (9-BBN),
metal hydride complexes such as diisobutylaluminum hydride
((iBu) 2A1H) , sodium borohydride (NaBH4) , lithium borohydride (LiBH4),
lithium aluminum hydride (LiAlH4) or the like. Preferably, the
reductant is lithium borohydride.

In the synthesis pathway A, a compound represented by the
general formula (10) (excluding compounds in which Rl is a carboxyl
group),

[0066]
[Formula 17]

OR H (10)
NHR4
[0067]

(In the formula (10), R5 represents a hydrogen atom or a general
protecting group for a phenolic hydroxyl group; and R1, R2, R4, and
Z are as defined above.),

[0068)

CA 02764126 2011-11-30

can be prepared by reacting the compound represented by the general
formula (8) and the compound represented by the general formula (9)
(excluding compounds in which R1 is a carboxyl group).

[0069]
[Formula 18]

R1 SH
(9)
R6

[0070]
(In the formula (9), R1 and R5 are as defined above.)
[0071]

The general protecting group for a phenolic hydroxyl group is not
especially limited as long as it protects a phenolic hydroxyl group.
For example, a methyl group, a benzyl group, a methoxymethyl group,
a tetrahydropyranyl group, a t-butyldimethylsilyl group, an acetyl
group, or a t-butoxycarbonyl group can be used (Step A-5).

For example, this reaction can be carried out in a reaction
solvent, such as toluene, N,N-dimethylformamide, 1,4-dioxane,
tetrahydrofuran, or diethyl ether, in the presence of an inorganic
or organic base using a catalyst at normal temperature to the ref lux
temperature. Examples of inorganic bases that can be used include
sodium carbonate or potassium t-butoxide. Examples of organic bases
that can be used include diisopropyethylamine. Further, examples
31

CA 02764126 2011-11-30

of the catalyst that can be used include palladium compounds, such
as tris(dibenzylideneacetone) dipalladium(0) or palladium(II)
acetate. Preferably, tris(dibenzylideneacetone) dipalladium(O)
is used.

A phosphine compound, such as
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene,
bis[2-(diphenylphosphino)phenyl] ether, or 1,1'-bis(di-t-butyl
phosphino)ferrocene, may be added to the reaction solvent as a
reaction accelerator.

In the synthesis pathway A, a compound represented by the
general formula (12) (excluding compounds in which R' is a carboxyl
group),

[0072]
[Formula 19]

R1 S Z

P::-O. R2 (12)
C?aRO(OR3)2
NFt4
[0073]

(in the formula (12) , R1, R2, R3, R , R5, and z are as defined above.) ,
[0074]

can be prepared by reacting the compound represented by the general
formula (10) (excluding compounds in which Rl is a carboxyl group)
and a compound represented by the general formula (11) (Step A-6).
[0075]

CA 02764126 2011-11-30
(Formula 20]

P(OR3)3 (11)
[0076]

(In the formula (11), R3 is as defined above.)
(0077]

For example, this reaction can be carried out in the presence
of carbon tetrabromide and pyridine, using no solvent or a solvent
such as methylene chloride, chlorof orm, acetonitrile, ethyl acetate,
tetrahydrofuran, or diethyl ether, at 0 C to normal temperature.

In the synthesis pathway A, a compound represented by the
general formula (1d) (excluding compounds in which R' is a carboxyl
group) can be prepared by subjecting the compound represented by
the general formula (12) (excluding compounds in which R'' is a carboxyl
group) to acid hydrolysis or treatment with a nucleophilic reagent,
such as trimethylsilyl bromide or trimethylsilyl iodide (Step A-7).

For the acid hydrolysis reaction, acid hydrolysis can be
carried out in an inorganic acid such as hydrochloric acid or
hydrobromic acid, or in a mixed solvent of an organic solvent such
as methanol or ethanol and an inorganic acid, at the ref lux temperature.
Further, a treatment using a nucleophilic reagent can be carried
out by reacting trimethylsilyl bromide or trimethylsilyl iodide at
0 C to normal temperature using acetonitrile or methylene chloride
as a preferred reaction solvent. Alternatively, the treatment with
33

CA 02764126 2011-11-30

a nucleophilic reagent can also be carried out by reacting
trimethylsilyl chloride and sodium bromide or trimethylsilyl
chloride and sodium iodide together.

In the synthesis pathway A, the compound represented by the
general formula (7) can also be prepared based on the following
synthesis pathway B, for example.

<Synthesis pathway B>
[0078]

[Formula 21]

Ab ` Z R3 A 3
A. ~N B-2 t(~41 N
N ( ) .~ N N
'4~ &RI *3
(13) B-i (14) R (15) R3
&3 Ab Z R2 Coo

(7) NHR4
[0079]

In the synthesis pathway B, an optically active compound
represented by the general formula (14),

[0080]
[Formula 221

Ab Z R3
N
R3 To,

CA 02764126 2011-11-30
[0081]

(In the formula (14), Ab, R3, and Z are as defined above.),
[0082]

can be prepared based on the same method as in Step A-2 using an
optically active compound represented by the general formula (13),
and the compound represented by the general formula (5) (Step B-1) .
[0083]

[Formula 23]
OR3

(13)
(00~
N,

3
[0084]

(In the formula (13), R3 is as defined above.)
[0085]

In the synthesis pathway B, an optically active compound
represented by the general formula (15),

[0086]
[Formula 24]

CA 02764126 2011-11-30
R3
=' = ( 15)
N
OR3
[0087)

(In the formula (15), Ab, R2, R3, and z are as defined above.),
[00881

can be prepared based on the same method as in Step A-1 using the
optically active compound represented by the general formula (14)
and the compound represented by the general formula (3) (Step B-2) .

In the synthesis pathway B, the compound represented by the
general formula (7) can be prepared based on the same method as in
Step A-3 using the compound represented by the general formula (15)
(Step B-3).

In the synthesis pathway A, the compound represented by the
general formula (10) (excluding compounds in which R1 is a carboxyl
group) can be prepared based on the following synthesis pathway C,
for example.

<Synthesis pathway C>
[0089)

[Formula 251

CA 02764126 2011-11-30
N
~ aN

R, s z TRIY R1 2 R3
(4) N
0-2
(16) (17) C7R3

R, ,~ S Z C-3 R1 S Z
RBI
C07R3R2
OH
4N4R4
(10) NHR4
[0090]

In the synthesis pathway C, an optically active compound
represented by the general formula (17) (excluding compounds in which
R1 is a carboxyl group),

[0091]
[Formula 26]

.r OR C)~ N (17)
OR3
[0092]

(In the formula (17), R', R2, R3, R5, and Z are as defined above.) ,
[0093]

can be prepared based on the same method as in Step A-2 using the
optically active compound represented by the general formula (4)
37

CA 02764126 2011-11-30

and a compound represented by the general formula (16) (excluding
compounds in which R1 is a carboxyl group) (Step C-1).

[00947
[Formula 27]

R z
(16)
Al

(0095]
(In the formula (16), R1, R5, Ac, and z are as defined above.)
(00967

In the synthesis pathway C, a compound represented by the
general formula (18) (excluding compounds in which R1 is a carboxyl
group), can be prepared based on the same method as in Step A-3 using
the compound represented by the general formula (17) (excluding
compounds in which R1 is a carboxyl group) (Step C-2).

[0097]
(Formula 28]

,,.2 (18)
s
R5 C02R
HR4
[0098]

(In the formula (18) , R1, R2, R3, R4, R5, and Z are as defined above.)
38

CA 02764126 2011-11-30
[0099]

In the synthesis pathway C, the compound represented by the
general formula (10) (excluding compounds in which R' is a carboxyl
group) can be prepared based on the same method as in Step A-4 using
the compound represented by the general formula (18) (excluding
compounds in which R' is a carboxyl group) (Step C-3).

In the synthesis pathway C, the compound represented by the
general formula (18) (excluding compounds in which R1 is a carboxyl
group) can be prepared based on the following synthesis pathway D,
for example.

<Synthesis pathway D>
[0100]

[Formula 29]

9
N% R, 3 \ Z
A I '
3 D-2
R5~ ' Ro OR N
(18) (19) OR3 ,
R3 R1 S,
O'O(OR tdD-3 2
- R5 CO R3
N ~
(20) TR*3 (18) NHR4
[0101]

In the synthesis pathway D, an optically active compound
represented by the general formula (19) (excluding compounds in which
R1 is a carboxyl group), can be prepared based on the same method
39

CA 02764126 2011-11-30

as in Step A-2 using the optically active compound represented by
the general formula (13) and the compound represented by the general
formula (16) (excluding compounds in which R1 is a carboxyl group)
(Step D-1).

[0102]
[Formula 30]

R, C s .z F
N YOnR0*3
[0103]

(In the formula (19), R1, R3, R5, and Z are as defined above.)
[01043

In the synthesis pathway D, an optically active compound
represented by the general formula(20) (excluding compounds in which
R1 is a carboxyl group), can be prepared based on the same method
as in Step A-1 using the optically active compound represented by
the general formula (19) (excluding compounds in which R1 is a carboxyl
group) and the compound represented by the general formula (3) (Step
D-2).

[0105]
[Formula 31]

CA 02764126 2011-11-30
R
N (20)
Nr-

CSR
[01063

(In the formula (20), R1, R2, R3, R5, and Z are as defined above.)
[01071

In the synthesis pathway D, the compound represented by the
general formula (18) (excluding compounds in which R1 is a carboxyl
group) can be prepared based on the same method as in Step A-3 using
the compound represented by the general formula (20) (excluding
compounds in which R1 is a carboxyl group) (Step D-3).

In the synthesis pathway A, among the compounds represented
by the general formula (10) , a compound in which R1 is a cyano group
or an acetyl group and R5 is a general protecting group for phenol,
specifically, a compound represented by the general formula (10a),
can be prepared by the following synthesis pathway E, for example.
[01083

[Formula 321

CA 02764126 2011-11-30
R18 z

HR'
[0109]

(In the formula (10a) , Rla represents an acetyl group or a cyano group
and Rya represents a general protecting group for a phenolic hydroxyl
group; and R2, R4, and Z are as defined above.)

[0110]

Rsa is not especially limited, as long as it protects a phenolic
hydroxyl group. For example, a methyl group, a benzyl group, a
methoxymethyl group, a tetrahydropyranyl group, a
t-butyldimethylsilyl group, an acetyl group, or a t-butoxycarbonyl
group can be used.

<Synthesis pathway E>
[0111]

[Formula 33]

CA 02764126 2011-11-30
Ab ` Z

R540 I SH (~3 t t' tt +o I I ~ ; 2 R2 E_2
H
(21) 1 (22) Nh

f2' S X E-3 H Z E-4
go W
H R w
(23) NNFY (24) M R;
3.
T(O,l I z ~~ -- .- R I r z ~1
H CH
(25) NW (108)
[0112]

In the synthesis pathway E, an optically active compound
represented by the general formula (22),

[0113]
[Formula 34]

R5bO 2
R2 (22)
1 O
HR4
[0114]

(In the formula (22), R5b represents a general protecting group for
a phenolic hydroxyl group; and R2, R4, and Z are as defined above.),
[0115]

can be prepared based on the same method as in Step A-5 using the
optically active compound represented by the general formula (8)
and a compound represented by the general formula (21).

CA 02764126 2011-11-30
[0116]

[Formula 35]

R5bO *;7H
bN
[0117]

(In the formula (21), Rsb is as defined above.)
[0118]

R5b is not especially limited, as long as it protects a phenolic
hydroxyl group. For example, a methyl group, a benzyl group, a
methoxymethyl group, a tetrahydropyranyl group, a
t-butyldimethylsilyl group, an acetyl group, or a t-butoxycarbonyl
group can be used (Step E-1).

in the synthesis pathway E, a compound represented by the
general formula (23), can be prepared by protecting the phenolic
hydroxyl group of the compound represented by the general formula
(22) (Step E-2).

[0119]
[Formula 36]

CA 02764126 2011-11-30
L
aa,~2 (23)
C7Rt H
NHR4
[0120)

(In the formula (23), R2, R4, RSa, RSb, and z are as defined above.)
[0121]

This reaction can be carried out by any technique that is
commonly used to protect a phenolic hydroxyl group. For example,
the reaction can be carried out in a solvent such as acetonitrile,
tetrahydrofuran, N,N-dimethylformamide, methylene chloride, or
chloroform, in the presence of an inorganic or organic base, by
reacting a compound represented by the general formula (22) with
a chloride or an acyl chloride. As the inorganic base, potassium
carbonate and the like canbe used. As the organic base, triethylamine,
diisopropylethylamine or the like can be used. Further, examples
of the chloride that can be used include methoxymethyl chloride,
t-butyldimethylsilyl chloride, and benzyl chloride. Examples of
the acyl chloride that can be used include acetyl chloride. Among
these, it is preferred to protect the phenolic hydroxyl group using
methoxymethyl chloride. In addition, the reaction can be carried
out by reacting at 0 C to normal temperature.

In the synthesis pathway E, a compound represented by the
general formula (24) , can be prepared by removing the RS' in the compound

CA 02764126 2011-11-30
represented by the general formula (23) (Step E-3).
[0122]

[Formula 37]

QH (24)
R'
NFR`
[0123]

(In the formula (24), R2, R4, RSa, and Z are as defined above.)
[0124]

The reaction is not especially limited, as long as the technique
is commonly used to remove a protecting group for a phenolic hydroxyl
group, and R 5a is not removed. An example will be described in which
R5b is a silyl protecting group, such as a t-butyldimethylsilyl group.
In this case, the deprotec tion reaction can be carried out in a reaction
solvent such as tetrahydrof uran, acetonitrile, or methylene chloride
using a fluorine compound, such as tetrabutylammonium fluoride or
hydrogen fluoride - pyridine, and preferably tetrabutylammonium
fluoride. This deprotection reaction can be carried out at from
0 C to the ref lux temperature, and preferably at 0 C.

In the synthesis pathway E, a compound represented by the
general formula (25), can be prepared by reacting the compound
represented by the general formula (24) with
N-phenyltrifluoromethanesulfonimide (Step E-4).

CA 02764126 2011-11-30
[0125]

[Formula 38]

TfONts Z
XR 2
(25)
C7R C3H
NHR4
[0126]

(In the formula (25), R2, R4, R5a, and Z are as defined above.)
[0127]

For example, this reaction can be carried out by reacting with
N-phenyltrifluoromethanesulfonimide in the presence of an organic
base such as pyridine, triethylamine or the like using a solvent
such as methylene chloride, chloroform, toluene or the like at 0 C
to 80 C, and preferably at normal temperature.

[0128]

In the synthesis pathway E, a compound represented by the
general formula (10a) can be prepared based on a known method using
zinc cyanide (e.g., Synth. Commun., 25, 3255-3261 (1995)) , or a known
method using a Heck reaction (e.g., J. Org., Chem., 55, 3654-3655
(1990)) from a compound represented by the general formula (2 5) (Step
E-5).

An example will be described in which Rla is a cyano group.
in this case, the reaction can be carried out in the presence of
zinc cyanide, in a reaction solvent such as toluene,
47

CA 02764126 2011-11-30
N,N-dimethylformamide, 1,4-dioxane, tetrahydrofuran or the like
using a catalyst at normal temperature to the ref lux temperature.
Examples of catalysts that can be used include palladium compounds
such as tet.rakistriphenylphosphine palladium(O) or tris
(dibenzylideneacetone) dipalladium(O), and preferably
tetrakistriphenylphosph.ine palladium(C). Further, a phosphine
compound, such as 1,l'-bis(diphenylphosphino)-ferrocene or
1,3-bis(diphenylphosphino)-propane, may be added to the reaction
solvent as a reaction accelerator.

Another example will be described in which R1a is an acetyl
group. In this case, the reaction can be carried out in the presence
of an organic base, using a catalyst and a reaction accelerator,
in a solvent such as toluene, N,N-dimethylformamide, 1,4-dioxane,
tetrahydrofuran or the like, by reacting with butyl vinyl ether.
As the organic base, triethylamine, diisopropylethylamine or the
like can be used. Further, as the catalyst, palladium (II) acetate
can be used. As the reaction accelerator,
1, 3-bis (diphenylphosphino) -propane may be used. The reaction can
be carried out at normal temperature to the ref lux temperature.

Among the compounds represented by the general formula (1),
a compound in which X is -CH2- or -CHF- and Y is a hydrogen atom,
specifically, a compound represented by the general formula (le)
(excluding compounds in which R' is a carboxyl group) , can be prepared
based on the following synthesis pathway F, for example.

CA 02764126 2011-11-30
[01291

[Formula 39]

R1 5 Z
R2
OH Po(OH)2 (1e)
~
,T T
[01301

(In the formula (le), R6 represents a hydrogen atom or a fluorine
atom; and R1, R2, and z are as defined above.)

[0131]
<Synthesis pathway F>
[0132]

[Formula 40]

R, F1 F F-2
RS ~RS CHO
(10) NHR4 (26) R"

R, Z F-3 R+
I R PO(OR3 )2 R2 / R OR3)2
29) R HN R8 (30) R'HM W
F-4 R,
z
H ffi PO(OH)2
(1e) t'2 W

in the synthesis pathway F, a compound represented by the
general formula (26) (excluding compounds in which R' is a carboxyl
group), can be prepared by oxidation of the compound represented
49

CA 02764126 2011-11-30

by the general formula (10) (excluding compounds in which R' is a
carboxyl group) (Step F-].).

[0133]
[Formula 41]

R2
CNO (26)
NHR4

[0134]
(In the formula (26), R1, R2, R4, R5, and Z are as defined above.)
[0135]

This reaction can be carried out using a generally used
oxidation method to generate aldehyde from alcohol. The oxidation
treatment can be carried out using a metal oxidant, such as a chromium
oxide - pyridine complex like pyridinium chlorochromate orpyridinium
dichromate, chromium oxide, silver carbonate, or manganese dioxide.
Alternatively, dimethyl sulfoxide oxidation using various dimethyl
sulfoxide activating agents, such as oxalyl chloride,
trifluoroacetic anhydride, acetic anhydride,
dicyclohexylcarbodiimide, or a sulfur trioxide - pyridine complex,
may be used.

In the synthesis pathway F, a compound represented by the
general formula (29) (excluding compounds in which R1 is a carboxyl
group),

CA 02764126 2011-11-30
[01.36]

[Formula 42]

R1 z
CCV4000 g2
PO(OR )2 (29)
RJR`
R4HN R6

(In the formula (29), R', R2, R3, R4, R5, R6, and z are as defined
above.),

[01371
can be prepared by, for example, reacting the compound represented
by the general formula (26) (excluding compounds in which R' is a
carboxyl group) and a compound represented by the general formula
(27), in a reaction solvent in the presence of a base.

[01381
[Formula 43]

0(OR3)2
(27)
O(OR3)2

[01393
(In the fourmula (27), R3 is as defined above.)
[01403

Further, the compound represented by the general formula (29)
51

CA 02764126 2011-11-30

(excluding compounds in which R' is a carboxyl group) can be prepared
by reacting the compound represented by the general formula (26)
(excluding compounds in which R1 is a carboxyl group) and a compound
represented by the general formula (28), in a reaction solvent in
the presence of chlorotrimethylsilane and a base (Step F-2).
[0141]

[Formula 44]

FBr2CPO(OR3)2 (28)
[0142]

(in the formula (28), R3 is as defined above.)
[0143]

Examples of the base that can be used in this reaction include
sodium hydride, potassium hydride, sodium alkoxide, potassium
alkoxide, or n-butyllithium, and preferably n-butyllithium. As the
reaction solvent, tetrahydrofuran, diethyl ether, or 1,4-dioxane
may be used. Further, the reaction temperature may be set to -78 C
to normal temperature.

In the synthesis pathway F, a compound represented by the
general formula (30) (excluding compounds in which R' is a carboxyl
group), can be prepared by reducing a compound represented by the
general formula (29) (excluding compounds in which R1 is a carboxyl
group) (Step F-3).

[0144]

CA 02764126 2011-11-30
[Formula 45]

S Z
2

R4HN R6
[0145]

(In the formula (30), R', R2, R3, R4, R5, R6, and z are as defined
above)

[0146]

For example, this reaction can be carried out in the presence
of a catalyst for catalytic hydrogenation, in a solvent such as ethanol,
methanol, tetrahydrof uran, N, N-dimethylf ormamide, or ethyl acetate,
under a normal pressure to increased hydrogen pressure at normal
temperature. Examples of catalysts for catalytic hydrogenation that
canbe used include palladium carbon, platinum carbon,platinum oxide,
rhodium carbon, or ruthenium carbon.

In the synthesis pathway F, a compound represented by the
general formula (1e) (excluding compounds in which R' is a carboxyl
group) can be prepared based on the same method as in Step A-7 using
the compound represented by the general formula (30) (excluding
compounds in which R1 is a carboxyl group) (Step F-4).

Among the compounds represented by the general formula (1),
a compound in which X is an oxygen atom and Y is an alkyl group having
1 to 6 carbon atoms, specifically, a compound represented by the
general formula (1f) (excluding compounds in which R1 is a carboxyl
53

CA 02764126 2011-11-30

group), can be prepared based on the following synthesis pathway
G, for example.

[0147]
[Formula 46]

Ri S Z

H OPO(OH)2
NH2
[0148]

(In the formula (1f), Ya represents an optionally substituted alkyl
group having 1 to 6 carbon atoms; and R1, R2, and z are as defined
above.)

[0149]
<Synthesis Pathway G>
[0150]

[Formula 47]

G=1 R` 2Ya G-2
(2R) Rd t3~) FtR"
12YISO G3
1'4Z RSja
(33) NHF14 (if) Nib

In the synthesis pathway G, a compound represented by the
general formula (31) (excluding compounds in which R1 is a carboxyl
group),

CA 02764126 2011-11-30
[0151]

[Formula 48]

R `' S Z R2 Ya

(31)
./ OR H'
NHR4
[0152]

(In the formula (31) , R1, R2, R4, R5, Ya, and Z are as defined above.) ,
[0153]

can be prepared by reacting the compound represented by the general
formula (26) (excluding compounds in which R1 is a carboxyl group)
and a compound represented by the general formula (32) (Step G-1).
[0154]

[Formula 49]

Ya-M (32)

(In the formula (32), M represents Li, MgCl, MgBr, or MgI; and Ya
is as defined above.)

[0155]

For example, this reaction can be performed at -78 C to normal
temperature using a solvent such as diethyl ether, 1,4-dioxane,
tetrahydrofuran or the like.

in the synthesis pathway G, a compound represented by the

CA 02764126 2011-11-30

general formula (33) (excluding compounds in which R1 is a carboxyl
group) , can be prepared by reacting the compound represented by the
general formula (31) (excluding compounds in which R' is a carboxyl
group) and the compound represented by the general formula (11) based
on the same method as in Step A-6 (Step G-2).

[0156]
[Formula 50]
4
R2ya 2ya
tJR5 PO(OR3)2 )
NHR4
[0157]

(In the formula (33) , R1, R2, R3, R4, R5, Ya, and z areas defined above. )
[0158]

In the synthesis pathway G, the compound represented by the
general formula (1f) (excluding compounds in which R1 is a carboxyl
group) can be prepared based on the same method as in Step A-7 using
the compound represented by the general formula (33) (excluding
compounds in which R1 is a carboxyl group) (Step G-3).

Among the compounds represented by the general formula (1),
a compound in which Xis -CFZ- and Y is a hydrogen atom, specifically,
a compound represented by the general formula (lg) (excluding
compounds in which R1 is a carboxyl group), can be prepared based
on the following synthesis pathway H, for example,

CA 02764126 2011-11-30
[01591

[Formula 51]

R2
C7H PO(OH)2 (19)
NH2FF
C01601

(In the formula (1g), R1, Rand z are as defined above.)
[0161]

<Synthesis Pathway H>
[0162]

[Formula 52]

F~ CM *S PO(CW)2 H-2
(26) R

Z e PO(OR3)2 H-3 RiNO~R Ft2 PO(OR3)2
(36) R~4 37)
t

'X'H'X POPH)2
(19) NH

In the synthesis pathway H, a compound represented by the
general formula (35) (excluding compounds in which R1 is a carboxyl
group),

[0163]

CA 02764126 2011-11-30
(Formula 53]

~ H
R, aX, z
ORt PO(OR), (35)
[0164]

(In the formula (35), R1, R2, R3, R4, RS, and Z are as defined above.) ,
[0165]

can be prepared by reacting the compound represented by the general
formula (26) (excluding compounds in which Rl is a carboxyl group)
and a compound representedby the general formula (34), in the presence
of a base (Step H-1).

[0166]
[Formula 54]

HF2CPO(OR3)2 (34)
[0167]

(In the formula (34), R3 is as defined above)
[0168]

Specifically, first,in a reaction solvent such as 1,4-dioxane,
tetrahydrofuran, diethyl ether or the like, the compound represented
by the general formula (34) is treated at -78 C using a base. Then,
the compound represented by the general formula (26) is reacted at
58

CA 02764126 2011-11-30

-78 C with the obtained anion of the compound represented by the
general formula(34)to obtain the compound represented bythe general
formula (35). Examples of the base that can be used include
n-butyllithium and lithium diisopropylamide, and lithium
diisopropylamide is preferred.

In the synthesis pathway H, a compound represented by the
general formula (36), can be prepared by halogenating a hydroxyl
group on the compound represented by the general formula (35) or
converting that hydroxyl group into a typical leaving group such
as a methanesulfonyloxy group (Step H-2).

[01691
[Formula 551

'0'0'50 = P (OR3)2 (36)
R4HN
[01701

(In the formula (36), R1, R2, R3, R4, R5, A , and Z are as defined
above.)

[01711

For example, this reaction can be carried out by, in a solvent
such as methylene chloride, chloroform, or toluene, in the presence
of an organic base, using a sulfonic acyl chloride or a sulfonic
acid anhydride at 0 C to 80 C, and preferably at normal temperature.
59

CA 02764126 2011-11-30

As the organic base, pyridine, triethylamine or the like can be used.
As the sulfonic acyl chloride, methanesulfonyl chloride or the like
can be used. Further, as the sulfonic acid anhydride,
methanesulfonic anhydride or the like can be used. In addition,
an alkali metal halide such as potassium iodide or sodium iodide
may be added to the solvent.

In the synthesis pathway H,.a compound represented by the
general formula (37) (excluding compounds in which R1 is a carboxyl
group) , can be prepared by removing the leaving group on the compound
represented by the general formula(36) (excluding compounds in which
R1 is a carboxyl group) (Step H-3).

[0172]
[Formula 56]

~2
ORS .r PO(OR3)2 (37)
R4HN

[0173]
(In the formula (37), R', R2, R3, R', R5, and Z are as defined above.)
[0174]

For example, this reaction can be carried out in a reaction
solvent such as N,N-dimethylformamide, dimethyl sulfide,
methylpyrrolidone or the like at 0 C to the ref lux temperature using
an alkyl borane derivative or a metal hydride complex. As the alkyl
borane derivative, borane or9-borabicyclo[3.3.1]nonane(9-BBN)can

CA 02764126 2011-11-30

be used. As the metal hydride complex, diisobutylaluminum hydride
((iBu) 2A1H) , sodium borohydride (NaBH4), lithiumborohydride (LiBH4),
lithium aluminum hydride (LiAlH4) or the like can be used. Among
these, it is preferred to perform this reaction using lithium
borohydride.

In the synthesis pathway H, the compound represented by the
general formula (1g) (excluding compounds in which R' is a carboxyl
group) can be prepared based on the same method as in Step A-7 using
the compound represented by the general formula (37) (excluding
compounds in which R1 is a carboxyl group) (Step H-4).

Among the compounds represented by the general formula (id) ,
a compound in which R1 is a carboxyl group, specifically, a compound
represented by the general formula (lh), can be prepared based on
the following synthesis pathway I, for example.

[01751
[Formula 57]
HOOC

~C(OH ' 2 (1h)
PO(OH)2
NH2
[0176]

(In the formula (lh), R2 and Z are as defined above.)
[0177]

<Synthesis Pathway I>
(0178]

[Formula 581

CA 02764126 2011-11-30
F3,,XS*, Z 1-1 - HOOQ,~S,,,,::~ ~Z
n(OH)2 H PO(C)H)2
R2 2
NH rv
(1i) (1h)
[01791

In the synthesis pathway I, a compound represented by the
general formula (1h),

[0180]
[Formula 591
HOQC ' , s z
,,~- ,. 2 (lh)
H OPO(OH)2
NH2
[01811

(In the formula (lh), R2 and z are as defined above.),
[01821

can be prepared by hydrolysis of a compound represented by
the general formula (1i), in the presence of a base (Step I-1).
[0183]

[Formula 601

F3C S Z
(ii)
R2 ~OPO(OH)2
NH2
[0184]

(In the formula (1i), R2 and Z are as defined above)
[0185]

For example, this reaction can be carried out in the presence
62

CA 02764126 2011-11-30

of a base such as sodium hydroxide or potassium hydroxide in an aqueous
solution or an aqueous alcohol solution at room temperature to the
ref lux temperature. The compound represented by the general formula
(1i) can be prepared based on the synthesis pathway A.

[0186]

Further, the synthesis method of the compound represent by
the general formula (16) can be carried out based on the method
described in WO 03029184, WO 03029205, WO 04026817, WO 04074297,
and WO 050444780 pamphlets.

[0187]

The diphenyl sulfide derivative according to the present
invention, or a pharmaceutically acceptable salt or hydrate thereof,
exhibits an excellent S1P3 antagonistic action. Therefore, a
medicine having at least one kind or more of such compounds as an
active ingredient is effective as a therapeutic or preventive medicine
for diseases for which it is known that an S1P3 antagonist is an
effective therapeutic or preventive medicine. Examples of diseases
for which it is known that an S1P3 antagonist is an effective
therapeutic or preventive medicine include

sepsis, respiratory tract contraction, bronchial asthma, chronic
obstructive pulmonary disease (COPD), pulmonary emphysema, tracheal
stenosis, diffuse panbronchiolitis, bronchitis resulting from
infection, connective tissue disease, or transplantation, diffuse
pulmonary hamartoangiomyomatosis, adult respiratory distress
syndrome (ARDS), interstitial pneumonitis, lung cancer, pneumonia
63

CA 02764126 2011-11-30

hypersensitivity, idiopathic interstitial pneumonia, fibrosis of
the lung, and cytokine storm caused by an influenza virus or RS virus
infection.

[0188]

Further, other than the above-described diseases, the medicine
according to the present invention is also effective for therapy
or prevention for diseases for which it is known that an S1P3
antagonistic action is effective. Examples of diseases for which
it is known that an S1P3 antagonistic action is effective include
arterial sclerosis, blood vessel intimal hypertrophy, solid tumors,
diabetic retinopathy, rheumatoid arthritis, cardiac arrest,
ischemia-reperfusion disorders, cerebral blood vessel spasms after
subarachnoid bleeding, angina pectoris or myocardial infarction
caused by coronary vessel spasms, glomerulonephritis, thrombosis,
lung disease caused by pulmonary edema such as ARDS, cardiac
arrhythmia, eye disease, eye hypertension, glaucoma, glaucomatous
retinopathy, optic neuropathy, and macula-lutea degeneration.
[0189]

The medicine according to the present invention may be
administered orally, or via a non-oral means, for example,
intrarectally, subcutaneously, intravenously, intramuscularly,
transdermally or the like.

[0190]

To use as a medicine, the compound according to the present
invention, or a pharmaceutically acceptable salt or hydrate thereof,
64

CA 02764126 2011-11-30

maybe in the form of any of a solid composition, a liquid composition,
or some other composition. The optimum form is selected as necessary.
The medicine composition according to the present invention can be
prepared by mixing the compound according to the present invention
with a pharmaceutically acceptable carrier. Specifically, the
composition according to the present invention can be prepared by
ordinary formulation techniques as a tablet, pill, capsule, granule,
powder, dispersion, liquid, emulsion, suspension, injection or the
like, by adding common diluents, fillers, binders, disintegrants,
coatings, sugar coatings, pH adjusting agents, dissolving agents,
or aqueous or non-aqueous solvents.

[0191]

The present invention will now be described based on the
following specific examples. However, the present invention is not
limited to these examples.

<Reference Example 1>
(2S,5R)-2-(4-bromo-2-chlorophenyl)ethyl-3,6-dimethoxy-2-methyl-
5-isopropyl-2,5-dihydropyrazine

[0192]
[Formula 61]

CA 02764126 2011-11-30
I
Me
N

1
Me

[0193)

Under an argon atmosphere, an n-butyllithium-hexane solution
(1.60 mol/L, 25.5 mL) was added at -78 C into a solution of
(5R)-3,6-dimethoxy-2-methyl-5-isopropyl-2,5-dihydropyrazine
(7.36 g) in tetrahydrofuran (160 mL) to form a reaction solution.
Next, this reaction solution was stirred at -78 C for 30 minutes.
Then, a solution of 4-bromo-2-chloro-l- (3 -iodoethyl) benzene (15.34
g) in tetrahydrofuran (26 mL) was added to the reaction solution,
and the reaction solution was stirred at -78 C for 30 minutes and
then at 0 C for 1 hour. Water was added to the reaction solution,
and the reaction solution was extracted with ethyl acetate. The
extract was washed with water and saturated brine in that order,
and then dried over anhydrous sodium sulfate. The anhydrous sodium
sulfate was removed by filtration, and then the solvent was removed
by distillation under reduced pressure. The resultant product was
purified by silica gel column chromatography (hexane : ethyl acetate
= 60 : 1) to obtain the target product (8.40 g) as a colorless oil.
1H NMR (CDC13, 400 MHz): 5 0.71 (3H, d, J = 6.7 Hz), 1.09 (3H, d,
J = 6.7 Hz), 1.35 (3H, s), 1.78 (1H, ddd, J = 12.8, 11.6, 4.9 Hz),

CA 02764126 2011-11-30

2.08 (1H, ddd, J = 12.8, 11.6, 4.9 Hz) , 2.21-2.31 (1H, m) , 2.35 (1H,
ddd, J = 13.4, 11.6, 4.9 Hz), 2.46 (1H, ddd, J = 13.4, 11.6, 4.9
Hz) , 3.68 (3H, s) , 3.69 (3H, s) , 4.00 (1H, d, J = 3.7 Hz) , 7.02 (1H,
d, J = 7.9 Hz), 7.27 (1H, dd, J = 7.9, 1.8 Hz), 7.47 (1H, d, J =
1.8 Hz).

ESIMS (+): 415 [M+H]+.
<Reference Example 2>
(2R,5R)-2-allyl-2-(4-bromo-2-chlorophenyl)ethyl-3,6-dimethoxy-5

-isopropyl-2,5-dihydropyrazine
[0194]

[Formula 62]

Br C
XNOM.
N
N '*r
Me
[0195]

The target product (6.04 g) was obtained as a colorless oil
by reacting
(5R)-2-allyl-3,6-dimethoxy-5-isopropyl-2,5-dihydropyrazine(3.64
g) in the same manner as in Reference Example I.

H NMR (CDC13, 400 MHz): 8 0.69 (3H, d, J = 6.7 Hz), 1.10 (3H, d,
J = 6.7 Hz) , 1.79 (1H, ddd, J = 12.8, 11.6, 4.9 Hz) , 2.02 (1H, ddd,
67

CA 02764126 2011-11-30

J = 12.8, 11.6, 4.9 Hz) , 2.27-2.48 (4H, m) , 2.54 (1H, dd, J = 13.4,
7.3 Hz), 3.69 (3H, s), 3.70 (3H, s), 3.95 (1H, d, J = 3.1 Hz), 4.97
(1H, dd, 10.4, 2.4 Hz), 5.01 (1H, d, J = 17.7 Hz), 5.61-5.72 (1H,
m), 7.01 (1H, d, J = 7.9 Hz), 7.27 (1H, dd, J = 7.9, 1.8 Hz), 7.47
(1H, d, J = 1.8 Hz).

ESIMS (+): 441 [M+H]+.
<Reference Example 3>
Methyl

(S)-4-(4-bromo-2-chlorophenyl)-2-t-butoxycarbonylamino-2-methyl
butyrate

[0196]
[Formula 63]

Br L;
cL'sco2Me
NHBoc
[0197]

0.5 mol/L hydrochloric acid (200 mL) was added to a solution
of the compound of Reference Example 1 (8.40 g) in 1,4-dioxane (400
mL) to form a first reaction solution. This first reaction solution
was stirred at normal temperature for 1 hour, and then left to stand
at normal temperature overnight. Then, the first reaction solution
was concentrated, neutralized with a saturated sodium hydrogen
68

CA 02764126 2011-11-30

carbonate aqueous solution, and extracted with ethyl acetate. The
extract was washed with water and saturated brine, and then dried
over anhydrous sodium sulfate. After the anhydrous sodium sulfate
was removed by filtration, the extract was concentrated, and the
resultant residue was dissolved in acetonitrile (16 mL).
Di-tert-butoxydicarbonate (11.0 g) was added to this solution to
forma second reaction solution. This second reaction solution was
stirred at normal temperature for 1 hour and then left to stand at
normal temperature overnight. Water was added to the second reaction
solution, and the second reaction solution was extracted with ethyl
acetate. The extract was washed with water and saturated brine in
that order, and then dried over anhydrous sodium sulfate. The
anhydrous sodium sulfate was removed by filtration, and then the
solvent was removed by distillation under reduced pressure. The
resultant product was purified by silica gel column chromatography
(hexane : ethyl acetate = 4 : 1) to obtain the target product (6.58
g) as a colorless oil.

lH NMR (CDC13, 400 MHz) : 1.45 (9H, s) , 1.58 (3H, s) , 2.09 (1H, ddd,
J = 13.4, 12.2, 5.5 Hz), 2.39 (1H, br s), 2.51 (1F1, td, J = 12.8,
4.9 Hz), 2.65 (1H, td, J = 12.8, 4.9 Hz), 3.75 (3H, s), 5.42 (1H,
br s), 7.04 (1H, d, J = 7.9 Hz), 7.30 (1H, dd, J = 7.9, 1.8 Hz),
7.48 (1H, d, J = 1.8 Hz).

ESIMS (+): 420 (M+H] '.
<Reference Example 4>
Methyl

CA 02764126 2011-11-30
(R)-2-allyl-4-(4-bromo-2-chlorophenyl)-2-t-butoxycarbonylamino
butyrate

[0198]
[Formula 64]

Br C1

2
IHBoc
[0199]

A solution of 50% trfluoroacetic acid - water (108 mL) was
added to the compound of Reference Example 2 (5.44 g) to form a first
reaction solution. This first reaction solution was stirred at
normal temperature for 1 hour, and then left to stand at normal
temperature overnight. Then, the first reaction solution was
neutralized with a saturated sodium hydrogen carbonate aqueous
solution, and extracted with ethyl acetate. The extract was washed
with water and saturated brine, and then dried over anhydrous sodium
sulfate. After the anhydrous sodium sulfate was removed by
filtration, the extract was concentrated, and the resultant residue
was dissolved in acetonitrile (86 mL). Di-tert-butoxydicarbonate
(11.0 g) was added to this solution to forma second reaction solution.
This second reaction solution was stirred at normal temperature for
1 hour and then left to stand at normal temperature overnight. Water

CA 02764126 2011-11-30

was added to the second reaction solution, and the second reaction
solution was extracted with ethyl acetate. The extract was washed
with water and saturated brine in that order, and then dried over
anhydrous sodium sulfate. The anhydrous sodium sulfate was removed
by filtration, and then the solvent was removed by distillation under
reduced pressure. The resultant product was purified by silica gel
column chromatography (hexane : ethyl acetate = 6 : 1) to obtain
the target product (6.16 g) as a colorless oil.

''H NMR (CDC13, 400 MHz): 5 1.45 (9H, s) , 2.08 (1H, ddd, J = 13.4,
11.0, 5.5 Hz), 2.39-2.51 (2H, m), 2.51-2.61 (1H, m), 2.67 (1H, td,
J = 12.8, 4.9 Hz), 3.00-3.14 (1H, m), 3.74 (3H, s), 5.07 (1H, d,
J = 4.9 Hz), 5.10 (1H, s), 5.52-5.69 (1H, m), 7.03 (1H, d, J = 7.9
Hz), 7.29 (1H, dd, J = 7.9, 1.8 Hz), 7.48 (1H, d, J = 1.8 Hz).
ESIMS (+): 446 [M+H]+.

<Reference Example 5>
(S)-4-(4-bromo-2-chlorophenyl)-2-t-butoxycarbonylamino-2-methyl
butan-1-ol

[0200]
[Formula 65]

6r C1

H
NH oc

CA 02764126 2011-11-30
[0201]

Lithium borohydride (259 mg) was added to a solution of the
compound of Reference Example 3 (1.00 g) in tetrahydrofuran (24 mL)
under ice cooling to form a reaction solution. Next, ethanol (2.4
mL) was added dropwise to the reaction solution, and the reaction
solution was then stirred for 2 hours under ice cooling. A 10% citric
acid aqueous solution was added to the reaction solution, and the
reaction solution was extracted with ethyl acetate. The extract
was washed with water and saturated brine in that order, and then
dried over anhydrous sodium sulfate. The anhydrous sodium sulfate
was removed by filtration, and then the solvent was removed by
distillation under reduced pressure. The resultant product was
purified by silica gel column chromatography (hexane : ethyl acetate

2 : 1) to obtain the target product (775 mg) as a white solid.
'H NMR (CDC13, 400 MHz) : 8 1.24 (3H, s) , 1.44 (9H, s) , 1.81 (1H, ddd,
J = 13.4, 12.2, 5.5 Hz), 2.05 (1H, ddd, J = 13.4, 12.2, 5.5 Hz),
2.67 (1H, ddd, J = 13.4, 12.2, 5.5 Hz), 2.74 (1H, ddd, J = 13.4,
12.2, 5.5 Hz), 3.63-3.74 (2H, m), 4.07 (1H, br s), 4.67 (1H, s),
7.11 (1H, d, J = 8.6 Hz), 7.31 (1H, dd, J = 8.6, 1.8 Hz), 7.50 (1H,
d, J = 1.8 Hz).

ESIMS (+): 392 [M+H]'.
<Reference Example 6>
(R)-2-[2-(4_bromo-2-chlorophenyl)ethyl]-2-t-butoxycarbonylamino

-4-penten-1-ol
[0202]

CA 02764126 2011-11-30
[Formula 661

B H
NHBoC
[0203]

The target product (3.20 g) was obtained as a white powder
by reacting the compound of Reference Example 4 (6.16 g) in the same
manner as in Reference Example 5.

1H NMR (CDC13, 400 MHz) : 3 1.43 (9H, s) , 1.80-1.94 (2H, m) , 2.32 (1H,
td, J = 14.1, 7.9 Hz), 2.44 (1H, dd, J = 14.1, 6.7 Hz), 2.63-2.77
(2H, m), 3.69-3.79 (2H, m), 4.09 (1H, br s), 4.72 (1H, s) , 5.19 (1H,
dd, J = 6.1, 1.8 Hz), 5.22 (1H, s), 5.80-5.91 (1H, s), 7.11 (1H,
d, J = 7.9 Hz), 7.31 (1H, dd, J = 7.9, 1.8 Hz), 7.49 (1H, d, J =
1.8 Hz).

ESIMS (+): 418 [M+H]`.
<Reference Example 7>
1-(Methoxymethoxy)-4-propylbenzene

[0204]
[Formula 67)

CA 02764126 2011-11-30
OMOM

[0205]

Ethyldiisopropylamine (10. 3 mL) and chloromethyl methyl ether
(4.5 mL) were added to a solution of 4-propylphenol (4.09 g) in
methylene chloride (60 mL) under ice cooling to form a reaction
solution. This reaction solution was stirred for 15 minutes under
ice cooling, and then left overnight at normal temperature. Water
was added to the reaction solution, and the reaction solution was
extracted with ethyl acetate. The extract was washed with water
and saturated brine in that order, and then dried over anhydrous
sodium sulfate. The anhydrous sodium sulfate was removed by
filtration, and then the solvent was removed by distillation under
reduced pressure. The resultant product was purified by silica gel
column chromatography (hexane : ethyl acetate = 9 : 1) to obtain
the target product (4.50 g) as a colorless oil.

1H NMR (CDC13, 400 MHz): S 0.93 (3H, t, J = 7.3 Hz), 1.55-1.67 (2H,
m) , 2.53 (2H, t, J = 7.3 Hz), 3.48 (3H, s) , 5.15 (2H, s) , 6.95 (2H,
dt, J = 8.6, 2.4 Hz), 7.09 (2H, dt, J = 8.6, 2.4 Hz).

EIMS (+) : 180 [M]+.
<Reference Example 8>
1-Cyclopropyl-4-(methoxymethoxy)benzene

CA 02764126 2011-11-30
[0206]

[Formula 681

QMOM
(0207]

The target product (958 mg) was obtained as a colorless oil
by reacting 4-cyclopropylphenol (1.00 g) in the same manner as in
Reference Example 7.

1H NMR (CDC13, 400 MHz): 8 0.59-0.62 (2H, m), 0.86-0.93 (2H, m),
1.80-1.90 (1H, m), 3.47 (3H, s), 5.14 (2H, s), 6.94 (2H, dt, J =
9.2, 2.4 Hz), 7.01 (2H, dt, J = 9.2, 2.4 Hz).

EIMS (+): 178 [M]+.
<Reference Example 9>
2-(Methoxymethoxy)-5-methylbenzenethiol
[0208]

[Formula 69]

SH
C MOM

CA 02764126 2011-11-30
[0209)

Under an argon atmosphere, N,N,N',N'-tetramethyl-
ethylenediamine(375p.L)and an n-butyllithium-hexane solution (1. 60
mol/L, 1.5 mL) were added under ice cooling to a solution of
1-(methoxymethoxy)-4-methylbenzene (304 mg) in diethyl ether (10
mL) to forma reaction solution. This reaction solution was stirred
for 1.5 hours at normal temperature. Then, sulfur (80 mg) was added
to the reaction solution under ice cooling, and the reaction solution
was stirred for 12 hours. 1 mol/L hydrochloric acid was added to
the reaction solution, and the reaction solution was extracted with
ethyl acetate. The extract was washed with water and saturated brine
in that order, and then dried over anhydrous sodium sulfate. The
anhydrous sodium sulfate was removed by filtration, and then the
solvent was removed by distillation under reduced pressure. The
resultant product was purified by silica gel column chromatography
(hexane : ethyl acetate = 9 : 1) to obtain the target product (200
mg) as a colorless oil.

1H NMR (CDC13, 400 MHz) : S 2.24 (3H, s) , 3.50 (3H, s) , 3.75 (1H, s) ,
5.21 (2H, s), 6.88 (1H, dd, J = 8.6, 1.2 Hz), 6.97 (1H, d, J = 8.6
Hz), 7.07 (1H, d, J = 1.2 Hz).

EIMS (+) : 184 [M] +.
<Reference Example 10>
5-Ethyl-2-(methoxymethoxy)benzenethiol
[0210]

CA 02764126 2011-11-30
(Formula 70]

~aomom
(0211)

The target product (600 mg) was obtained as a colorless oil
by reacting 1-ethyl-4- (methoxymethoxy) benzene (665 mg) in the same
manner as in Reference Example 9.

1H NMR (CDC13, 400 MHz): 5 1.19 (3H, t, j = 7.9 Hz), 2.54 (2H, q,
J = 7.9 Hz), 3.51 (3H, s), 3.76 (1H, s), 5.22 (2H, s), 6.91 (1H,
dd, J = 7.9, 1.8 Hz), 7.00 (1H, d, J = 7.9 Hz), 7.09 (1H, d, J =
1.8 Hz).

EIMS (+) : 198 [M]+.
<Reference Example 11>
2-(Methoxymethoxy)-5-propylbenzenethiol
(0212]

[Formula 71]

OMOM

CA 02764126 2011-11-30
[0213]

The target product (687 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 7 (720 mg) in the same
manner as in Reference Example 9.

1H NMR (CDC13, 400 MHz) : 8 0.92 (3H, t, j = 7.3 Hz) , 1.55-1.65 (2H.
m) , 2.47 (2H, t, J = 7.3 Hz) , 3.51 (3H, s) , 3.76 (1H, s) , 5.21 (2H,
s), 6.88 (1H, dd, J = 8.6, 1.8 Hz), 6.99 (1H, d, J = 8.6 Hz), 7.07
(1H, d, J = 1.8 Hz).

EIMS (+): 212 [M]t.
<Reference Example 12>
2-(Methoxymethoxy)-5-isopropylbenzenethiol
[0214]

[Formula 72]

e%# i
10000
OMOM
[0215]

The target product (646 mg) was obtained as a colorless oil
by reacting 1- (methoxymethoxy) -4-isopropylbenzene (720 mg) in the
same manner as in Reference Example 9.

1H NMR (CDC13, 400 MHz) : 8 1.21 (6H, t, j = 6.7 Hz) , 2.74-2.83 (1H.
m) , 3.51 (3H, s), 3.78 (1H, s), 5.22 (2H, s) , 6.94 (1H, dd, J = 8.6,
2.4 Hz), 7.01 (1H, d, J = 8.6 Hz), 7.12 (1H, d, J = 2.4 Hz).

CA 02764126 2011-11-30
EIMS (+): 212 [M];.

<Reference Example 13>
5-Cyclopropyl-2-(methoxymethoxy)benzenethiol
[02161

[Formula 73]

SH
OMM
[0217]

The target product (210 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 8 (450 mg) in the same
manner as in Reference Example 9.

1H NMR (CIDC13, 400 MHz) : 8 0.61 (2H, dt, J = 6.1, 4. 9 Hz) , 0.86-0.92
(2H, m), 1.75-1.86 (1H. m), 3.50 (3H, s), 3.76 (1H, s), 5.20 (2H,
s), 6.80 (1H, dd, J = 8.6, 2.4 Hz), 6,98 (1H, d, J = 8.6 Hz), 6.98
(1H, d, J = 2.4 Hz).

EIMS (+): 210 [M]`.
<Reference Example 14>
6-Ethoxy-1,3-benzoxathiol-2-one
[0218]

CA 02764126 2011-11-30
[Formula 74]

N1,400-00 S
ao
>=O

[0219]

Potassium carbonate (533 mg) and ethyl iodide (160 L) were
added to a solution of 6-hydroxy-1,3-benzoxathiol-2-one (336 mg)
in N,N-dimethylformamide (10 mL) to form a reaction solution. This
reaction solution was stirred for 4 hours at normal temperature.
Water was added to the reaction solution. The precipitated crystals
were filtrated off, thoroughly washed with water and diisopropyl
ether, and then dried under reduced pressure to obtain the target
product (245 mg) as a white powder.

'H NMR (CDC13i 400 MHz): 5 1.42 (3H, t, J = 6.7 Hz), 4.02 (2H, q,
J = 6.7 Hz), 6.84 (1H, dd, J = 8.6, 2.4 Hz), 6.91 (1H, d, J = 2.4
Hz), 7.18 (1H, d, J = 8.6 Hz).

EIMS (+) : 196 [M]`.
<Reference Example 15>
6-Isopropoxy-1,3-benzoxathiol-2-one
[0220]

CA 02764126 2011-11-30
[Formula 75]

Y S
>=O
ad,
[0221]

Potassium carbonate (829 mg) and isopropyl iodide (300 L)
were added to a solution of 6-hydroxy-l,3-benzoxathiol-2-one (504
mg) in N,N-dimethylformamide (15 mL) to form a reaction solution.
This reaction solution was stirred for 4 hours at normal temperature,
and then for 8 hours at 40 C . Water was added to the reaction solution,
and the reaction solution was extracted with ethyl acetate. The
extract was washed with water and saturated brine in that order,
and then dried over anhydrous sodium sulfate. The anhydrous sodium
sulfate was removed by filtration, and then the solvent was removed
by distillation under reduced pressure. The resultant product was
purified by silica gel column chromatography (hexane: ethyl acetate
= 4 : 1) to obtain the target product (300 mg) as a colorless oil.
1H NMR (CDC13, 400 MHz): 6 1.33 (6H, d, J = 6.1 Hz), 4.44-4.53 (1H,
m), 6.83 (1H, dd, J = 9.2, 2.4 Hz), 6.91 (1H, d, J = 2.4 Hz), 7.17
(1H, d, J = 9.2 Hz).

EIMS (+): 210 [M]r.
<Reference Example 16>
5-Ethoxy-2-hydroxybenzenethiol

CA 02764126 2011-11-30
[0222]

[Formula 76]
[0223]

Under an argon atmosphere, lithium aluminum hydride (119 mg)
was added under ice cooling to a solution of the compound of Reference
Example 14 (245 mg) in tetrahydrofuran (12.5 mL) to form a reaction
solution. This reaction solution was stirred for 30 minutes under
ice cooling . Then, l mol /L hydrochloric acid was added to the reaction
solution, and the reaction solution was extracted with ethyl acetate.
The extract was washed with water and saturated brine in that order,
and then dried over anhydrous sodium sulfate. The anhydrous sodium
sulfate was removed by filtration, and the solvent was removed by
distillation under reduced pressure to obtain the target product
(210 mg) as a colorless oil.

1H NMR (CDC13, 400 MHz): 5 1.38 (3H, t, J = 7.3 Hz), 3.10 (1H, s),
3.96 (2H, q, J = 7.3 Hz), 5.73 (1H, s), 6.78 (1H, dd, J = 9.2, 3.1
Hz), 6.87 (1H, d, J = 9.2 Hz), 6.98 (1H, d, J = 3.1 Hz)

EIMS (+): 170 [M] +.
<Reference Example 17>
2-Hydroxy-5-isopropoxybenzenethiol

CA 02764126 2011-11-30
[0224]

[Formula 771

CJ SH
~C(OH
(0225]

The target product (120 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 15 (300 mg) in the
same manner as in Reference Example 16.

1H NMR (CDC13, 400 MHz) : 8 1.29 (6H, d, J = 6.1 Hz), 3.09 (1H, s),
4.33-4.43 (1H, m), 5.74 (1H, s), 6.78 (1H, dd, J = 9.2, 3.1 Hz),
6.86 (1H, d, J = 9.2 Hz), 7.00 (1H, d, J = 3.1 Hz).

ELMS (+) : 184 [M] +.
<Reference Example 18>
(2-Methoxy-5-trifluoromethylphenylthio)ethoxymethane-l-thione
(0226]

[Formula 78]

F3C OEt
''~ one

CA 02764126 2011-11-30
[0227]

Under an argon atmosphere, water (13 mL) and concentrated
hydrochloric acid (6.9 mL) were added under ice cooling to a solution
of 2-methoxy-5-trifluoromethylaniline (2.50 g) in methanol (13 mL)
to form a first reaction solution. This first reaction solution
was stirred under ice cooling for. 1.0 minutes, and sodium nitrite
(1.26 g) was added thereto and then the first reaction solution was
stirred under ice cooling for 1 hour. The first reaction solution
was slowly added dropwise to an aqueous solution (13 mL) of potassium
ethylxanthate (4.19 g) heated at 65 C to form a second reaction
solution. The second reaction solution was stirred at 65 C for 1
hour. The temperature of this second reaction solution was returned
to normal temperature. Then ice water was added to the second reaction
solution, and the second reaction solution was extracted with ethyl
acetate. The extract was washed with water and saturated brine in
that order, and then dried over anhydrous sodium sulfate. The
anhydrous sodium sulfate was removed by filtration, and then the
solvent was removed by distillation under reduced pressure. The
resultant product was purified by silica gel column chromatography
(hexane : ethyl acetate = 20 : 1) to obtain the target product (913
mg) as a colorless oil.

1H NMR (CDC13, 400 MHz): 6 1.31 (3H, t, J = 7.3 Hz), 3.92 (3H, s)
4.60 (2H, q, J = 7.3 Hz), 7.04 (1H, d, J = 8.6 Hz), 7.71 (1H, dd,
j = 8.6, 2.4 Hz), 7.73 (1H, d, J = 2.4 Hz).

EIMS (+): 296 [M]+.

CA 02764126 2011-11-30
<Reference Example 19>

2-Methoxy-5-tri fluoromethylbenzenethiol
(0228]

[Formula 79]

F3C SH
L.kOMe
[0229]

The target product (400 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 18 (600 mg) in the
same manner as in Reference Example 16.

1H NMR (CDC13, 400 MHz): 6 3.93 (1H, s) , 3.95 (3H, s), 6.90 (1H, d,
J = 8.6 Hz), 7.38 (1H, dd, J = 8.6, 2.4 Hz), 7.51 (1H, d, J = 2.4
Hz).

EIMS (+): 296 [M]'.
<Reference Example 20>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-m
ethylphenylthio)phenyl]-2-methylbutan-l-ol
[0230]

CA 02764126 2011-11-30
[Formula 80]

mom "
NH G
[0231]

Under an argon atmosphere,
4, 5-bis (diphenylphosphino) -9, 9 -dime thyl xanthene (52 mg) was added
to a solution of a tris(dibenzylideneacetone)
dipalladium (0) -chloroform adduct (47 mg) in 1,4-dioxane (1 mL) to
forma reaction solution. This reaction solution was heated to ref lux
for 15 minutes. To the reaction solution, a solution of the compound
of Reference Example 5 (178 mg) in 1,4-dioxane (0.7 mL),
ethyldiisopropylamine (148 L), and a solution of the compound of
Reference Example 9 (100 mg) in 1,4-dioxane (0.6 mL) were added in
this order, and the reaction solution was heated to reflux for 3
hours. Water was added to the reaction solution, and the reaction
solution was extracted with ethyl acetate. The extract was washed
with water and saturated brine in that order, and then dried over
anhydrous sodium sulfate. The solvent was removed by distillation
under reduced pressure, and the resultant product was purified by
silica gel column chromatography (hexane : ethyl acetate = 4 : 1)
to obtain the target product (188 mg) as a colorless oil.

CA 02764126 2011-11-30

1H NMR (CDC13, 400 MHz) : 8 1.24 (3H, s) , 1.44 (9H, s) , 1.80 (1H, ddd,
J = 13.4, 12.2, 5.5 Hz), 2.05 (1H, ddd, J = 13.4, 12.2, 5.5 Hz),
2.25 (3H, s), 2.67 (1H, td, J = 12.8, 5.5 Hz), 2.74 (1H, td, J =
12.8, 5.5 Hz), 3.40 (3H, s), 3.62-3.74 (2H, m), 4.09 (1H, br s),
4.68 (1H, s), 5.16 (2H, s), 7.02-7.07 (3H, m), 7.11 (1H, dd, J =
7.9, 1.8 Hz), 7.14 (1H, d, J = 7.9 Hz), 7.26 (1H, d, J = 1.8 Hz).
ESIMS (+): 496 [M+H]+.

<Reference Example 21>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-ethyl-2-methoxymet
hoxyphenylthio)phenyl]-2-methylbutan-l-ol
[0232]

[Formula 81]

MQM H
100
NHB
[0233]

The target product (205 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 10 (100 mg) and the
compound of Reference Example 5 (165 mg) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz): 8 1.17 (3H, t, J = 7.3 Hz), 1.24 (3H, s),
1.44 (911, s) , 1.79 (1H, ddd, J = 13.4, 12.2, 5.5 Hz) , 2.04 (1H, ddd,
87

CA 02764126 2011-11-30

J = 13.4, 12.2, 5.5 Hz), 2.55 (2H, q, J = 7.3 Hz), 2.67 (1H, td,
J = 12.8, 4.9 Hz), 2.74 (1H, td, J = 12.8, 4.9 Hz), 3.40 (3H, s),
3.62-3.75 (2H, m) , 4.09 (113, brs) , 4.68 (1H, s) , 5.17 (2H, s) , 7.06-7.12
(4H, m), 7.14 (1H, d, J = 7.9 Hz), 7.25 (lH, d, J = 1.8 Hz).
ESIMS (+): 510 [M+H]".

<Reference Example 22>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-p
ropylphenylthio)phenyl]-2-methylbutan-l-o1
[0234]

[Formula 82]

H
wom
NHBoc
[0235]

The target product (152 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 11 (110 mg) and the
compound of Reference Example 5 (170 mg) in the same manner as in
Reference Example 20.

'H NMR (CDC13, 400 MHz): 8 0.90 (3H, t, j = 7.3 Hz), 1.24 (3H, s),
1.44 (9H, s) , 1.51-1.61 (2H, m), 1. 79 (1H, ddd, J = 13.6, 12.2, 5.5
Hz), 2.04 (1H, ddd, J = 13.6, 12.2, 5.5 Hz), 2.48 (2H, q, J = 7.3
Hz), 2.67 (1H, td, J = 12.8, 4.9 Hz), 2.74 (1H, td, J = 12.8, 4.9
88

CA 02764126 2011-11-30

Hz), 3.40 (3H, s), 3.63-3.74 (2H, m), 4.09 (1H, br s), 4.67 (1H,
s), 5.16 (2H, s), 7.08 (3H, s), 7.10 (1H, d, J = 1.8 Hz), 7.13 (1H,
d, J = 7.9 Hz), 7.24 (1H, d, J = 1..8 Hz).

ESIMS (+): 524 [M+H]'.
<Reference Example 23>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-i
sopropylphenylthio)phenyl]-2-methylbutan-l-ol
[02361

[Formula 83]

S CI
mom OH
NI-oc
[0237]

The target product (264 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 12 (139 mg) and the
compound of Reference Example 5 (200 mg) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz): 5 1.19 (6H, t, j = 6.7 Hz), 1.24 (3H, s),
1.44 (9H, s) , 1.79 (1H, ddd, J = 13.4, 12.2, 5.5 Hz) , 2.03 (1H, ddd,
J = 13.4, 12.2, 5.5 Hz) , 2.67 (1H, td, J = 12.8, 4.9 Hz) , 2.74 (1H,
td, J = 12.8, 4.9 Hz), 2.78-2.87 (1H. m), 3.39 (3H, s), 3.63-3.74
(2H, m) , 4.08 (1H, br s), 4.67 (1H, s) , 5.16 (2H, s) , 7.06-7.16 (5H,
89

CA 02764126 2011-11-30
m), 7.24 (1H, d, J = 1.8 Hz).

ESIMS (+): 524 [M+H]+.
<Reference Example 24>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-cyclopropyl-2-meth

oxymethoxy-phenylthio)phenyl]-2-methylbutan-l-ol
[0238]

[Formula 84]

NHBoc
[0239]

The target product (262 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 13 (129 mg) and the
compound of Reference Example 5 (200 mg) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz) : 8 0.58 (2H, dt, J = 6.1, 4.9 Hz), 0.86-0.92
(2H, m), 1.24 (3H, s), 1.44 (9H, s), 1.74-1.85 (2H. m), 1.98-2.08
(1H, m), 2.67 (1H, td, J = 12.8, 4.9 Hz), 2.74 (1H, td, J = 12.8,
4.9 Hz) , 3.38 (3H, s) , 3.63-3.74 (2H, m) , 4.06 (1H, br s) , 4.67 (1H,
s), 5.15 (2H, s), 6.96 (1H, dd, J = 7.9, 1.8 Hz), 7.00 (1H, d, J
= 1.8 Hz) , 7.05 (1H, d, J = 7.9 Hz) , 7.09 (1H, dd, J = 7.9, 1.8 Hz) ,
7.13 (1H, d, J = 7.9 Hz), 7.24 (1H, d, J = 1.8 Hz).

ESIMS (+): 522 [M+H]+.

CA 02764126 2011-11-30
<Reference Example 25>

(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-hydroxy-5-methoxyp
henylthio)phenyl]-2-methylbutan-l-ol
[02401

[Formula 851

19
C3H ~OH
NHBoo
[0241]

The target product (233 mg) was obtained as a colorless oil
by reacting 2-hydroxy-5-methoxybenzenethiol (100 mg) and the
compound of Reference Example 5 (209 mg) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz) : 8 1.23 (3H, s) , 1.43 (9H, s) , 1.78 (1H, ddd,
J = 13.4, 12.2, 4.9 Hz), 2.00 (1H, ddd, J = 13.4, 12.2, 4.9 Hz),
2.59-2. 76 (2H, m), 3.61-3.73 (2H, m), 3.77 (3H, s), 4.06 (1H, br
s) , 4.65 (1H, s) , 6.03 (1H, s) , 6.91 (1H, dd, J = 7.9, 1.8 Hz), 6.97
(1H, dd, J = 7.9, 3.1 Hz), 7.01 (1H, d, J = 7.9 Hz), 7.02 (1H, d,
J = 3.1 Hz), 7.06 (1H, d, J = 1.8 Hz), 7.11 (1H, d, J = 7.9 Hz).
ESIMS (+) : 468 [M+H]'.

<Reference Example 26>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-ethoxy-2-hydroxyph
enylthio)phenyl]-2-methylbutan-l-o1

CA 02764126 2011-11-30
[0242]

[Formula 86]

OH
'GC~
S
NHBoc
[0243]

The target product (204 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 16 (104 mg) and the
compound of Reference Example 5 (200 mg) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz): S 1.22 (3H, s), 1.39 (3H, t, J = 7.3 Hz).
1.43 (9H, s), 1.77 (1H, ddd, J = 13.4, 12.2, 4.9 Hz) , 2.00 (1H, ddd,
j = 13.4, 12.2, 4.9 Hz), 2.64 (1H, td, J = 12.8, 4.9 Hz), 2.71 (1H,
td, J = 12.8, 4.9 Hz), 3.65 (1H, dd, J = 11.6, 4.9 Hz), 3.60 (1H,
dd, J = 11.6, 7.3 Hz), 3.97 (2H, q, J = 7.3 Hz), 4.10 (1H, br s),
4.66 (1H, s), 6.03 (1H, s), 6.91 (1H, dd, J = 7.9, 1.8 Hz), 6.96
(1H, dd, J = 8.6, 2.4 Hz), 7.00 (1H, d, J = 8.6 Hz), 7.02 (1H, d,
J = 2.4 Hz), 7.06 (1H, d, J = 1.8 Hz), 7.10 (1H, d, J = 7.9 Hz).
ESIMS (+): 482 [M+H]+.

<Reference Example 27>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-hydroxy-5-isopropo
xyphenylthio)phenyl]-2-methylbutan-l-ol

CA 02764126 2011-11-30
[0244]

[Formula 87]

H OH
NHB0c
[0245]

The target product (252 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 17 (113 mg) and the
compound of Reference Example 5 (200 mg) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz): 6 1.22 (3H, s), 1.30 (6H, d, J = 6.1 Hz),
1.43 (9H, S), 1.77 (1H, ddd, J = 13.4, 11.6, 5.5 Hz), 2.00 (1H, ddd,
J = 13.4, 11.6, 5.5 Hz), 2.59-2.75 (2H, m), 3.61-3.73 (2H, m), 4.07
(1H, br s), 4.35-4.46 (1H, m), 4.65 (1H, s), 6.04 (1H, s), 6.91 (1H,
dd, J = 7.9, 2.4 Hz), 6.95 (1H, dd, J = 8.6, 2.4 Hz), 6.99 (1H, d,
J = 8.6 Hz), 7.04 (1H, d, J = 2.4 Hz), 7.06 (1H, d, J = 1.8 Hz),
7.10 (1H, d, J = 7.9 Hz).

ESIMS (+): 496 [M+H]+.
<Reference Example 28>
(S)-2-t-butoxycarbonylamino-4-[4-(5-benzyloxy-2-hydroxyphenylth

io)-2-chlorophenyl]-2-methylbutan-l-o1
[0246]

CA 02764126 2011-11-30
[Formula 88]

S CI

~, 0 NH CH
0)
lac
[0247]

The target product (132 mg) was obtained as a colorless oil
by reacting 5-benzyloxy-2-hydroxybenzenethiol (142 mg, Z. J. Song
et al . , Proc Natl Acad Sci USA, 101 (16), 5776 (2004)) and the compound
of Reference Example 5 (200 mg) in the same manner as in Reference
Example 20.

1H NMR (CDC13, 400 MHz) : 8 1.23 (3H, s) , 1.43 (9H, s) , 1.78 (1H, ddd,
J = 13.4, 12.2, 4.9 Hz), 2.01 (1H, ddd, J = 13.4, 12.2, 4.9 Hz),
2.59-2.78 (2H, m), 3.62-3.74 (2H, m), 4.04 (1H, br s), 4.65 (1H,
s) , 5.01 (2H, s) , 6.04 (1H, s) , 6.90 (1H, dd, J = 7.9, 1.8 Hz) , 7.00
(1H, dd, J = 7.9, 2.4 Hz) , 7.04 (1H, dd, J = 7.9, 2.4 Hz) , 7.06 (1H,
d, J = 2. 4 Hz) , 7.10 (1H, d, J = 7. 9 Hz) , 7 .11 (1H, d, J = 2. 4 Hz),
7.30-7.45 (5H, m).

ESIMS (+): 544 [M+H]'.
<Reference Example 29>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-chloro-2-methoxyph

enylthio)phenyl]-2-methylbutan-l-ol
94

CA 02764126 2011-11-30
[0248]

[Formula 89]

c J P c,

O
NHBr,~
[0249]

The target product was obtained as a colorless oil by reacting
the compound of Reference Example 5 (200 mg) and the compound described
in Example 1(1) of WO 05018529 pamphlet
(5-chloro-2-methoxybenzenethiol, 107 mg) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz) 1.26 (3H, s) , 1.45 (9H, s) , 1.83 (1H, ddd,
J = 13.4, 11.6, 5.5 Hz), 2.08 (1H, ddd, J = 13.4, 11.6, 5.5 Hz),
2.71 (1H, td, J = 12.8, 4.9 Hz), 2.78 (1H, td, J = 12.8, 4.9 Hz),
3.68 (1H, dd, J = 11.6, 5.5 Hz), 3.72 (1H, dd, J = 11.6, 7.3 Hz),
3.86 (3H, s), 4.10 (1H, br s) , 4.69 (1H, s) , 6.04 (1H, s) , 6.81 (1H,
d, J = 9.2 Hz), 6.97 (1H, d, J = 2.4 Hz), 7.15-7.24 (3H, m), 7.36
(1H, d, J = 1.2 Hz).

ESIMS (+): 486 [M+H]t.
<Reference Example 30>
(S)-2-t-butoxycarbonylamino-4-[4-(2-t-butoxycarbonyloxy-5-chlor

ophenylthio)-2-chlorophenyl]-2-methylbutan-l-ol

CA 02764126 2011-11-30
(0250]

[Formula 901

CI S U

QBoc OH
NHBOc
[0251]

Under an argon atmosphere, a boron tribromide - methylene
chloride solution (1 mol/L, 1.4 mL) was added under ice cooling to
a solution of the compoundof Reference Example 29 (228 mg) inmethylene
chloride (5mL) to f orm a f irst reaction solution. This first reaction
solution was stirred under ice cooling for 1 hour. Then, a saturated
sodium hydrogen carbonate aqueous solution was added to the first
reaction solution, and the first reaction solution was extracted
with ethyl acetate. The extract was washed with water and saturated
brine in that order, and then dried over anhydrous sodium sulfate.
After the anhydrous sodium sulfate was removed by filtration, the
solvent was removed by distillation under reduced pressure, and the
resultant residue was dissolved in an acetonitrile (5 mL) / methanol
(0.2 mL) mixed solvent. Triethylamine (197 L) and
di-tert-butoxydicarbonate (307 mg) were added to the solution to
forma second reaction solution. This second reaction solution was
stirred at normal temperature for 2 hours and then left to stand
96

CA 02764126 2011-11-30

overnight. Then, water was added to the second reaction solution,
and the second reaction solution was extracted with ethyl acetate.
The extract was washed with water and saturated brine in that order,
and then dried over anhydrous sodium sulfate. The anhydrous sodium
sulfate was removed by filtration, and then the solvent was removed
by distillation under reduced pressure. The resultant product was
purified by silica gel column chromatography (hexane : ethyl acetate
= 2 : 1) to obtain the target product (188 mg) as a colorless oil.
'H NMR (CDC13, 400 MHz) : 3 1.25 (3H, s) , 1.44 (9H, s) , 1.53 (9H, s) ,
1.82 (1H, ddd, J = 13.4, 12.2, 4.9 Hz), 2.07 (1H, ddd, J = 13.4,
12.2, 4.9 Hz), 2.70 (1H, td, J = 12.8, 4.9 Hz), 2.77 (1H, td, J =
12.8, 4.9 Hz), 3.67 (1H, dd, J = 11.6, 5.5 Hz), 3.72 (iH, dd, J =
11.6, 7.3 Hz), 4.11 (1H, br s), 4.69 (1H, s), 7.11 (1H, d, J = 8.6
Hz), 7.14 (1H, d, J = 2.4 Hz), 7.21 (2H, d, J = 1.2 Hz), 7.23 (1H,
dd, J = 8.6, 2.4 Hz), 7.39 (1H, d, J = 1.2 Hz).

CIMS (+) : 572 [M+H] +.
<Reference Example 31>
(S)-2-t-butoxycarbonylamino-4-[4-(2-t-butoxycarbonyloxy-5-trifl
uoromethylphenylthio)-2-chlorophenyl]-2-methylbutan-l-ol
[02521

CA 02764126 2011-11-30
[Formula 91]

C"(Boc
"ac
-I BOC

[0253]

A diphenyl sulfide was obtained by reacting the compound of
Reference Example 19 (127 mg) and the compound of Reference Example
(200 mg) in the same manner as in Reference Example 20. Then,
the target product (160 mg) was obtained as a colorless oil by reacting
this diphenyl sulfide in the same manner as in Reference Example
30.

1H NMR (CDC13, 400 MHz) : 6 1.25 (3H, s), 1.44 (9H, s), 1.54 (9H, s) ,
1.83 (1H, ddd, J = 14,1, 12.2, 5.5 Hz), 2.07 (1H, ddd, J = 14.1,
12.2, 5.5 Hz), 2.65-2.82 (2H, m), 3.67 (1H, dd, J = 11.6, 5.5 Hz),
3.72 (1H, dd, J = 11.6, 7.3 Hz) , 4.06 (1H, br s) , 4.68 (1H, s) , 7.20
(2H, d, J = 1.8 Hz) , 7.31 (1H, d, J = 7.9 Hz) , 7.40 (1H, d, J = 1.8
Hz), 7.46 (1H, d, J = 1.8 Hz), 7.54 (1H, dd, J = 7.9, 1.8 Hz).
ESIMS (+): 606 [M+H]+.

<Reference Example 32>
(R)-2-allyl-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxy-5-
trifluoromethylphenylthio)phenyl]butan-l-ol
[0254]

CA 02764126 2011-11-30
[Formula 92]

F3
S C1
CIC(MI , e H
NHc
[0255]

The target product (1.60 g) was obtained as a colorless oil
by reacting the compound of Reference Example 19 (750 mg) and the
compound of Reference Example 6 (1.26 g) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz): 8 1.47 (9H, s), 1.92 (1H, dd, J = 6.1, 1.8
Hz), 1.95 (1H, dd, J = 6.1, 1,8 Hz), 2.37 (1H, dd, J = 14.1, 7.9
Hz), 2.48 (1H, dd, J = 14.1, 6.7 Hz), 2.73-2.82 (2H, m), 3.71-3.84
(2H, m), 3.95 (3H, s), 4.12(1H, br s)4.76 (1H, s), 5.22 (1H, d, J
= 2.4 Hz), 5.25 (1H, s), 5.86-5.96 (1H, m), 6.98

(1H, d, J = 8.6 Hz), 7.19 (1H, dd, J = 7.9, 1.8 Hz), 7.24 (1H, d,
J = 7.9 Hz), 7.35 (1H, d, J = 1.8 Hz), 7.36 (1H, t, J = 1.8 Hz),
7.53 (1H, dd, J = 8.6, 1.8 Hz).

ESIMS (+): 546 [M+H]`.
<Reference Example 33>
(R)-2-allyl-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-hydroxy-5-

trifluoromethylphenylthio) phenyl]butan-l-ol
99

CA 02764126 2011-11-30
[0256]

[Formula 93]
F3

C* I
H
NHBoc

[0257]

Under an argon atmosphere, a boron tribromide - methylene
chloride solution (1 mol/L, 8.5 mL) was added under ice cooling to
a solution of the compound of Reference Example 32 (1.55g) inmethylene
chloride (30 mL) to form a first reaction solution. This first
reaction solution was stirred under ice cooling for 1 hour. Then,
a saturated sodium hydrogen carbonate aqueous solution was added
to the first reaction solution, and the first reaction solution was
extracted with ethyl acetate. The extract was washed with water
and saturated brine in that order, and then dried over anhydrous
sodium sulfate. After the anhydrous sodium sulfate was removed by
filtration, the solvent was removed by distillation under reduced
pressure, and the resultant residue was dissolved in an acetonitrile
(30 mL) solvent. Di-tert-butoxydicarbonate (930 mg) was added to
this solution to form a second reaction solution. This second
reaction solution was stirred at normal temperature for 2 hours and
then left to stand overnight. Then, water was added to the second
100

CA 02764126 2011-11-30

reaction solution, and the second reaction solution was extracted
with ethyl acetate. The extract was washed with water and saturated
brine in that order, and then dried over anhydrous sodium sulfate.
The anhydrous sodium sulfate was removed by filtration, and then
the solvent was removed by distillation under reduced pressure. The
resultant product was purified by silica gel column chromatography
(hexane : ethyl acetate = 2 : 1) to obtain the target product (940
mg) as a colorless oil.

'H NMR (CDC13, 400 MHz) : 8 1.43 (9H, s) , 1.82-1.88 (2H, m), 2.31 (1H,
dd, J = 14.1, 8.6 Hz), 2.43 (1H, dd, J = 14.1, 6.7 Hz), 2.69 (2H,
dt, J = 11.0, 6.7 Hz), 3.67-3.78 (2H, m), 4.07(1H, br s)4.71 (1H,
s), 5.18 (1H, dd, J = 6.1, 1.8 Hz), 5.21 (1H, s), 5.78-5.90 (1H,
m), 6.74 (1H, s), 6.91 (1H, dd, J = 7.9, 2.4 Hz), 7.08 (1H, d, J
= 2.4 Hz), 7.15 (2H, t, J = 7.9 Hz) , 7.63 (1H, dd, J = 7.9, 1.8 Hz),
7.80 (1H, d, J = 2.4 Hz).

ESIMS (+): 532 [M+H]`.
<Reference Example 34>
(S)-2-t-butoxycarbonylamino-4-[4-(2-t-butoxycarbonyloxy-5-trif1

uoromethylphenylthio)-2-chlorophenyl]-2-propylbutan-l-ol
[0258]

101

CA 02764126 2011-11-30
[Formula 941

F3 q I)C
NHBac
'OC

[0259]

A palladium on activated carbon/ethylene diamine complex (100
mg) was added to a solution of the compound of Example 33 (473 mg)
in ethyl acetate (9 mL) to form a first reaction solution. This
first reaction solution was stirred at normal temperature for 20
hours under hydrogen purging. The first reaction solution was
filtrated through Celite, and the solvent was removedbydistillation.
The resultant residue was dissolved in acetonitrile (9 mL).
Triethylamine (145 L) and di-tert-butoxydicarbonate (284 mg) were
added to this solution to form a second reaction solution. This
second reaction solution was stirred at normal temperature for 2
hours and then left to stand overnight. Then, water was added to
the second reaction solution, and the second reaction solution was
extracted with ethyl acetate. The extract was washed with water
and saturated brine in that order, and then dried over anhydrous
sodium sulfate. The anhydrous sodium sulfate was removed by
filtration, and then the solvent was removed by distillation under
reduced pressure. The resultant product was purified by silica gel
102

CA 02764126 2011-11-30

column chromatography (hexane : ethyl acetate = 2 : 1) to obtain
the target product (434 mg) as a colorless oil.

IH NMR (CDC13, 400 MHz): 8 0.96 (3H, t, J = 7.3 Hz), 1.30-1.42 (2H,
m), 1.44 (9H, s), 1.54 (9H, s), 1.54-1.60 (2H, m), 1.83 (1H, ddd,
J = 13.4, 12.2, 5.5 Hz), 1.94 (lH, ddd, J = 13.4, 12.2, 5.5 Hz),
2.62-2.77 (2H, m) , 3.74(2H, d, J = 6.1 Hz) , 4.10 (1H, br s) 4.62 (1H,
s) , 7.20 (2H, s) , 7.30 (1H, d, J = 8.6 Hz) , 7.39 (1H, s) , 7.47 (1H,
d, J = 1.8 Hz), 7.53 (1H, dd, J = 8.6, 1.8 Hz).

ESIMS (+): 634 [M+H]4.
<Reference Example 35>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-m

ethylphenylthio)phenyl]-l-dimethoxyphosphoryloxy-2-methylbutane
(0260]

[Formula 95]

PO(CO)2
mom
NHBoc
[0261]

Carbon tetrabromide (130 mg) and trimethyl phosphite (46' L)
were added under ice cooling to a solution of the compound of Example
20 (97 mg) in pyridine (0.5 mL) to form a reaction solution. This
reaction solution was stirred for 2 hours under ice cooling. Then,
water was added to the reaction solution, and the reaction solution
103

CA 02764126 2011-11-30

was extracted with ethyl acetate. The extract was washed with water
and saturated brine in that order, and then dried over anhydrous
sodium sulfate. The anhydrous sodium sulfate was removed by
filtration, and then the solvent was removed by distillation under
reduced pressure. The resultant product was purified by silica gel
column chromatography (hexane : ethyl acetate = 1 : 1) to obtain
the target product (84 mg) as a colorless oil.

1H NMR (CDC13, 400 MHz) : S 1.35 (3H, s) , 1.44 (9H, s), 1.76 (1H, add,
J = 14.1, 12.2, 6.1 Hz) , 2.05-2.13 (1H, in), 2.25 (3H, s), 2.62-2.75
(2H, m), 3.40 (3H, s), 3.78 (6H, d, J = 11.0 Hz), 4.02 (1H, dd, J
= 9.8, 5.5 Hz), 4.22 (1H, dd, J = 9.8, 5.5 Hz), 4.62 (1H, s), 5.16
(2H, s), 7.02-7.07 (3H, m), 7.11 (2H, d, J = 1.8 Hz), 7.25 (1H, d,
J = 1.8 Hz).

ESIMS (+): 604[M+H] +.
<Reference Example 36>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-ethyl-2-methoxymet

hoxyphenylthio)phenyl]-1-dimethoxyphosphoryloxy-2-methylbutane
[0262]

[Formula 96]

MCM f PC(OMe)2
0000 SIC
NHBoc
104

CA 02764126 2011-11-30
[0263]

The target product (114 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 21 (100 mg) in the
same manner as in Reference Example 35.

''H NMR (CDC13, 400 MHz): 8 1.17 (3H, t, J = 7.3 Hz), 1.35 (3H, s),
1.44 (9H, s) , 1.75 (1H, ddd, J = 13.4, 11.6, 5.5 Hz) , 1.98-2.15 (1H,
m) , 2.55 (2H, q, J = 7.3 Hz), 2.61-2.76 (2H, m) , 3.39 (3H, s) , 3.78
(6H, d, J = 11.0 Hz), 4.02 (1H, dd, J = 9.8, 5.5 Hz), 4.23 (1H, dd,
J = 9.8, 5.5 Hz), 4.62 (1H, s), 5.16 (2H, s), 7.05-7.13 (511, m),
7.24 (1H, d, J = 1.2 Hz).

ESIMS (+): 618 [M+H]+.
<Reference Example 37>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-p

ropylphenylthio)phenyl]-1-dimethoxyphosphoryloxy-2-methylbutane
[0264]

[Formula 97]

S CI

OPO(OMe)2
mom
NHBoc
[0265]

The target product (112 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 22 (100 mg) in the
same manner as in Reference Example 35.

105

CA 02764126 2011-11-30

~H NMR (CDC13, 400 MHz): 8 0.90 (3H, t, J = 7.3 Hz), 1.35 (3H, s),
1. 44 (9H, s) , 1.51-1. 61 (2H, m) , 1.75 (1H, ddd, J = 13. 4, 11.6, 5.5
Hz), 1.97-2.13 (1H, m), 2.48 (2H, t, J = 7.3 Hz), 2.61-2.76 (2H,
m), 3.39 (3H, s), 3.78 (6H, d, J = 11.0 Hz), 4.02 (1H, dd, J = 9.8,
5.5 Hz), 4.23 (1H, dd, J = 9.8, 5.5 Hz), 4.62 (1H, s), 5.16 (2H,
s), 7.05-7.13 (5H, m), 7.24 (1H, d, J = 1.2 Hz).

ESIMS (+): 632 [M+H]+.
<Reference Example 38>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-i

sopropylphenylthio)phenyl]-1-dimethoxyphosphoryloxy-2-methylbut
ane

[0266]
[Formula 98]

S CI

/ aPO(OMe)2
MC~M
.001,
NHBoc
[0267]

The target product (86 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 23 (132 mg) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz): 8 1.19 (6H, t, J = 7.3 Hz), 1.35 (3H, s),
1.44 (9H, s ) , 1.75 (1H, ddd, J = 13.4, 12.2, 5. 5 Hz) , 2.00-2.15 (1H,
m), 2.61-2.76 (2H, m), 2.76-2.87 (1H. m), 3.39 (3H, s), 3.78 (6H,
106

CA 02764126 2011-11-30

d, J = 11.0 Hz), 4.02 (1H, dd, J = 9.8, 5.5 Hz), 4.23 (1H, dd, J
= 9.8, 5.5 Hz) , 4.62 (1H, s) , 5.16 (2H, s) , 7.05-7. 13 (5H, m) , 7.23
(1H, d, J = 1.8 Hz).

ESIMS (+): 632 [M+H]'.
<Reference Example 39>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-cyclopropyl-2-meth

oxymethoxyphenylthio)phenyl]-1-dimethoxyphosphoryloxy-2-methylb
utane

[0268]
[Formula 99]

A,C

(OMe)2
NHBcc

[0269]

The target product (144 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 24 (122 mg) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz) : 6 0.58 (2H, dt, J = 6.7, 4.9 Hz) , 0.80-0.90
(2H, m), 1.35 (3H, s), 1.44 (9H, s), 1.69-1.85 (2H, m), 1.96-2.14
(1H, m), 2.61-2.76 (2H, m), 3.38 (3H, s) , 3.78 (6H, d, J = 11.0 Hz) ,
4.02 (1H, dd, J = 9.8, 5.5 Hz) , 4.22 (1H, dd, J = 9.8, 5.5 Hz) , 4.62
(1H, s), 5.14 (2H, s), 6.96 (1H, dd, J = 8.6, 1.8 Hz), 7.00 (1H,
d, J = 1.8 Hz), 7.03-7.13 (3H, m), 7.23 (1H, d, J = 1.8 Hz).

107

CA 02764126 2011-11-30
ESIMS (+): 630 [M+H]+.

<Reference Example 40>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-hydroxy-5-methoxyp
henylthio)phenyl]-1-dimethoxyphosphoryloxy-2-methylbutane
[0270]

[Formula 100]

l o p S 1

a
/ N OPO(OMO
NHBoc
[0271]

The target product (89 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 25 (110 mg) in the
same manner as in Reference Example 35.

IH NMR (CDC13, 400 MHz) : 8 1.34 (3H, s) , 1.43 (9H, s) , 1.73 (1H, ddd,
J = 13.4, 11.6, 5.5 Hz) , 1.99-2.09 (1H, m), 2.64 (1H, td, j = 13.4,
5.5 Hz), 2.68 (1H, td, J = 13.4, 5.5 Hz), 3.77 (3H, s), 3.77 (3H,
d, J = 11.0 Hz), 3.78 (3H, d, J = 11.0 Hz), 4.00 (1H, dd, J = 9.8,
4.9 Hz), 4.20 (1H, dd, J = 9.8, 4.9 Hz), 4.61 (1H, s), 6.05 (1H,
s) , 6.91 (1H, dd, J = 7.9, 1.8 Hz), 6.97 (1H, dd, J = 8.6, 3.1 Hz) ,
7.01 (1H, d, J = 7.9 Hz), 7.02 (1H, d, J = 3.1 Hz), 7.06 (1H, d,
J = 1.8 Hz), 7.08 (1H, d, J = 8.6 Hz).

ESIMS (+): 576 [M+H]+.
<Reference Example 41>

108

CA 02764126 2011-11-30
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-etboxy-2-hydroxyph
enylthio)phenyl]-1-dimethoxyphosphoryloxy-2-methylbutane
[0272]

[Formula 1011

.~ O OMe)2
NH$oc
[0273]

The target product (108 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 26 (146 mg) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz): 6 1.34 (3H, s) , 1.39 (3H, t, J = 7.3 Hz).
1 .43 (9H, s) , 1.73 (1H, ddd, J = 14.1, 12.2, 5.5 Hz) , 1.99-2.09 (1H,
m), 2.64 (1H, td, J = 13.4, 5.5 Hz), 2.68 (1H, td, J = 13.4, 5.5
Hz) , 3 .77 (3H, d, J = 11.0 Hz) , 3 .78 (3H, d, J = 11.0 Hz) , 3 .97 (2H,
q, J = 7.3 Hz), 4.00 (1H, dd, J = 9.8, 4.9 Hz), 4.20 (1H, dd, J =
9.8, 4.9 Hz), 4.61 (1H, s), 6.04 (1H, s), 6.91 (1H, dd,J = 7.9,
1.8 Hz), 6.96 (1H, dd, J = 8.6, 2.4 Hz), 7.00 (1H, d, J = 8.6 Hz),
7.02 (1H, d, J = 2.4 Hz), 7.05 (1H, d, J = 1.8 Hz), 7.08 (1H, d,
J = 7.9 Hz).

ESIMS (+): 590 [M+H]+.
<Reference Example 42>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-hydroxy-5-isopropo

109

CA 02764126 2011-11-30
xyphenylthio)phenyl]-l-dimethoxyphosphoryloxy-2-methylbutane
[0274]

[Formula 102]

Q CI

H C ~1 P PM02
NHBac
(0275]

The target product (90 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 27 (129 mg) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz): 8 1.30 (6H, d, J = 6.1 Hz), 1.34 (3H, s),
1.43 (9H, s), 1.73 (1H, ddd, J = 13.4, 11.0, 6.1 Hz), 1.98-2.10 (1H,
m), 2.59-2.75 (2H, m), 3.77 (3H, d, J = 11.0 Hz), 3.78 (3H, d, J
= 11.0 Hz), 4.00 (1H, dd, J = 9.8, 4.9 Hz), 4.20 (1H, dd, J = 9.8,
4.9 Hz), 4.36-4.46 (1H, m), 4.60 (1H, s), 6.04 (1H, s), 6.90 (1H,
dd, J = 7.9, 2.4 Hz), 6.95 (1H, dd, J = 9.2, 2.4 Hz), 6.99 (1H, d,
J = 9.2 Hz), 7.04 (1H, d, J = 2.4 Hz), 7.06 (1H, d, J = 2.4 Hz),
7.08 (1H, d, J = 7.9 Hz).

ESIMS (+): 604 [M+H]+.
<Reference Example 43>
(S)-4-[4-(5-benzyloxy-2-hydroxyphenylthio)-2-chlorophenyl]-2-t-

butoxycarbonylamino-l-dimethoxyphosphoryloxy-2-methylbutane
110

CA 02764126 2011-11-30
[0276]

(Formula 1031
;41 1
S. 0

=~ H .= OPO(OMe)2
NHBoc
(0277]

The target product (158 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 28 (132 mg) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz) : 8 1.34 (3H, s) , 1.43 (9H, s) , 1.74 (1H, ddd,
j = 13.4, 12.2, 5.5 Hz), 1.95-2.14 (1H, m), 2.59-2.73 (2H, m), 3.77
(3H, d, J = 11.0 Hz), 3.78 (3H, d, J = 11.0 Hz), 4.00 (1H, dd, J
= 9.8, 4.9 Hz), 4.21 (1H, dd, J = 9.8, 4.9 Hz), 4.61 (1H, s), 5.01
(2H, s), 6.06 (1H, s), 6.89 (1H, dd, J = 7.9, 1.8 Hz), 7.00 (iH,
d, J = 9.2 Hz), 7.04 (2H, dd, J = 9.2, 1.8 Hz), 7.08 (1H, d, J =
7.9 Hz), 7.10 (1H, d, J = 2.4 Hz), 7.30-7.43 (5H, m).

ESIMS (+): 652 [M+H]+.
<Reference Example 44>
(S)-2-t-butoxycarbonylamino-4-[4-(2-t-butoxycarbonyloxy-5-chlor

ophenylthio)-2-chlorophenyl]-1-dimethoxyphosphoryloxy-2-methylb
utane

[0278]

111

CA 02764126 2011-11-30
[Formula 104]

CI S

OPO(OMe)2
NHBoc

[0279]

The target product (105 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 30 (94 mg) in the same
manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz) : 8 1.36 (3H, s) . 1.45 (9H, s) , 1.53 (9H, s) ,
1.78 (1H, ddd, J = 13 . 4 , 11.6, 5.5 Hz) , 2.05-2. 17 (1H, m) , 2. 6 5 - 2.79
(2H, m), 3.78 (3H, d, J = 11.0 Hz), 3.79 (3H, d, J = 11.0 Hz), 4.03
(1H, dd, J = 9.8, 4.9 Hz), 4.24 (1H, dd, J = 9.8, 4.9 Hz), 4.64 (1H,
s), 7.11 (1H, d, J = 8.6 Hz), 7.14 (1H, d, J = 2.4 Hz), 7.16-7.21
(2H, m), 7.23 (1H, dd, J = 8.6, 2.4 Hz), 7.39 (1H, d, J = 2.4 Hz).
ESIMS (+): 680 [M+H]'.

<Reference Example 45>
(S)-2-t-butoxycarbonylamino-4-[4-(2-t-butoxycarbonyloxy-5-trif1
uoromethylphenylthio)-2-chlorophenyl]-1-dimethoxyphosphoryloxy-
2-methylbutane

[0280]

112

CA 02764126 2011-11-30
[Formula 105]

F3C,,CCS3IOCCI
oc OPO(OMe)2
NHBoc
[0281]

The target product (80 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 31 (90 mg) in the same
manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz) : 6 1.36 (3H, s) . 1.44 (9H, s) , 1.54 (9H, s) ,
1.78 (1H, ddd, J = 13.4, 12.2, 5.5 Hz), 2.06-2.18 (1H, m), 2.65-2.80
(2H, m) , 3.78 (3H, d, J = 11.0 Hz) , 3.79 (3H, d, J = 11.0 Hz) , 4.03
(1H, dd, J = 9.8, 4.9 Hz) , 4.23 (1H, dd, J = 9.8, 4.9 Hz), 4.63 (1H,
s), 7.19 (2H, d, J = 1.2 Hz), 7.30 (1H, d, J = 8.6 Hz), 7.39 (1H,
d, J = 1.2 Hz), 7.46 (1H, d, J = 1.8 Hz), 7.53 (1H, dd, J = 8.6,
1.8 Hz).

ESIMS (+): 714 [M+H]+.
<Reference Example 46>
(R)-2-allyl-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-hydroxy-5-

trifluoromethylphenylthio)phenyl]-1-dimethoxyphosphoryloxybutan
e

[0282]

113

CA 02764126 2011-11-30
[Formula 106]

FCC ~ ,~ S CC~O(OM02
"' ON NHBoc

[0283]

The target product (350 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 33 (473 mg) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz) : 8 1.43 (9H, s), 1.75-1.78 (1H, M), 1.90-2.03
(1H, m) , 2.41-2.55 (2H, m), 2.64-2.75 (2H, m), 3.78 (6H, d, J = 11.0
Hz), 4.08 (1H, dd, J = 9.8, 4.9 Hz), 4.19 (1H, dd, J = 9.8, 4.9 Hz),
4.58 (1H, s), 5.18 (1H, s) , 5.21 (1H, d, J = 6.1 Hz), 5.73-5.87 (1H,
m), 6.80 (1H, s), 6.90 (1H, dd, J = 7.9, 1.8 Hz), 7.09 (1H, d, J
= 1.8 Hz), 7.12 (1H, d, J = 7.9 Hz), 7.15(1H, d, J = 8.6 Hz), 7.63
(1H, dd, J = 8.6, 1.8 Hz), 7.80 (1H, d, J = 1.8 Hz).

ESIMS (+): 640 [M+H]+.
<Reference Example 47>
(S)-2-t-butoxycarbonylamino-4-[4-(2-t-butoxycarbonyloxy-5-trifl

uoromethylphenylthio)-2-chlorophenyl]-1-dimethoxyphosphoryloxy-
2-propylbutane

[0284]

114

CA 02764126 2011-11-30
(Formula 107]

F S CI

Boc t~F''C(OMe)2
NHBoc
[0285]

The target product (222 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 34 (209 mg) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz) : 8 0.96 (3H, t, j = 7.3 Hz) , 1.31-1.42 (3H,
m). 1.44 (9H, s), 1.54 (9H, s), 1.59-1.72 (1H, m), 1.75-1.85 (1H,
m) , 1.95-2.11 (1H, m) , 2.66-2.74 (2H, m), 3.78 (3H, d, J = 11.0 Hz) ,
3.79 (3H, d, J = 11.0 Hz) , 4.11 (1H, dd, J = 9.8, 4.9 Hz) , 4.25 (1H,
dd, J = 9.8, 4.9 Hz), 4.51 (1H, s), 7.19 (2H, d, J = 1.8 Hz), 7.30
(1H, d, J = 7.9 Hz), 7.39 (1H, s), 7.46 (1H, d, J = 1.8 Hz), 7.53
(1H, dd, J = 7.9, 1.8 Hz).

ESIMS (+); 742 [M+H]+.
[Example 1]

[0286]
(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-methylphenylthio)phenyl]
-2-methylbutylphosphoric acid monoester

(0287]

115

CA 02764126 2011-11-30
[Formula 108]

NH2
[0288]

I odotr imethyls i lane (97 L) was added dropwise under ice
cooling and under an argon atmosphere to a solution of the compound
of Reference Example 35 (84 mg) in acetonitrile (1.4 mL) to form
a reaction solution. This reaction solution was stirred under ice
cooling for 30 minutes. Then water (10 mL) was added to the reaction
solution, and the reaction solution was then further stirred under
ice cooling for 30 minutes. The precipitated crystals were then
filtrated off, thoroughly washed with water and ethyl acetate, and
then dried to obtain the target product (42 mg) as a white powder.
Optical rotation: [a]D24-8.32 (c 0.50, McOH).

lH NMR (DMSO-d6-dTFA, 400 MHz) : 5 1.27 (3H, s) , 1.69-1.85 (2H, m),
2.17 (3H, s) , 2.66 (2H, t, J = 8.6 Hz), 3.85 (1H, dd, J = 11.0, 4.9
Hz), 3.92 (1H, dd, J=11.0, 4.9Hz), 6.86 (1H, d, J=8.6Hz), 7.01-7.10
(4H, m), 7.25 (1H, d, J = 8.6 Hz).

HRESIMS (+) : 432.07951 (432.08013 calcd. for C18H24C1NO5PS)
Elemental analysis: measured C 47.06%, H 5.35%, N 2.95%, calcd. for
C18H23C1NOSPS. 1.5 H2O: C 47.11%, H 5.71%, N 3.05%.

116

CA 02764126 2011-11-30
[Example 2]

[0289]
(S)-2-amino-4-[2-chloro-4-(5-ethyl-2-hydroxyphenylthio)phenyl]-
2-methylbutylphosphoric acid monoester

[0290]
(Formula 109]

N
[0291]

The target product (65 mg) was obtained as a white powder by
reacting the compound of Reference Example 36 (114 mg) in the same
manner as in Example 1.

Optical rotation: [a]D25-9.94 (c 0.50, MeOH).

1H NMR (DMSO-d6-dTFA, 400 MHz) : S 1.09 (3H, t, J = 7.3 Hz) , 1.27 (3H,
s), 1.67-1.86 (2H, m), 2.47 (2H, q, J = 7.3 Hz), 2.66 (2H, t, J =
8.6 Hz), 3.86 (1H, dd, J = 11.0, 4.9 Hz), 3.93 (1H, dd, J = 11.0,
4.9 Hz), 6.88 (1H, d, J = 7.9 Hz), 7.04 (2H, dd, J = 7.9, 1.8 Hz),
7.07-7.14 (2H, m), 7.25 (1H, d, J = 8.6 Hz).

HRESIMS (+): 446.09569 (446.09578 calcd. for C19H26C1NO5PS)
Elemental analysis: measured C 48.89%, H 5.66%, N 2.75%, calcd. for
C19H25C1NO5PS. H2O C 49.19%, H 5.87%, N 3.02%.

[Example 31

117

CA 02764126 2011-11-30
[0292]

(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-propylphenylthio)phenyl]
-2-methylbutylphosphoric acid monoester

[0293]
[Formula 1101

OH =r ~t~H
NH2
[0294]

The target product (60 mg) was obtained as a white powder by
reacting the compound of Reference Example 37 (112 mg) in the same
manner as in Example 1.

Optical rotation: [a]D25 -9.86 (c 0.50, McOH).

1H NMR (DMSO-d6-dTFA, 400 MHz) : 8 0.82 (3H, t, J = 7.3 Hz) , 1.27 (3H,
s) , 1.43-1.56 (2H, m) , 1.68-1.85 (2H, m) , 2.42 (2H, t, j = 7.3 Hz),
2.65 (1H, d, J = 7.9 Hz), 2.67 (1H, d, J = 7.9 Hz), 3.86 (1H, dd,
j = 11.0, 4.9 Hz), 3.92 (1H, dd, J = 11.0, 4.9 Hz), 6.88 (1H, d,
J = 7.9 Hz), 7.00-7.11 (4H, m), 7.24 (1H, d, J = 8.6 Hz).

HRESIMS (+) : 460.11106 (460.11143 calcd. for C20H28C1NO5PS)
[Example 41

[0295]
(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-isopropylphenylthio)phen
yl]-2-methylbutylphosphoric acid monoester

118

CA 02764126 2011-11-30
[02961

[Formula 111]

%q S 04
-*Iola
ON X OPd(tJhi)x
NH2
[0297]

The target product (40 mg) was obtained as a white powder by
reacting the compound of Reference Example 38 (86 mg) in the same
manner as in Example 1.

Optical rotation: [a]L125 -8.12 (c 0.50, McOH).

1H NMR (DMSO-d6-dTFA, 400 MHz) : 8 1.12 (6H, d, J = 6.7 Hz) , 1.27 (3H,
s), 1.67-1.87 (2H, m), 2.66 (2H, t, J = 8.6 Hz), 2.74-2.81 (1H, m),
3.86 (1H, dd, J = 11.0, 4.9 Hz), 3.93 (1H, dd, J = 11.0, 4.9 Hz),
6.89 (1H, d, J = 7.9 Hz), 7.03 (1H, dd, J = 7.9, 1.8 Hz), 7.05 (1H,
s), 7.11-7.17(2H, m), 7.25 (1H, d, J = 7.9 Hz).

HRESIMS (+): 460.11124 (460.11143 calcd. for C20H28C1NO5PS)
Elemental analysis: measured C 50.13%, H 5.88%, N 2.72%, calcd. for
C20H27C1NO5PS. H2O C 50.26%, H 6.12%, N 2.93%.

[Example 5]
[0298]
(S)-2-amino-4-[2-chloro-4-(5-cyclopropyl-2-hydroxyphenylthio)ph
enyl]-2-methylbutylphosphoric acid monoester

[0299]

119

CA 02764126 2011-11-30
[Formula 112]

17
OcOcOPO(OH)2
OH
N
[0300]

The target product (99 mg) was obtained as a white powder by
reacting the compound of Reference Example 39 (144 mg) in the same
manner as in Example 1.

Optical rotation: [a]o5 -8.15 (c 0.50, NeOH).

1H NMR (DMSO-d6-dTFA, 400 MHz): 8 0.52 (2H, dt, J = 6.1, 4.3 Hz),
0.80-0.86 (2H, m), 1.28 (3H, s), 1.67-1.86 (3H, m), 2.66 (2H, t,
J = 8.6 Hz) , 3.86 (1H, dd, J = 11.0, 4.9 Hz) , 3.93 (1H, dd, J = 11.0,
4.9 Hz), 6.85 (1H, d, J = 8.6 Hz), 6.96 (1H, dd, J = 8.6, 2.4 Hz),
7.01 (1H, d, J = 2.4 Hz), 7.04 (1H, dd, J = 7.9, 1.8 Hz), 7.05 (1H,
s), 7.11-7.17(2H, m), 7.25 (1H, d, J = 7.9 Hz).

HRESIMS (+) : 458.09631 (458.09578 calcd. for C20H26ClNO5PS).
[Example 6]

[0301]
(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-methoxyphenylthio)phenyl
1-2-methylbutylphosphoric acid monoester

[0302]

120

CA 02764126 2011-11-30
[Formula 113]

HNH PO(OH)2
NH2
[0303]

The target product (50 mg) was obtained as a white powder by
reacting the compound of Reference Example 40 (89 mg) in the same
manner as in Example 1.

Optical rotation: [a]D25 -8.31 (c 0.50, McOH).

1H NMR (DMSO-d6-dTFA, 400 MHz): 8 1.23 (3H, s), 1.64-1.84 (2H, m),
2.66 (2H, t, J = 7.9 Hz), 3.63 (3H, s), 3.76 (2H, d, J = 11.0 Hz),
6.73 (1H, d, J = 3.1 Hz), 6.83 (1H, dd, J = 8.6, 3.1 Hz), 6.87 (1H,
d, J = 8.6 Hz), 7.08 (1H, dd, J = 7.9, 1.8 Hz), 7.12 (1H, d, J =
1.8 Hz), 7.28 (1H, d, J = 7.9 Hz).

HRESIMS (+) : 448.07498 (448.07505 calcd. for C18H24C1NO6PS).
Elemental analysis: measured C 45.69%, H 5.08%, N 2.96%, calcd. for
C18H23C1NO6PS. 1.2 H2O C 46.06%, H 5.45%, N 2.98%.

[Example 7]
[0304]
(S)-2-amino-4-[2-chloro-4-(5-ethoxy-2-hydroxyphenylthio)phenyl]
-2-methylbutylphosphoric acid monoester

[0305]

121

CA 02764126 2011-11-30
[Formula 114]

2
[0306]

The target product (72 mg) was obtained as a white powder by
reacting the compound of Reference Example 41 (108 mg) in the same
manner as in Example 1.

Optical rotation: [a]D25 -8.50 (c 0.50, MeOH).

1H NMR (DMSO-d6-dTFA, 400 MHz) : 8 1.23 (3H, t, J = 6.7 Hz) , 1.28 (3H,
s), 1.70-1.88 (2H, m), 2.68 (2H, t, j = 7.9 Hz), 3.87 (2H, q, J =
6.7 Hz), 3.88 (1H, dd, J = 11.0, 4.9 Hz), 3.93 (1H, dd, J = 11.0,
4.9 Hz), 6.71 (1H, d, J = 2.4 Hz), 6.83 (1H, dd, J = 9.2, 2.4 Hz),
6.86 (1H, d, J = 9.2 Hz), 7.11 (1H, dd, J = 7.9, 1.8 Hz), 7.13 (1H,
d, J = 1.8 Hz), 7.28 (1H, d, J = 7.9 Hz).

HRESIMS (+): 462.09023 (462.09070 calcd. for C19H26C1NO6PS)
Elemental analysis: measured C 47.55%, H 5.32%, N 2.55%, calcd. for
C19H25C1NO6PS. H2O C 47.55%, H 5.67%, N 2.92%.

[Example 8]
[0307]
(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-isopropoxyphenylthio)phe
nyl]-2-methylbutylphosphoric acid monoester

[0308]

122

CA 02764126 2011-11-30
[Formula 1151

Y"*CCH a
NH2
[0309]

The target product (41 mg) was obtained as a white powder by
reacting the compound of Reference Example 42 (90 mg) in the same
manner as in Example 1.

Optical rotation: [a]D25 -8.51 (c 0.50, MeOH).

'. H NMR (DMSO-d6-dTFA, 400 MHz) : 8 1.16 (6H, d, J = 6.1 Hz) , 1.28 (3H,
s), 1.71-1.86 (2H, m), 2.68 (2H, t, J = 8.6 Hz), 3.88 (1H, dd, J
= 11.0, 4.9 Hz), 3.93 (1H, dd, J = 11.0, 4.9 Hz), 4.31-4.40 (1H,
m), 6.69 (1H, d, J = 3.1 Hz), 6.81 (1H, dd, J = 8.6, 3.1 Hz), 6.85
(1H, d, J = 8.6 Hz), 7.12 (1H, dd, J = 7.9, 1.8 Hz), 7.14 (1H, d,
J = 1.8 Hz), 7.28 (1H, d, J = 7.9 Hz).

HRESIMS (+) : 476.10592 (476.10635 calcd. for C20H28C1NO6PS) .
Elemental analysis: measured C 49.21%, H 5.61%, N 2.72%, calcd. for
C20H27C1NO6PS. 0.7H20 C 49.17%, H 5.86%, N 2.87%.

[Example 91
[0310]
(S)-2-amino-4-[4-(5-benzyloxy-2-hydroxyphenylthio)-2-chiorophen
yl]-2-methylbutylphosphoric acid monoester

[0311]

123

CA 02764126 2011-11-30
[Formula 1161

H PQOH~
NH2
[0312]

The target product (26 mg) was obtained as a white powder by
reacting the compound of Reference Example 43 (59 mg) in the same
manner as in Example 1.

Optical rotation: [a]D25 -9.77 (c 0.50, MeOH).

1H NMR (DMSO-d6-dTFA, 400 MHz): 5 1.27 (3H, s), 1.68-1.86 (2H, m),
2.67 (2H, t, J = 8.6 Hz) , 3.86 (1H, dd, J = 11.0, 4.9 Hz) , 3.92 (1H,
dd, J = 11.0, 4.9 Hz), 4.95 (2H, s), 6.78 (1H, d, J = 3.1 Hz), 6.85
(1H, dd, J = 9.2, 3.1 Hz), 6.85 (1H, d, J = 9.2 Hz), 7.08 (1H, dd,
J = 7.9, 1.8 Hz), 7.12 (1H, d, J = 1.8 Hz), 7.28-7.31 (6H, m).
HRESIMS (+) : 524.10694 (524.10635 calcd. for C24H28C1NO6PS) .

Elemental analysis: measured C 52.62%, H 5.05%, N 2.50%, calcd. for
C24H27C1NO6PS. 1.2H20 C 52.83%, H 5.43%, N 2.57%.

[Example 10]
[0313]

(S)-2-amino-4-[2-chloro-4-(5-chloro-2-hydroxyphenylthio)phenyl]
-2-methylbutylphosphoric acid monoester

[0314]

124

CA 02764126 2011-11-30
[Formula 117]

S I
NPO{ OH,
NH2
[0315]

The target product (56 mg) was obtained as a white powder by
reacting the compound of Reference Example 44 (105 mg) in the same
manner as in Example 1.

Optical rotation: [a]D25 -8.28 (c 0.50, MeOH).

1H NMR (DMSO-d6-dTFA, 400 MHz) : 5 1.29 (3H, s), 1.70-1.90 (2H, m),
2.70 (2H, t, J = 8.6 Hz) , 3. 8 7 (1H, dd, J = 11.0, 4.9 Hz) , 3. 9 4 (1H,
dd, J = 11.0, 4.9 Hz), 6.94 (1H, d, J = 8.6 Hz), 7.08 (1H, d, J =
2.4 Hz) , 7.19 (1H, dt, J = 7.9, 1.8 Hz) , 7.23 (1H, dt, J = 8.6, 2.4
Hz), 7.26 (1H, t, J = 8.6 Hz), 7.33 (iH, d, J = 7.9 Hz).

HRESIMS (+) : 452.02541 (452.02551 calcd. for C17H21C12NO5PS)
Elemental analysis: measured C 43.98%, H 4.46%, N 3.15%, calcd. for
C17H2OCl2NO5PS. 0.5 H2O C 44.26%, H 4.59%, N 3.04%.

[Example 11]
[0316]

(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-trifluoromethylphenylthi
o)phenyl]-2-methylbutylphosphoric acid monoester

[0317]

125

CA 02764126 2011-11-30
[Formula 1181

r H _ PO(OH)2
NH2
[03181

The target product (30 mg) was obtained as a white powder by
reacting the compound of Reference Example 45 (71 tng) in the same
manner as in Example 1.

Optical rotation: [a]D24 -7.12 (c 0.50, MeOH).

1H NMR (DMSO-d6-dTFA, 400 MHz): 8 1.28 (3H, s), 1.71-1.88 (2H, m),
2.70 (2H, t, j = 8.6 Hz), 3.87 (1H, dd, J = 11.0, 4.9 Hz), 3.94 (1H,
dd, J = 11.0, 4.9 Hz), 7.09 (1H, d, J = 8.6 Hz), 7.18 (1H, dd, J
= 8.6, 1.8 Hz), 7.27 (1H, d, J = 1.8 Hz), 7.33 (1H, d, J = 8.6 Hz),
7.39 (1H, d, J = 1.8 Hz), 7.55 (1H, d, J = 8.6 Hz).

HRESIMS (+) : 486.05108 (486.05187 calcd. for C18H21C1F3NO5PS)
Elemental analysis: measured C 42.61%, H 4.00%, N 2.76%, calcd. for
C,8H?0C1F3NO5PS. H2O C 42.91%, H 4.40%, N 2.78%.

[Example 12]
[0319]

(R)-2-allyl--2-amino-4-[2-chloro-4-(2-hydroxy-5-trifluoromethylp
henylthio)phenyl]-butylphosphoric acid monoester

[03201

126

CA 02764126 2011-11-30
[Formula 119]

F3C C10HO" C' I
PQOH~
NH2

[0321]

The target product (89 mg) was obtained as a white powder by
reacting the compound of Reference Example 46 (156 mg) in the same
manner as in Example 1.

Optical rotation: [a]D25 -7.41 (c 0.50, MeOH).

'H NMR (DMSO-d6-dTFA, 400 MHz) : 8 1.65-1.73 (2H, m), 2.39 (2H, d,
J = 7.3 Hz), 2,66-2.74 (2H, m), 3.42-3.53 (2H, m), 5.19 (1H, dd,
J = 10.4, 1.8 Hz), 5.24 (1H, dd, J = 17.1, 1.8 Hz), 5.55 (1H, s),
5.75-5.88 (1H, m), 7.10 (1H, d, J = 8.6 Hz), 7.18 (1H, dd, J = 8.6,
1.8 Hz), 7.25 (1H, d, J = 1.8 Hz), 7.31 (1H, d, J = 8.6 Hz), 7.36
(1H, d, J = 1.8 Hz), 7.55 (1H, d, J = 8.6 Hz), 7.94 (3H, br s).
HRESIMS (+) : 512.06693 (512.06752 calcd. for C20H23C1F3NO5PS)

Elemental analysis: measured C 45.80%, H 4.08%, N 2.61%, calcd. for
C2DH22C1F3NO5PS. 0. 5H20 C 46.12%, H 4.45%, N 2.69%.

[Example 13]
[0322]

(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-trifluoromethylphenylthi
o)phenyl]-2-propylbutylphosphoric acid monoester

[0323]

127

CA 02764126 2011-11-30
[Formula 120]

F3C C, ej
NH2
[0324]

The target product (50 mg) was obtained as a white powder by
reacting the compound of Reference Example 47 (122 mg) in the same
manner as in Example 1.

Optical rotation: [a]D25-7.67 (c 0.50, McOH).

1H NMR (DMSO-d6-dTFA, 400 MHz) ; 6 0.90 (3H, t, J = 7.3 Hz) , 1.26-1.40
(2H, m), 1.54-1.66 (2H, m), 1.70-1.82 (2H, m), 2,61-2.75 (2H, m),
3.86-3.97 (2H, m) , 7.10 (1H, d, J = 7.9 Hz), 7.19 (1H, dd, J = 7.9,
1.8 Hz), 7.27 (1H, d, J = 1.8 Hz), 7.34 (1H, d, J = 7.9 Hz), 7.40
(1H, d, J = 1.8 Hz), 7.56 (1H, d, J = 8.6 Hz).

HRESIMS (+) 514.08255 (514.08317 calcd. for C20H25C1F3NO5PS)
<Reference Example 48>

2-Chloro-4-(2-methoxyphenylthio)benzaldehyde
[0325]

128

CA 02764126 2011-11-30
[Formula 121]

Me CHO
[0326]

The target product (5.50 g) was obtained as a colorless oil
by reacting 2-methoxybenzenethiol (2.80 g) and
2-chloro-4-fluorobenzaldehyde (3.17 g) based on the same experiment
operations as in Reference Example 1 of WO 03029205 pamphlet.

1H NMR (CDC13, 400 MHz) : 8 3.83 (3H, s) , 6.98-7.08 (4H, m) , 7.49 (1H,
td, J = 7.9, 1.2 Hz), 7.52 (1H, dd, J = 7.9, 1.2 Hz), 7.74 (1H, d,
J = 7.9 Hz), 10.34 (1H, s).

EIMS (+) : 278 [M]+.
<Reference Example 49>
[2-Chloro-4-(2-methoxyphenylthio)phenyl]acetaldehyde
[0327]

[Formula 1221

S G1
Ome

[0328]

129

CA 02764126 2011-11-30

The target product (5.60 g) was obtained as a colorless oil
by reacting the compound of Reference Example 48 (5.50 g) based on
the same experiment operations inReference Example 326 of WO 04074297
pamphlet.

1H NMR (CDC13, 400 MHz): 8 3.80 (2H, d, J = 1.8 Hz), 3.86 (3H, s),
6.92-6.99 (2H, m), 7.12 (2H, d, J = 1.8 Hz), 7.29 (2H, dd, J = 7.9,
1.8 Hz), 7.35 (1H, td, J = 7.9, 1.8 Hz), 9.73 (1H, t, J = 1.8 Hz).
ELMS (+) : 292 [M]+.

<Reference Example 50>
2-Chloro-l-(2-iodoethyl)-4-(2-methoxyphenylthio)benzene
[03291

[Formula 1231
{
me o I
a si

[03301

The target product (5.40 g) was obtained as a colorless oil
by reacting the compound of Reference Example 49 (5.60 g) based on
the same experiment operations as in Reference Example 327 of WO
04074297 pamphlet.

1H NMR (CDC13, 400 MHz) : 6 3.25 (2H, ddd, J = B. 6, 6.7, 1.8 Hz) , 3.34
(2H, ddd, J = 8.6, 6.7, 1.8 Hz), 3.86 (3H, s), 6.90-6.96 (2H, m),
7.10 (1H, dd, J = 7.9, 1.8 Hz) , 7.14 (1H, d, J - 7.9 Hz) , 7.21-7.26
130

CA 02764126 2011-11-30
(2H, m), 7.32 (1H, td, J = 7.9, 1.8 Hz).
EIMS (+) : 404 [M]+.

<Reference Example 51>
(2S,5R)-2-[2-chloro-4-(2-methoxyphenylthio)phenyl)ethyl-3,6-dim
ethoxy-2-methyl-5-isopropyl-2,5-dihydropyrazine
[0331]

[Formula 1241

S CI
Ome
ao'Me N
N

OMe
[0332]

The target product (843 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 50 (1.23 g) in the
same manner as in Reference Example 1.

'H NMR (CDC13, 400 MHz) : 8 0.71 (3H, d, J = 6.7 Hz), 1.09 (3H, d,
J = 6.7 Hz), 1.36 (3H, s), 1.80 (1H, ddd, J = 12.8, 11.6, 4.9 Hz),
2.11 (1H, ddd, J = 12.8, 11.6, 4.9 Hz) , 2.21-2.33 (1H, m) , 2.38 (1H,
ddd, J = 12.8, 11.6, 4.9 Hz), 2.46 (1H, ddd, J = 12.8, 11.6, 4.9
Hz), 3.68 (3H, s), 3.69 (3H, s), 3.87 (3H, s), 3.99 (1H, d, J = 3.7
Hz), 6.85-6.93 (2H, m), 7.07 (1H, d, J = 7.9 Hz), 7.11 (1H, dd, J
= 7.9, 1.8 Hz), 7.12 (1H, dd, J = 7.9, 1.8 Hz) , 7.21-7.26 (2H, m).
131

CA 02764126 2011-11-30
ESIMS (+): 475 [M+H].

<Reference Example 52>
Methyl

(S)-4-[2-chloro-4-(2--methoxyphenylthio)phenyl]-2-t-butoxycarbon
ylamino-2-methyl butyrate

[0333]
[Formula 125]

S CI

Me C'CO2Me
NHBoc
[0334]

The target product (457 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 51 (542 mg) in the
same manner as in Reference Example 3.

1H NMR (CDC13, 400 MHz) : 6 1.45 (9H, s) , 1.58 (3H, s) , 2.11 (1H, ddd,
J = 13.4, 11.6, 4.9 Hz), 2.38 (1H, br s), 2.52 (1H, td, J = 12.8,
4.9 Hz), 2.67 (1H, td, J = 12.8, 4.9 Hz), 3.74 (3H, s), 3.86 (3H,
s), 5.41 (1H, br s), 6.91 (2H, td, J = 7.9, 1.8 Hz), 7.08 (1H, d,
J = 7. 9 Hz) , 7.10 (1H, d, J = 1. 8 Hz) , 7. 14 (1H, td, J = 7. 9, 1. 8Hz) ,
7.28 (2H, td, J = 7.9, 1.8 Hz).

ESIMS (+) : 480 [M+H] `.
<Reference Example 53>

132

CA 02764126 2011-11-30
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxyphenylthio)
phenyl]-2-methylbutan-l-ol

[0335]
[Formula 1261

NHBoc
aOW

[0336]

The target product (457 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 52 (542 mg) in the
same manner as in Reference Example 5.

1H NMR (CDC13, 400 MHz) : 8 1.25 (3H, s) , 1.44 (9H, s) , 1.81 (1H, ddd,
J = 13.4, 12.2, 4.9 Hz), 2.06 (1H, ddd, J = 13.4, 12.2, 4.9 Hz),
2.68 (1H, td, J = 12.8, 4.9 Hz), 2.75 (1H, td, J = 12.8, 4.9 Hz),
3.63-3.75 (2H, m) , 3.87 (3H, s) , 4.09 (1H, br s) , 4.68 (1H, s), 6.91
(2H, td, J = 7.3, 1.2 Hz) , 7.09-7.18 (3H, m) , 7.28 (1H, dd, J = 7.3,
1.2 Hz),.

ESIMS (+): 452 [M+H]+.
<Reference Example 54>
(S)-2-t-butoxycarbonylamino-4-[4-(2-t-butoxycarbonyloxyphenylth

io)-2-chlorophenyl]-1-dimethoxyphosphoryloxy-2-methylbutane
[0337]

133

CA 02764126 2011-11-30
[Formula 127]

Bic PO(We
HBoc
[0338]

An 0-butoxycarbonyl compound (363 mg) was obtained by reacting
the compound of Reference Example 53 (457 mg) in the same manner
as in Reference Example 30. Then, the target product (169 mg) was
obtained as a colorless oil by reacting this 0-butoxycarbonyl compound
(199 mg) in the same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz) : 5 1.35 (3H, s), 1.44 (9H, s), 1.52 (9H, s),
1.76 (1H, ddd, J = 13.4, 11.0, 5.5 Hz), 2.02-2.16 (1H, m) , 2.61-2.77
(2H, m), 3.78 (3H, d, J = 11.0 Hz), 3.79 (3H, d, J = 11.0 Hz), 4.02
(1H, dd, J = 9.8, 5.5 Hz) , 4.22 (1H, dd, J = 9.8, 5.5 Hz) , 4.63 (1H,
s) , 7.09-7.17 (2H, m), 7.19 (2H, d, J = 7.9 Hz), 7.27-7.35 (3H, m) .
ESIMS (+): 646 [M+H]+.

[Example 14]
[0339]

(S)-2-amino-4-[2-chloro-4-(2-hydroxyphenylthio)phenyl]-2-methyl
butylphosphoric acid monoester

[0340]

134

CA 02764126 2011-11-30
[Formula 128]

O
COH PO(OH)2
NH2
[0341]

The target product (80 mg) was obtained as a white powder by
reacting the compound of Reference Example 54 (199 mg) in the same
manner as in Example 1.

Optical rotation: [a]D27 -9.06 (c 0.50, McOH).

1H NMR (DMSO-d6-dTFA, 400 MHz): 8 1.26 (3H, s), 1.66-1.85 (2H, m),
2.65 (2H, t, j = 8.6 Hz), 3.84 (1H, dd, J = 11.0, 5.5 Hz) , 3.91 (1H,
dd, J = 11.0, 5.5 Hz), 6.81 (1H, t, j = 7.3 Hz), 6.94 (1H, d, J =
7.3 Hz), 7.06 (1H, d, J = 7.3 Hz, 7.07 (1H, s), 7.21 (2H, d, J =
7.9 Hz), 7.25 (1H, d, J = 7.9 Hz).

HRESIMS (+): 418.06445 (418.06448 calcd. for C17H22C1NO5PS)
Elemental analysis: measured C 47.91%, H 4.94%, N 3.21%, calcd. for
C17H21C1NO5PS. 0.4H20 C 48.04%, H 5.17%, N 3.30%.

<Reference Example 55>
6-t-butyldimethylsilyloxy-1,3-benzoxathiol-2-one
[0342]

135

CA 02764126 2011-11-30
(Formula 129]

TBSt
I %'Z .S
(0343]

Imidazole (972 mg) and t-butylchlorodimethylsilane (2.15 g)
were added to a solution of 6-hydroxy-1, 3-benzoxathiol-2-one (2.00
g) in N,N-dimethylformamide (60 mL) to form a reaction solution.
This reaction solution was stirred for 4 hours at normal temperature.
Water was added to the reaction solution, and the reaction solution
was extracted with ethyl acetate. The extract was washed with water
and saturated brine in that order, and then dried over anhydrous
sodium sulfate. The anhydrous sodium sulfate was removed by
filtration, and then the solvent was removed by distillation under
reduced pressure. The resultant residue was purified by silica gel
column chromatography (hexane : ethyl acetate = 20 : 1) to obtain
the target product (3.00 g) as a colorless oil.

1H NMR (CDC13, 400 MHz) : 6 0.20 (6H, s), 0.98 (9H, s), 6.77 (1H, dd,
J = 8.6, 2.4 Hz), 6.87 (1H, d, J = 2.4 Hz), 7.14 (1H, d, J = 8.6
Hz).

CIMS (+): 283 [M+H]*.
<Reference Example 56>
5-t-butyldimethylsilyloxy-2-hydroxybenzenethiol

136

CA 02764126 2011-11-30
[0344]

[Formula 130]

TBSO H
C(OH
[0345]

The target product (2.72 g) was obtained as a colorless oil
by reacting the compound of Reference Example 55 (3.00 g) in the
same manner as in Reference Example 16.

1H NMR (CDC13, 400 MHz) : 8 0.16 (6H, s) , 0.97 (9H, s) , 3.06 (1H, s) ,
5.73 (1H, s), 6.71 (1H, dd, J = 8.6, 2.4 Hz), 6.81 (1H, d, J = 2.4
Hz), 6.93 (1H, d, J = 8.6 Hz).

CIMS (+): 257 [M+H]
<Reference Example 57>
(S)-2-t-butoxycarbonylamino-4-[4-(5-t-butyldimethylsilyloxy-2-h
ydroxyphenylthio)-2-chlorophenyl]-2-methylbutan-l-ol
(0346]

137

CA 02764126 2011-11-30
[Formula 131]

`CBS O CI

H
NHBoc

[0347]

The target product (633 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 5 (800 mg) and the
compound of Reference Example 56 (627 mg) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz) : 6 0.17 (6H, s) , 0.97 (9H, s) , 1.23 (3H, s) ,
1.43 (9H, s) , 1.77 (1H, ddd, J = 13. 4, 12.2, 5.5 Hz) , 2.00 (1H, ddd,
J = 13.4, 12.2, 5.5 Hz) , 2.58-2.76 (2H, m), 3.65 (1H, dd, J = 11.6,
4.9 Hz), 3.70 (1H, dd, J = 11,6, 7.3 Hz), 4.07 (1H, br s), 4.65 (1H,
s), 6.05 (1H, s), 6.90 (2H, td, J = 8.6, 2.4 Hz), 6.94 (1H, d, J
= 8.6 Hz), 6.98 (1H, d, J = 2.4 Hz), 7.03 (1H, d, J = 2.4 Hz), 7.10
(1H, d, J = 8.6 Hz).

ESIMS (+): 568 [M+H]+
<Reference Example 58>
(S)-2-t-butoxycarbonylamino-4-[4-(5-t-butyldimethylsilyloxy-2-m

ethoxymethoxyphenylthio)-2-chlorophenyl]-2-methylbutan-l-o1
[0348]

138

CA 02764126 2011-11-30
[Formula 132)

TBSO CI

NHBoc
[03491

Potassium carbonate (185 mg) and chioromethyl methyl ether
(102 L) were added under ice cooling to a solution of the compound
of Reference Example 57 (760 mg) in acetone (13 .4 mL) to form a reaction
solution. This reaction solution was stirred for 3 hours under ice
cooling. Water was added to the reaction solution, and the reaction
solution was extracted with ethyl acetate. The extract was washed
with water and saturated brine in that order, and then dried over
anhydrous sodium sulfate. The anhydrous sodium sulfate was removed
by filtration, and then the solvent was removed by distillation under
reduced pressure. The resultant residue was purified by silica gel
column chromatography (hexane : ethyl acetate = 3 : 1) to obtain
the target product (639 mg) as a colorless oil.

1H NMR (CDC13, 400 MHz) : 5 0.08 (6H, s) , 0.91 (9H, s) , 1.25 (3H, s) ,
1.44 (9H, s) , 1.80 (1H, ddd, J = 13.4, 12.2, 5.5 Hz) , 2.02 (1H, ddd,
J = 13.4, 12.2, 5.5 Hz) , 2.69 (1H, td, J = 12.8, 4.9 Hz), 2.76 (1H,
td, J = 12.8, 4.9 Hz), 3.45 (3H, s), 3.67 (1H, dd, J = 11.6, 4.9
Hz), 3.72 (1H, dd, J = 11,6, 7.3 Hz), 4.08 (1H, br s), 4.68 (1H,
s), 5.13 (2H, s), 6.53 (1H, d, J = 3.1 Hz), 6.68 (1H, dd, J = 9.2,
139

CA 02764126 2011-11-30

3.1 Hz), 7.00 (1H, d, J = 9.2 Hz), 7.18 (2H, d, J = 1.8 Hz), 7.32
(1H, d, J = 1.8 Hz).

ESIMS (+) 612 [M+H]+.
<Reference Example 59>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-hydroxy-2-methoxym

ethoxyphenylthio)phenyl]-2-methylbutan-l-ol
[0350]

[Formula 133]

HO S ,p C

N!OM N
Nloe
[0351]

A tetrabutyl ammonium f luoride - tetrahydrofuran solution (1.0
mol/L, 1.0 mL) was added under ice cooling to a solution of the compound
of Reference Example 58 (639 mg) in tetrahydrofuran (10 mL) to form
a reaction solution. This reaction solution was stirred for 30
minutes under ice cooling. Water was added to the reaction solution,
and the reaction solution was extracted with ethyl acetate. The
extract was washed with water and saturated brine in that order,
and then dried over anhydrous sodium sulfate. The anhydrous sodium
sulfate was removed by filtration, and then the solvent was removed
by distillation under reduced pressure. The resultant residue was
140

CA 02764126 2011-11-30

purified by silica gel column chromatography (hexane : ethyl acetate
= 1 : 1) to obtain the target product (486 mg) as a colorless oil.
1H NMR (DMSO-d6, 400 MHz): 51.17 (3H, s) , 1.37 (9H, s) , 1.65-1.77
(1H, m), 1.81-1.95 (1H, m), 2.60 (2H, t, j = 8.6 Hz), 3.29 (3H, s),
3.36-3.41 (2H, m) , 4.72 (1H, t, J = 5.5 Hz) , 5.08 (2H, s) , 6.28 (1H,
s), 6.42 (1H, d, J = 3.1 Hz), 6.63 (1H, dd, J = 9.2, 3.1 Hz), 6.98
(1H, d, J = 9.2 Hz), 7.22 (1H, dd, J = 7.9, 1.8 Hz), 7.28 (1H, d,
J = 1.8 Hz), 7.30 (1H, d, J = 7.9 Hz), 9.24 (1H, S).

ESIMS (+) :498 [M+H] `.
<Reference Example 60>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-t
rifluoromethanesulfonyloxyphenylthio)phenyl]-2-methylbutan-l-ol
[0352]

[Formula 134]

H
inn
NHBoc
[0353]

Under an argon atmosphere, triethylamine (273 L) and
N-phenyltrifluoromethanesulfonimide (366 mg) were added under ice
cooling to a solution of the compound of Reference Example 59 (486
mg) in methylene chloride (5 mL) to form a reaction solution. This
reaction solution was stirred for 2 hours at normal temperature.
141

CA 02764126 2011-11-30

Water was added to the reaction solution, and the reaction solution
was extracted with ethyl acetate. The extract was washed with water
and saturated brine in that order, and then dried over anhydrous
sodium sulfate. The anhydrous sodium sulfate was removed by
filtration, and then the solvent was removed by distillation under
reduced pressure. The resultant residue was purified by silica gel
column chromatography (hexane : ethyl acetate = 2 : 1) to obtain
the target product (586 mg) as a colorless oil.

''H NMR (CDC13, 400 MHz) : 5 1.26 (3H, s), 1.45 (9H, s), 1.84 (1H, ddd,
j = 14.1, 12.2, 5.5 Hz), 2.06 (1H, ddd, J = 14.1, 12.2, 5.5 Hz),
2.74 (1H, td, J = 12.8, 4.9 Hz), 2.81 (1H, td, J = 12.8, 4.9 Hz),
3.47 (3H, s), 3.68 (1H, dd, J = 11.6, 4.9 Hz), 3.73 (1H, dd, J =
11,6, 7.3 Hz), 4.08 (1H, br s), 4.69 (1H, s), 5.24 (2H, s), 6.78
(1H, d, J = 3.1 Hz), 7.05 (1H, dd, J = 9.2, 3.1 Hz), 7.16 (1H, d,
J = 9.2 Hz), 7.27 (2H, d, j = 1.8 Hz), 7.43 (1H, d, J = 1.8 Hz).
ESIMS (+): 630 [M+H]+.

<Reference Example 61>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-cyano-2-methoxymet
hoxyphenylthio)phenyl]-2-methylbutan-l-ol
[0354]

142

CA 02764126 2011-11-30
[Formula 1.35)

MOM
NHBoc
[0355]

Under an argon atmosphere,
1,1'-bis(diphenylphosphino)-ferrocene (17.6 mg) was added into a
solution of tetrakistriphenylphosphine palladium(0) (36.7 mg) in
N,N-dimethylformamide (1.6 mL) to form a reaction solution. This
reaction solution was stirred for 10 minutes at normal temperature.
Then, zinc cyanide (74.5 mg) and the compound of Reference Example
60 (200 mg) were added to the reaction solution, and the reaction
solution was stirred for 4 hours at 80 C. Then,
tetrakistriphenylphosphine palladium(0) (36.7 mg),
1,1'-bis (diphenylphosphino) - ferrocene (17.6 mg), and zinc cyanide
(74.5 mg) were further added to the reaction solution, and the reaction
solution was stirred for 2 hours at 80 C. Water was added to the
reaction solution, and the reaction solution was extracted with ethyl
acetate. The extract was washed with water and saturated brine in
that order, and then dried over anhydrous sodium sulfate. The
anhydrous sodium sulfate was removed by filtration, and then the
solvent was removed by distillation under reduced pressure. The
resultant residue was purified by silica gel column chromatography
143

CA 02764126 2011-11-30

(hexane : ethyl acetate = 2 : 1) to obtain the target product (97
mg) as a colorless oil.

'H NMR (CDC13, 400 MHz) : S 1.26 (3H, s) , 1.45 (9H, s) , 1.84 (1H, ddd,
J = 14.1, 12.2, 5.5 Hz), 2.07 (1H, ddd, J = 14.1, 12.2, 5.5 Hz),
2.69-2.85 (2H, m), 3.47 (3H, s), 3.66-3.75 (2H, m), 4.06 (1H, br
s), 4.69 (1H, s), 5.24 (2H, s), 6.78 (1H, d, J = 3.1 Hz), 7.05 (1H,
dd, J = 9.2, 3.1 Hz), 7.16 (1H, d, J = 9.2 Hz), 7.27 (2H, s), 7.43
(1H, d, J = 1.2 Hz).

ESIMS (+): 490 [M+H]'.
<Reference Example 62>
(S)-4-[4-(5-acetyl-2-methoxymethoxyphenylthio)-2-chlorophenyl]-

2-t-butoxycarbonylamino-2-methylbutan-l-ol
(0356]

[Formula 136]
0

m
NHBoc
[0357]

Under an argon atmosphere,
1,3'-bis(diphenylphosphino)-propane (26.2 mg) was added to a
solution of palladium(II) acetate (7.1 mg) in N,N-dimethylformamide
(1.5 mL) to form a reaction solution. This reaction solution was
144

CA 02764126 2011-11-30

stirred for 30 minutes at normal temperature. Then, triethylamine
(44 L), butylvinyl ether (205 L), and a solution of the compound
of Reference Example 60 (200 mg) in N,N-dimethylformamide (3.5 mL)
were added to the reaction solution, and the reaction solution was
stirred for 18 hours at 80 C. A saturated ammonium chloride aqueous
solution was added to the reaction solution, and the reaction solution
was extracted with ethyl acetate. The extract was washed with water
and saturated brine in that order, and then dried over anhydrous
sodium sulfate. The anhydrous sodium sulfate was removed by
filtration, and then the solvent was removed by distillation under
reduced pressure. The resultant residue was purified by silica gel
column chromatography (hexane : ethyl acetate = 2 : 1) to obtain
the target product (117 mg) as a colorless oil.

1H NMR (CDC13, 400 MHz) : 8 1.25 (3H, s) , 1.44 (9H, s) , 1.81 (1H, ddd,
J = 14.1, 12.2, 5.5 Hz), 2.04 (1H, ddd, J = 14.1, 12.2, 5.5 Hz),
2.50 (3H, s), 2.64-2.81 (2H, m), 3.38 (3H, s), 3.63-3.74 (2H, m),
4.07 (1H, br s) , 4.68 (1H, s), 5.27 (2H, s), 7.13-7.21 (3H, m) , 7.31
(1H, d, J = 1.8 Hz), 7.82 (1H, d, J = 1.8 Hz), 7.87 (1H, dd, J =
8.6, 1.8 Hz).

ESIMS (+) : 524 [M+fi]' .
<Reference Example 63>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-cyano-2-methoxymet

hoxyphenylthio)phenyl]-1-dimethoxyphosphoryloxy-2-methylbutane
[0358]

145

CA 02764126 2011-11-30
(Formula 137]

NC 10(mom S
PO(OMe)2
NHBoc

[0359]

The target product (62 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 61 (97 mg) in the same
manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz) : 8 1.38 (3H, s) , 1.45 (9H, s) , 1.82 (1H, ddd,
J = 13.4, 12.2, 5.5 Hz), 2.07-2.21 (1H, m), 2.69-2.84 (2H, m), 3.45
(3H, s), 3.79 (3H, d, J = 11.0 Hz), 3.80 (3H, d, J = 11.0 Hz), 4.04
(1H, dd, J = 9.8, 5.5 Hz) , 4.25 (1H, dd, J = 9.8, 5.5 Hz), 4.65 (1H,
s), 5.29 (2H, s), 7.15 (1H, d, J = 1.8 Hz), 7.17 (1H, d, J = 8.6
Hz) , 7.25-7.27 (2H, m) , 7.42 (1H, t, J = 1. 8 Hz), 7.46 (1H, J = 8.6,
1.8 Hz).

ESIMS (+): 615 [M+H]'.
<Reference Example 64>
(S)-4-[4-(5-acetyl-2-methoxymethoxyphenylthio)-2-chlorophenyl]-

2-t-butoxycarbonylamino-l-dimethoxyphosphoryloxy-2-methylbutane
[0360]

146

CA 02764126 2011-11-30
[Formula 1381

MOM PO(OMe)2
NHBoc
[03611

The target product (112 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 62 (117 mg) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz) : 8 1.36 (3H, s) , 1.44 (9H, s) , 1.76 (1H, ddd,
j = 13.4, 12.2, 5.5 Hz) , 1.98-2.17 (1H, m), 2.50 (3H, s), 2.64-2.76
(2H, m), 3.38 (3H, s), 3.78 (3H, d, J = 11.0 Hz), 3.79 (3H, d, J
= 11.0 Hz), 4.02 (1H, dd, J = 9.8, 4.9 Hz), 4.23 (1H, dd, J = 9.8,
4.9 Hz), 4.63 (1H, s), 5.26 (2H, s), 7.16 (2H, S), 7.20 (1H, t, J

7.9 Hz), 7.29 (1H, s), 7.84 (1H, d, J = 1.8 Hz), 7.88 (1H, dd,
J = 8.6, 1.8 Hz).

ESIMS (+): 632 [M+H]+.
[Example 151

(03621
(S)-2-amino-4-[2-chloro-4-(5-cyano-2-hydroxyphenylthio)phenyll-
2-methylbutylphosphoric acid monoester

[03631

147

CA 02764126 2011-11-30
[Formula 139]

NC S a

OH PC(OH)2
NH2
[0364]

The target product (30 mg) was obtained as a white powder by
reacting the compound of Reference Example 63 (62 mg) in the same
manner as in Example 1.

Optical rotation: [aID5 -6.74 (c 0.50, MeOH).

1H NMR (DMSO-d6-dTFA, 400 MHz) 8 1.27 (3H, s), 1.69-1.87 (2H, m),
2.70 (2H, t, J = 8.6 Hz) , 3.86 (1H, dd, J = 11.0, 4.9 Hz) , 3.93 (1H,
dd, J = 11.0, 4.9 Hz), 7.04 (1H, d, J = 7.9 Hz), 7.20 (1H, dd, J
= 7.9, 1.2 Hz), 7.26-7.44 (2H, m), 7.48 (1H, d, J = 1.8 Hz), 7.63
(1H, dd, J = 7.9, 1.8 Hz).

HRESIMS (+): 443.05981 (443.05973 calcd. for C18H21C1N2O5PS)
[Example 16]

[03651
(S)-4-[4-(5-acetyl-2-hydroxyphenylthio)-2-chlorophenyl]-2-amino
-2-methylbutylphosphoric acid monoester

[03661

148

CA 02764126 2011-11-30
[Formula 140]

aH PO(OH )2
NFfj
[0367]

The target product (69 mg) was obtained as a white powder by
reacting the compound of Reference Example 64 (112 mg) in the same
manner as in Example 1.

Optical rotation: [a]o5 -9.53 (c 0.50, MeOH).

1H NMR (DMSO-d6-dTFA, 400 MHz): S 1.27 (3H, s) , 1.68-1.86 (2H, m),
2.43 (3H, s), 2.68 (2H, t, J = 8.6 Hz), 3.85 (1H, dd, J = 11.0, 4.9
Hz) , 3.92 (1H, dd, J = 11.0, 4.9 Hz), 7.01 (1H, d, J = 7.9 Hz), 7.13
(1H, dd, J = 7.9, 1.2 Hz), 7.19 (1H, d, J = 1.8 Hz), 7.29 (1H, d,
J = 7.9 Hz), 7.77 (1H, d, J = 1.8 Hz), 7.85 (1H, dd, J = 7.9, 1.8
Hz).

HRESIMS (+): 460.07487 (460.07505 calcd. for C19H24C1NO6PS)
Elemental analysis: measured C 47.44%, H 5.07%, N 2.59%, calcd. for
C19H23C1NO6PS. H2O C 47.75%, H 5.27%, N 2.93%.

<Reference Example 65>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-hydroxy-5-trifluor
omethylphenylthio)phenyl]-2-methylbutan-l-o1
[0368]

149

CA 02764126 2011-11-30
[Formula 1411

F3C S Cl

H
NHBoc

[0369]

A diphenyl sulfide was obtained by reacting the compound of
Reference Example 19 (400 mg) and the compound of Reference Example
(554 mg) in the same manner as in Reference Example 20. Then,
the target product (480 mg) was obtained as a colorless oil by reacting
this diphenyl sulfide in the same manner as in Reference Example
33.

1H NMR (CDC13, 400 MHz) : 8 1.22 (3H, s) , 1.43 (9H, s) , 1.79 (1H, ddd,
J = 13.4, 12.2, 5.5 Hz), 2.01 (1H, ddd, J = 13.4, 12.2, 5.5 Hz),
2.60-2.76 (2H, m), 3.61-3.73 (2H, m), 4.00 (1H, br s), 4.65 (1H,
s), 6.74 (1H, s), 6.91 (1H, dd, J = 7.9, 1.8 Hz), 7.09 (1H, d, J
= 1.8 Hz), 7.12-7.18 (2H, m), 7.63 (1H, dd, J = 8.6, 2.4 Hz), 7.80
(1H, d, J = 2.4 Hz).

ESIMS (+); 506 [M+H]+.
<Reference Example 66>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-t

rifluoromethylphenylthio)phenyl]-2-methylbutan-l-o1
[0370]

150

CA 02764126 2011-11-30
[Formula 142]

F3C
Cl
M40aM H
N0C
I~Xhw
HB
[0371]

The target product (302 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 65 (300 mg) in the
same manner as in Reference Example 58.

2H NMR (CDC13, 400 MHz) : 8 1.25 (3H, s) , 1.44 (9H, s) , 1.82 (1H, ddd,
J = 13.4, 12.2, 5.5 Hz), 2.06 (1H, ddd, J = 13.4, 12.2, 5.5 Hz),
2.66-2.83 (2H, m), 3.41 (3H, s), 3.65-3.74 (2H, m), 4.03 (1H, br
s), 4.67 (1H, s), 5.25 (2H, s), 7.16-7.24 (3H, m), 7.35 (1H, d, J
= 1.8 Hz) , 7.36 (1H, d, J = 1.8 Hz) , 7.47 (iH, dd, J = 8.6, 1.8 Hz) .
ESIMS (+): 550 [M+H]+.

<Reference Example 67>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-t
rifluoromethylphenylthio)phenyl]-2-methylbutan-l-al
[0372]

151

CA 02764126 2011-11-30
[Formula 1433

F3C S CI
OMOM
NHBoc
[03733

Under an argon atmosphere, a solution of dimethyl sulfide in
methylene chloride (1 mL) was added at -78 C to a solution of oxalyl
chloride (67.5 L) in methylene chloride (5 mL) to form a reaction
solution. This reaction solution was stirred at -78 C for 10 minutes.
Then, a solution of the compound of Reference Example 66 (213 mg)
in methylene chloride (2 mL) was added to the reaction solution,
and the reaction solution was stirred at -78 C for 1.5 hours.
Triethylamine (378 L) was added to the reaction solution, and then
stirred for 30 minutes under ice cooling. Then, a saturated ammonium
chloride aqueous solution was added to the reaction solution, and
the reaction solution was extracted with ethyl acetate. The extract
was washed with water and saturated brine in that order, and then
dried over anhydrous sodium sulfate. The anhydrous sodium sulfate
was removed by filtration, and then the solvent was removed by
distillation under reduced pressure. The resultant residue was
purified by silica gel column chromatography (hexane: ethyl acetate
= 3 : 1) to obtain the target product (201 mg) as a colorless oil.
152

CA 02764126 2011-11-30

1H NMR (CDC13, 400 MHz) : S 1.41 (3H, s) , 1.46 (9H, s) , 1.91-2.08 (1H,
m) , 2.16--2.31 (1H, m) , 2.57 (1H, ddd, J = 13.4, 12.2, 4.9 Hz) , 2.68
(1H, ddd, J = 13.4, 12.2, 4.9 Hz), 3.41 (3H, s), 5.24 (1H, br s),
5.25 (2H, s), 7.14 (1H, d, J = 7.9 Hz), 7.17 (1H, dd, J = 7.9, 1.8
Hz), 7.21 (1H, d, J = 7.9 Hz), 7.33 (1H, d, J = 1.8 Hz), 7.38 (1H,
d, J = 1.8 Hz), 7.48 (1H, dd, J = 7.9, 1.8 Hz), 9.40 (1H, S).
ESIMS (-): 546 [M-H]+.

<Reference Example 68>
Dimethyl

(S)-3-t-butoxycarbonylamino-5-[2-chloro-4-(2-methoxymethoxy-5-t
rifluoromethylphenylthio)phenyl]-3-methyl-l-pentenylphosphonate
[0374]

[Formula 144]
F3c

X Z PO(OMe)2
Comm
NHBoc
[0375]

Under an argon atmosphere, an n-butyllithium-hexane solution
(1.60 mol/L, 25.5 mL) was added at -78 C to a solution of tetramethyl
methylenediphosphonate (111 mg) in tetrahydrofuran (3 mL) to form
a reaction solution. This reaction solution was stirred at -78 C
for 30 minutes. Then, a solution of the compound of Reference Example
67 (111 mg) in tetrahydrofuran (1 mL) was added to the reaction solution.
153

CA 02764126 2011-11-30

The reaction solution was stirred at -78 C for 2 hours, and then
stirred for 2 hours at normal temperature. Then, a saturated ammonium
chloride aqueous solution was added to the reaction solution, and
the reaction solution was extracted with ethyl acetate. The extract
was washed with water and saturated brine in that order, and then
dried over anhydrous sodium sulfate. The anhydrous sodium sulfate
was removed by filtration, and then the solvent was removed by
distillation under reduced pressure. The resultant residue was
purified by silica gel column chromatography (hexane: ethyl acetate
= 2 : 1, and then ethyl acetate) to obtain the target product (160
mg) as a colorless oil.

IH NMR (CDC13, 400 MHz) : 5 1.44 (3H, s) , 1.46 (9H, s) , 1.90 (1H, td,
J = 12.2, 4.9 Hz), 2.06-2.17 (1H, m), 2.61-2.78 (2H, m), 3.41 (3H,
s), 3.73 (6H, d, J = 11.0 Hz), 4.65 (1H, br s), 5.25 (2H, s), 5.70
(1H, t, J = 17.7 Hz), 6.82 (1H, dd, J = 22.6, 17.7 Hz), 7.16-7.18
(2H, m), 7.21 (1H, d, J = 8.6 Hz), 7.34 (1H, d, J = 1.8 Hz), 7.34
(1H, d, J = 1.8 Hz), 7.48 (1H, dd, J = 8.6, 1.8 Hz).

ESIMS (+): 654 [M+H]`.
<Reference Example 69>
Dimethyl

(S)-3-t-butoxycarbonylamino-5-[2-chloro-4-(2-methoxymethoxy-5-t
rifluoromethylphenylthio)phenyl]-3-methyl-l-pentylphosphonate
[0376]

154

CA 02764126 2011-11-30
[Formula 145]

F3C S = CI

PO(OMe)2
IC(omom-
NHBcc
[0377]

10% Palladium-active carbon (200 mg) was added to a solution
of the compound of Reference Example 68 (160 mg) in ethyl acetate
(16 mL) to form a reaction solution. This reaction solution was,
under hydrogen purging, stirred for 20 hours at normal temperature.
The reaction solution wasfiltrated using Celite, and then the solvent
was removed by distillation. The resultant residue was purified
by silica gel column chromatography (hexane : ethyl acetate = 2
1) to obtain the target product (150 mg) as a colorless oil.

1H NMR (CDC13, 400 MHz) : 8 1.30 (3H, s), 1.46 (9H, s) , 1.77-1.86 (4H,
m), 2.04-2.15 (1H, m), 2.16-2.28 (1H, m), 2.66-2.82 (2H, rn), 3.45
(3H, s), 3.79 (6H, d, J = 11.0 Hz), 4.44 (1H, br s), 5.30 (2H, s),
7.17-7.28 (3H, m), 7.38-7.42 (2H, m), 7.52 (1H, dd, J = 7.9, 2.4
Hz).

ESIMS (+): 656 [M+H].
[Example 17]

[0378]
(S)-3-amino-5-[2-chloro-4-(2-hydroxy-5-trifluoromethylphenylthi
o)phenyl]-3-methylpentylphosphonic acid

155

CA 02764126 2011-11-30
[0379]

[Formula 146]

OH )2
PO(OH
NH2
[0380]

The target product (19 mg) was obtained as a white powder by
reacting the compound of Reference Example 69 (70 mg) in the same
manner as in Example.

Optical rotation: [a]p25 -14.62 (c 0.11, MeOH).

1H NMR (DMSO-d6-dTFA, 400 MHz): 6 1.23 (3H, s), 1.37-1.52 (2H, m),
1.64-1.86 (4H, m) , 2.56-2.70 (2H, m), 7.08 (2H, d, J = 8.6 Hz), 7.21
(1H, s), 7.24-7.33 (1H, m), 7.41 (1H, s), 7.51 (1H, d, J = 8.6 Hz) .
HRESIMS (+) : 484.07277 (484.07260 calcd. for C,9H23C1F3NO4PS)

<Reference Example 70>
(2S,5R)-2-(4-bromo-2-chlorophenyl)ethyl-3,6-dimethoxy-2-propyl-
5-isopropyl-2,5-dihydropyrazine

[0381]

156

CA 02764126 2011-11-30
[Formula 147]

CIII come
Me
[0382]

The target product (8.01 g) was obtained as a colorless oil
by reacting
(5R)-3,6-dimethoxy-2-propyl-5-isopropyl-2,5-dihydropyrazine

(5.21 g) in the same manner as in Reference Example 1.

H NMR (CDCl3, 400 MHz): 8 0.70 (3H, d, J = 6.7 Hz), 0.86 (3H, t,
J = 7.3 Hz), 1.11 (3H, d, J = 6.7 Hz), 1.15-1.30 (2H, m), 1.49-1.62
(1H, m) , 1.71-1.84 (2H, m), 1.98 (1H, td, J = 12.4, 4.8 Hz) , 2.29-2.47
(3H, m), 3.69 (3H, s), 3.70 (3H, s), 3.95 (1H, d, J = 3.0 Hz), 7.01
(1H, d, J = 7.9 Hz), 7.27 (1H, dd, J = 7.9, 1.8 Hz), 7.46 (1H, d,
J = 1.8 Hz).

ESIMS (+):443 [M+H]+.
<Reference Example 71>
(2S,5R)-2-(4-bromo-2-chlorophenyl)ethyl-2-butyl-3,6-dimethoxy-5
-isopropyl-2,5-dihydropyrazine

[0383]

157

CA 02764126 2011-11-30
[Formula 148]

Me
Me
[0384]

The target product (2.52 g) was obtained as a colorless oil
by reacting
(5R)-2-butyl-3,6-dimethoxy-5-isopropyl-2,5-dihydropyrazine(3.83
g) in the same manner as in Reference Example 1.

'H NMR (CDC13, 400 MHz) : 8 0.71 (3H, d, J = 6.7 Hz), 0.85 (3H, t,
J = 7.3 Hz), 0.94-1.06 (1H, m), 1.11 (3H, d, J = 6.7 Hz), 1.12-1.30
(3H, m), 1.57-1.64 (1H, m), 1.74-1.84 (2H, m), 1.98 (1H, dt, J =
12.2, 4.9 Hz), 2.30-2.45 (3H, m), 3.70 (3H, s), 3.71 (3H, s), 3.95
(1H, d, J = 3.7 Hz), 7.00 (1H, d, J = 8.6 Hz), 7.25-7.29 (1H, m),
7.46 (1H, d, J = 1.8 Hz).

EIMS (+): 456 [M]+
Reference Example 72>
Methyl

(S)-4-(4-bromo-2-chlorophenyl)-2-t-butoxycarbonylamino-2-propyl
butyrate

[0385]

158

CA 02764126 2011-11-30
[Formula 149]

Br,
,, C1

C02Me
NHBoc
[0386]

The target product (35.6 g) was obtained as a colorless oil
by reacting the compound of Reference Example 70 (53.4 g) in the
same manner as in Reference Example 4.

1H NMR (CDC13, 400 MHz) : 6 0.89 (3H, t, J = 7.3 Hz) . 0.96-1.10 (1H,
m), 1.25-1.39 (1H, m), 1.46 (9H, s), 1.69 (1H, ddd, J = 13.9, 11.5.
4.8 Hz), 1.99-2.10 (1H, m), 2.20-2.35 (1H, m), 2.42 (1H, ddd, J =
13.9, 11.5, 4.8 Hz) , 2.49-2.60 (1H, m) , 2.64 (1H, td, J = 13.9, 4.8
Hz), 3.74 (3H, s), 5.62 (1H, br s), 7.03 (1H, d, J = 8.5 Hz), 7.29
(1H, dd, J = 8.5, 1.8 Hz), 7.48 (1H, J = 1.8 Hz).

ESIMS (+): 448 [M+H]+.
<Reference Example 73>
Methyl

(S)-4-(4-bromo-2-chlorophenyl)-2-t-butoxycarbonylamino-2-butylb
utyrate

[0387]
[Formula 150]

Br. U

C1 Me
NM tc

159

CA 02764126 2011-11-30
[0388]

The target product (1.26 g) was obtained as a colorless oil
by reacting the compound of Reference Example 71 (2.52 g) in the
same manner as in Reference Example 4.

1H NMR (CDC13, 400 MHz) : 6 0.87 (3H, t, J = 6.7 Hz), 0.95-1.02 (1H,
m), 1.22-1.32 (3H, m), 1.46 (9H, s), 1.65-1.75 (1H, m), 2.00-2.10
(1H, m), 2.22-2.34 (1H, m), 2.38-2.48 (1H, m), 2.50-2.70 (2H, m),
3.75 (3H, s), 5.62 (1H, br), 7.02 (1H, d, J = 8.6 Hz), 7.28 (1H,
dd, J = 8.6, 2.4 Hz), 7.47 (1H, d, J = 2.4 Hz).

ESIMS (+): 462 [M+H]t.
<Reference Example 74>
(S)-2-[2-(4-bromo-2-chlorophenyl)ethyl]-2-t-butoxycarbonylamino

pentan-l-ol
[0389]
[Formula 151]

Br CI

OH
NHBoC

[0390]

The target product (28.6 g) was obtained as a white powder
by reacting the compound of Reference Example 72 (35.6 g) in the
same manner as in Reference Example 5.

1H NMR (CDC13, 400 MHz) : 5 0.96 (3H, t, j = 7.3 Hz) , 1.29-1.42 (2H,
m) , 1.44 (9H, s) , 1.53-1.62 (2H, m) , 1.81 (1H, ddd, J = 13.9, 11.5,
160

CA 02764126 2011-11-30

5.4 Hz) , 1.93 (1H, ddd, J = 13.9, 11.5, 5.4 Hz) , 2. 59 -2. 7 5 (2H, m)
3.73 (2H, d, J = 6.7 Hz), 4.15 (1H, br s), 4.62 (1H, br s), 7.11
(1H, d, J = 7.9 Hz), 7.31 (1H, dd, J = 7.9, 1.8 Hz), 7.49 (1H, d,
J = 1.8 Hz).

ESIMS (+): 420 [M+H]+.
<Reference Example 75>
(S)-2-[2-(4-bromo-2-chlorophenyl)ethyl]-2-t-butoxycarbonylamino

hexan-1-ol
[0391]
(Formula 1521
Br CI

H
NHBoc

[0392]

The target product (1.17 g) was obtained as a white powder
by reacting the compound of Reference Example 73 (1.26 g) in the
same manner as in Reference Example 5.

iH NMR (CDC13, 400 MHz): 8 0.93 (3H, t, J =7.3 Hz), 1.30-1.40 (4H,
m) , 1.44 (9H, s) , 1. 58-1.62 (2H, m) , 1. 78-1.86 (1H, m) , 1.88-1 .94
(1H, m) , 2.60-2.72 (2H, m) , 3.73 (2H, d, J = 6.2 Hz) , 4.13 (1H, br) ,
4.62 (1H, br), 7.10 (1H, d, J = 8.6 Hz), 7.31 (1H, dd, J = 8.6, 2.4
Hz), 7.49 (1H, d, j = 2.4 Hz).

ESIMS (+) : 434 [M+H]
<Reference Example 76>

161

CA 02764126 2011-11-30
5-t-Butyl-2-(methoxymethoxy)benzenethiol
(0393)

[Formula 153]

,>LsH
OMOM
[0394]

The target product (3.78 g) was obtained as a colorless oil
by reacting 1-t-butyl-4- (methoxymethoxy) benzene (10. 0 g) in the same
manner as in Reference Example 9.

'H NMR (CDC13, 400 MHz) : 6 1.27 (9H, s) , 3.51 (3H, s) , 3.78 (1H, br) ,
5.22 (2H, s), 7.01 (1H, d, J = 8.6 Hz), 7.09 (1H, dd, J = 8.6, 1.8
Hz), 7.26 (1H, d, J = 1.8 Hz).

EIMS (+) : 226 [M] 4.
<Reference Example 77>
2-(Methoxymethoxy)-5-phenylbenzenethiol
[0395]

[Formula 154]
1
'- SEA
MOM
(0396]

The target product (4.58 g) was obtained as a colorless oil
by reacting 4-(methoxymethoxy)biphenyl (10.0 g) in the same manner
162

CA 02764126 2011-11-30
as in Reference Example 9.

'H NMR (CDC13, 400 MHz) : 8 3.53 (3H, s) , 3.86 (1H, s) , 5.29 (2H, s) ,
7.15 (1H, d, J=8.6Hz) , 7.27-7.38 (2H, m) , 7.39-7.45 (2H, m) , 7.48-7.56
(3H, m).

EIMS (+): 246 [M)+
<Reference Example 78>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-p
ropylphenylthio)phenyl]-2-propylbutan-l-ol
[0397]

[Formula 1551

mom H
NHBM
[0398]

The target product (2.90 g) was obtained as a colorless oil
by reacting the compound of Reference Example 11 (2.00 g) and the
compound of Reference Example 74 (1.21 g) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz): 8 0.90 (3H, t, J = 7.3 Hz), 0.95 (3H, t,
J = 7.3 Hz), 1.30-1.40 (2H, m), 1.44 (9H, s), 1.55-1.60 (2H, m),
1.74-1.84 (2H, m) , 1.88-1.94 (2H, m) , 2.48 (2H, t, J=7.3 Hz) , 2.60-2.70
(2H, m) , 3.39 (3H, s) , 3.73 (2H, d, J = 6.1 Hz) , 4.16 (1H, br) , 4.62
(1H, br), 5.16 (2H, s), 7.05-7.08 (3H, m) , 7.09-7.10 (1H, m), 7.11-7.13
(1H, m), 7.22-7.24 (1H, m).

163

CA 02764126 2011-11-30
ESIMS (+): 552 [M+H]+.

<Reference Example 79>
(S)-2-t-butoxycarbonylamino-4-(2-chloro-4-(2-methoxymethoxy-5-i
sopropylphenylthio)phenyl]-2-propylbutan-l-ol
(0399]

[Formula 1561

S CI

-*I-IaOMOM H
NHBoc
[0400]

The target product (770 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 12 (350 mg) and the
compound of Reference Example 74 (584 mg) in the same manner as in
Reference Example 20.

''H NMR (CDC13, 400 MHz): 8 0.95 (3H, t, J = 7.3 Hz), 1.18 (6H, d,
J = 6.7 Hz), 1.30-1.40 (2H, m), 1.44 (9H, s), 1.45-1.60 (2H, m),
1.75-1.83 (1H, m), 1.86-1.95 (1H, m), 2.59-2.71 (2H, m), 2.82 (1H,
sept, J = 6.7 Hz), 3.39 (3H, s), 3.70-3.72 (2H, m), 4.19 (1H, br),
4.63 (1H, br), 5.16 (2H, s), 7.08-7.16 (5H, m), 7.23 (1H, d, J =
1.8 Hz).

ESIMS (+) : 552 [M+H]+.
<Reference Example 80>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-cyclopropyl-2-meth

oxymethoxyphenylthio)phenyl]-2-propylbutan-l-ol
164

CA 02764126 2011-11-30
[0401]

(Formula 157]

CI
mlom OH
NHS W
[0402]

The target product (760 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 13 (350 mg) and the
compound of Reference Example 74 (584 mg) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz) : 6 0.55-0.60 (2H, m) , 0.85-0.92 (2H, m) , 0.95
(3H, t, J = 7.3 Hz), 1.30-1.40 (2H, m), 1.44 (9H, s), 1.52-1.60 (2H,
m), 1.72-1.85 (2H, m), 1.91 (1H, dt, J = 11.6, 5.5 Hz), 2.60-2.75
(2H, m), 3.38 (3H, s) , 3.73 (2H, d, J = 6.1 Hz) , 4.18 (1H, br) , 4.63
(1H, br), 5.15 (2H, s), 6.96 (1H, dd, J = 8.6, 2.4 Hz), 6.99 (1H,
d, J = 2.4 Hz), 7.02-7.12 (3H, m), 7.23 (1H, d, J = 1.8 Hz).
ESIMS (+): 550 [M+H]+.

<Reference Example 81>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-t-butyl-2-methoxym
ethoxyphenylthio)phenyl]-2-propylbutan-l-ol
[0403]

165

CA 02764126 2011-11-30
[Formula 1581

r
OH
NHBoc

[0404]

The target product (532 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 76 (269 mg) and the
compound of Reference Example 74 (428 mg) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz): 8 0.95 (3H, t, J = 7.3 Hz), 1.26 (9H, s),
1.28-1.40 (2H, m), 1.43 (9H, s), 1.50-1.61 (2H, m), 1.72-1.82 (1H,
m), 1.84-1.96 (1H, m), 2.58-2.72 (2H, m), 3.38 (3H, s), 3.72 (2H,
d, J = 6.7 Hz), 4.18 (1H, br), 4.62 (1H, br), 5.16 (2H, s), 7.06
(1H, dd, J = 8.6, 1.8 Hz), 7.07-7.14 (2H, m), 7.21 (1H, d, J = 1.8
Hz), 7.30 (1H, dd, J = 8.6, 3.0 Hz), 7.33 (1H, d, J = 3.0 Hz).
ESIMS (+): 566 [M+H]`.

<Reference Example 82>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-b
iphenylthio)phenyl]-2-propylbutan-l-o1
[0405]

166

CA 02764126 2011-11-30
[Formula 1591

IOM off
NHBoc
(04061

The target product (453 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 77 (118 mg) and the
compound of Reference Example 74 (428 mg) in the same manner as in
Reference Example 20.

''H NMR (CDC13, 400 MHz) : 3 0.95 (3H, t, j = 7.3 Hz) , 1.28-1.40 (2H,
m), 1.43 (9H, s), 1.54-1.62 (2H, m), 1.75-1.85 (1H, m), 1.88-1.96
(1H, m), 2.60-2.74 (2H, m), 3.42 (3H, s), 3.72 (2H, d, j = 6.1 Hz),
4.16 (1H, br) , 4.62 (1H, br) , 5.24 (2H, s) , 7.14-7.16 (2H, m) , 7.22
(1H, d, J = 8.6 Hz) , 7,28-7.34 (2H, m) , 7.34-7.42 (2H, m) , 7.44-7.50
(4H, m).

ESIMS (+): 586 [M+H]t.
<Reference Example 83>
(S)-2-t-butoxycarbonylamino-4-[4-(5-benzyloxy-2-hydroxyphenylth

io)-2-chlorophenyl]-2-propylbutan-l-ol
[0407]

167

CA 02764126 2011-11-30
[Formula 1601

ON"0000 C,

aH aH
NHBoc
[0408]

The target product (705 mg) was obtained as a colorless oil
by reacting 5-benzyloxy-2-hydroxybenzenethiol (161 mg) and the
compound of Reference Example 74 (583 mg) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz): 5 0.95 (3H, t, J = 7.3 Hz), 1.20-1.40 (2H,
m), 1.43 (9H, s), 1.52-1.58 (2H, m), 1.72-1.94 (2H, m), 2.55-2.70
(2H, m), 3.71 (2H, d, J = 6.7 Hz), 4.15 (1H, br), 4.61 (1H, br),
5.00 (2H, s), 6.06 (1H, s), 6.89 (1H, dd, J = 8.5, 1.8 Hz), 6.99
(1H, d, J = 8.5 Hz) , 7.02-7.07 (2H, m) , 7.08-7.12 (2H, m) , 7.30-7.42
(5H, m).

ESIMS (+): 572 [M+H]'.
<Reference Example 84>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxy-5-trifluor

omethylphenylthio)phenyl]-2-butylbutan-l-ol
[0409]

168

CA 02764126 2011-11-30
[Formula 161]

F3C

H
WP'
N HBoc
[0410]

The target product (479 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 19 (230 mg) and the
compound of Reference Example 75 (400 mg) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz) : 8 0.94 (3H, t, j = 7.3 Hz) , 1.20-1.40 (4H,
m), 1.44 (9H, s), 1.57-1.62 (2H, m), 1.78-1.88 (1H, m), 1.90-2.00
(1H, m) , 2.60-2.78 (2H, m) , 3.74 (2H, d, J = 6.7 Hz) , 3.92 (3H, s) ,
4.20 (1H, br), 4.63 (lH, br), 6.95 (1H, d, J = 8.6 Hz), 7.15-7.21
(2H, m), 7.31-7.35 (2H, m), 7.47-7.54 (1H, m).

ESIMS (+) : 562 [M+H]+.
<Reference Example 85>
(S)-2-t-butoxycarbonylamino-4-[4-(2-t-butoxycarbonyloxy-5-trifl

uoromethylphenylthio)-2-chlorophenyl]-2-butylbutan-l-ol
[0411]

[Formula 162]

BOC
NC" CH
N I3oc

169

CA 02764126 2011-11-30
[0412]

The target product (319 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 84 (440 mg) in the
same manner as in Reference Example 30.

1H NMR (CDC13, 400 MHz): 5 0.93 (3H, t, J = 7.3 Hz), 1.20-1.40 (4H,
m), 1.44 (9H, s), 1.54 (9H, s), 1.56-1.62 (2H, m), 1.77-1.87 (1H,
m) , 1.90-1.97 (1H, m), 2.60-2.80 (2H, m) , 3.73 (2H, d, J = 6.1 Hz),
4.13 (1H, br), 4.62 (1H, br), 7.20 (2H, m), 7.30 (1H, d, J = 8.6
Hz), 7.39 (1H, s), 7.47 (1H, d, J = 1.8 Hz), 7.53 (1H, dd, J = 8.6,
1.8 Hz).

ESIMS (+) : 648 [M+H]+.
<Reference Example 86>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-p

ropylphenylthio)phenyl]-1-dimethoxyphosphoryloxy-2-propylbutane
[0413]

[Formula 163]

A114t1A POPM02
iHc
[0414]

The target product (1.90 g) was obtained as a colorless oil
by reacting the compound of Reference Example 78 (2.90 g) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz): 8 0.90 (3H, t, J = 7.3 Hz), 0.95 (3H, t,
J = 7,3 Hz), 1.30-1.40 (2H, m), 1.43 (9H, s), 1.52-1.60 (2H, m),
170

CA 02764126 2011-11-30

1.61-1.71 (2H, m), 1.72-1.82 (1H, m), 1.85-2.05 (1H, m), 2.48 (2H,
t, J - 7.3 Hz), 2.66 (2H, t, J - 8.6 Hz), 3.39 (3H, s), 3.77 (3H,
d, J - 11.0 Hz), 3.78 (3H, d, J = 11.0 Hz), 4.06-4.14 (1H, m), 4.23
(1H, dd, J = 10.3, 4.9 Hz), 4.50 (1H, br), 5.16 (2H, s), 7.06-7.12
(5H, m), 7.22 (1H, d, J = 1.8 Hz).

ESIMS (+): 660 [M+HJ+.
<Reference Example 87>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-i

sopropylphenylthio)phenyl]-1-dimethoxyphosphoryloxy-2-propylbut
ane

[0415]
[Formula 164]

MO PCXOMe)2
NHBW
[0416]

The target product (687 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 79 (650 mg) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz): 6 0.95 (3H, t, J = 7.2 Hz), 1.18 (6H, d,
J = 6.8 Hz), 1.20-1.40 (4H, m), 1.44 (9H, s), 1.58-1.82 (2H, m),
2.66 (2H, t, J = 8.4 Hz), 2.82 (1H, sept, J = 6.8 Hz), 3.39 (3H,
s) , 3.77 (3H, d, J = 11.0 Hz), 3.78 (3H, d, J - 11.0 Hz), 4.08-4.14
(1H, m), 4.20-4.26 (1H, m), 4.50 (1H, br), 5.16 (2H, s), 7.02-7.20
171

CA 02764126 2011-11-30

(5H, m), 7.23 (1H, d, J = 1.6 Hz).
ESIMS (+): 660 [M+H]+.

<Reference Example 88>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-cyclopropyl-2-meth
oxymethoxyphenylthio)phenyl]-1-dimethoxyphosphoryloxy-2-propylb
utane

[0417]
[Formula 165]

MC1M'C QMe)2
~NHBQc
[0418]

The target product (622 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 80 (710 mg) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz) : S 0.55-0.60 (2H, m) , 0.85-0.90 (2H, m) , 0.95
(3H, t, J = 7.3 Hz) , 1.30-1.40 (2H, m) , 1. 4 4 (9H, s) , 1.54-1.68 (2H,
m), 1.72-1.84 (2H, m), 1.95-2.05 (1H, m), 2.66 (2H, t, J = 8.6 Hz),
3.38 (3H, s), 3.77 (3H, d, J = 11.0 Hz), 3.78 (3H, d, J = 11.0 Hz),
4.11 (1H, dd, J = 9.7, 4.9 Hz), 4.23 (1H, dd, J = 9.7, 4.9 Hz) , 4.50
(1H, br), 5.14 (2H, s), 6.96 (1H, dd, J = 8.6, 2.4 Hz), 6.99 (1H,
d, J = 2.4 Hz), 7.02-7.12 (3H, m), 7.23 (1H, d, J = 1.8 Hz).
ESIMS (+): 658 (M+H]+.

172

CA 02764126 2011-11-30
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(5-t-butyl-2-methoxym
ethoxyphenylthio)phenyl]-1-dimethoxyphosphoryloxy-2-propylbutan
e

[04191
[Formula 166]

MOM Pa(OMe
NHBoc
[0420]

The target product (308 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 81 (400 mg) in the
same manner as in Reference Example 35.

'H NMR (CDC13, 400 MHz): 8 0.95 (3H, t, J = 7.3 Hz), 1.26 (9H, s),
1.30-1.40 (2H, m), 1.44 (9H, s), 1.60-1.80 (3H, m), 1.90-2.05 (1H,
m) , 2.62-2.76 (2H, m), 3.38 (3H, s) , 3.77 (3H, d, J = 11.0 Hz) , 3.78
(3H, d, J = 11.0 Hz) , 4.09-4.14 (1H, m) , 4.23 (1H, dd, J = 9.8, 4.0
Hz), 4.49 (1H, br), 5.16 (2H, s), 7.05 (1H, dd, J = 8.0, 1.8 Hz),
7.08-7.12 (2H, m) , 7.21 (1H, d, J = 1.8 Hz), 7.30 (1H, dd, J = 8.6,
2.4 Hz), 7.33 (1H, d, J = 2.4 Hz).

ESIMS (+): 674 (M+H)+.
<Reference Example 90>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-b

iphenylthio)phenyl]-1-dimethoxyphosphoryloxy-2-propylbutane
[0421)

173

CA 02764126 2011-11-30
[Formula 1671

Mom C?nC(C}Me)2
4NHBOC
[0422]

The target product (421 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 82 (360 mg) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz) : 8 0.95 (3H, t, j = 7.3 Hz) , 1.30-1.38 (2H,
m), 1.43 (9H, m), 1.60-1.70 (2H, m), 1.72-1.82 (1H, m), 1.85-2.02
(1H, m) , 2.62-2.72 (2H, m), 3.42 (3H, s), 3.77 (3H, d, J = 11.0 Hz),
3.78 (3H, d, J = 11.0 Hz), 4.09-4.14 (1H, m), 4.20-4.28 (1H, m),
4.50 (1H, br), 5.23 (2H, s), 7.10-7.17 (2H, m), 7.20-7.25 (1H, m),
7.28-7.34 (2H, m), 7.36-7.43 (2H, m), 7.45-7.51 (4H, m).

ESIMS (+): 694 [M+H]+.
<Reference Example 91>
(S)-4-[4-(5-benzyloxy-2-hydroxyphenylthio)-2-chlorophenyl]-2-t-

butoxycarbonylamino-l-dimethoxyphosphoryloxy-2-propylbutane
[0423]

(Formula 168]

.~` H Pgome}2
NHBoc
174

CA 02764126 2011-11-30
[0424]

The target product (445 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 83 (450 mg) in the
same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz); 8 0.94 (3H, t, J = 7.3 Hz), 1.28-1.38 (2H,
m), 1.43 (9H, s), 1.58-1.68 (2H, m), 1.72-1.82 (1H, m), 1.90-2.05
(1H, m), 2.58-2.68 (2H, m), 3.77 (3H, d, J = 11.0 Hz), 3.78 (3H,
d, J = 11.0 Hz) , 4.02-4.12 (1H, m) , 4.22 (1H, dd, J = 10.4, 5.5 Hz) ,
5.00 (2H, s), 6.07 (1H, s), 6.88 (1H, dd, J = 8.0, 2.4 Hz), 6.99
(1H, d, J = 8.0 Hz), 7.02-7.12 (4H, m), 7.30-7.38 (1H, m) , 7.35-7.42
(4H, m).

ESIMS (+); 680 [M+H]+.
<Reference Example 92>
(S)-2-t-butoxycarbonylamino-4-[4-(2-t-butoxycarbonyloxy-5-trifl

uoromethylphenylthio)-2-chlorophenyl]-1-dimethoxyphosphoryloxy-
2-butylbutane

[0425]
[Formula 1691

lop
PBoC
[04261

The target product (258 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 85 (260 mg) in the
175

CA 02764126 2011-11-30

same manner as in Reference Example 35.

1H NMR (CDC13, 400 MHz): 6 0.93 (3H, t, J = 7.3 Hz), 1.27-1.40 (4H,
m), 1.44 (9H, s), 1.54 (9H, s), 1.62-1.70 (2H, m), 1.77-1.84 (1H,
m) , 1.96-2.04 (1H, m) , 2.64-2.74 (2H, m) , 3.77 (3H, d, J = 11.0 Hz) ,
3.78 (3H, d, J = 11.0 Hz) , 4.08-4.14 (1H, m) , 4.20-4.27 (1H, m) , 7.19
(2H, m), 7.30 (1H, d, J = 8.6 Hz), 7.39 (1H, s), 7.47 (1H, d, J =
1.8 Hz), 7.53 (iH, dd, J = 8.6, 1.8 Hz).

ESIMS (+): 756 [M+H]".
[Example 18]

(0427]
(S)-2-amino-4-(2-chloro-4-(2-hydroxy-5-propylphenylthio)phenyl)
-2-propylbutyiphosphoric acid monoester

(0428]
[Formula 170]
.10
14
(aM
[0429]

The compound of Reference Example 86 (1.90 g) was dissolved
in 10% hydrogen chloride - methanol (19 mL) to form a first reaction
solution. This first reaction solution was stirred at 40 C for 3
hours. The solvent in the first reaction solution was removed by
distillation under reduced pressure, and the resultant residue was
dissolved in acetonitrile (20 mL). Under an argon atmosphere,
176

CA 02764126 2011-11-30

iodotrimethylsilane (1.85 mL) was added dropwise under ice cooling
to the acetonitrile solution to form a second reaction solution.
This second reaction solution was stirred under ice cooling for 30
minutes. Water (80 mL) was added to the second reaction solution,
and the reaction solution was stirred under ice cooling for a further
30 minutes. The oily product was then separated by decantation.
The obtained oily product was dried, and then recrystallized in
acetonitrile-tetrahydrofuran. The solid obtained from the
recrystallization was dissolved in a 0.5 mol/L sodium hydroxide
aqueous solution (10.0 mL) , and 1.0 mol/L hydrochloric acid was then
added to the resultant solution to adjust the pH to 3. The
precipitated solid was removed by filtration and then dried to obtain
the target product (1.00 g) as a white powder.

Optical rotation: [a]D26 -2.00 (c 0.51, McOH).

1H NMR (DMSO-d6, 400 MHz): 6 0.82 (3H, t, J = 7.3 Hz), 0.88 (3H, t,
J = 7.3 Hz), 1.20-1.35 (2H, m), 1.43-1.60 (4H, m), 1.64-1.76 (2H,
m) , 2.41 (2H, t, J = 7.3 Hz), 2.54-2.66 (2H, m) , 3.65-3.85 (2H, M),
6.87 (1H, d, J = 8.6 Hz) , 6.98-7.04 (2H, m) , 7.05-7.09 (2H, m) , 7.24
(1H, d, J = 8.6 Hz).

HRESIMS (+): 488.1430 (488.1427 calcd. for C22H32C1NO5PS)
Elemental analysis: measured C 52.49%, H 6.24%, N 2.79%, calcd. for
C22H31C1NO5PS. 0.75H20 C 52.69%, H 6.53%, N 2.79%.

[Example 19]
[0430]

(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-isopropylphenylthio)phen
177

CA 02764126 2011-11-30

yl]-2-propylbutyiphosphoric acid monoester
[0431]

[Formula 171]

` S I ~` cl PQOH~
NH2
[0432]

The target product (376 mg) was obtained as a white powder
by reacting the compound of Reference Example 87 (640 mg) in the
same manner as in Example 1.

Optical rotation: [a]D25 -2.08 (c 0.50, MeOH).

1H NMR (DMSO-d6, 400 MHz): 5 0.87 (3H, t, J = 7.3 Hz), 1.11 (6H, d,
J = 6.7 Hz), 1.20-1.40 (2H, m), 1.45-1.60 (2H, m), 1.68 (2H, br),
2.59 (2H, br), 2.76 (1H, sept, J = 6.7 Hz), 3.70-3.82 (2H, m), 6.89
(1H, d, J = 8.4 Hz), 6.95-7.05 (2H, m) , 7.10-7.15 (2H, m) , 7.24 (1H,
d, J = 8.4 Hz).

HRESIMS (+): 488.1433 (488.1427 calcd. for C22H32ClNO5PS)
Elemental analysis: measured C 51.93%, H 6.38%, N 2.69%, calcd. for
C22H31C1NO5PS. H2O C 52.22%, H 6.18%, N 2.77%.

[Example 20]
[0433]

(S)-2-amino-4-[2-chloro-4-(5-cyclopropyl-2-hydroxyphenylthio)ph
enyl]-2-propylbutylphosphoric acid monoester

[04341

178

CA 02764126 2011-11-30
[Formula 172]

CH PO(OH)2
[0435]

The target product (314 mg) was obtained as a white powder
by reacting the compound of Reference Example 88 (622 mg) in the
same manner as in Example 1.

Optical rotation: (a]D26 -1.68 (c 0.50, MeOH).

1H NMR (DMSO-d6, 400 MHz): 8 0.50-0.58 (2H, m), 0.80-0.87 (2H, m),
0.91 (3H, t, J=7.4Hz) , 1.28-1.40 (2H, m) , 1.50-1.63 (2H, m) , 1.68-1.76
(2H, m), 1.78-1.88 (1H, m), 2.58-2.70 (2H, m), 3.70-3.84 (2H, m),
6.87 (1H, d, J = 8.4 Hz) , 6.97 (1H, dd, J = 8.4, 1.8 Hz), 7.00-7.07
(3H, m), 7.28 (1H, d, J = 8.0 Hz).

HRESIMS (+): 486.1276 (486.1271 calcd. for C22H30C1N05PS).
Elemental analysis : measured C 52.06%, H 5.97%, N 2.64%, calcd. for
C22H29C1NO5PS. 1.2H2O C 52.06%, H 5.76%, N 2.76%.

[Example 21]
[0436]

(S)-2-amino-4-[4-(5-t-butyl-2-hydroxyphenylthio)-2-chiorophenyl
1-2-propylbutylphosphoric acid monoester

[0437]

179

CA 02764126 2011-11-30
[Formula 173]

S CI
[0438]

The target product (157 mg) was obtained as a white powder
by reacting the compound of Reference Example 89 (308 mg) in the
same manner as in Example 1.

Optical rotation: [a]D26 -1.94 (c 0.53, McOH).

1H NMR (DMSO-d6, 400 MHz) : 8 0.87 (3H, t, J = 7.3 Hz) , 1.19 (9H, s) ,
1.20-1.35 (2H, m), 1.45-1.60 (2H, m), 1.62-1.74 (2H, m), 2.55-2.70
(2H, m) , 3.60-3.85 (2H, m), 6.90 (1H, d, J = 8.6 Hz), 6.96-7.04 (2H,
m), 7.22-7.30 (3H, m).

HRESIMS (+): 502.1582 (502.1584 calcd. for C23H34C1NO5PS)
Elemental analysis: measured C 53.85%, H 6.44%, N 2.55%, calcd. for
C23H33C1NO5PS. 0.5H20 C 54.06%, H 6.71%, N 2.74%.

[Example 22]
[0439]

(S)-2-amino-4-(2-chloro-4-(2-hydroxy-5-biphenylthio)phenyl]-2-p
ropylbutylphosphoric acid monoester

[0440]

180

CA 02764126 2011-11-30
[Formula 1741

aa,.OH OPqOH)2
NH2
[0441]

The target product (219 mg) was obtained as a white powder
by reacting the compound of Reference Example 90 (400 mg) in the
same manner as in Example 1.

Optical rotation: [a]D26 -1.32 (c 0.50, McOH).

1H NMR (DMSO-d6, 400 MHz) : 8 0.86 (3H, t, j = 7.3 Hz) , 1.20-1.35 (2H,
m), 1.45-1.62 (2H, m) , 1.64-1.76 (2H, m) , 2.58-2.68 (2H, m), 3.68-3.80
(2H, m) , 7.03-7.10 (2H, m), 7.13 (1H, d, J = 1.8 Hz) , 7.22-7.30 (2H,
m), 7.38 (2H, t, J = 8.0 Hz), 7.48-7.58 (4H, m).

HRESIMS (+) : 522.1275 (522.1271 calcd. for C25H30C1NO5PS)
Elemental analysis: measured C 56.58%, H 5.52%, N 2.38%, calcd. for
C25H29C1NO5PS. 0.5H20 C 56.55%, H 5.69%, N 2.64%.

[Example 231
[0442]

(S)-2-amino-4-[4-(5-benzyloxy-2-hydroxyphenylthio)-2-chlorophen
yl)-2-propylbutylphosphoric acid monoester

[0443]

181

CA 02764126 2011-11-30
[Formula 175]

H PO(OH)2
NH2
(0444]

The target product (210 mg) was obtained as a white powder
by reacting the compound of Reference Example 91 (430 mg) in the
same manner as in Example 1.

Optical rotation: [a]D26 -2.15 (c 0.50, MeOH).

1H NMR (DMSO-d6, 400 MHz) : 8 0.88 (3H, t, j =7.3 Hz) , 1.22-1.38 (2H,
m), 1.46-1.62 (2H, m) , 1.66-1.80 (2H, m), 2.56-2.70 (2H, m), 3.70-3.82
(2H, m), 4.96 (2H, s), 6.80 (1H, d, J = 3.1 Hz), 6.85 (1H, d, J =
8.6 Hz), 6.90 (1H, dd, J = 8.6, 3.1 Hz), 7.07 (1H, dd, J = 8.0, 1.8
Hz), 7.11 (1H, d, J = 1.8 Hz), 7.24-7.38 (6H, m).

HRESIMS (+) : 552.1384 (552.1377 calcd. for C26H32ClNO6PS)
Elemental analysis: measured C 54.89%, H 5.53%, N 2.52%, calcd. for
C26H31C1NO6PS. H2O C 54.78%, H 5.48%, N 2.46%.

[Example 24]
[0445)

(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-trifluoromethylphenylthi
o)phenyl]-2-butylbutylphosphoric acid monoester

(0446]

182

CA 02764126 2011-11-30
[Formula 176]

F3 (3

2
[0447]

The target product (90 mg) was obtained as a white powder by
reacting the compound of Reference Example 92 (250 mg) in the same
manner as in Example 1.

Optical rotation: [a]D26 -1.76 (c 0.50, McOH).

''H NMR (DMSO-d6, 400 MHz) : 6 0.86 (3H, t, J = 7.3 Hz) , 1.20-1.32 (4H,
m), 1.50-1.64 (2H, m) , 1. 68-1.80 (2H, m), 2.60-2.70 (2H, m), 3.70-3.80
(2H, m), 7.11 (2H, m), 7.22 (1H, d, J = 1.8 Hz), 7.30 (1H, d, J =
8.6 Hz), 7.40 (1H, d, J = 1.8 Hz), 7.52 (1H, dd, J = 8.6, 1.8Hz).
HRESIMS (+) : 528.0985 (528.0988 calcd. for C21H27C1F3NO5PS) .

Elemental analysis: measured C 47.06%, H 4.92%, N 2.22%, calcd. for
C21H26C1F3NO5PS. 1/2H2O C 46.98%, H 5.07%, N 2.61%.

[Example 25]
[0448]

(S)-2-amino-4-[2-chloro-4-(2-hydroxy-5-carboxyphenylthio)phenyl
1-2-propylbutylphosphoric acid monoester

[0449]

183

CA 02764126 2011-11-30
[Formula 177]

Q'P0(0H)2
NH2
[0450]

A 1 mol/L potassium hydroxide aqueous solution (2.00 ml) was
added to the compound of Example 13 (100 mg) , and the resultant solution
was stirred at 50 C for 2 hours. The reaction solution was cooled
to room temperature, and 3 mol/L hydrochloric acid was added to adjust
the pH to 6, whereby a solid precipitated. The precipitated solid
was removed by filtration to obtain the target product (75.0 mg)
as a white powder.

Optical rotation: (a] D25 -5.00 (c 0.10, DMSO).

1H NMR (DMSO-d6, 400 MHz) : 5 0.87 (3H, t, J = 6.7 Hz) , 1.30 (2H, br) ,
1.53 (2H, br), 1.68 (2H, br) , 2.62 (2H, br), 3.70-3.75 (2H, m), 6.90
(1H, d, J = 8.0 Hz), 7.02-7.06 (1H, m), 7.15 (1H, d, J = 1.8 Hz),
7.26 (1H, d, J = 8.6 Hz), 7.65-7.72 (2H, m).

HRESIMS (+) : 490.0857 (490.0856 calcd. for C20H26C1NO7PS
Elemental analysis: measured C 44.94%, H 4.89%, N 2.51%, calcd. for
C20H25C1NO7PS. 0.7NaCl C 45.25%, H 4.75%, N 2.64%.

<Reference Example 93>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxy-5-trifluor
omethylphenylthio)phenyl]-2-propylbutan-l-ol
[0451]

184

CA 02764126 2011-11-30
[Formula 178]

E: f%

OH
NHBoc

[0452]

The target product (19.2 g) was obtained as a colorless oil
by reacting the compound of Reference Example 19 (11.0 g) and the
compound of Reference Example 74 (18.5 g) in the same manner as in
Reference Example 20.

1H NMR (CDC13, 400 MHz): 8 0.96 (3H, t, J = 7.3 Hz), 1.29-1.43 (2H,
m), 1.44 (9H, s), 1.59 (2H, dd, J = 7.3, 1.8 Hz), 1.83 (1H, ddd.
J = 13.9, 12.1, 5.4 Hz), 1.96 (1H, ddd, J = 13.9, 12.1, 5.4 Hz),
2.63-2.78 (2H, m) , 3.75 (2H, d, J = 6.7 Hz), 3.92 (3H, s), 4.17 (1H,
br s), 4.64 (1H, s), 6.96 (1H, d, J = 8.5 Hz), 7.17 (1H, dd, J =
8.5, 1.8 Hz) , 7.17 (1H, d, J = 8.5 Hz), 7.32-7.35 (2H, m), 7.51 (1H,
dd, J = 8.5, 1.8 Hz)

ESIMS (+):548 [M+H]'.
<Reference Example 94>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4--(2-hydroxy-5-trifluor
omethylphenylthio)phenyl]-2-propylbutan-l-ol
[0453]

185

CA 02764126 2011-11-30
[Formula 1791

17 3%,' CI

IO(HI NHBac
[0454]

The target product (3.34 g) was obtained as a colorless oil
by reacting the compound of Reference Example 93 (3.69 g) in the
same manner as in Reference Example 33.

1H NMR (CDC13, 400 MHz) : 8 0.95 (3H, t, J = 7.3 Hz) , 1.28-1.41 (2H,
m), 1.44 (9H, s), 1.59 (2H, dd, J = 7.3, 1.8 Hz), 1.79 (1H, ddd.
J = 13.9, 11.6, 5.5 Hz), 1.90 (1H, ddd, J = 13.9, 11.6, 5.5 Hz),
2.57-2.72 (2H, m), 3.72 (2H, d, J = 6.7 Hz), 4.16 (1H, br s), 4.61
(1H, br s), 6.77 (1H, s), 6.91 (1H, dd, J = 7.9, 1.8 Hz), 7.09 (1H,
d, J = 1.8 Hz), 7.13 (1H, d, J = 7.9 Hz), 7.16 (1H, j = 7.9 Hz),
7.63 (1H, dd, J = 7.9, 1.8 Hz), 7.80 (1H, d, J = 1.8 Hz).

ESIMS (+): 534 [M+H]'.
<Reference Example 95>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-t

rifluoromethylphenylthio)phenyl]-2-propylbutan-l-ol
[0455]

186

CA 02764126 2011-11-30
[Formula 180]

FaC ,,
S Cl
1(mom
NHBoc
[0456]

The target product (1.05 g) was obtained as a colorless oil
by reacting the compound of Reference Example 94 (1.00 g) in the
same manner as in Reference Example 58.

1H NMR (CDC13, 400 MHz) : 6 0.96 (3H, t, J = 7.3 Hz) , 1.30-1.43 (2H,
m) , 1.44 (9H, s) , 1.54-1.60 (2H, m) , 1.82 (1H, ddd, J = 13.9, 12.1,
5.4 Hz), 1.94 (1H, ddd, J = 13.9, 12.1, 5.4 Hz), 2.63-2.78 (2H, m),
3.40 (3H, s), 3.74 (2H, d, J = 3. 0 Hz) , 4.16 (1H, br s) 4.63 (1H, s) ,
5.25 (2H, s), 7.16-7.23 (3H, m), 7.36 (2H, dd, J = 8.5, 1.8 Hz),
7.47 (1H, dd, J = 8.5, 1.8 Hz).

ESIMS (+): 578 [M+H]+.
<Reference Example 96>
(S)-2-t-butoxycarbonylamino-4-[2-chloro-4-(2-methoxymethoxy-5-t

rifluoromethylphenylthio)phenyl]-2-propylbutan-l-al
[0457]

[Formula 181]
F3C

I I *N- (C100" ec
10C100"O .0 t HO
NHBoc

187

CA 02764126 2011-11-30
[0458]

The target product (940 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 95 (1.05 g) in the
same manner as in Reference Example 67,

1H NMR (CDC13, 400 MHz): 6 0.96 (3H, t, J = 7.3 Hz), 1.30-1.43 (2H,
m), 1.46 (9H, s), 1.53-1.64 (2H, m), 1.89-2.30 (2H, m), 2.56 (1H,
ddd, J = 13.4, 12.2, 4.9 Hz), 2.65 (1H, ddd, J = 13.4, 12.2, 4.9
Hz), 3.41 (3H, s), 5.23 (1H, br s), 5.25 (2H, s), 7.12 (1H, d, J
= 7.9 Hz) , 7.19 (1H, dd, J = 7.9, 1.8 Hz) , 7.22 (1H, d, J = 7.9 Hz) ,
7.31 (1H, d, J = 1.8 Hz), 7.38 (1H, d, J = 1.8 Hz), 7.48 (1H, dd,
J = 7.9, 1.8 Hz), 9.34 (1H, S).

ESIMS (+): 576 [M+H]+.
<Reference Example 97>
Dimethyl

(S)-3-t-butoxycarbonylamino-5-[2-chloro-4-(2-methoxymethoxy-5-t
rifluoromethylphenylthio)phenyl]-3-propyl-l-pentenylphosphonate
[0459]

[Formula 182]

F3C 00 Y"t 7-*11
mom
NHBoc
[0460]

The target product (662 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 96 (940 mg) in the
188

CA 02764126 2011-11-30
same manner as in Reference Example 68.

1H NMR (CDC13, 400 MHz) : 5 0.95 (3H, t, j = 7.3 Hz) , 1.30-1.44 (2H,
m), 1.46 (9H, s), 1.52-1.65 (2H, m), 1.92 (1H, td, J = 12.2, 4.9
Hz), 2.02-2.16 (1H, m), 2.62-2.80 (2H, m), 3.41 (3H, s), 3.72 (6H,
d, J = 11.0 Hz), 4.63 (1H, br s), 5.22 (2H, s), 5.73 (1H, t, J =
17.7 Hz), 6.81 (1H, dd, J = 22.6, 17.7 Hz), 7.16-7.19 (2H, m), 7.22
(1H, d, J = 8.6 Hz), 7.35 (1H, d, J = 1.8 Hz), 7.36 (1H, d, J = 1.8
Hz), 7.50 (1H, dd, J = 8.6, 1.8 Hz).

ESIMS (+): 682 [M+H].
<Reference Example 98>
Dimethyl

(S)-3-t-butoxycarbonylamino-5-[2-chloro-4-(2-methoxymethoxy-5-t
rifluoromethylphenylthio)phenyl]-3-propylpentyiphosphonate
[0461]

[Formula 183]
F3C

NHB
[0462]

The target product (290 mg) was obtained as a colorless oil
by reacting the compound of Reference Example 97 (662 mg) in the
same manner as in Reference Example 69.

1H NMR (CDC13, 400 MHz): 8 0.95 (3H, t, J = 7.3 Hz), 1.26-1.40 (2H,
m), 1.44 (9H, s), 1.52-1.65 (2H, m), 1.65-1.86 (4H, m), 2.04-2.28
189

CA 02764126 2011-11-30

(2H, m) , 2.60-2.72 (2H, m) , 3.41 (3H, s) , 3.74 (6H, d, J = 11.0 Hz) ,
4.27 (1H, br s), 5.25 (2H, s), 7.10-7.28 (3H, m), 7.38-7.42 (2H,
m), 7.52 (1H, dd, J = 7.9, 2.4 Hz).

ESIMS (+): 684 [M+H]+.
[Example 26]

[0463]
(S)-3-amino-5-[2-chloro-4-(2-hydroxy-5-trifluoromethylphenylthi
o)phenyl]-3-propylpentyiphosphonic acid

[0464]
[Formula 184]

0 '0
H PO(OH)2
NH2
[0465]

The target product (30 mg) was obtained as a white powder by
reacting the compound of Reference Example 98 (250 mg) in the same
manner as in Example 1.

Optical rotation: [a]D25 -17.81 (c 0.10, McOH).

1H NMR (DMSO-d6, 400 MHz) : 6 0.95 (3H, t, J = 7.3 Hz), 1.22-1.58
(6H, m), 1.64-1.86 (4H, m), 2.52-2.78 (2H, m), 7.09 (2H, d, j = 8.6
Hz), 7.23 (1H, s), 7.25-7.33 (1H, m), 7.41 (1H, s), 7.49 (1H, d,
J = 8.6 Hz).

ESIMS (+): 512 (M+H]+.
[0466]

190

CA 02764126 2011-11-30

Results supporting the effectiveness of the compounds
illustrated as examples will now be shown in Experiment Examples
1, 2, 3 and 4.

[0467)
<Experiment Example 1> Suppression effect of test compound against
cellular calcium mobilization of human S1 P3 receptor -expression cell
by SiP (sphingosine 1-phosphoric acid)

Human S1P3 receptor-expression CHO cells were subcultured in
a Ham's F-12 culture medium containing 10% fetal bovine serum, and
300 gg/mL of Geneticin. The human S1P3receptor-expression CHO cells
were subjected to 0.25 % trypsinization, then recovered from the
dish, and floated in a Ham's F-12 culture medium containing 10% fetal
bovine serum, and 300 gg/mL of Geneticin. After that, the human
S1P3 receptor-expression CHO cells were disseminated into a 96-well
black clear bottom plate (BD Falcon Biocoat) so that 7 x 104 / 100
L / well of the human S1P3 receptor-expression CHO cells were
disseminated. And then, the human S1P3 receptor-expression CHO
cells were cultivated for one night at 37 C under 5% CO2. The next
day, the wells were washed 3 times with 100 L of PBS containing
0.1 % fatty acid-free bovine serum albumin (BSA). The culture medium
was exchanged with a Ham's F-12 culture medium containing 0.1% BSA,
and then starved of serum for 6 hours in an CO2 incubator at 37 C.
[0468]

The culture medium was thrown away after the 6 hours. Then,
50 L/well of a Fluo3 loading buffer was added, and the cultures
191

CA 02764126 2011-11-30

were cultivated for further 1 hour. The Fluo3 loading buffer was
prepared as follows. First, equal amounts of Fluo3-AM (Dojindo)
and pluronic F-127(20% DMSO solution, invitrogen) weremixed. Next,
the mixture of Fluo3-AM and pluronic F-127 was added to a Hanks-HEPES
buffer (balanced salt solution containing 20 mM HEPES (pH 7.4) , 0.1%
BSA (fatty acid-free) , and 2.5 mM probenecid) to form a Fluo3 loading
buffer having a final Fluo3-AM concentration of 4 M.

After incubating for 1 hour, the cultures were washed 3 times
with 100 gL of the Hanks-HEPES buffer. Then, 100 gL of the same
buffer in which a test compound (125 nM, 1.25 M, 12.5 M) or DMSO
had been dissolved was added to the cultures, and then incubated
for 30 minutes at 37 C in a microplate spectrophotofluorometer (FLEX
Station) (Molecular Device Co., Ltd.). Then, 25 L of SiP prepared
at 5 times the concentration of the final concentration by serial
dilution (final concentration of 0.1 nM, 1 nM, 10 riM, 100 nM, and
1 M) was added, and the fluorescence based on the Fluo3 due to calcium
mobilization was detected and measured at an excitation wavelength
of 485 remand a detection wavelength of 525 nm using the same apparatus.
Based on the measurement data, the increase in fluorescence was
calculated by subtracting the minimum fluorescence intensity from
the maximum fluorescence intensity. The measured increase in
fluorescence was used to perform a curve approximation of the
relationship between the S1P concentration and the increase in
fluorescence using PRISM 4 software (GraphPad). Based on the results,
the EC50 value of the compound-untreated and the EC50 value of the
192

CA 02764126 2011-11-30

compound-treated at each concentration were calculated. Schild Plot
analysis was performedbased on these values to determine dissociation
constant Kd value. The results are shown in Table 1. In Table 1,
1, 000 nmol/L > Kd value ;-> 100 nmol/L is indicated as "+, " 100 nmol/L
> Kd value Z 10 nmol/L is indicated as "++,N 10 nmol/L > Kd value

1 nmol/L is indicated as "+++, N and 1 nmol/L > Kd value is indicated
as "++++.N

[0469]
[Table 1]

Example S1 P3 Example SiP3 Example S1P3
Number Number Number
I ++ 10 ++ 18 +++
2 ++ 11 +++ 19 +++
3 ++ 12 ++++ 20 +++
4 +++ 13 ++++ 21 ++
+++ 14 ++ 22 ++++
6 + 15 + 23 +++
7 ++ 16 ++
8 ++ 17 +
9 +++

[0470]
<Experiment Example 2> Intracellular calcium mobilization
derivative test of test compound against human S1Pl
receptor-expression cell

Human SiP receptor-expression CHO cells (hS1P1
receptor-expression CHO cells, hSlP3 receptor- expression CHO cells,
and hS1P4 receptor-expression CHO cells) subcultured in a Ham's F-12
culture medium containing 10% fetal bovine serum, and 200 g/mL of
Geneticin were disseminated into a 96-well black clear bottom plate
193

CA 02764126 2011-11-30

(coaster) so that 4 x 10 cells/well of the human SiP
receptor-expression CHO cells were disseminated. The human S1P3
receptor-expression CHO cells were then cultivated for one night
at 37 C under 5% CO2. A Calcium Screening Kit reagent (Dojindo) was
added as a Ca2*binding fluorescence indicator, and then the cultures
were cultivated for 60 minutes at 37 C under 5% CO2. After cultivation,
the f luoroesence intensity was measured at an excitation wavelength
of 485 nm and a detection wavelength of 525 nm using a microplate
spectrophotofluorometer (FLEX Station) (Molecular Device Co.,Ltd.).
S1P prepared in a culture medium so that the concentration would
be 10 times that of the final concentration, or a test compound (final
DMSO concentration 0.1%) was added 18 seconds after the start of
the fluorescence measurement, and the fluorescence intensity was
continuously measured every 1.5 seconds until 100 seconds after the
addition. Based on the measurement data, the increase in
fluorescence was calculated by subtracting the minimum fluorescence
intensity from the maximum fluorescence intensity. The percentage
increase in fluorescence (%) of the test compound was calculated
based on a difference of 100% between the increase in fluorescence
when the solvent was added and the increase in fluorescence when
acted on by 10-6 M S1P. The EC50 value was determined using PRISM
software (GraphPad) as the intracellular calcium mobilization
derivative action of the test compound.

(04711

The EC50 values of the compounds of Example 13 and Example
194

CA 02764126 2011-11-30

15 were larger than 10 mol/L. Further, an evaluation of the
antagonistic action of the S1P1 receptor using the method of
Experiment Example 1 showed that the Kd values of the compounds of
Example 13 and Example 15 were larger than 100 mol/L.

[04721
<Experiment Example 3> LPS induced sepsis model

The experiment was carried out with reference to the method
described in Non-Patent Literature 5 (F. Nissen et al., Nature, 452,
654 (2008)). A solution of 1 mg/ml LPS (lipopolysaccharide) in
physiological saline was intraperitoneally administered (10 ml/kg)
to a C57BL/6J mouse (Charles River, male, 7 to 8 weeks). The test
compound in an amount of 10 mg/kg was intravenously administered
twice, 15 minutes before LPS administration and 2 hours after LPS
administration. The mouse was dissected 18 hours after LPS
administration, and the mesenteric lymph nodes and lungs were excised.
The mesenteric lymph nodes were dissolved in 200 1 and the lungs
in 1 ml of a dissolving buffer (30 mM Tris (pH7.4), 150 mM NaCl,
0.1% TritonX-100, 2 mM CaC12, and 2 mM MgC12) . Non-dissolved matter
was removed by centrifugal separation. The IL-1J3 in the tissue
solutions was measured using an IL-l(3 ELISA Kit (Thermo Co., Ltd.) .
The results are shown in Table 2. In Table 2, compounds having a
suppression rate of 50% or more are indicated as "+++, compounds
having a suppression rate of 50% > suppression rate Z 30% are indicated
as "++,,v and compounds having a suppression rate of 30% > suppression
rate Z 20% are indicated as "+."

195

CA 02764126 2011-11-30

The suppression rate was calculated using the following
calculation equation.

(0473]
[Equation 1]

a = (1-X/Y)x100

X: IL-1(3 PRODUCTION AMOUNT WHEN 10 mg/kg OF TEST COMPOUND WAS
ADMINISTERED TO RESPECTIVE TISSUE

Y: IL-113 PRODUCTION AMOUNT WHEN TEST COMPOUND WAS NOT ADMINISTERED
TO RESPECTIVE TISSUE

a: SUPPRESSION RATE ($)
[0474]

[Table 2]

Example Suppression Rate Suppression Rate
Number for Lung for Mesenteric
L Nodes
11 ++ ++
12 ++ +
13 +++ ++
[0475]

<Experimental Example 4> cecal ligation and puncture sepsis model
This model is widely used as a model for polymicrobial abdominal
sepsis caused by leakage of intestinal bacteria. The experiment
was carried out with reference to the method described in Non-Patent
Literature 9 (D. Rittirsch et al., Nature Protocols, 4, 31 (2009)) .
Long-Evans rats were used (Nihon SLC, male 9 weeks). The abdominal
portion of the rats was cut open under isoflurane anesthesia, and
the cecum was removed. The cecum was ligated with silk thread, and
196

CA 02764126 2011-11-30

3 holes were opened in the tip portion of the cecum using an 18 G
syringe needle. After the treatment, the cecum was returned to the
body, and the wound was sutured. The rats were then returned to
their cages, and observed for 4 days to determine the survival rate.
The test compound (0.1 mg/kg/hr) was continuously administered from
a cannula stuck in the femoral vein from 2 hours after the CLP treatment.

The group administered with the compound of Example 22 was
found to have a survival curve that had shifted to the right as compared
with the medium administration group (survival lengthening action).
Further, although the survival rate after 4 days was 40% for the
medium group, an improvement in the survival rate to 70% was found
for the compound of Example 22. These results suggest that the
compound of Example 22 is effective against sepsis.

[0476]

Based on the above results, it is clear that despite exhibiting
an excellent antagonistic action against the human SiP3 receptor,
the compound of the present invention exhibits a weak or no
antagonistic action or agonistic action against the S1P1 receptor.
Further, it was also confirmed that the compound of the present
invention exhibits an excellent suppressive effect against sepsis.
INDUSTRIAL APPLICABILITY

[0477]

According to the present invention, a diphenyl sulfide
derivative can be provided that has an excellent S1P3 antagonistic
activity. The inventive compound is effective as a preventive or
197

CA 02764126 2011-11-30

a therapeutic medicine for respiratory tract contraction, bronchial
asthma, chronic obstructive pulmonary disease (COPD), pulmonary
emphysema, tracheal stenosis, diffuse panbronchiolitis, bronchitis
resulting from infection, connective tissue disease, or
transplantation, diffuse pulmonary hamartoangiomyomatosis, adult
respiratory distress syndrome (ARDS), interstitial pneumonitis,
lung cancer, pneumonia hypersensitivity, idiopathic interstitial
pneumonia, fibrosis of the lung, or cytokine storm caused by an
influenza virus or RS virus infection, arterial sclerosis, blood
vessel intimal hypertrophy, solid tumors, diabetic retinopathy,
rheumatoid arthritis, cardiac arrest, ischemia-reperfusion
disorders, cerebral blood vessel spasms after subarachnoid bleeding,
angina pectoris or myocardial infarction caused by coronary vessel
spasms, glomerulonephritis, thrombosis, lung disease caused by
pulmonary edema such as ARDS, cardiac arrhythmia, eye disease, eye
hypertension, glaucoma, glaucomatous retinopathy, optic neuropathy,
and macula-lutea degeneration.

198

FAQs

How do I find out if something is patented in Canada? ›

Welcome to CIPO's Canadian Patent Database. This database lets you access 154 years of patent descriptions and images. You can search, retrieve and study more than 2,520,000 patent documents. You can now access a wider variety of patent-specific documents on the Canadian Patents Database page.

Read On ›

How often is the Canadian patent Database updated? ›

Canada shares weekly patent data with patent offices and commercial clients for integration in various search databases.

Learn More ›

How do I verify a patent number? ›

There are two very good online databases to search for patent numbers:

USPTO Patent Number Search. Enter the patent number you would like to access. Requires a TIFF file viewer to view the patent images. ...
Google Patents. Enter a patent number and you will be able to access the PDF version of the patent.

How do you check if a patent already exists? ›

Patents may be searched using the following resources:

Patent Public Search.
Global Dossier.
Patent Application Information Retrieval (PAIR)
Public Search Facility.
Patent and Trademark Resource Centers (PTRCs)
Patent Official Gazette.
Common Citation Document (CCD)
Search International Patent Offices.

More items...

Oct 18, 2018

Show Me More ›

Are Canadian patents valid in the US? ›

A Canadian patent, trademark or industrial design does not secure your rights outside of Canada. You should consider obtaining IP protection in the countries where you plan on doing business, including selling products over the Internet and/or manufacturing products overseas.

Tell Me More ›

Are US patents valid in Canada? ›

Are US patents valid in other countries? No, patents are granted in a jurisdiction, which can be a country or a region. In the case of the United States, a granted U.S. patent can only apply in the United States.

Get More Info Here ›

Do Canadian patents expire? ›

Patent protection applies in the country or region that issues the patent. In Canada, a patent lasts for 20 years from the date that you file it.

Discover More Details ›

How much does it cost to renew a Canadian design patent? ›

1640

Patent Year	Official renewal fee for Standard patent (CAD)	Official renewal fee Small Entity patent (CAD)
1
2	100	50
3	100	50
4	100	50

15 more rows

Keep Reading ›

How do I get a copy of a Canadian patent? ›

If you want a copy of published documents associated with your patent, you can download them from the Canadian Patents Database. You can also download a PDF containing all of the documents for your patent including a copy of the patent, the cover page, the description, the claims and the drawings.

Can you look up patents by number? ›

Patent Number Searching

If you know the patent number use either Google Patents or the USPTO website to find the patent. Enter the patent number without commas and when using the USPTO website the patent number must be seven numbers in length (add preceding zero's if necessary).

Learn More ›

Can you look up old patent numbers? ›

United States Patent and Trademark Office (www.uspto.gov)

Patent searching can be done directly on the USPTO's web site. Full text and images are available for patents from 1976 to the most recent Tuesday. Images, current classification numbers and patent numbers only are available for patents from 1790 to 1975.

Are patent numbers public information? ›

Can I obtain a patent and keep my invention secret? No. Patents are granted by patent offices in exchange for a full disclosure of the invention. In general, the details of the invention are then published and made available to the public at large.

Read The Full Story ›

How long is a patent good for? ›

154 to provide that the term of a patent (other than a design patent) begins on the date the patent issues and ends on the date that is twenty years from the date on which the application for the patent was filed in the United States or, if the application contains a specific reference to an earlier filed application ...

Show Me More ›

Do patents expire? ›

Generally speaking, the term of a U.S. utility patent begins on the date the patent issues and expires on the date that is 20 years from the date that the application was originally filed.

Keep Reading ›

How do I know if my invention is patentable? ›

Go to the official website of the U.S. Patent and Trademark Office. Use the "Full-Text and Image Database" search to verify any present patent applications and pictures. You can find filed applications and pictures for patents filed after 1975.

Who issues patents in Canada? ›

To apply for a patent, a patent application needs to be filed with the patent office. The patent application is then examined by patent examiners.

Tell Me More ›

What is the life of patent in Canada? ›

For patent applications filed on or after October 1, 1989, the term of the patent is 20 years from the date of filing of the application. The right of a patent is conditional on the payment of the annual maintenance fees. For more information about patents, visit CIPO's Web site.

Get More Info ›

What is the Canadian version of USPTO? ›

The Canadian Intellectual Property Office (CIPO) is a special operating agency of Innovation, Science and Economic Development Canada.

Show Me More ›

What Cannot be patented in Canada? ›

Some things that cannot be patented include:

"disembodied ideas, concepts or discoveries"
"scientific principles and abstract theorems"
"methods of medical treatment or surgery"
"higher life forms"
"forms of energy"
"features of solely intellectual or aesthetic significance"
"printed matter"

Get More Info ›

Do Canadian patents apply in other countries? ›

From: Canadian Intellectual Property Office

Obtaining a Canadian patent does not protect your invention in another country. If you want this protection, you'll have to apply for a foreign patent.

Know More ›

Does a patent in Canada protect your intellectual property in other countries? ›

Many countries extend copyright protection automatically to original literary, artistic, dramatic or musical works created in Canada. However, a Canadian patent, or a Canadian trademark or industrial design registration, does not provide protection abroad.

Know More ›

Is it worth buying expired patents? ›

Benefits of Expired Patents for Sale

When someone purchases an expired patent, it is less work for the buyer, as all work has already been completed by the previous owner. This allows the new owner to focus on improving the quality and functionality of the invention.

Can you copy a patent after 20 years? ›

After a patent has been in place for 20 years for utility patents and 14 years for design and plant patents, the invention becomes part of the public domain. This means the invention no longer has patent protection and is no longer off limits, so anyone can make, use, or sell the invention without infringement.

Discover More ›

What happens after 20 years with patents? ›

A patent becomes public domain (free for use by the public) upon its expiration, which is defined as 20 years from the patent's earliest non-provisional filing date. MPEP §201.04. The 20-year patent term applies to utility and plant patents.

Read On ›

How long do Canadian design patents last? ›

You may obtain protection for the design of the entire finished article, or part of it. In Canada, this exclusive right can last for up to 15 years.

Learn More Now ›

Is it hard to get a patent in Canada? ›

Preparing and filing a patent application is not easy and can be time consuming. Think about hiring a licensed patent agent to help you. Patent agents understand patent and intellectual property laws and the application process. Make sure your patent agent is licensed.

Show Me More ›

Do Canadian design patents have maintenance fees? ›

Maintenance of Patent Applications in Canada. According to the Patent Act and Patent Rules, an applicant who files a Canadian patent application must pay maintenance fees to maintain the application.

What is Section 52 of the Canadian patent Rules? ›

52 The Federal Court has jurisdiction, on the application of the Commissioner or of any person interested, to order that any entry in the records of the Patent Office relating to the title to a patent be varied or expunged.

What is Section 56 of the Canadian patent Rules? ›

56 (1) Subject to subsection (2), if — before the claim date of a claim in a patent — a person, in good faith, committed an act that would otherwise constitute an infringement of the patent in respect of that claim, or made serious and effective preparations to commit such an act, it is not an infringement of the ...

Get More Info ›

Can you do a free patent search? ›

The U.S. Patent & Trademark Office provides free patent searching of its public databases. Google Patents. A relatively easy database to search which PDF images available. Limited to U.S. patents and U.S. published applications.

How do you get a patent everything you need to know? ›

Step 1: Get ready to apply. Contact Patents. ...
Step 2: File your application. Filing a patent application on your own. ...
Step 3: Application prosecution. Filing a patent application on your own. ...
Step 4: Receive your patent. Inventors Assistance Center. ...
Step 5: Maintain legal protection of your patent. Pay your maintenance fees.

Jan 31, 2019

How can I search for inventions that have already been patented? ›

One way of checking whether or not your product or idea has already been invented and patented by somebody else is to consult the EPO's free search service Espacenet. The database contains more than 140 million patent documents ‒ both published patent applications and granted patents ‒ from around the world.

How do I search a patent before 1976? ›

Patents produced before 1976 can be tricky to find. If you have the patent number, or are searching for all the patents produced with a specific classification number, you can do this on the USPTO web site. Click on "Quick Search" in the green Issued Patents box.

Know More ›

How do I get a copy of an old patent? ›

You must have a USPTO.gov account to order certified copies of patent and trademark documents. To view your previous order history, link your USPTO.gov account to your online document ordering system account. Only online orders (not fax or email orders) are viewable online.

Know More ›

Can the public view patents? ›

Hence, after a U.S. patent application is filed, it is published by the U.S. Patent Office for the public to view even if it hasn't been granted as a patent yet. Published patent applications can also be searched on the U.S. Patent Office website.

Get More Info ›

Can you see who owns a patent? ›

Go to the USPTO's online database. In the pull-down field menu, select "Assignee Name" if searching for company name or "Inventor Name" if searching for Inventor name. Type in the name of the inventor or company, keeping in mind that you might need to try several times.

View Details ›

Can anyone see a patent? ›

Legal documents that determine patent ownership affect the public's rights and obligations directly; therefore, they should be available to the public, just like any law or regulation affecting the public.

Keep Reading ›

What is considered the most valuable patent of all time? ›

In the last century, the invention of the bell's valve has become the most valuable patent in history. It was patented in 1878 and was used to transmit human voice. Bell was lucky to receive a patent for this invention within three weeks after filing his patent application.

Get More Info ›

What is the average cost for a patent? ›

The average cost of a patent is between $15,500 to $28,000. The average patent costs depend on various factors: Whether you file a provisional or nonprovisional patent application first; Whether you expedite your patent application; and.

View Details ›

Does a patent lose value? ›

Because a patent has a definite life and known value, under U.S. accounting practices, it must be amortized. The process of amortization decreases the value of the patent, converting each annual decrease into an expense. A patent's useful life is 20 years because that is when it terminates.

Read On ›

Can you prevent a patent from expiring? ›

The only way to extend protection is to invent and patent an improvement to the originally patented invention. A new patent will only protect the improvement, however, and not the original invention, which others will be free to copy and use when the original patent expires.

Discover More Details ›

Can I buy an abandoned patent? ›

If you would like to purchase an expired patent, this IS possible. In order to do so, you must first contact the patent owner or the attorney on record for that patent.

Get More Info ›

Can you claim an abandoned patent? ›

Fortunately, after a patent application has been declared abandoned, it can usually be revived by completing a few simple steps: Filing a petition to revive abandoned patent application. Paying a revival fee and any other fees that are owed. Correcting the issue that resulted in the abandonment.

See Details ›

What Cannot be patented in USA? ›

What can't be patented?

literary, dramatic, musical or artistic works,
a way of doing business, playing a game or thinking,
a method of medical treatment or diagnosis,
a discovery, scientific theory or mathematical method,
the way information is presented,
some computer programs or mobile apps,

More items...

Dec 17, 2018

Discover More ›

What are the 3 basic requirements for a patent? ›

Patent applications: the three criteria

Novelty. This means that your invention must not have been made public – not even by yourself – before the date of the application.
Inventive step. This means that your product or process must be an inventive solution. ...
Industrial applicability.

View Details ›

What are the 5 requirements of a patent? ›

The five primary requirements for patentability are: (1) patentable subject matter, (2) utility, (3) novelty, (4) nonobviousness, and (5) enablement.

Read On ›

Does Canada have a patent system? ›

In Canada, a patent lasts for 20 years from the date that you file it. Patents can have a great deal of value. You can sell them, license them or use them as assets to attract funding from investors.

Learn More ›

How do you check if something is trademarked or patented? ›

You may conduct a free online search of the USPTO database at the Public Search Facility (Madison East, 1st Floor; 600 Dulany Street, Alexandria, Virginia) between 8:00 a.m. and 8:00 p.m. USPTO personnel may not conduct trademark searches for the public. Private trademark search firms will conduct searches for a fee.

Read The Full Story ›

How do I download a Canadian patent? ›

Instructions

Find the document code within the CIPO correspondence letter that was sent to you ("Congratulations on obtaining a Canadian patent!"). ...
Enter your code in the "Document code" box below.
Click "Download Patent" to download your patent.
Click on "Download Specification" to download your patent specification.

Jul 20, 2021

Tell Me More ›

Who controls patents in Canada? ›

In Canada, patent applications are made to the Canadian Intellectual Property Office (CIPO). The CIPO website explains how to draft a patent and what goes into a patent application.

What is the grace period for patents in Canada? ›

The Canadian grace period covers any disclosure within one year of the filing date of a patent application in Canada. If a patent application is not filed in Canada within one year of the first public disclosure of an invention anywhere in the world, then the right to obtain patent protection in Canada is forfeited.

Get More Info ›

How do you know which names are trademarked in Canada? ›

The Canadian Trademark Database is accessible online as well as in person at the Canadian Intellectual Property Office (CIPO). This Canadian trademarks database is the most accurate source of information on which registered trademarks in Canada.

Do Canadian trademarks apply in the US? ›

While many prudent Canadian businesses will already own a trademark registration, trademark protection is territorial. A Canadian trademark registration does not protect that trademark in the U.S. – for that, a U.S. trademark registration is critical.

Explore More ›

What are the two types of patent infringement in Canada? ›

Generally, two types of patent infringement can be alleged in Canada, "direct" infringement and "indirect" infringement. The party alleging infringement bears the burden of proof to demonstrate on a balance of probabilities (see below) that the defendant's actions infringe the patent.

View Details ›

How do you know if a product is patentable? ›

In order for your invention to qualify for patent eligibility, it must cover subject matter that Congress has defined as patentable. According to USPTO this means the subject matter should be any "new and useful" process, machine, manufacture or composition of matter.

Keep Reading ›

Can I use a dead trademark? ›

A Dead Trademark means that the owner of a trademark has lost federal legal rights to it. This means anyone can register the logo or phrase for their use. Many businesses take advantage of this by assuming ownership of the Dead Trademark so they can rebrand without putting much effort into creating something new.

What tiny marks can tell you if something is trademarked? ›

The symbol ® is a notice of registered trademark ownership. It is used to advise the public that a trademark or service mark is registered, providing notice of the legal ownership status of the mark with which it is used. The ® symbol should be used only in connection with registered trademarks or service marks.

Know More ›