DESCRIPTION
Therapeutic or Prophylactic Agent for Multiple Sclerosis
Technical Field
[0001]
The present invention relates to a therapeutic or prophylactic agent for
multiple sclerosis comprising as an effective ingredient a glycine derivative or a
pharmaceutically acceptable salt thereof.
Background Art
[0002]
Multiple sclerosis is a nervous demyelinating disease with unknown etiology
which is clinically characterized in that recurrence and remission are repeated and
often develops into progressive physical disability. It is known that multiple
sclerosis causes visual impairment, loss of equilibrium sense, poorly coordinated
movement, slurred speech, tremor, numbness, extreme fatigue, alteration of
intellectual functions, muscular weakness, paresthesia, and blindness (see, for
example, Non-patent Literature 1 and Non-patent Literature 2).
[0003]
Although it is said that there are 2,500,000 patients suffering from multiple
sclerosis in the world, there may be a lot of potential patients who are not diagnosed
yet due to the difficulty to detect the symptoms. In addition, multiple sclerosis is a
chronic disease which repeats recurrence and remission and QOL of the patients is
often seriously disturbed. Therefore, the importance of therapy for multiple
sclerosis is increasing year by year.
[0004]
Multiple sclerosis cannot be cured and the therapy is performed mainly by
means of symptomatic treatments. Such treatments alleviate or prevent the
symptoms, but cannot cure demyelination which is the basic defect of this disease.

Corticosteroids which shorten the period of recurrence are used in the present therapy.
However, a long-term improvement effect on the course of the disease has not been
observed. In some cases, immunosuppressants, cyclophosphamide, leflunomide and
cyclosporin A arc used for progressive multiple sclerosis. However, it is known
that such agents have a teratogenicity and carcinogenicity.
[0005]
Thus, the therapeutic agents for multiple sclerosis now existing are not fully
satisfactory with respect to effects thereof and reduction of side effects.
[00061
On the other hand, Patent Literature 1 discloses a use of the compounds
represented by the below-described Formula:
[0007]

[0008]
(wherein Ar represents substituted phenylene or the like, L represents -S(0)2 - or the
like, X represents a bond or the like, Z represents COOH or the like. R2. R3, R4
represent hydrogen or the like, R5 represents -C2-10 alkenyl-aryl-NRdRe-heteroaryl-
(Rd, Re represent hydrogen, C2-10 alkyl) or the like, R6 represents hydrogen or the
like), which can be considered to be structurally similar to the compounds contained
as an effective ingredient in the present invention, for the therapy of multiple
sclerosis. However, the compounds whose use mentioned above is disclosed in the
publication are structurally different from the compounds in the present invention in
that the disclosed compounds are characterized in that Ar in the Formula is
substituted with aminoalkyl structure (R1 LN(R2)CHR4 -: R1 represents substituted
aryl or the like). Moreover, concrete data which demonstrate the effectiveness
against multiple sclerosis are not described, therefore the publication foils to fully

disclose that the compounds really have the therapeutic effect. Activity thereof
when orally administered is also not disclosed.
[0009]
In claims of Patent Literatures 2 and 3, use of the compounds represented by
the Formula:
[0010]
X-Y-Z-Aryl-A-B
[0011]
(wherein X represents a 6-membered aromatic ring comprising two nitrogen atoms or
the like, Y represents -Co-6 alkylene-NC1-10 alkyl-Co-6 alkylene- or the like. Z
represents a bond or the like, Aryl represents a 6-membered aromatic ring, A
represents -(CH2 )C=C(CH2 )n - (n=0-6) or the like, B represents -CH(substituted
aryl-carbonylamino)C02H or the like), which includes the compounds in the present
invention, as a bone resorption inhibitor and platelet aggregation inhibitor is
described. However, the compounds in the present invention are not concretely-
described in the publications. The use disclosed therein is also totally different
from that of in the present invention. The therapeutic or prophylactic effect on
multiple sclerosis according to the present invention is not suggested at all.
[0012]
Patent Literature 4, which was published after the priority date of the present
application, discloses a compound which is contained as an effective ingredient in the
present invention and use thereof for therapy of inflammatory bowel disease.
However, the therapeutic or prophylactic effect on multiple sclerosis according to the
present invention is not disclosed at all.
[0013]
Patent Literature 1: WO 99/26923
Patent Literature 2: WO 95/32710 "

Patent Literature 3: WO 94/12181
Patent Literature 4: WO 2006/068213
Non-patent Literature 1: Proc. Soc. Exp. Biol. Med., 175, 282-294 (1984)
Non-patent Literature 2: Immunol. Rev., 100, 307-332 (1987)
Disclosure of the Invention
Problems Which the Invention Tries to Solve
[0014]
In view of the fact that development of a compound useful as a
pharmaceutical for therapy or prophylaxis of multiple sclerosis is strongly demanded,
an object of the present invention is to provide a therapeutic or prophylactic agent for
multiple sclerosis comprising as an effective ingredient a low molecular compound
or a pharmaceutically acceptable salt thereof which is useful as a pharmaceutical.
Means for Solving the Problem
[0015]
The present inventors intensively studied to discover that the glycine
derivatives represented by Formula (I) and pharmaceutically acceptable salts thereof
have high oral absorbability and excellent in vivo stability, and are effective for the
therapy or prophylaxis of multiple sclerosis, to complete the present invention.
[0016]
That is, the present invention provides a therapeutic or prophylactic agent for
multiple sclerosis comprising as an effective ingredient a compound of the Formula
(I):
[0017]


[0018]
[wherein
R' represents hydrogen or C1-C5 alkyl;
Xs independently represent fluoro, chloro, bromo, iodo or C1-C3 alkyl;
V represents -CH=CH- or -OC-;
Y represents Formula (II) or Formula (III):

[0020]
(wherein
R represents C1-C5 alkyl or C1-C3 alkoxy;
R3 represents hydrogen or C1-C5 alkyl;
m represents an integer of 0 to 3;
n represents 0 or 1;
p represents an integer of 0 to 2;
W represents -O- or -N(R4)-
(wherein
R4 represents hydrogen, C1-C5 alkyl, C3-C6 alkenyl, phenyl, benzyl, benzyl
substituted with one or two R5s, tetrahydropyranyl, -(CH2)q-O-CH3, pyridylmethyl,
-(CH2)q-CN. C4-C7 cycloalkylmethyl or thiazol-4-ylmethyl;

R5 represents hydroxy or C1-C3 alkoxy; and
q represents an integer of 1 to 3))]
or a pharmaceutically acceptable salt thereof. The present invention also provides a
use of the compound of the Formula (I) described above or the pharmaceutically
acceptable salt thereof for the production of a therapeutic or prophylactic agent for
multiple sclerosis. The present invention further provides a therapeutic or
prophylactic method for multiple sclerosis, comprising administering to a patient an
effective amount of the compound of the Formula (1) described above or the
pharmaceutically acceptable salt thereof.
Effects of the Invention
[0021]
The therapeutic or prophylactic agents for multiple sclerosis according to the
present invention have excellent oral absorbability and in vivo stability, and therefore
exhibit high therapeutic or prophylactic effects.
Best Mode for Carrying out the Invention
[0022]
The term "alkyl" means a straight or branched hydrocarbon chain. Examples
of alkyl include methyl, ethyl, propyl, isopropyl, butyl and isobutyl.
[0023]
The term "alkoxy" means an oxygen atom having an alkyl substituent.
Examples of alkoxy include methoxy, ethoxy, propoxy, isopropoxy and butoxy.
[0024]
The term "alkenyl" means a straight or branched hydrocarbon chain having
one or more carbon-carbon double bond. Examples of alkenyl include vinyl, allyl,
isopropenyl, butenyl and pentenyl.
[0025]
The term "cycloalkyl" means a monocyclic saturated hydrocarbon ring.

Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.
[0026]
The term "cycloalkylmethyl" means a methyl group having a cycloalkyl group.
Examples of cycloalkylmethyl include cyclopropylmethyl, cyclobutylmethyl,
cyclopentylmethyl and cyclohexylmethyl.
[0027]
For the compounds represented by the above-described Formula (I), preferred
modes are as follows:
As the "X", fluoro, chloro or methyl is preferred, and chloro or methyl is
especially preferred.
As the "V", -CH=CH- is preferred, and trans form -CH=CH- is especially
preferred.
As the "R1", hydrogen, methyl or tert-butyl is preferred, and hydrogen is
especially preferred.
Among the "Y" structures represented by the above-described Formula (II) or
(HI):
As the "W", -N(R4)- is preferred.
As the "R2" when it exists, methyl, ethyl or methoxy is preferred, and methyl
is especially preferred.
As the "R3", hydrogen or C1-C3 alkyl (e.g., methyl, ethyl or propyl) is
preferred, and methyl is especially preferred.
As the "R4", C1-C3 alkyl (e.g., methyl, ethyl, propyl or isopropyl), cyanoethyl,
tetrahydropyranyl or phenyl is preferred, and methyl or isopropyl is especially
preferred.
As the "m", an integer of 0 to 2 is preferred, and 0 is especially preferred.
As the "n", 0 is preferred.
As the "p". 1 or 2 is preferred, and 1 is especially preferred.

The wavy line in Formulae (II) and (III) represents the moiety in the
compound, to which the group represented by Formula (II) or (III) is bound.
[0028]
Especially,
(1) the compounds represented by Formula (I), wherein
V is -CH=CH-;
when Y is represented by Formula (II), m is 0;
when Y is represented by Formula (III), p is 1;
and pharmaceutically acceptable salts thereof are preferred. Among these.
(2) the compounds represented by Formula (I), wherein
R1 is hydrogen;
when Y is represented by Formula (II). W is -N(R4)- and R4 is C1-C3 alkyl,
cyanoethyl, tetrahydropyranyl or phenyl;
when Y is represented by Formula (III), n is 0 and R3 is C1-C3 alkyl:
and pharmaceutically acceptable salts thereof are more preferred. Among these,
(3) the compounds represented by Formula (I), wherein
Xs are independently chloro or methyl;
V is trans form -CH=CH-;
when Y is represented by Formula (II), W is -N(R4)- and R4 is methyl or
isopropyl;
when Y is represented by Formula (III), R3 is methyl,
and pharmaceutically acceptable salts thereof are more preferred.
[0029]
Examples of the pharmaceutically acceptable salts of the compounds
represented by Formula (1) include, for acidic group(s) such as carboxylic group in
the formula, ammonium salt; salts of alkaline metals such as sodium and potassium:
salts of alkaline earth metals such as calcium and magnesium; aluminum salt; zinc

salt; salts of organic amines or the like such as triethylamine, ethanolamine.
morpholine, piperidine and dicyclohexylamine; and salts of basic amino acids such as
arginine and lysine. In cases where a basic group(s) exist(s) in the formula,
examples of the pharmaceutically acceptable salts of the compounds include, for the
basic group(s), salts of inorganic acids such as hydrochloric acid, sulfuric acid, nitric
acid, hydrobromic acid and phosphoric acid; salts of organic carboxylic acids such as
acetic acid, lactic acid, citric acid, maleic acid, benzoic acid, oxalic acid, glutaric acid,
malic acid, tartaric acid, fumaric acid, mandelic acid and succinic acid; and salts of
organic sulfonic acids such as methanesulfonic acid,/?-toluenesulfonic acid and
camphor sulfonic acid.
[0030]
The structure represented by Formula (I) includes optical isomers represented
by Formula (IV) and (V) about the asymmetric carbon atom, and the present
invention includes these isomers and mixtures thereof. Further, in the structure
represented by Formula (I), in cases where Y has an asymmetric carbon atom, the
present invention include the isomers based on the asymmetric carbon atom and
mixtures thereof.

[0032]
Among the ingredients of the therapeutic or prophylactic agents for multiple
sclerosis according to the present invention, preferred ingredients include compounds
represented by Formula (la) or (lb). Specific examples of the compounds

represented by Formula (la) or (lb) are shown in Tables 1 to 5.

The symbol "-lBu" shown in Tables 4 and 5 represents terl-buty\ group; the
symbol "-nPr" shown in Tables 1, 3 and 4 represents normal propyl group; and the
symbol "-'Pr" shown in Tables 1, 3 and 4 represents isopropyl group. The
compounds shown in Tables 1 to 5 include pharmaceutical!)' acceptable salts thereof.
[0039]
Among the ingredients of the therapeutic or prophylactic agents lor multiple
sclerosis according to the present invention, preferred ingredients also include
compounds represented by Formula (Ic) or (Id). Specific examples of the
compounds represented by Formula (Ic) or (Id) are shown in Tables 6 and 7.

[0042]
The symbol "-'Bu" shown in Table 7 represents tert-butyl group, and the
symbol "bond" shown in Tables 6 and 7 represents valence bond. The compounds
shown in Tables 6 and 7 include pharmaceutically acceptable salts thereof.
[0043]
The compounds represented by Formula (I) or the pharmaceutically
acceptable salts thereof which are comprised as an effective ingredient in the
therapeutic or prophylactic agents for multiple sclerosis according to the present
invention can be synthesized by the following steps:
[0044]
Conversion of the compounds represented by the following Formula (VI) :
[0045]

(wherein the symbols have the same meanings as described above):
[0046]
to a pharmaceutically acceptable salt thereof can be attained by a conventional
method using a base (inorganic base such as sodium hydroxide; organic base such as
triethylamine; or basic amino acid such as lysine) or an acid (acetic acid; inorganic
acid such as nitric acid or sulfuric acid; organic acid such as acetic acid or maleic
acid; organic sulfonic acid such as p-toluenesulfonic acid; acidic amino acid such as
aspartic acid).
[0047]
The compounds of Formula (VI) can be obtained by converting the esterified

carboxyl group CO2R16 of a compound of the Formula (VII):
[0048]

[0049]
[wherein R16 represents C1-C5 alkyl, and the other symbols have the same meanings
as described above]
to carboxyl group.
[0050]
Conversion of the esterified carboxyl group CO2R16 to carboxyl group may be
attained by a conventional method such as hydrolysis using a base (alkaline metal
hydroxide such as lithium hydroxide, sodium hydroxide, barium hydroxide) or an
acid (such as hydrochloric acid), or acid (e.g. trifluoroacetic acid) treatment. The
amount of the base used is usually 0.9 to 100 equivalents, preferably 0.9 to 10.0
equivalents with respect to the compound of Formula (VII). The amount of the acid
used is usually 1.0 equivalent with respect to the compound of Formula (VII) to an
excess amount used as a solvent, preferably 1.0 to 100 equivalents with respect to the
compound of Formula (VII).
[0051]
Examples of the solvent include aprotic bipolar solvents such as DMF and
DMSO: ether solvents such as diethyl ether, THF and DME: alcoholic solvents such
as methanol and cthanol; halogen-containing solvents such as dichloromethane.
chloroform, carbon tetrachloride and 1,2-dichloroethane; water; and mixtures thereof.
Among these, DMF, THF, methanol, ethanol or water is preferably used. The

reaction temperature is thought to be -30°C to 200°C. In case of hydrolysis using a
base, the reaction temperature is preferably -20°C to 60°C, more preferably -10°C to
40°C. In case of hydrolysis using an acid, the reaction temperature is preferably 0°C
to 180°C, more preferably 0°C to 100°C. The reaction time may appropriately be
selected depending on the conditions such as reaction temperature, and satisfactory
results are usually obtained by selecting a reaction time between about 1 minute and
30 hours.
[0052]
The compounds of Formula (VII) may be synthesized by the following
method (Method A or Method B) depending on the structure of "V" (V has the same
meaning as described above) in Formula (I):
[0053]
Method A: In cases where V is -CH=CH-
[0054]

[0055]
[wherein Z1 represents a leaving group such as halogen atom or
trifiuoromethanesulfonyloxy group, and the other symbols have the same meanings
as described above]
[0056]
The compounds of Formula (VIIa) can be synthesized by coupling a
compound of Formula (Villa) with a compound of Formula (IX).
[0057]

The coupling reaction is carried out in the presence of a palladium catalyst
and a base, and in the presence or absence of a phosphine ligand, in an appropriate
solvent.
[0058J
The compound of Formula (IX) is used in an amount of 1.0 to 10 equivalents,
preferably 1.0 to 4.0 equivalents with respect to the compound of Formula (Villa).
[0059]
Examples of the palladium catalyst include palladium acetate,
tetrakis(triphenylphosphine)palladium, bis(triphenylphosphine)palladiumdichloride,
palladium(II) chloride, bis(dibenzylideneacetone)palladium, and
bis(diphenylphosphino) ferrocene palladium dichloride. Among these, palladium
acetate, tetrakis(triphenylphosphine)palladium or
bis(dibenzylideneacetone)palladium is preferably used. The amount of the
palladium catalyst used is 0.001 to 1 equivalent, preferably 0.01 to 0.2 equivalents
with respect to the compound of Formula (Villa).
[0060]
Examples of the base include potassium carbonate, potassium phosphate,
cesium carbonate, sodium hydroxide, potassium hydroxide, sodium-tert-butoxide,
triethylamine, diisopropylamine, diisopropylethylamine and ^-butylamine. Among
these, potassium carbonate or potassium phosphate is preferably used. The amount
of the base used is 1 to 10 equivalents, preferably 1 to 4 equivalents with respect to
the compound of Formula (Villa).
[0061]
Examples of the phosphine ligand include triphenylphosphine, tris(2-
methylphenyl)phosphine, tributylphosphine, triethylphosphine. tri-tert-
butylphosphine, 2.2'-bis(diphenylphosphino)-l, 1 '-biphenyl, 2-(d\-tert-
butylphosphino)-1,1'-binaphthyl, 2-(dicyclohexylphosphino)diphenyl and 9,9-

dimethyl-4,5-bis(diphenylphosphino)xanthene. Among these, tributylphosphine,
tris(2-methylphenyl)phosphineor 9,9-dimethyl-4,5-bis(diphenylphosphino)xanlhene
is preferably used. The amount of the phosphine ligand used is 0.001 to 1
equivalent, preferably 0.01 to 0.2 equivalents with respect to the compound of
Formula (Villa).
[0062]
Examples of the solvent include aprotic bipolar solvents such as DMF,
DMSO and NMP; ether solvents such as THF, DME and dioxane; alcoholic solvents
such as methanol and ethanol; halogen-containing solvents such as dichloromethane.
chloroform, carbon tetrachloride and 1,2-dichloroethane; aromatic solvents such as
benzene, toluene and xylene; and water. Among these, DMF, dioxane or water is
preferably used. The reaction temperature is thought to be 0°C to 200°C, and is
preferably 40°C to 160°C, more preferably 60°C to 140°C. The reaction time may
appropriately be selected depending on the conditions such as reaction temperature,
and satisfactory results are usually obtained by selecting a reaction time between
about 30 minutes and 30 hours.
[0063]
Method B: In cases where V is -CII=CII-
[0064]

[0065]
[wherein the symbols have the same meanings as described above]
[0066]

The compounds of Formula (Vllb) can be synthesized by the coupling
reaction between a compound of Formula (VIIIb) and a compound of Formula (IX).
The coupling reaction is carried out in the presence of a palladium catalyst, a base,
and a copper catalyst, in the presence or absence of a phosphine ligand, in an
appropriate solvent.
[0067]
The compound of Formula (IX) is used in an amount of 1.0 to 10 equivalents,
preferably 1.0 to 4.0 equivalents with respect to the compound of Formula (VHIb).
[0068]
Examples of the palladium catalyst include palladium acetate,
tetrakis(triphenylphosphine)palladium, bis(triphenylphosphine)palladium dichloride.
palladium(II) chloride, bis(dibenzylideneacetone)palladium, and
bis(diphenylphosphino) ferrocene palladium dichloride. Among these,
tetrakis(triphenylphosphine)palladium or bis(triphenylphosphine)palladium
dichloride is preferably used. The amount of the palladium catalyst used is 0.001 to
1 equivalent, preferably 0.005 to 0.2 equivalents with respect to the compound of
Formula (VIIIb).
[0069]
Examples of the base include potassium carbonate, potassium phosphate,
cesium carbonate, sodium hydroxide, potassium hydroxide, sodium-tert-butoxide,
diethylamine, triethylamine, diisopropylamine, diisopropylethylamine and n-
butylamine. Among these, diethylamine, triethylamine, diisopropylamine or
diisopropylethylamine is preferably used. The amount of the base used is 1
equivalent with respect to the compound of Formula (VIIIb) to an excess amount
used as a solvent, preferably 4 equivalents with respect to the compound of Formula
(VIIIb) to an excess amount.
[0070]

Examples of the copper catalyst include copper powder, copper iodide and
copper bromide, and copper iodide is preferably used. The copper catalyst is used
in an amount of 0.001 to 0.5 equivalents, preferably 0.01 to 0.4 equivalents with
respect to the compound of Formula (VIIIb).
[0071]
Examples of the phosphine ligand include triphenylphosphine, tris(2-
methylphenyl)phosphine, tetrakis(triphenyl)phosphine, tributylphosphine,
triethylphosphine. tri-tert-butylphosphine, 2,2'-bis(diphenylphosphino)-l,1-biphenyl,
2-(di-tert-butylphosphino)-l ,l'-binaphthyl, 2-(dicyclohexylphosphino)diphenyl and
9,9-dimethyl-4.5-bis(diphenylphosphino)xanthene. Preferably, no phosphine ligand
is used, or tris(2-methylphenyl)phosphine or tetrakis(triphenyl)phosphine is
preferably used. When the phosphine ligand is used, the amount thereof is 0.001 to
0.5 equivalents, preferably 0.005 to 0.4 equivalents with respect to the compound of
Formula (VIIIb).
[0072]
Examples of the solvent include aprotic bipolar solvents such as DMF.
DMSO and NMP; ether solvents such as THF, DME and dioxanc; alcoholic solvents
such as methanol and ethanol; halogen-containing solvents such as dichloromethanc.
chloroform, carbon tetrachloride and 1,2-dichloroethane; aromatic solvents such as
benzene, toluene and xylene; organic amine solvents such as diethylamine,
triethylamine and diisopropylamine; and water; as well as mixtures thereof.
Preferably, THF, diethylamine, diisopropylamine, triethylamine or a mixture thereof
is used. The reaction temperature is thought to be -40°C to 200°C. and is preferably
-20°C to 100°C, more preferably -10°C to 60°C. The reaction time may
appropriately be selected depending on the conditions such as reaction temperature,
and satisfactory results are usually obtained by selecting a reaction time between
about 1 minute and 30 hours.

[0073]
The compounds of Formula (Villa) and the compounds of Formula (VII lb)
can be synthesized by the following method:
[0074]

[0075]
[wherein Z2 represents -CH=CH2 or -C=CH, and the other symbols have the same
meanings as described above]
[0076]
The compounds of Formula (Villa) and the compounds of Formula (VHIb)
can be synthesized by the following methods (Method C, Method D or Method E):
[0077]
Method C:
The compounds can be synthesized by condensing a compound of Formula
(X) with Rl6-OH (wherein R16 represents the same meaning as described above).
The condensation reaction may be carried out in the presence of a condensing agent,
in the presence or absence of a base, in an appropriate solvent. Examples of the
condensing agent include dicyclohexylcarbodiimide, BOP reagent and EDC.
Preferably, EDC or BOP reagent is used. In cases where the base exists, examples
of the base include triethylamine, diisopropylethylamine, pyridine, 4-
dimethylaminopyridine and N-mefhylmorpholine. Preferably, triethylamine. 4-
dimethylaminopyridine or diisopropylethylamine is used. The amount of R '-OH
used is 1.0 to 100 equivalents, preferably 1.0 to 4.0 equivalents with respect to the
compound of Formula (X). The condensing agent is used in an amount of 1.0 to 20

equivalents, preferably 1.0 to 5.0 equivalents with respect to the compound of
Formula (X). The base is used in an amount of 1.0 to 100 equivalents, preferably
1.0 to 40 equivalents with respect to the compound of Formula (X).
[0078]
Examples of the solvent include aprotic bipolar solvents such as DMF,
DMSO and NMP; ether solvents such as THF, DME and dioxane; and halogen-
containing solvents such as dichloromethane, chloroform and 1,2-dichloroethane.
Preferably, THF or dichloromethane is used. The reaction temperature is thought to
be -40°C to 100°C, and is preferably 0°C to 60°C. The reaction time may
appropriately be selected depending on the conditions such as reaction temperature,
and satisfactory results are usually obtained by selecting a reaction time between
about 1 minute and 30 hours.
[0079J
Method D:
The compounds can be synthesized by condensing a reactive derivative of a
compound of Formula (X) with Ri6-OH (wherein R16 represents the same meaning as
described above) in the presence of a base. Examples of the reactive derivative of
the compound of Formula (X) include acid halides (such as acid chloride).
Examples of the base include organic amine bases such as triethylamine, pyridine and
diisopropylethylamine; and inorganic bases such as sodium carbonate, potassium
carbonate and sodium hydrogen carbonate. Preferably, triethylamine, pyridine or
diisopropylethylamine is used. The amount of R!6-OH is 1.0 to 100 equivalents,
preferably 1.0 to 20 equivalents with respect to the compound of Formula (X). The
base is used in an amount of 1.0 to 100 equivalents, preferably 1.0 to 4.0 equivalents
with respect to the compound of Formula (X).
|0080]
Examples of the solvent include aprotic bipolar solvents such as DMF,

DMSO and NMP; ether solvents such as THF, DME and dioxane; halogen-
containing solvents such as dichloromethane, chloroform and 1,2-dichloroethane;
and aromatic solvents such as benzene and toluene. Preferably, dichloromethane or
THF is used. The reaction temperature is thought to be -10°C to 100°C, and is
preferably 0°C to 40°C. The reaction time may appropriately be selected depending
on the conditions such as reaction temperature, and satisfactory results are usually
obtained by selecting a reaction time between about 1 minute and 30 hours.
[0081]
Method E:
In cases where R16 is methyl or ethyl, the compounds can be synthesized by
adding a compound of Formula (X) to a mixture of methanol or ethanol and thionyl
chloride. The amount of methanol or ethanol used is usually 1.0 equivalent with
respect to the compound of Formula (X) to an excess amount used as a solvent, and
is preferably 10 equivalents with respect to the compound of Formula (X) to an
excess amount used as a solvent. The amount of thionyl chloride used is usually 1
equivalent to an excess amount used as a solvent. The reaction temperature is
thought to be -50°C to 60°C, and is preferably -20°C to 40°C. The reaction time
may appropriately be selected depending on the conditions such as reaction
temperature, and satisfactory results are usually obtained by selecting a reaction time
between about 1 minute and 60 hours.
[0082]
The compounds of Formula (X) can be synthesized by the following method:


[wherein the symbols have the same meanings as described above]
[0085]
The compounds can be synthesized by condensing a compound of Formula
(XI) with a compound of Formula (XII) in the presence of a base in an appropriate
solvent. The compounds of Formulae (XI) and (XII) arc normally available
materials. The compound of Formula (XII) is used in an amount of 1.0 to 20
equivalents, preferably 1.0 to 4.0 equivalents with respect to the compound of
Formula (XI).
[0086]
Examples of the base include organic amine bases such as triethylamine,
pyridine and diisopropylethylamine; and inorganic bases such as sodium hydroxide,
potassium hydroxide, sodium carbonate, potassium carbonate and sodium hydrogen
carbonate. Preferably, triethylamine, pyridine, diisopropylethylamine, sodium
hydroxide or potassium hydroxide is used. The base is used in an amount of 1.0 to
100 equivalents, preferably 1.0 to 50 equivalents with respect to the compound of
Formula (XI).
[0087]
Examples of the solvent include aprotic bipolar solvents such as DMF,
DMSO and NMP; ether solvents such as THF, DME and dioxane; halogen-
containing solvents such as dichloromethane, chloroform and 1,2-dichloroethane;
and water, as well as mixtures thereof. Preferably, dichloromethane, THF. dioxane,
water or a mixture thereof is used. The reaction temperature is thought to be -10°C
to 100°C, and is preferably 0°C to 40°C. The reaction time may appropriately be
selected depending on the conditions such as reaction temperature, and satisfactory-
results are usually obtained by selecting a reaction time between about 1 minute and
60 hours.
[0088]

The compounds of Formula (IX) can be synthesized by the following method
(Method F to Method H) depending on the structure of "Y" (Y has the same meaning
as described above) in Formula (IX):
[0089]
Method F:
In cases where Y is represented by the following Formula (Ha):
[0090]

[0091]
| wherein the symbols have the same meanings as described above]
the compounds of Formula (IX) can be synthesized by the following method:
[0092]

[0093]
[wherein the symbols have the same meanings as described above]
[0094]
Step 1:
The compounds of Formula (XV) may be synthesized by condensing a
compound of Formula (XIII) and a compound of Formula (XIV) in the presence of an
acid in an appropriate solvent. Examples of the acid include acetic acid,
trifluoroacetic acid, methanesulfonic acid, p-loluenesulfonic acid and camphor
sulfonic acid. Preferably, acetic acid is used. The compounds of Formula (XIII)
are normally available materials and the compounds of Formula (XIV) are normally

available materials or may be synthesized by a known method. The compound of
Formula (XIV) is used in an amount of 1.0 to 10 equivalents, preferably 1.0 to 4.0
equivalents with respect to the compound of Formula (XIII). The acid is used in an
amount of 1.0 equivalent to an excess amount used as a solvent, preferably in an
amount of 1.0 to 20 equivalents.
[0095]
Examples of the solvent include ether solvents such as THF, DME and
dioxane; halogen-containing solvents such as dichloromethane and carbon
tetrachloride; aromatic solvents such as benzene, toluene and xylene; and aprotic
bipolar solvents such as DMF and DMSO. Preferably, DME or dioxane is used.
The reaction temperature is thought to be 0°C to 160°C, and is preferably 40°C to
100°C. The reaction time may appropriately be selected depending on the
conditions such as reaction temperature, and satisfactory results are usually obtained
by selecting a reaction time between about 1 hour and 30 hours.
[0096]
Step 2:
The compounds of Formula (IXa) can be synthesized by adding a base to a
compound of Formula (XV) in an appropriate solvent and further adding R -Z
(wherein the symbols have the same meanings as described above) to conduct
condensation. The compound of R4-Z' is used in an amount of 1.0 to 10
equivalents, preferably 1.0 to 4.0 equivalents with respect to the compound of
Formula (XV).
[0097]
Hxamples of the base include metal hydrides such as sodium hydride and
potassium hydride; inorganic bases such as potassium carbonate, sodium carbonate
and cesium carbonate; organic metals such as butyllithium; and organic amines such
as DBU, diisopropylethylamine and triethylamine. Preferably, sodium hydride is

used. The base is used in an amount of 1.0 to 50 equivalents, preferably 1.0 to 20
equivalents with respect to the compound of Formula (XV). Examples of the
solvent include ether solvents such as THF, DME and dioxane; halogen-containing
solvents such as dichloromethane and carbon tetrachloride; aromatic solvents such as
benzene, toluene and xylene; and aprotic bipolar solvents such as DMF and DMSO.
Preferably, DMF or THF is used. The reaction temperature is thought to be -78°C
to 160°C, and is preferably -20°C to 40°C. The reaction time may appropriately be
selected depending on the conditions such as reaction temperature, and satisfactory
results are usually obtained by selecting a reaction time between about 15 minutes
and 30 hours.
[0098]
Method G:
In cases where Y is represented by the following Formula (IIb):
[0099]

[0100]
[wherein the symbols have the same meanings as described above]
the compounds of Formula (IX) can be synthesized by the following method:
[0101]

[0102]
[wherein the symbols have the same meanings as described above]
[0103]

The compounds can be synthesized by condensing a compound of Formula
(XVI) and a compound of Formula (XIV) in the presence of a base in an appropriate
solvent. The compounds of Formula (XVI) are normally available materials. The
amount of the compound of Formula (XIV) used is 1.0 to 10 equivalents, preferably
1.0 to 4.0 equivalents with respect to the compound of Formula (XVI).
[0104]
Examples of the base include metal hydrides such as sodium hydride and
potassium hydride; inorganic bases such as potassium carbonate, sodium carbonate
and cesium carbonate; organic metals such as butyllithium; and organic amines such
as DBU, diisopropylethylamine and triethylamine. Preferably, potassium carbonate
or sodium carbonate is used. The amount of the base used is 1.0 to 50 equivalents,
preferably 1.0 to 20 equivalents with respect to the compound of Formula (XVI).
[0105]
Examples of the solvent include ether solvents such as THF, DME and
dioxane; halogen-containing solvents such as dichloromethane and carbon
tetrachloride; aromatic solvents such as benzene, toluene and xylene; and aprotic
bipolar solvents such as DMF and DMSO. Preferably, DMF is used. The reaction
temperature is thought to be 0°C to 200°C, and is preferably 20°C to 140°C. The
reaction time may appropriately be selected depending on the conditions such as
reaction temperature, and satisfactory results are usually obtained by selecting a
reaction time between about 30 minutes and 30 hours.
[0106]
Method H:
In cases where Y is represented by the following Formula (Ilia):
[0107]


[0108]
[wherein the symbols have the same meanings as described above]
the compounds of Formula (IX) can be synthesized by the following method:

[wherein the symbols have the same meanings as described above]
[0111]
Steps 1 and 2:
By lithiation of a compound of Formula (XVII) in an appropriate solvent, a
compound of Formula (XVIII) is obtained. By subsequently reacting this
compound without isolation with a compound of Formula (XIX), a compound of
Formula (XX) is obtained. The compounds of Formula (XVII) are normally
available materials. Examples of the lithiating agent include lithium, w-butyllithium.
sec-butyl lithium and tert-butyllithium. Preferably, n-butyllithium is used. The
amount of the lithiating agent used is 1.0 to 4.0 equivalents, preferably 1.0 to 2.2
equivalents with respect to the compound of Formula (XVII). The compounds of
Formula (XIX) are normally available materials. The amount of the compound of
Formula (XIX) used is 1.0 to 10 equivalents, preferably 1.0 lo 4.0 equivalents with
respect to the compound of Formula (XVII).
[0112]

Examples of the solvent include ether solvents such as ether, dioxane and
THF, and preferably, THF is used. The reaction temperature in Step 1 is thought to
be -100°C to 0°C, and is preferably -78°C to 0°C. The reaction temperature in Step
2 is thought to be -100°C to 40°C, and is preferably -78°C to 40°C. The reaction
time may appropriately be selected depending on the conditions such as reaction
temperature, and in Step 1, about 5 minutes to 2 hours is usually appropriate, and in
Step 2, satisfactory results are usually obtained by selecting a reaction time between
about 5 minutes and 24 hours.
[0113]
Step 3
The compounds of Formula (IXc) can be synthesized by adding a base to a
compound of Formula (XX) in an appropriate solvent, and subsequently adding R3-
Z (wherein the symbols have the same meanings as described above) to conduct
condensation. The compounds represented by R -Z are normally available
materials. The amount of R3-Z1 used is 1.0 to 20 equivalents, preferably 1.0 to 4.0
equivalents with respect to the compound of Formula (XX).
[0114]
Examples of the base include metal hydrides such as sodium hydride and
potassium hydride; inorganic bases such as potassium carbonate, sodium carbonate
and cesium carbonate; organic metals such as butyllithium; and organic amines such
as DBU, diisopropylethylamine and triethylamine. Preferably, sodium hydride is
used. The amount of the base used is 1.0 to 50 equivalents, preferably 1.0 to 20
equivalents with respect to the compound of Formula (XX).
[0115]
Examples of the solvent include ether solvents such as THF, DME and
dioxane; halogen-containing solvents such as dichloromethane and carbon
tetrachloride; aromatic solvents such as benzene, toluene and xylene; and aprotic

bipolar solvents such as DMF and DMSO. Preferably, DMF or THF is used. The
reaction temperature is thought to be -78°C to 100°C, and is preferably -20°C to 40°C.
The reaction time may appropriately be selected depending on the conditions such as
reaction temperature, and satisfactory results are usually obtained by selecting a
reaction time between about 15 minutes and 30 hours.
[0116J
The workup and purification of each step in the synthesis of the compounds
represented by Formula (I) or pharmaceutically acceptable salts thereof can be carried
out by an ordinary method. That is, as the workup, separation extraction, filtration
or the like may be employed. As the purification, column chromatography, thin
layer chromatography, recrystallization, reprecipitation, distillation or the like may be
employed.
[0117]
The therapeutic or prophylactic agents for multiple sclerosis according to the
present invention have a glycine structure substituted with an allyl group or propargyl
group, a substructure represented by Formula (II) or (III), and the 2,6-di-substituted
benzoyl structure shown in Formula (I). These compounds have more excellent oral
absorbability and in vivo stability than those disclosed in W099/26923 mentioned in
Background Art, and therefore exhibit therapeutic or prophylactic effects on multiple
sclerosis when orally administered.
[0118]
The oral absorbability and in vivo stability of the therapeutic or prophylactic
agents of the present invention may be confirmed by using as an index bioavailability
(BA) and clearance of the compound administered to an animal, but the method for
confirmation is not restricted thereto.
[0119]
The excellent effect of the therapeutic or prophylactic agents of the present

invention on multiple sclerosis may be evaluated based on the effect to reduce the
increase in the score of nerve paralysis by using mouse experimental autoimmune
encephalomyelitis (EAE) model described in the literature "Int. Immunol.. 9, 1243-
1251 (1997)". Experimental autoimmune encephalomyelitis is the animal model in
which neuropathy such as hind limb paralysis due to demyelination in central nerve
system is induced by immunizing an experimental animal with myelin basic protein
or a partial peptide thereof. The symptoms and pathological findings are similar to
those observed in human, and thus the model is widely used for the study of
pharmacological effects of therapeutic or prophylactic agents for multiple sclerosis.
However, the evaluation method is not restricted thereto.
[0120J
The therapeutic or prophylactic agents according to the present invention
suppress neuropathy such as hind limb paralysis induced by myelin basic protein or a
partial peptide thereof, and therefore can be used for ameliorating multiple sclerosis,
more specifically, acute multiple sclerosis, generalized multiple sclerosis, brainstem
multiple sclerosis, multiple sclerosis in the spinal cord, asymptomatic multiple
sclerosis or the like. Examples of the causative substance of multiple sclerosis
include myelin proteins such as proteolipid protein (PLP); myelin oligodendrocyte
glycoprotein (MOG); myelin basic protein (MBP); myelin-associated glycoprotein
(MAG), myelin-associated oligodendrocytic basic protein (MBOP); citrullinated
MBP (MBP C8 isomer in which six arginines are converted to citrullines by
deimination), cyclic nucleotide phosphodiesterase (CNPase), alpha-B crystal 1 in and
the like. Infection by measles virus, herpes simplex virus, EB virus, retrovirus,
human herpesvirus 6 or the like may also cause multiple sclerosis.
[0121|
Further, the therapeutic or prophylactic agents according to the present
invention suppress nerve paralysis due to demyelination in central nerve system, and

therefore can be used for acute disseminated encephalomyelitis,
adrenoleukodystrophy, adrenomyeloneuropathy, tropical spastic paraparesis, and
Leber's hereditary optic atrophy, which are the primary demyelinating disease similar
to multiple sclerosis.
[0122]
Further, the therapeutic or prophylactic agents according to the present
invention can be used for ameliorating abnormality of sensation (e.g., numbness,
pain), vision (e.g., diplopia, partial blindness, loss of central vision, vertigo),
movement (weakness, painful tonic spasm, rigidity, tremor) and nerve (e.g.. elation,
depressed state, mental disorders), which are the symptom accompanied by multiple
sclerosis, as well as poorly coordinated movement (e.g., loss of equilibrium sense,
slurred speech), extreme fatigue, alteration of intellectual functions, muscular
weakness, paresthesia, Uthoff s phenomenon, voiding dysfunction and constipation.
[0123]
The effect of the therapeutic or prophylactic agents according to the present
invention on multiple sclerosis can be explained by the inhibition of leukocyte
functions (e.g., adhesion and growth of cells) and inhibition of production of
inflammatory mediators (e.g., cytokines and chemical mediators) by leukocytes. As
the leukocytes, neutrophils, monocytes, lymphocytes, which are originated from
peripheral blood, or established cell lines thereof or the like, are used. For the
evaluations of the leukocyte functions and the inflammatory mediators, the methods
described in, for example, Current Protocols in Immunology (John Wiley & Sons.
Inc) are used, but the evaluation methods are not restricted thereto.
[0124]
The therapeutic or prophylactic agents for multiple sclerosis according to the
present invention can be used as a pharmaceutical for mammals (e.g., mouse, rat.
hamster, rabbit, dog, monkey, bovine, ovine, human and the like).

[0125]
When using the compound clinically, the drug may be the free form or a salt
thereof per se, or an additive(s) such as a vehicle, stabilizer, preservative, buffering
agent, solubilizer, emulsifier, diluent and/or isotonic agent, may be admixed
appropriately. The drug may be produced by a conventional method by
appropriately using these pharmaceutical carriers. Examples of the administration
mode include oral preparations such as tablets, capsules, granules, powders and
syrups; parenteral preparations such as inhalants, injection solutions, suppositories
and liquids; or topical preparations such as ointments, creams and patches.
Further, known sustained-release preparations are also included.
[0126]
The therapeutic or prophylactic agent for multiple sclerosis according to the
present invention preferably contains the above-described effective ingredient in an
amount of 0.001 to 90% by weight, more preferably 0.01 to 70% by weight.
Although the dose may be selected depending on the symptoms, age, body weight,
sex, administration method and the like, in case of an injection solution, a dose of
0.01 mg to 25 g, preferably 0.1 mg to 10 g, and in case of an oral preparation, a dose
of 0.1 mg to 50 g, preferably 1 mg to 20 g, in terms of the effective ingredient,
respectively, may be administered to an adult per day. Although the dosing period
and dosing intervals of the therapeutic or prophylactic agent of the present invention
may be altered depending on various situations, administration method such as
divided administration, daily administration, intermittent administration, short-term
high-dose administration, repeated administration or the like may be employed. For
example, in case of an oral administration, the agent may preferably be administered
once or dividedly in several times a day (especially two to three times a day). The
agent may also be administered by intravenous drip infusion over a long time.
[0127]

Examples of the pharmaceutically acceptable carrier or diluent include
binders (syrups, gelatin, gum Arabic, sorbitol, polyvinyl chloride, tragacanth and the
like), vehicles (sucrose, lactose, corn starch, calcium phosphate, sorbitol, glycine and
the like), lubricants (magnesium stearate, polyethylene glycol, talc, silica and the
like).
[0128]
The therapeutic or prophylactic agent for multiple sclerosis according to the
present invention may be used alone, or may be blended with or used together with
other drug(s) for therapy or prophylaxis of the disease, or for alleviation or
suppression of symptoms, or for complementation or enhancement of the
prophylactic or therapeutic effect, or for decreasing the dose.
[0129]
Examples of the drugs which may be used together in therapy or prophylaxis
of multiple sclerosis include corticosteroids (prednisolone, methylprednisolone and
the like), immunosuppressants (methotrexate, azathiopurine, cyclophosphamide,
cyclosporin A, tacrolimus, mizoribine, leflunomide and the like), interferon
preparations (interferon a, interferon [3 and the like), copolymer I, immunoglobulins,
mitoxantrone, glatiramer acetate, T cell receptor vaccines, adhesion molecule
inhibitors, analgesics (indomethacin, diclofenac and the like), muscle relaxants
(tizanidine, eperisone, afloqualone, baclofen, diazepam, dantrolene sodium and the
like), anticonvulsants (carbamazepine, phenytoin, clonazepam, amitriptyline and the
like), therapeutic agents for urinary incontinence or urinary frequency (oxybutynin.
propiverine, flavoxate, distigmine and the like) and the like.
[0130]
Examples
The present invention will now be described more concretely by way of
examples thereof. Abbreviations used herein mean the following compounds.

Abbreviations:
THF: tetrahydrofuran
DMF: dimethylformamide
Boc: /err-butoxycarbonyl
DMF: dimethylformamide
NMP: N-methylpyrrolidone
DME: dimethoxyethane
DMSO: dimethylsulfoxide
BOP: benzotriazol-l-yloxytris(dimethylamino)-phosphonium hexafluoro phosphate
EDC: 1 -ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride
DBU: 1,8-diazabicyclo[5.4.0]undeca-7-ene
HOBT: 1 -hydroxybenzotriazole
[0131]
Reference Example 1
Tetrahydro-4-(4-iodophenyl)-2H-pyran-4-ol
[0132]

[0133]
Under an argon atmosphere, a solution of n-butyllithium (2.59M, in hexane)
(8.76 ml) was added dropwise to a solution of 1,4-diiodobenzene (7.49 g) in
anhydrous THF (50 ml) at -78°C, and the resulting mixture was stirred at -78"C for
30 minutes. Tetrahydro-4H-pyran-4-one (2.09 ml) was added dropwise thereto and
the resulting mixture was stirred at -78°C for 1.5 hours. After stirring the mixture
for another 3 hours at room temperature, water was added, and the resulting mixture
was extracted with ethyl acetate. Organic layer was washed once with water and

once with saturated brine, and dried over anhydrous sodium sulfate. After removing
anhydrous sodium sulfate by filtration, the filtrate was concentrated. The residue
was recrystallized from cyclohexane/ethyl acetate mixed solvent to obtain tetrahydro-
4-(4-iodophenyl)-2H-pyran-4-ol (4.12 g).
[0134]
Reference Example 2
Tetrahydro-4-(4-iodophenyl)-4-methoxy-2H-pyran
[0135]

[0136]
Under an argon atmosphere, a solution of tetrahydro-4-(4-iodophenyl)-2H-
pyran-4-ol (2.99 g) in anhydrous DMF (10 ml) was added dropwise to a suspension
of sodium hydride (60 wt %) (432 mg) in anhydrous DMF (20 ml) at room
temperature, and the resulting mixture was stirred at room temperature for 75
minutes. Methyl iodide (0.92 ml) was added dropwise to the reaction solution and
the resulting mixture was stirred at room temperature for 8.5 hours. Water was
added thereto and the resulting mixture was extracted twice with ethyl acetate.
Organic layers were washed with saturated brine, and dried over anhydrous sodium
sulfate. After removing anhydrous sodium sulfate by filtration, the filtrate was
concentrated. The residue was purified by column chromatography (silica gel,
eluent: cyclohexane/ethyl acetate = 10/1) to obtain tetrahydro-4-(4-iodophenyl)-4-
methoxy-2H-pyran (2.87 g). NMR(H1 CDC13):5 1.90-2.02(411, m). 2.97(3H. s).
3.80-3.88(411, m), 7.14(111, d, J=8.5Hz), 7.70(1H, d, J=8.5Hz)
[0137J
Reference Example 3

4-Ethoxy-tetrahydro-4-(4-iodophenyl)-2H-pyran
[0138]

[0139]
In the same manner as in Reference Example 2, tetrahydro-4-(4-iodophenyl)-
2H-pyran-4-ol (302 mg) and iodoethane (0.119 ml) were reacted to obtain 4-ethoxy-
tetrahydro-4-(4-iodophenyl)-2H-pyran (269 mg).
[0140]
Reference Example 4
4-(4-Bromophenyl)-tetrahydro-4-(methoxymethyl)-2H-pyran
[0141]

[0142]
(1) Under an argon atmosphere, sodium hydride (60 wt %) (240 mg) was added
to a solution of 2-(4-bromophenyl) acetic acid methyl ester (916 mg) in anhydrous
DMF (20 ml) at 0°C, and the resulting mixture was stirred at 0°C for 30 minutes.
To the reaction solution, bis(2-bromoethyl)ether (0.70 mL) was added, and the
resulting mixture was stirred at 0°C for 1 hour. Sodium hydride (60 wt %) (240 mg)
was added thereto and the resulting mixture was further stirred at 0°C for 1 hour.
Saturated aqueous ammonium chloride solution was added thereto, and the resulting
mixture was extracted with ethyl acetate. Organic layer was washed once with
saturated brine, and dried over anhydrous sodium sulfate. After removing
anhydrous sodium sulfate by filtration, the filtrate was concentrated. The residue

was purified by column chromatography (silica gel, eluent: hexane/ethyl acetate =
4/1) to obtain 4-(4-bromophenyl)-tetrahydro-2H-pyran-4-carboxylic acid methyl ester
(472 mg).
(2) Under an argon atmosphere, a solution of 4-(4-bromophenyl)-tetrahydro-2/1/-
pyran-4-carboxylic acid methyl ester (472 mg) obtained in (1) in anhydrous THF (8.7
mL) was added to a 1.0 M solution of diisobutylaluminum hydride in hexane (8.7
mL) at 0°C, and the resulting mixture was stirred at 0°C for 30 minutes. After
adding methanol (2.1 ml) to the reaction solution, IN hydrochloric acid (8.7 mL) was
added thereto and the resulting mixture was extracted 4 times with diethyl ether.
Organic layers were washed with saturated brine, and dried over anhydrous sodium
sulfate. After removing anhydrous sodium sulfate by filtration, the filtrate was
concentrated to obtain (4-(4-bromophenyl)-tetrahydro-2H-pyran-4-yl)methanol (405
mg)-
(3) Under an argon atmosphere, sodium hydride (60 wt %) (88 mg) was added to
a solution of (4-(4-bromophenyl)-tetrahydro-2H-pyran-4-yl)methanol (397 mg)
obtained in (2) in anhydrous DMF (1.5 mL) at 0°C, and the resulting mixture was
stirred at 0°C for 30 minutes. Methyl iodide (0.13 ml) was added dropwise to the
reaction solution, and the resulting mixture was stirred at room temperature for 2
hours. Saturated brine was added thereto and the resulting mixture was extracted 3
times with ethyl acetate. Organic layers were washed with saturated brine and dried
over anhydrous sodium sulfate. After removing anhydrous sodium sulfate by
filtration, the filtrate was concentrated. The residue was purified by column
chromatography (silica gel, eluent: hexane/ethyl acetate = 5/1) to obtain 4-(4-
bromophenyl)-tetrahydro-4-(methoxymethyl)-2H-pyran (399 mg).
[0143]
Reference Example 5
4-(4-Iodophenyl)-4-methoxyoxepane

[0144]

[0145]
(1) Under an argon atmosphere, a solution of M-butyllithium (1.50 M, in hexane)
(0.5 ml) was added dropwise to a solution of 1,4-diiodobenzene (250 mg) in
anhydrous THF (3 ml) at -78°C, and the resulting mixture was stirred at -78°C for 30
minutes. Oxepan-4-one (151 mg) was added to the reaction solution, and the
resulting mixture was stirred at -78°C for 1 hour. Saturated aqueous ammonium
chloride solution was added thereto, and the resulting mixture was extracted with
ethyl acetate. Organic layer was washed with saturated brine, and dried over
anhydrous sodium sulfate. After removing anhydrous sodium sulfate by filtration,
the filtrate was concentrated. The residue was purified by column chromatography
(silica gel, eluent: hexane/ethyl acetate = 5/1) to obtain 4-(4-iodophenyl)oxepan-4-ol
(89 mg).
(2) Under an argon atmosphere, a solution of 4-(4-iodophenyl)oxepan-4-ol (86
mg) obtained in (1) in anhydrous DMF (1.0 mL) was added to a suspension of
sodium hydride (60 wt %) (16 mg) in anhydrous DMF (1.0 ml) at 0°C, and the
resulting mixture was stirred at 0°C for 30 minutes. Methyl iodide (0.025 ml) was
added dropwise to the reaction solution, and the resulting mixture was stirred at room
temperature for 10 hours. Saturated brine was added thereto, and the resulting
mixture was extracted 3 times with ethyl acetate. Organic layers were washed with
saturated brine and dried over anhydrous sodium sulfate. After removing anhydrous
sodium sulfate by filtration, the filtrate was concentrated. The residue was purified
by column chromatography (silica gel, eluent: hexane/ethyl acetate = 6/1) to obtain 4-
(4-iodophenyl)-4-methoxyoxepane (63 mg).

[0146]
Reference Example 6
2-(4-Iodophenoxy)pyrimidine
[0147]

[0148]
Under an argon atmosphere, potassium carbonate (207 mg) was added to a
solution of 4-iodophenol (220 mg) and 2-chloropyrimidine (114 mg) in DMF (1 ml),
and the resulting mixture was stirred at 120°C for 3 hours. The reaction solution
was cooled to room temperature and saturated brine was added thereto, followed by
extracting the resulting mixture with chloroform. Organic layer was washed with
saturated brine and dried over anhydrous sodium sulfate. After removing anhydrous
sodium sulfate by filtration, the filtrate was concentrated. The residue was purified
by column chromatography (silica gel. eluent: hexane/ethyl acetate = 3/1) to obtain 2-
(4-iodophenoxy)pyrimidine (288 mg). NMR(H1? CDC13):S 6.97-7.00(2H, m),
7.06(1 H, t, J=4.6Hz), 7.73-7.75(2H, m), 8.57(2H, d, J=4.6Hz)
[0149]
Reference Example 7
2-(4-Iodophenoxy)-5-ethylpyrimidine
[0150]

[0151]
In the same manner as in Reference Example 6, 4-iodophenol (220 mg) and 5-
ethyl-2-chloropyrimidine (0.121 ml) were reacted in the presence of potassium
carbonate (415 mg) to obtain 2-(4-iodophenoxy)-5-ethylpyrimidine (256 mg).

[0152]
Reference Example 8
2-(4-Iodophenoxy)-4-methoxypyrimidine
[0153]

[0154]
Under an argon atmosphere, sodium hydride (108 mg) was added to a
solution of 4-iodophenol (220 mg) and 2-chloro-4-methoxypyrimidine (168 mg) in
anhydrous DMF (10 ml), and the resulting mixture was stirred at 125°C for 9 hours.
The reaction solution was cooled to room temperature and water was added thereto,
followed by extracting the resulting mixture with ethyl acetate. Organic layer was
washed with saturated brine and dried over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was purified by column chromatography (silica gel, eluent: hexane/ethyl
acetate = 5/1) to obtain 2-(4-iodophenoxy)-4-methoxypyrimidine (312 mg).
[0155]
Reference Example 9
2-(4-Iodophenoxy)-4,6-dimethoxypyrimidinc
[0156]

[0157]
In the same manner as in Reference Example 8, 4-iodophenol (223 mg) and 2-
chloro-4,6-dimethoxypyrimidine (192 mg) were reacted by using sodium hydride to
obtain 2-(4-iodophenoxy)-4,6-dimethoxypyrimidine (322 mg).

[0158]
Reference Example 10
2-(4-Iodophenoxy)-4,6-dimethylpyrimidine
[0159]

[0160]
(1) Under an argon atmosphere, phosphorus oxychloride (2 ml) was added to 4,6-
dimethyl-2-hydroxypyrimidine (400 mg), and the resulting mixture was stirred for 1
hour while heating the mixture to reflux. The reaction solution was cooled to room
temperature, and the cooled solution was added in small portions to water. After
adding sodium hydrogen carbonate in small portions to the resulting mixture to
neutralize it, the mixture was extracted with ethyl acetate. Organic layer was
washed twice with water and once with saturated brine, and dried over anhydrous
sodium sulfate. After removing anhydrous sodium sulfate by filtration, the filtrate
was concentrated to obtain 2-chloro-4,6-dimethylpyrirnidine (350 mg).
(2) In the same manner as in Reference Example 8, 4-iodophenol (223 mg) and 2-
chloro-4,6-dimethylpyrimidine (157 mg) were reacted by using sodium hydride to
obtain 2-(4-iodophenoxy)-4,6-dimethylpyrimidine (226 mg).
[0161]
Reference Example 11
N-(4-Iodophenyl)pyrimidin-2-amine
[0162]

[0163]
Under an argon atmosphere, 2-chloropyrimidine (16.5 g) and acetic acid (11.7

ml) were added to a solution of 4-iodoaniline (30 g) in dioxane (500 ml), and the
resulting mixture was stirred for 13 hours while heating the mixture to reflux. The
reaction solution was cooled to room temperature and saturated aqueous sodium
hydrogen carbonate solution was added thereto, followed by extracting the resulting
mixture 4 times with ethyl acetate. Organic layers were washed 3 times with
saturated brine and dried over anhydrous sodium sulfate. After removing anhydrous
sodium sulfate by filtration, the filtrate was concentrated. The residue was purified
by column chromatography (silica gel, eluent: cyclohexane/ethyl acetate = 4/1).
The product was recrystallized from dichloromethane/hexane mixed solvent to obtain
N-(4-iodophenyl)pyrimidin-2-amine (22.27 g). NMR(H', CDC13):5 6.73(1 H. t.
J=4.6Hz), 7.18(1H, brs), 7.40-7.42(2H, m), 7.59-7.61(2H, m), 8.41(2H, d, J=4.6Hz)
[0164]
Reference Example 12
N-(4-Bromophenyl)pyrimidin-2-amine
[0165]

[0166]
Under an argon atmosphere, 2-chloropyrimidine (2.68 g) and acetic acid (1.97
ml) were added to a solution of 4-bromoaniline (4.03 g) in dioxane (200 ml), and the
resulting mixture was stirred for 3 hours while heating the mixture to reflux.
Saturated aqueous sodium hydrogen carbonate solution was added in small portions
to the reaction solution to neutralize it, and the resulting mixture was concentrated.
Ethyl acetate (500 ml) was added to the residue and the resulting mixture was stirred
at room temperature for 10 minutes, followed by removing insoluble matter by
filtration. The filtrate was washed with saturated aqueous sodium hydrogen
carbonate solution and with saturated brine, and dried over anhydrous sodium sulfate.

After removing anhydrous sodium sulfate by filtration, the filtrate was concentrated.
The residue was recrystallized from ethyl acetate to obtain N-(4-
bromophenyl)pyrimidin-2-amine (3.02 g).
[0167]
Reference Example 13
N-(4-Iodophenyl)-N-methylpyrimidin-2-amine
[0168]

[0169]
Under an argon atmosphere, a solution of N-(4-iodophenyI)pyrimidin-2-
amine (1.47 g) in anhydrous DMF (10 ml) was added dropwise to a suspension of
sodium hydride (218 mg) in anhydrous DMF (8 ml), and the resulting mixture was
stirred at room temperature for 75 minutes. Methyl iodide (0.37 ml) was added
dropwise to the reaction solution and the resulting mixture was stirred at room
temperature for another 1 hour. Water was added thereto and the resulting mixture
was extracted with ethyl acetate. Organic layer was washed twice with water and
once with saturated brine, and dried over anhydrous sodium sulfate. After removing
anhydrous sodium sulfate by filtration, the filtrate was concentrated. The residue
was recrystallized from hexane to obtain N-(4-iodophenyl)-N-methylpyrimidin-2-
amine (1.38 g). NMR(H1, CDC13):8 3.49(3H, S), 6.58(1 H, t, J=4.6Hz), 7.07-
7.09(2H, m), 7.67-7.69(2H, m), 8.32(2H, d, J=4.6Hz)
[01701
Reference Example 14
N-(4-Iodophenyl)-N-ethylpyrimidin-2-arnine
[0171]


[0172]
Under an argon atmosphere, a solution of N-(4-iodophenyl)pyrimidin-2-
amine (255 mg) in anhydrous DMF (1 ml) was added dropwise to a suspension of
sodium hydride (40 mg) in anhydrous DMF (1 ml), and the resulting mixture was
stirred at room temperature for 75 minutes. Ethyl iodide (0.10 ml) was added
dropwise to the reaction solution and the resulting mixture was stirred at room
temperature overnight. Saturated brine was added thereto and the resulting mixture
was extracted with chloroform. Organic layer was washed with saturated brine and
dried over anhydrous sodium sulfate. After removing anhydrous sodium sulfate by
filtration, the filtrate was concentrated. The residue was purified by column
chromatography (silica gel, eluent: cyclohexane/chloroform = 1/10) to obtain N-(4-
iodophenyl)-N-ethylpyrimidin-2-amine (264 mg). NMR^CDCl.,)^ 1.23(3H, t.
J=6.8Hz), 4.01(211, q, J=6.8Hz), 6.57(1H, t, J=4.9Hz), 7.03-7.06(211, m), 7.70-
7.74(2H, m), 8.32(2H, d, J=4.9Hz)
[0173]
Reference Example 15
N-(4-Iodophenyl)-N-(2-methoxyethyl)pyrimidin-2-amine
[0174]

[0175]
Under an argon atmosphere, 2-bromoethylmethyl ether (0.14 ml) and sodium
hydride (48 mg) were added to a solution of N-(4-iodophenyl)pyrimidin-2-amine

(300 mg) in anhydrous DMF (1 ml), and the resulting mixture was stirred at room
temperature overnight. Saturated brine was added to the reaction solution and the
resulting mixture was extracted with chloroform, followed by drying the organic
layer over anhydrous sodium sulfate. After removing anhydrous sodium sulfate by
filtration, the filtrate was concentrated. The residue was purified by column
chromatography (silica gel, eluent: hexane/chloroform = 1/10) to obtain N-(4-
iodophenyl)-N-(2-methoxyethyl)pyrimidin-2-amine (233 mg). NMR(H', CDC1;0:8
3.32(3H, s), 3.63(2H, t, J=5.9Hz), 4.14(2H, t, J=5.9Hz), 6.59(1H, t, J=4.9Hz), 7.09-
7.13(2H, m), 7.70-7.73(2H, m), 8.32(2H, d, J=4.9Hz)
[0176]
Reference Example 16
N-(4-Iodophenyl)-N-isopropylpyrimidin-2-amine
[0177]

[0178]
Under an argon atmosphere, a solution of N-(4-iodophenyl)pyrimidin-2-
amine (8.00 g) in anhydrous DMF (50 ml) was added dropwise to a suspension of
sodium hydride (1.08 g) in anhydrous DMF (200 ml), and the resulting mixture was
stirred at room temperature for 80 minutes. To the reaction solution, 2-iodopropane
(4.03 ml) was added, and the resulting mixture was stirred at room temperature for 2
hours. Sodium hydride (1.08 g) was added to the reaction solution and thereafter 2-
iodopropane (4.03 ml) was added dropwise thereto, followed by stirring the resulting
mixture at room temperature for 6 hours. Water was added to the reaction solution
and the resulting mixture was extracted twice with ethyl acetate. Organic layers
were washed 4 times with water and once with saturated brine, and dried over

anhydrous sodium sulfate. After removing anhydrous sodium sulfate by filtration,
the filtrate was concentrated. The residue was purified by column chromatography
(silica gel, eluent: cyclohexane—»cyclohexane/ethyI acetate = 50/1). The obtained
solid was recrystallized from hexane to obtain N-(4-iodophenyl)-N-
isopropylpyrimidin-2-amine (7.06 g). NMR(H', CDC13):S 1.14(3H, s), 1.16(3H, S),
5.12-5.19(1 H, m), 6.52(1H, t, J=4.6Hz). 6.88-6.92(2H. m), 7.74-7.77(2H, m),
8.29(2H, d, J=4.6Hz)
[0179]
Reference Example 17
(4-Acetoxybenzyl)-(4-iodophenyl)pyrimidin-2-ylamine

Under an argon atmosphere, a solution of N-(4-iodophenyl)pyrimidin-2-
amine (68 mg) in anhydrous DMF (1.0 ml) was added dropwise to a solution of
sodium hydride (16 mg) in anhydrous DMF (1.0 ml), and the resulting mixture was
stirred at room temperature for 30 minutes. To the reaction solution, a solution of
4-acetoxybenzyl chloride (0.04 ml) in anhydrous DMF (0.5 ml) was added, and the
resulting mixture was stirred at room temperature overnight. Saturated brine was
added to the reaction solution and the resulting mixture was extracted with
chloroform. Organic layer was dried over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was purified by column chromatography (silica gel, eluent: hexane/ethyl
acetate = 6/1) to obtain (4-acetoxybenzyl)-(4-iodophenyl)pyrimidin-2-ylamine (67
mg). NMR(H', CDC13):8 2.28(3H, S), 5.22(2H, S), 6.63(1 H, t, J=4.6Hz), 6.98-

7.02(4H, m), 7.26(2H, d, J=8.1Hz), 7.64-7.67(2H, m), 8.34(2H, d, J=4.6Hz)
[0182]
Reference Example 18
3-(N-(4-lodophenyl)-N-(pyrimidin-2-yI)amino)propanenilrile

Under an argon atmosphere, a solution of N-(4-iodophenyl)pyrimidin-2-
amine (100 mg) in anhydrous DMF (1.0 ml) was added to a suspension of sodium
hydride (16 mg) in anhydrous DMF (1.0 ml), and the resulting mixture was stirred at
room temperature for 30 minutes. To the reaction solution, a solution of 3-
bromopropionitrile (0.04 ml) in anhydrous DMF (0.5 ml) was added, and the
resulting mixture was stirred at room temperature overnight. Saturated brine was
added to the reaction solution and the resulting mixture was extracted with
chloroform. Organic layer was dried over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was purified by column chromatography (silica gel, eluent:
hexane/chloroform = 1/10) to obtain 3-(N-(4-iodophenyl)-N-(pyrimidin-2-
yl)amino)propanenitrile (94 mg). NMR(H', CDC13):5 2.82(2H, t, J=6.8Hz),
4.24(2H, t, J=6.8Hz), 6.68(1H, t, J=4.9Hz), 7.06-7.10(2H, m), 7.74-7.78(2H, m),
8.35(2H, d, J=4.9Hz)
[0185]
Reference Example 19
N-Benzyl-N-(4-iodophenyl)pyrimidin-2-amine
[0186]


Under an argon atmosphere, N-(4-iodophenyl)pyrimidin-2-amine (75 mg) was
added to a suspension of sodium hydride (16 mg) in anhydrous DMF (3 ml), and the
resulting mixture was stirred at room temperature for 30 minutes. The reaction
solution was cooled to 0°C and benzyl bromide (0.039 ml) was added thereto,
followed by stirring the resulting mixture at room temperature for 1 hour. Saturated
brine was added to the reaction solution and the resulting mixture was extracted 3
times with ethyl acetate. Organic layers were washed 3 times with water and dried
over anhydrous sodium sulfate. After removing anhydrous sodium sulfate by
filtration, the filtrate was concentrated. The residue was purified by column
chromatography (silica gel, eluent: ethyl acetate/hexane = 1/15) to obtain N-benzyl-
N-(4-iodophenyl)pyrimidin-2-amine (67 mg).
[0188]
Reference Example 20
N-(4-Bromophenyl)-N-(4-methoxybenzyl)pyrimidin-2-amine

Under an argon atmosphere, N-(4-bromophenyl)pyrimidin-2-amine (75 mg)
was added to a suspension of sodium hydride (16 mg) in anhydrous DMF (3 ml), and
the resulting mixture was stirred at room temperature for 30 minutes. The reaction
solution was cooled to 0°C and 4-methoxybenzyl chloride (0.045 ml) was added

thereto, followed by stirring the resulting mixture at room temperature for 1 hour.
Saturated brine was added to the reaction solution and the resulting mixture was
extracted 3 times with ethyl acetate. Organic layers were washed 3 times with water
and once with saturated brine, and dried over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was purified by column chromatography (silica gel, eluent: ethyl
acetate/hexane = 1/10) to obtain N-(4-bromophenyl)-N-(4-methoxybenzyl)pyrimidin-
2-amine(107 mg).
[0191]
Reference Example 21
N-(4-Bromophenyl)-N-butylpyrimidin-2-amine

Under an argon atmosphere, N-(4-bromophcnyl)pyrimidin-2-amine (160 mg)
was added to a suspension of sodium hydride (33 mg) in anhydrous DMF (3 ml), and
the resulting mixture was stirred at room temperature for 30 minutes. The reaction
solution was cooled to 0°C and rc-butyl iodide (0.080 ml) was added thereto, followed
by stirring the resulting mixture at room temperature for 1 hour. Saturated brine
was added to the reaction solution and the resulting mixture was extracted 3 times
with ethyl acetate. Organic layers were washed 3 times with water and once with
saturated brine, and dried over anhydrous sodium sulfate. After removing
anhydrous sodium sulfate by filtration, the filtrate was concentrated. The residue
was purified by column chromatography (silica gel, eluent: ethyl acetate/hexane -
1/10) to obtain N-(4-bromophenyl)-N-butylpyrimidin-2-amine (194 mg).

[0194]
Reference Example 22
N-(4-Bromophenyl)-N-((pyridin-3-yl)methyl)pyrimidin-2-amine

Under an argon atmosphere, N-(4-bromophenyl)pyrimidin-2-amine (200 mg)
was added to a suspension of sodium hydride (48 mg) in anhydrous DMF (8 ml), and
the resulting mixture was stirred at room temperature for 30 minutes. The reaction
solution was cooled to 0°C, and 3-chloromethylpyridine hydrochloride (600 mg) and
triethylamine (1.00 ml) were added thereto, followed by stirring the resulting mixture
at room temperature for 2 hours. Saturated brine was added to the reaction solution
and the resulting mixture was extracted 3 times with ethyl acetate. Organic layers
were washed 3 times with water and once with saturated brine, and dried over
anhydrous sodium sulfate. After removing anhydrous sodium sulfate by filtration,
the filtrate was concentrated. The residue was purified by column chromatography
(silica gel, eluent: ethyl acetate/hexane = 3/1) to obtain N-(4-bromophenyl)-N-
((pyridin-3-yl)methyl)pyrimidin-2-amine (232 mg). NMR(H', CDC13):8 5.23(2H, s),
6.65(1H, I, J=4.6Hz), 7.08-7.12(2H, m). 7.20(1 H, dd, J=7.8, 4.9Hz), 7.46-7.50(2H,
m). 7.59-7.61(11-1. m), 8.35(2H, d, J=4.6Hz), 8.49(1H, dd, J=4.9, 1.5Hz), 8.54(1H. d,
J=2.2Hz)
[0197]
Reference Example 23
N-(4-Bromophenyi)-N-((miazol-4-yl)methyl)pyrimidin-2-amine
[0198]


Under an argon atmosphere, N-(4-bromophenyl)pyrimidin-2-amine (200 mg)
was added to a suspension of sodium hydride (48 mg) in anhydrous DMF (8 ml), and
the resulting mixture was stirred at room temperature for 30 minutes. The reaction
solution was cooled to 0°C, and thiazoylmethyl chloride hydrochloride (816 mg) and
triethylamine (1.00 ml) were added thereto, followed by stirring the resulting mixture
at room temperature for 2 hours. Saturated brine was added to the reaction solution
and the resulting mixture was extracted 3 times with ethyl acetate. Organic layers
were washed 3 times with water and once with saturated brine, and dried over
anhydrous sodium sulfate. After removing anhydrous sodium sulfate by filtration,
the filtrate was concentrated. The residue was purified by column chromatography
(silica gel, eluent: ethyl acetate/hexane = 1/3) to obtain N-(4-bromophenyl)-N-
((thiazol-4-yl)methyl)pyrimidin-2-amine (190 mg).
[0200]
Reference Example 24
N-(4-Iodophenyl)-4-methoxy-N-methylpyrimidin-2-amine

(1) Under an argon atmosphere, sodium hydride (100 mg) was added to a
solution of 4-iodoaniline (220 mg) and 2-chloro-4-methoxypyrimidine (145 mg) in
anhydrous DMF (10 ml), and the resulting mixture was stirred at 125°C for 21 hours.
The reaction solution was cooled to room temperature and water was added thereto.

followed by extracting the resulting mixture with ethyl acetate. Organic layer was
washed twice with water and once with saturated brine, and dried over anhydrous
sodium sulfate. After removing anhydrous sodium sulfate by filtration, the filtrate
was concentrated. The residue was purified by column chromatography (silica gel,
eluent: hexane/ethyl acetate = 10/1) to obtain N-(4-iodopherryl)-4-methoxypyrirnidin-
2-amine (46 mg).
(2) Under an argon atmosphere, N-(4-iodophenyl)-4-methoxypyrimidin-2-amine
(46 mg) was added to a suspension of sodium hydride (11 mg) in anhydrous DMF (5
ml), and the resulting mixture was stirred at room temperature for 5 minutes.
Methyl iodide (0.0096 ml) was added dropwise to the reaction solution, and the
resulting mixture was stirred at room temperature for 23 hours. Water was added
thereto and the resulting mixture was extracted with ethyl acetate. Organic layer
was washed twice with water and once with saturated brine, and dried over
anhydrous sodium sulfate. After removing anhydrous sodium sulfate by filtration,
the filtrate was concentrated. The residue was purified by column chromatography
(silica gel, eluent: hexane/ethyl acetate = 3/1) to obtain N-(4-iodophenyl)-4-methoxy-
N-methylpyrimidin-2-amine (46 mg).
[0203]
Reference Example 25
N-(4-Iodophenyl)-4,6-dimethoxy-N-methylpyrimidin-2-amine

(1) In the same manner as in Reference Example 24-( 1), 4-iodoaniline (220 mg)
and 2-chloro-4,6-dimethoxypyrimidine (200 mg) were reacted in the presence of

sodium hydride to obtain N-(4-iodophenyl)-4,6-dimethoxypyrimidin-2-amine (141
mg).
(2) In the same manner as in Reference Example 24-(2), sodium hydride (16 mg).
N-(4-iodophenyl)-4,6-dimethoxypyrimidin-2-amine (140 mg) and methyl iodide
(0.024 ml) were reacted to obtain N-(4-iodophenyl)-4,6-dimethoxy-N-
methylpyrimidin-2-amine (118 mg).
[0206]
Reference Example 26
N-(4-Bromophenyl)-N-phenylpyrimidin-2-amine

Under an argon atmosphere, a suspension of N-(4-bromophenyl)pyrimidin-2-
amine (250 mg). copper iodide (1.9 mg), trans-l,2-cyclohexanediamine (0.015 ml)
and sodium-terr-butoxide (144 mg) in dioxane (1 ml) was stirred at 110°C for 22
hours in a pressure-resistant test tube. The reaction solution was filtered and the
filtrate was concentrated. The residue was purified by thin layer chromatography
(silica gel, mobile phase: chloroform/ethyl acetate = 5/1) to obtain N-(4-
bromophenyl)-N-phenylpyrimidin-2-amine (56 mg).
[0209]
Reference Example 27
Trifluoromethanesulfonic acid 4-[pyrimidin-2-yl-(tetrahydropyran-4-yl)aminoJphenyl
ester
[0210]


(1) Acetic acid (0.856 ml) was added to a solution of p-anisidine (1.23 g) and 2-
chloropyrimidine (1.72 g) in dioxane (20 ml), and the resulting mixture was stirred
overnight while heating the mixture to reflux. After cooling the reaction solution to
room temperature., 1 N aqueous sodium hydroxide solution was added thereto and the
resulting mixture was extracted 3 times with chloroform, followed by drying the
organic layers over anhydrous sodium sulfate. After removing anhydrous sodium
sulfate by filtration, the filtrate was concentrated. The residue was reprecipitated
from chloroform/hexane mixed solvent to obtain N-(4-methoxyphenyl)pyrimidin-2-
amine (1.39 g).
(2) Under an argon atmosphere, p-toluenesulfonyl chloride (3.82 g) was added to
a solution of tetrahydro-2H-pyran-4-ol (2.00 g) in pyridine (50 ml) at 0°C, and the
resulting mixture was stirred at room temperature overnight. After adding 1 N
hydrochloric acid thereto, the resulting mixture was extracted 3 times with ethyl
acetate. Organic layers were washed 3 times with water and dried over anhydrous
sodium sulfate. After removing anhydrous sodium sulfate by filtration, the filtrate
was concentrated. The residue was purified by column chromatography (silica gel.
eluent: ethyl acetate/hexane = 1/6) to obtain tetrahydro-2H-pyran-4-yl-4-
methylbenzenesulfonate (2.92 g).
(3) Under an argon atmosphere, N-4-(methoxyphenyl)pyrimidin-2-amine (402
mg) was added to a suspension of sodium hydride (120 mg) in anhydrous DMF (15
ml), and the resulting mixture was stirred at room temperature for 30 minutes. A
solution of tetrahydro-2H-pyran-4-yl-4-methylbenzenesulfonate (2.92 g) in
anhydrous DMF (10 ml) was added to the reaction solution, and the resulting mixture

was stirred at 50°C for 24 hours. Sodium hydride (120 mg) was further added to the
reaction solution, and the resulting mixture was stirred at 50°C for 24 hours. The
solution was cooled to room temperature, and saturated brine was added thereto,
followed by extracting the resulting mixture 3 times with ethyl acetate. Organic
layers were washed 3 times with water and dried over anhydrous sodium sulfate.
After removing anhydrous sodium sulfate by filtration, the filtrate was concentrated.
The residue was purified by column chromatography (silica gel, eluent: ethyl
acetate/hexane = 1/3) to obtain N-(tetrahydro-2H-pyran-4-yl)-N-(4-
methoxyphenyl)pyrimidin-2-amine (342 mg).
(4) A mixture of N-(tetrahydro-2H-pyran-4-yl)-N-(4-methoxyphenyl)pyrimidin-
2-amine (342 mg) and pyridine hydrochloride (2.00 g) was heated to 170°C and
stirred for 1 hour. Pyridine hydrochloride (1.00 g) was further added to the reaction
solution, and the resulting mixture was stirred at 170°C for 1 hour. The reaction
solution was allowed to cool to room temperature and saturated brine was added
thereto, followed by extracting the resulting mixture 3 times with ethyl acetate.
Organic layers were washed 3 times with water and dried over anhydrous sodium
sulfate. After removing anhydrous sodium sulfate by filtration, the filtrate was
concentrated. To a suspension of the obtained residue and potassium carbonate
(498 mg) in THF (15 ml), N-phenylbis(trifluoromelhanesulfonimide) (514 mg) was
added, and the resulting mixture was stirred at room temperature for 5 hours.
Saturated brine was added to the reaction solution, and the resulting mixture was
extracted 3 times with ethyl acetate, followed by drying the organic layers over
anhydrous sodium sulfate. After removing anhydrous sodium sulfate by filtration,
the filtrate was concentrated. The residue was purified by column chromatography
(silica gel, eluent: ethyl acetate/hexane = 1/5) to obtain trifluoromethanesulfonic acid
4-lpyrimidin-2-yl-(tetrahydropyran-4-yl)amino]phenyl ester (228 mg). NMR(H].
CDC13):8 1.52-1.61 (2H, m), 1.83-1.87(2H, m), 3.57(2H, dt, .1=12.0, 1.6Hz). 4.00(2H,

dd, J=l 1.6, 4.8Hz), 4.97-5.03(lH, m), 6.59(1H, t, J=4.8Hz), 7.19-7.23(2H, m), 7.34-
7.37(2H, m), 8.29(2H, d, J=4.8Hz)
[0212]
Reference Example 28
N-(4-lodophenyl)-N-(3-methyl-2-butenyl)pyrimidin-2-amine

In the same manner as in Reference Example 13. N-(4-iodophenyl)pyrimidin-
2-amine (150 mg) and l-bromo-3-methyl-2-butene (90 mg) were reacted in the
presence of sodium hydride to obtain N-(4-iodophenyl)-N-(3-methyl-2-
butenyl)pyrimidin-2-amine (175 mg).
[0215]
Reference Example 29
N-(Cyclopropylmethyl)-N-(4-iodophenyl)pyrimidin-2-amine

In the same manner as in Reference Example 13. N-(4-iodophenyi)pyrimidin-
2-amine (150 mg) and cyclopropylmethyl bromide (82 mg) were reacted in the
presence of sodium hydride to obtain N-(cyclopropylmethyl)-N-(4-
iodophenyl)pyrimidin-2-amine (162 mg).
[0218]

Reference Example 30
N-(4-Iodophenyl)-N-isobutylpyrimidin-2-amine

In the same manner as in Reference Example 13, N-(4-iodophenyl)pyrimidin-
2-amine (150 mg) and isobutyl bromide (83 mg) were reacted in the presence of
sodium hydride to obtain N-(4-iodophenyl)-N-isobutylpyrimidin-2-amine (167 mg).
[0221]
Reference Example 31
N-(4-Iodophenyl)-N-propylpyrimidin-2-amine
In the same manner as in Reference Example 13, N-(4-iodophenyl)pyrimidin-
2-amine (300 mg) and propyl bromide (0.11 ml) were reacted in the presence of
sodium hydride to obtain N-(4-iodophenyl)-N-propylpyrimidin-2-amine (348 mg).
[0224]
Reference Example 32
N-(4-Iodophenyl)-N-((pyridin-4-yl)methyl)pyrimidin-2-amine
[0225]


In the same manner as in Reference Example 22, N-(4-iodophenyl)pyrimidin-
2-amine (300 mg) and 4-bromomethylpyridine hydrobromide (307 mg) were reacted
in the presence of sodium hydride to obtain N-(4-iodophenyl)-N-((pyridin-4-
yl)methyl)pyrimidin-2-amine (93 mg).
[0227]
Reference Example 33
N-(4-Iodophenyl)-N-((pyridin-2-yl)methyl)pyrimidin-2-amine

In the same manner as in Reference Example 22, N-(4-iodophenyl)pyrimidin-
2-amine (300 mg) and 2-bromomethylpyridine hydrobromide (307 mg) were reacted
in the presence of sodium hydride to obtain N-(4-iodophenyl)-N-((pyridin-2-
yl)methyl)pyrimidin-2-amine (282 mg).
[0230]
Reference Example 34
2-Amino-5-[4-(methyl-pyrimidin-2-ylamino)phenyl|pent-4-enoic acid methyl ester
[0231]


(1) Allylglycine (1.82 g) was dissolved in IN aqueous sodium hydroxide solution
(60 ml), and di-tert-butyl dicarbonate (4.15 g) was added thereto, followed by stirring
the resulting mixture at room temperature for 11 hours. Di-tert-butyl dicarbonate
(4.15 g) was further added to the reaction solution and the resulting mixture was
stirred at room temperature overnight. After adding water to the reaction solution
and washing it with ether, the solution was acidified with 3N hydrochloric acid.
The solution was extracted twice with ethyl acetate. Organic layers were washed
with saturated brine and dried over anhydrous sodium sulfate. After removing
anhydrous sodium sulfate by filtration, the filtrate was concentrated. The residue
was recrystallized from hexane/ethyl acetate mixed solvent to obtain N-Boc-
allylglycine(3.09g).
(2) Trimethylsilyldiazomethane (2N, in hexane) (12 ml) was added dropwise to a
solution of N-Boc-allylglycine (3.09 g) in dichloromethanetertethanol (2/1) mixture
(30 ml), and the resulting mixture was stirred at room temperature for 3 hours.
After concentrating the reaction solution, the residue was purified by column
chromatography (silica gel, eluent: cyclohexane/ethyl acetate = 20/1 → 10/1) to obtain
N-Boc-allylglycine methyl ester (3.11 g). NMR(H1, CDC13):8 2.43-2.55(2H, m),
3.72(3H, s), 4.34-4.39(lH. brm), 5.02(1 H, brs), 5.09-5.13(2H, m), 5.62-5.72(111. m)
(3) Under an argon atmosphere, palladium acetate (54 mg) and tris(2-
mcthylphenyl)phosphine (70 mg) were added to a suspension of N-Boc-allylglycine
methyl ester (1.03 g), N-(4-iodophenyl)-N-methylpyrimidin-2-amine (1.40 g) and
potassium carbonate (933 mg) in DMF (15 ml), and the resulting mixture was stirred

at 80°C overnight. The reaction solution was allowed to cool to room temperature
and ethyl acetate was added thereto. The solution was washed 3 times with water
and once with saturated brine, and dried over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was purified by column chromatography (silica gel, eluent: cyclohexane/ethyl
acetate = 20/1→4/l). The obtained solid was recrystallized from hexane/ethyl
acetate mixed solvent to obtain 2-tert-butoxycarbonylamino-5-[4-(methyl-pyrimidin-
2-ylamino)phenyl]pent-4-enoic acid methyl ester (976 mg).
(4) Trifluoroacetic acid (2 ml) was added to a solution of 2-tert-
butoxycarbonylamino-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid
methyl ester (976 mg) in dichloromethane (20 ml), and the resulting mixture was
stirred at room temperature for 4.5 hours. Water was added to the reaction solution,
and sodium hydrogen carbonate was added thereto in small portions to neutralize it.
followed by extracting the resulting solution twice with chloroform. Organic layers
were washed with saturated brine and dried over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated to
dryness to obtain 2-amino-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic
acid methyl ester (599 mg). NMR(H1, CDC13):6 2.58-2.64(2H, brm), 3.48-3.58(1 H.
brm), 3.52(3H, s), 3.75(3H, s), 6.07-6.14(lH, m), 6.49(1H, d, J=5.6Hz), 6.57(1H. t.
J=4.6Hz), 7.26(2H, d, J=8.1Hz), 7.38(2H, d, J=8.3Hz), 8.34(2H, d, J=4.6Hz)
[0233]


[0235]
(1) Allylglycine (10.11 g) was dissolved in a mixed solvent of 2N aqueous
sodium hydroxide solution (176 ml) and dioxane (175 ml), and 2,6-diehlorobenzoyl
chloride (15.10 ml) was added dropwise thereto, followed by stirring the resulting
mixture at room temperature for 10 hours. After concentrating the reaction solution
to remove dioxane, water (150 ml) was added thereto and the resulting solution was
washed with ether. Aqueous layer was acidified by adding 3N hydrochloric acid in
small portions thereto, and extracted 3 times with ethyl acetate. Organic layers were
washed with saturated brine and dried over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was recrystallized from hexane/ethyl acetate mixed solvent to obtain 2-(2,6-
dichlorobenzamido)pent-4-enoic acid (23.48 g).
(2) Under an argon atmosphere, a solution of 2-(2,6- dichlorobenzamido)pent-4-
enoic acid (23.48 g) in methanol (250 ml) was cooled to 0°C. Thionyl chloride
(10.70 ml) was added dropwise to the solution while keeping the reaction
temperature not higher than 10°C, and thereafter the resulting mixture was stirred at
room temperature for 5.5 hours. Water was added to the reaction solution, and
sodium hydrogen carbonate was added thereto in small portions to neutralize it.
After concentrating the reaction solution to remove methanol, aqueous layer was
extracted twice with ethyl acetate. Organic layers were washed 3 times with water
and once with saturated brine, and dried over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was recrystallized from hexane/ethyl acetate mixed solvent to obtain 2-(2.6-
dichlorobenzamido)pent-4-enoic acid methyl ester (21.70 g). NMR(II!, CDC1.,):8
2.55-2.61(111, m), 2.68-2.75(lH, m), 3.72(3H, s), 4.88(111, td, .1=7.8, 5.4Hz). 5.06-
5.13(2H, m), 5.64-5.74(lH, m), 6.33(1H, brd, J=7.1Hz), 7.17-7.26(3H, m)
[0236]

Reference Example 36
2-(2,6-Difluorobenzamido)pent-4-enoic acid methyl ester

(1) In the same manner as in Reference Example 35-(l), allylglycine (285 mg)
was reacted with 2,6-difluorobenzoyl chloride (0.374 ml) to obtain 2-(2,6-
difluorobenzamido)pent-4-enoic acid (471 mg).
(2) In the same manner as in Reference Example 35-(2), thionyl chloride (0.242
ml) was reacted with a solution of 2-(2,6-difluorobenzamido)pent-4-enoic acid (471
mg) in methanol (20 ml) to obtain 2-(2,6-difluorobenzamido)pent-4-enoic acid
methyl ester (430 mg).
[0239J
Reference Example 37
2-(2,6-Dichlorobenzamido)pent-4-ynoic acid methyl ester

(1) Propargylglycine (5.00 g) was dissolved in a mixed solvent of 2N aqueous
sodium hydroxide solution (100 ml) and dioxane (100 ml), and 2,6-dichlorobenzoyl
chloride (9.45 ml) was added dropwise thereto at 0°C, followed by stirring the
resulting mixture at room temperature overnight. Water was added to the reaction
solution, and the solution was washed with ether. Aqueous layer was acidified by

adding 3N hydrochloric acid in small portions thereto and extracted with ethyl
acetate, and organic layer was dried over anhydrous sodium sulfate. After removing
anhydrous sodium sulfate by filtration, the filtrate was concentrated to dryness to
obtain 2-(2,6-dichlorobenzamido)pent-4-ynoic acid.
(2) Under an argon atmosphere, a solution of thionyl chloride (5.25 ml) in
absolute methanol (200 ml) was cooled to 0°C. To the solution, a solution of 2-
(2,6-dichlorobenzamido)pent-4-ynoic acid obtained in (1) in methanol (50 ml) was
added dropwise, and the resulting mixture was stirred at room temperature overnight.
Water was added to the reaction solution, and the solution was extracted with ethyl
acetate. Organic layer was washed with saturated aqueous sodium hydrogen
carbonate solution and dried over anhydrous sodium sulfate. After removing
anhydrous sodium sulfate by filtration, the filtrate was concentrated. A mixed
solvent of ethyl acetate/ether (1/1) (300 ml) was added to the residue, and insoluble
matter was removed by filtration. The filtrate was concentrated and the residue was
recrystallized twice from hexane/ethyl acetate mixed solvent to obtain 2-(2,6-
dichlorobenzamido)pent-4-ynoic acid methyl ester (9.61 g).
[0242]
Reference Example 38
(S)-2-(2,6-Dichlorobenzamido)pent-4-enoic acid methyl ester

(1) In a mixed solvent of 2N aqueous sodium hydroxide solution (27.4 ml) and
THF (55 ml), L-allylglycine (5.22 g) was dissolved, and 2,6-dichlorobenzoyl chloride
(7.79 ml) was added dropwise thereto, followed by stirring the resulting mixture at

room temperature for 1 hour. After concentrating the reaction solution to remove
methanol, water (100 ml) was added thereto and the resulting mixture was washed
with ether. Aqueous layer was acidified by adding 3N hydrochloric acid in small
portions thereto, and extracted 3 times with ethyl acetate. Organic layers were
washed with saturated brine and dried over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was recrystallized from hexane/ethyl acetate mixed solvent to obtain (S)-2-
(2,6-dichlorobenzamido)pent-4-enoic acid (11.65 g). NMR(H'. CDC13):5 2.66-
2.73(1H, m), 2.80-2.86(lH, m), 4.99(1H, td, J=7.6, 5.6Hz)., 5.18-5.25(2H, m). 5.76-
5.86(1H, m), 6.39(1H, brd, J=7.6Hz), 7.25-7.34(3H, m)
(2) Under an argon atmosphere, a solution of (S)-2-(2,6-dichlorobenzamido)pent-
4-enoic acid (11.46 g) in methanol (120 ml) was cooled to 0°C. Thionyl chloride
(5.22 ml) was added dropwise to the solution while keeping the reaction temperature
not higher than 10°C, and thereafter the resulting mixture was stirred at room
temperature for 3 hours. Water was added to the reaction solution, and sodium
hydrogen carbonate was added thereto in small portions lo neutralize it. After
concentrating the reaction solution to remove methanol, aqueous layer was extracted
twice with ethyl acetate. Organic layers were washed 3 times with water and once
with saturated brine, and dried over anhydrous sodium sulfate. After removing
anhydrous sodium sulfate by filtration, the filtrate was concentrated. The residue
was recrystallized from hexane/ethyl acetate mixed solvent to obtain (S)-2-(2,6-
dichlorobenzamido)pent-4-enoic acid methyl ester (11.68 g). NMR(l-l', CDC13):8
2.62-2.69(lH, m), 2.76-2.83(1H, m), 3.79(3H, s), 4.96(1H, td, .1=7.8. 5.4Hz), 5.14-
5.21 (2H, m), 5.72-5.82(lH, m), 6.42(1 H, brd, J=6.8Hz). 7.25-7.34(3H, m)
Example 1
[0245]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-hydroxytetrahydropyran-4-yl)-phenyl]pent-4-


Under an argon atmosphere, palladium acetate (4.7 mg) and tris(2-
methylphenyl)phosphine (6.1 mg) were added to a suspension of 2-(2,6-
dichlorobenzamido)pent-4-enoic acid methyl ester (60.4 mg), tetrahydro-4-(4-
iodophenyl)-2H-pyran-4-ol (60.8 mg) and potassium carbonate (41.5 mg) in DMF (4
ml), and the resulting mixture was stirred at 80°C for 6 hours. After cooling the
reaction solution to room temperature, ethyl acetate was added thereto and the
solution was washed 3 times with water and once with saturated brine, followed by
drying the organic layer over anhydrous sodium sulfate. After removing anhydrous
sodium sulfate by filtration, the filtrate was concentrated. The residue was purified
by column chromatography (silica gel, eluent: cyclohexane/chloroform = 2/1→ 1/4)
and thereafter by thin layer chromatography (silica gel, developing solvent:
cyclohexane/ethyl acetate = 1/1) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-
hydroxytetrahydropyran-4-yl)-phenyl]pent-4-enoic acid methyl ester (56 mg).
Example 2
[0248]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-hydroxytetrahydropyran-4-yl)-phenyl]pent-4-
enoic acid
[0249]


To a solution of (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-
hydroxytetrahydropyran-4-yl)-phenyl]pent-4-enoic acid methyl ester (56 mg) in THF
(1.76 ml), 0.1N aqueous sodium hydroxide solution (1.76 ml) was added, and the
resulting mixture was stirred at room temperature for 2 hours. After washing the
reaction solution with ether, aqueous layer was acidified by adding 1N hydrochloric
acid thereto and extracted twice with ethyl acetate. Organic layers were washed
with saturated brine and dried over anhydrous sodium sulfate. After removing
anhydrous sodium sulfate by filtration, the filtrate was concentrated to dryness to
obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-hydroxytetrahydropyran-4-yl)-
phenyl]pent-4-enoic acid (48 mg).
Example 3
[0251]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-methoxytetrahydropyran-4-yl)-phenyl]pent-
4-enoic acid methyl ester


In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (2.11 g) was reacted with tetrahydro-4-(4-iodophenyl)-4-methoxy-
2H-pyran (2.22 g) in the presence of potassium carbonate (1.45 g), palladium acetate
(81.7 mg) and tris(2-methylphenyl)phosphine (106.2 mg) to obtain (E)-2-(2,6-
dichlorobenzamido)-5-[4-(4-methoxytetrahydropyran-4-yl)-phenyl]pent-4-enoic acid
methyl ester (2.74 g). Column chromatography (silica gel, eluent:
cyclohexane/chloroform = 2/1→cyclohexane/ethyl acetate =: 10/1→
cyclohexane/ethyl acetate = 4/1) was used for purification.
Example 4
[0254J
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-methoxytetrahydropyran-4-yl)-phenyl]pent-
4-enoic acid

In the same manner as in Example 2, 0.1N aqueous sodium hydroxide
solution (63.1 ml) was added to a solution of (E)-2-(2,6-dichlorobenzamido)-5-f4-(4-
methoxytetrahydropyran-4-yl)-phenyl]pent-4-enoic acid methyl ester (2.70 g) in THF
(63.1 ml) to hydrolyze it to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-
methoxytetrahydropyran-4-yl)-phenyl]pent-4-enoic acid (2.50 g).
Example 5
[0257]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-ethoxytetrahydropyran-4-yl)-phenyl]pent-4-

enoic acid methyl ester

In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (60.4 mg) was reacted with tetrahydro-4-(4-iodophenyl)-4-ethoxy-
2H-pyran (66.4 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-
ethoxytetrahydropyran-4-yl)-phenyl]pent-4-enoic acid methyl ester (68.9 mg).
Column chromatography (silica gel, eluent: cyclohexane/chloroform = 2/1—>
cyclohexane/ethyl acetate = 4/1) was used for purification.
Example 6
[0260]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-ethoxytetrahydropyran-4-yl)-phenyl]pcnt-4-
enoic acid

In the same manner as in Example 2. (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-
cthoxytetrahydropyran-4-yl)-phenyl]pent-4-enoic acid methyl ester (68.9 mg) was

hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-ethoxytetrahydropyran-
4-yl)-phenyl]pent-4-enoic acid (59.8 mg).
Example 7
[0263]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(tetrahydro-4-(methoxymethyl)-2H-pyran-4-
yl)phenyl]pent-4-enoic acid methyl ester

In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (78 mg) was reacted with tetrahydro-4-(4-bromophenyl)-4-
(methoxymethyl)-2H-pyran (88 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-
(tetrahydro-4-(methoxymethyl)-2H-pyran-4-yl)phenyl]pent-4-enoic acid methyl ester
(97 mg). Column chromatography (silica gel, eluent: hexane/ethyl acetate = 2/1)
was used for purification.
Example 8
[0266]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(tetrahydro-4-(methox)'methyl)-2H-pyran-4-
yl)phenyl]pent-4-enoic acid sodium salt
[0267]


To a solution of (E)-2-(2,6-dichlorobenzamido)-5-[4-(tetrahydro-4-
(methoxymethyl)-2H-pyran-4-yl)phenyl]pent-4-enoic acid methyl ester (86 mg) in
THF (1.7 ml), 0.1N aqueous sodium hydroxide solution (1.7 ml) was added, and the
resulting mixture was stirred at room temperature for 2 hours. After washing the
reaction solution with ether, aqueous layer was concentrated to obtain (E)-2-(2,6-
dichlorobenzamido)-5-[4-(tetrahydro-4-(methoxymethyl)-2H-pyran-4-
yl)phenyl]pent-4-enoic acid sodium salt (66 mg).
Example 9
[0269]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-methoxyoxepan-4-yl)phenyljpent-4-enoic
acid methyl ester

In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (46 mg) was reacted with 4-(4-iodophenyl)-4-methoxyoxepane (60
mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-methoxyoxepan-4-

yl)phenyl]pent-4-enoic acid methyl ester (66 mg). Column chromatography (silica
gel, eluent: hexane/ethyl acetate = 2/1) was used for purification.
Example 10
[0272]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-methoxyoxepan-4-yl)phenyl]pent-4-enoic
acid sodium salt

In the same manner as in Example 8, (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-
methoxyoxepan-4-yl)phenyl]pent-4-enoic acid methyl ester (57 mg) was hydrolyzed
to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-methoxyoxepan-4-yl)phenyl|pent-4-
enoic acid sodium salt (18 mg).
Example 11
[0275]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyrimidin-2-yloxy)phenyl]pent-4-enoic acid
methyl ester


In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (54.4 mg) was reacted with 2-(4-iodophenoxy)pyrimidine (59.2 mg)
to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(pyrimidin-2-yloxy)phenyl]pent-4-
enoic acid methyl ester (53.0 mg). Thin layer chromatography (silica gel,
developing solvent: cyclohexane/ethyl acetate = 10/1) was used for purification.
Example 12
[0278]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyrimidin-2-yloxy)phenyl]pent-4-enoic acid

In the same manner as in Example 2, (E)-2-(2,6-dichlorobenzamido)-5-|4-
(pyrimidin-2-yloxy)phenyl]pent-4-enoic acid methyl ester (97.2 mg) was hydrolyzed
to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(pyrimidin-2-yloxy)phenyl]penl-4-
enoic acid (77.5 mg).
Example 13
[0281]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(5-ethylpyrimidin-2-yloxy)phenyl]pent-4-enoic
acid methyl ester


[0283]
In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (50.0 mg) was reacted with 2-(4-iodophenoxy)-5-ethylpyrirnidine
(54.1 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(5-ethylpyrimidin-2-
yloxy)phenyl]pent-4-enoic acid methyl ester (62.2 mg). Column chromatography
(chloroform) and thin layer chromatography (cyclohexane/ethyl acetate =1/2) were
used for purification.
Example 14
[0284]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(5-ethylpyrimidin-2-yloxy)phenyl]pent-4-enoic
acid sodium salt

In the same manner as in Example 8, (E)-2-(2,6-dichlorobenzamido)-5-[4-(5-
ethylpyrimidin-2-yloxy)phenyl]pent-4-enoic acid methyl ester (62.2 mg) was
hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(5-ethylpyrimidin-2-
yloxy)phenyl]pent-4-enoic acid sodium salt (45.1 mg).
Example 15
[0287]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-methoxypyrimidin-2-yloxy)phenyl]pcnt-4-
enoic acid methyl ester
[0288]


In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (60 mg) was reacted with 2-(4-iodophenoxy)-4-methoxypyrimidine
(78 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-methoxypyrimidin-2-
yloxy)phenyl]pent-4-enoic acid methyl ester (32 mg). Column chromatography
(silica gel, eluent: hexane/ethyl acetate = 2/1—^cyclohexane/chloroform = 1/1—>1/10
—»0/l) was used for purification.
Example 16
[0290]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-methoxypyrimidin-2-yloxy)phenyl]pent-4-
enoic acid

In the same manner as in Example 2. (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-
methoxypyrimidin-2-yloxy)phenyl]pent-4-enoic acid methyl ester (30 mg) was
hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(4-methoxypyrimidin-2-
yloxy)phenyl]pent-4-enoic acid (28 mg).
Example 17
[0293]

(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4,6-dimethoxypyrimidin-2-yloxy)phenyl]pent-
4-enoic acid methyl ester

In the same manner as in Example 1. 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (30 mg) was reacted with 2-(4-iodophenoxy)-4,6-
dimethoxypyrimidine (40 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(4,6-
dimethoxypyrimidin-2-yloxy)phenyl]pent-4-enoic acid methyl ester (30 mg).
Column chromatography (silica gel, eluent: hexane/ethyl acetate = 2/1) was used for
purification.
Example 18
[0296]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4,6-dimethoxypyrimidin-2-yloxy)phenyl]pent-
4-enoic acid

In the same manner as in Example 2, (E)-2-(2,6-dichlorobenzamido)-5-[4-
(4,6-dimethoxypyrimidin-2-yloxy)phenyl]pent-4-enoic acid methyl ester (29 mg) was
hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(4,6-dimethoxypyrimidin-

2-yloxy)phenyl]pent-4-enoic acid (11 mg).
Example 19
[0299]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(4,6-dimethylpyrimidin-2-yloxy)phenyl]penl-4-
enoic acid methyl ester

In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (47 mg) was reacted with 2-(4-iodophenoxy)-4,6-
dimethylpyrimidine (56 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-|4-(4,6-
dimethylpyrimidin-2-yloxy)phenyl]pent-4-enoic acid methyl ester (44 mg). Column
chromatography (silica gel, eluent: hexane/ethyl acetate = 2/1) was used for
purification.
Example 20
[0302]
(E)-2-(2.6-Dichlorobenzamido)-5-[4-(4,6-dimethylpyrimidin-2-yloxy)phenyl]pent-4-
enoic acid


In the same manner as in Example 2, (E)-2-(2,6-dichlorobenzamido)-5-[4-
(4,6-dimethylpyrimidin-2-yloxy)phenyl]pent-4-enoic acid methyl ester (44 mg) was
hydrolyzed to obtain (E)-2-(2,6-diehlorobenzamido)-5-[4-(4,6-dimethylpyrimidin-2-
yloxy)phenyl]pent-4-enoic acid (36 mg).
Example 21
[0305]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid methyl ester

Under an argon atmosphere, palladium acetate (170.8 mg) and tris(2-
methylphenyl)phosphine (222.2 mg) were added to a suspension of 2-(2,6-
dichlorobenzamido)pent-4-enoic acid methyl ester (2.20 g), N-(4-iodophenyl)-N-
methylpyrimidin-2-amine (2.27 g) and potassium carbonate (1.51 g) in DMF (20 ml),
and the resulting mixture was stirred at 80°C for 3 hours. After cooling the reaction
solution to room temperature, ethyl acetate was added to the reaction solution, and
the resulting mixture was washed twice with water and once with saturated brine,
followed by drying the organic layer over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was purified by column chromatography (silica gel, eluent:
cyclohexane/chloroform = 1/1—'•chloroform). The obtained crudely purified
product was purified again by column chromatography (silica gel, eluent:

cyclohexane/ethyl acetate = 4/1→2/l) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-
(methyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (2.38 g).
Example 22
[0308]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid

To a solution of (E)-2-(2,6-dichlorobenzamido)-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester (2.38 g) in THF (20 ml), 0.5N
aqueous sodium hydroxide solution (14.7 ml) was added, and the resulting mixture
was stirred at room temperature for 1 hour. Water (120 ml) was added to the
reaction solution and the resulting mixture was washed with ether. Aqueous layer
was acidified by adding IN hydrochloric acid thereto and extracted twice with ethyl
acetate. Organic layers were washed with saturated brine and dried over anhydrous
sodium sulfate. After removing anhydrous sodium sulfate by filtration, the filtrate
was concentrated to dryness to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(methyl-
pyrimidin-2-ylamino)phenyl]pent-4-enoic acid (2.06 g).
Example 23
[0311]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(ethyl-pyrimidin-2-ylamino)phenyl]pcnt-4-
enoic acid methyl ester


In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (214 mg) was reacted with N-(4-iodophenyl)-N-ethylpyrimidin-2-
amine (254 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(ethyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester (258 mg). Column chromatography
(silica gel, eluent: hexane/ethyl acetate = 4/1-^hexane/ethyl acetate = 2/1—>
hexane/ethyl acetate = 1/2) was used for purification.
Example 24
[0314]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(ethyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid

In the same manner as in Example 2, (E)-2-(2,6-dichlorobenzamido)-5-|4-
(ethyl-pyrimidin-2--ylarnino)phenyl]pent-4-enoic acid methyl ester (258 mg) was
hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(ethyl-pyrimidin-2-

ylamino)phenyl]pent-4-enoic acid (206 mg).
Example 25
[0317]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-((2-methoxyethyl)-pyrimidin-2-
ylamino)phenyl]pcnt-4-enoic acid methyl ester

In the same, manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (89 mg) was reacted with N-(4-iodopheny])-N-(2-
methoxyethyl)pyrimidin-2-arnine (116 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-
5-[4-((2-methoxyethyl)-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester
(120 mg). Column chromatography (silica gel, eluent: cyclohexane/chloroform =
2/1—>cyclohexane/chloroform = 1/1—>cyclohexane/chloroform = 1/2) was used for
purification.
Example 26
[0320]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-((2-methoxyethyl)-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid sodium salt
[0321]


In the same manner as in Example 8, (E)-2-(2,6-dichlorobenzamido)-5-[4-((2-
methoxyethyl)-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (117 mg)
was hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-((2-methoxyethyl)-
pyrimidin-2-ylamino)phenyl]pent-4-enoic acid sodium salt (96 mg).
Example 27
[0323]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(isopropyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid methyl ester

Under an argon atmosphere, palladium acetate (316.7 mg) and tris(2-
methylphenyl)phosphine (411.8 mg) were added to a suspension of 2-(2,6-
dichlorobenzamido)pcnt-4-enoic acid methyl ester (4.09 g), N-(4-iodophenyl)-N-
isopropylpyrimidin-2-amine (4.59 g) and potassium carbonate (2.80 g) in DMF (50
ml), and the resulting mixture was stirred at 80°C for 6 hours. Alter cooling the

reaction solution to room temperature, ethyl acetate was added thereto and the
resulting mixture was washed twice with water and once with saturated brine,
followed by drying the organic layer over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was purified by column chromatography (silica gel, eluent:
cyclohexane/chloroform = 2/1→ 1/2). The obtained crudely purified product was
purified again by column chromatography (silica gel, eluent: cyclohexane/ethyl
acetate = 10/1→4/l) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(isopropyi-
pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (4.65 g).
Example 28
[0326]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(isopropyl-pyrimidin-2-ylamino)phenylJpent-4-
enoic acid

To a solution of (E)-2-(2,6-dichlorobenzamido)-5-[4-(isopropyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester (5.63 g) in THF (165 ml), 0.1N
aqueous sodium hydroxide solution (165 ml) was added, and the resulting mixture
was stirred at room temperature for 1 hour. Water (200 ml) was added to the
reaction solution and the resulting mixture was washed with ether. Aqueous layer
was acidified by adding IN hydrochloric acid thereto and extracted twice with ethyl
acetate. Organic layers were washed with saturated brine and dried over anhydrous

sodium sulfate. After removing anhydrous sodium sulfate by filtration, the filtrate
was concentrated to dryness to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-
(isopropyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid (4.68 g).
Example 29
[0329]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-((4-hydroxy-benzyl)-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid

(1) In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (45.6 mg) was reacted with (4-iodophenyl)-(4-
acetoxybenzyl)pyrimidin-2-ylamine (67.4 mg) to obtain (E)-2-(2,6-
dichlorobenzamido)-5-[4-((4-acetoxy-benzyl)-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid methyl ester (61.7 mg). Column chromatography (silica gel, eluent: ethyl
acetate/hexane = 1/3→3/l) and thin layer chromatography (silica gel, developing
solvent: cyclohexane/ethyl acetate = 1/5) were used for purification.
(2) To a solution of (E)-2-(2,6-dichlorobenzamido)-5-|4-((4-acetoxy-benzyl)-
pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (61.7 mg) in THE (3.0
ml), 0.1N aqueous sodium hydroxide solution (3.0 ml) was added, and the resulting
mixture was stirred at room temperature for 2 hours. Water (50 ml) was added to
the reaction solution and the resulting mixture was washed with ether. Aqueous

layer was acidified by adding IN hydrochloric acid thereto and extracted twice with
ethyl acetate. Organic layers were washed with saturated brine and dried over
anhydrous sodium sulfate. After removing anhydrous sodium sulfate by filtration,
the filtrate was concentrated to dryness to obtain (E)-2-(2,6-dichIorobenzamido)-5-
[4-((4-hydroxy-benzyl)-pyrimidin-2-ylamino)phenylJpent-4-enoic acid (43 mg).
Example 30
[0332]
(E)-5-[4-((2-Cyano-ethyl)-pyrimidin-2-yl-amino)-phenyl]-2-(2,6-
dichlorobenzamido)-pent-4-enoic acid methyl ester

In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (73 mg) was reacted with 3-(N-(4-iodophenyl)-N-(pyrimidin-2-
yl)amino)propanenitrile (94 mg) to obtain (E)-5-[4-((2-cyano-ethyl)-pyrimidin-2-yl-
amino)-phenyl]-2-(2,6-dichlorobenzamido)-pent-4-enoic acid methyl ester (100 mg).
Column chromatography (silica gel, eluent: cyclohexane/chloroform = 2/1→ 1/2→
1/5) was used for purification.
Example 31
[0335]
(E)-5-[4-((2-Cyano-ethyl)-pyrimidin-2-yl-amino)-phenyl]-2-(2,6-
dichlorobenzamido)-pent-4-enoic acid sodium salt


In the same manner as in Example 8, (E)-5-[4-((2-cyano-ethyI)-pyrimidin-2-
yl-amino)-phenyl]-2-(2,6-dichlorobenzamido)-pent-4-enoic acid methyl ester (90 mg)
was hydrolyzed to obtain (E)-5-[4-((2-cyano-ethyl)-pyrimidin-2-yl-amino)-phenylJ-2-
(2,6-dichlorobenzamido)-pent-4-enoic acid sodium salt (68 mg).
Example 32
[0338]
(E)-5-[4-(Benzyl-pyrimidin-2-yl-amino)-phenyl]-2-(2,6-dichlorobenzamido)-pent-4-
enoic acid methyl ester

In the same manner as in Example 1. 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (59.3 mg) was reacted with N-benzyl-N-(4-iodophenyl)pyrimidin-
2-amine (67.0 mg) to obtain (E)-5-[4-(benzyl-pyrimidin-2-yl-amino)-phenyl]-2-(2,6-

dichlorobenzamido)-pent-4-enoic acid methyl ester (58.7 mg). Column
chromatography (silica gel, eluent: chloroform/cyclohexane = 2/1 -*chloroform) and
thin layer chromatography (silica gel, developing solvent: cyclohexane/ethyl acetate
= 2/1) were used for purification.
Example 33
[0341]
(E)-5-[4-(Benzyl-pyrimidin-2-yl-amino)-phenyl]-2-(2,6-dichlorobenzamido)-pent-4-
enoic acid sodium salt
In the same manner as in Example 8, (E)-5-[4-(benzyl-pyrimidin-2-yl-amino)-
phenyl]-2-(2,6-dichlorobenzamido)-pent-4-enoic acid methyl ester (58.7 mg) was
hydrolyzed to obtain (E)-5-[4-(benzyl-pyrimidin-2-yl-amino)-phenyl]-2-(2,6-
dichlorobenzamido)-pent-4-enoic acid sodium salt (33.5 mg).
Example 34
[0344]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-((4-methoxy-benzyl)-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester
[0345]


In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (83.3 mg) was reacted with N-(4-methoxybenzyl)-N-(4-
bromophenyl)pyrimidin-2-amine (102.4 mg) to obtain (E)-2-(2,6-
dichlorobenzami do)-5 - [4-((4-methoxy-benzyl)-pyrimidin-2-ylamino)phenyl] pen t-4-
enoic acid methyl ester (102.4 mg). Column chromatography (silica gel, eluent:
hexane/ethyl acetate = 3/1→1/3) and thin layer chromatography (silica gel,
developing solvent: cyclohexanc/ethyl acetate = 1/5) were used for purification.
Example 35
[0347]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-((4-methoxy-benzyl)-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid sodium salt

In the same manner as in Example 8, (E)-2-(2,6-dichlorobenzamido)-5-[4-((4-

methoxy-benzyl)-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (102.4
mg) was hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-((4-methoxy-
benzyl)-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid sodium salt (76.4 mg).
Example 36
[0350]
(E)-5-(4-Butyl-pyrimidin-2-yl-amino)-pheny]]-2-(2,6-dichlorobenzamido)-pent-4-
enoic acid methyl ester

In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (90.4 mg) was reacted with N-(4-bromophenyl)-N-butylpyrimidin-
2-amine (92.0 mg) to obtain (E)-5-(4-butyl-pyrimidin-2-yl-amino)-phenyl]-2-(2,6-
dichlorobenzamido)-pent-4-enoic acid methyl ester (78.8 mg). Column
chromatography (silica gel, eluent: chloroform/cyclohexane = 2/1 →chloroform) and
thin layer chromatography (silica gel, developing solvent: cyclohexane/ethyl acetate
= 2/1) were used for purification.
Example 37
[0353]
(I.v)-5-(4-Butyl-pyrimidin-2-yl-amino)-phenyl]-2-(2,6-dichlorobenzamido)-pent-4-
enoic acid sodium salt
[0354]


In the same manner as in Example 8, (E)-5-(4-butyl-pyrimidin-2-yl-amino)-
phenyl]-2-(2,6-dichlorobenzamido)-pent-4-enoic acid methyl ester (78.8 mg) was
hydrolyzed to obtain (E)-5-(4-butyl-pyrimidin-2-yl-amino)-phenyl]-2-(2,6-
dichlorobenzamido)-pent-4-enoic acid sodium salt (42.0 mg).
Example 38
[0356]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyridin-3-ylmethyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester

In the same manner as in Example 1. 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (1060.0 mg) was reacted with N-(4-bromophenyl)-N-((pyridin-3-
yl)methyl)pyrimidin-2-amine (120.0 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-
[4-(pyridin-3-ylmethyl-pyrimidin-2-ylamino)phenyl]pent-4-cnoic acid methyl ester

(147.8 mg). Column chromatography (silica gel, eluent: chloroform-*
chloroform/ethyl acetate =1/1) and thin layer chromatography (chloroform/ethyl
acetate =1/1) were used for purification.
Example 39
[0359]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyridin-3-ylmethyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid sodium salt

In the same manner as in Example 8, (E)-2-(2,6-dichlorobenzamido)-5-[4-
(pyridin-3-ylmethyl-pyrimidin-2-ylamino)pheny]]pent-4-cnoic acid methyl ester
(147.8 mg) was hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(pyridin-3-
ylmethyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid sodium salt (114.4 mg).
Example 40
[0362]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyrimidin-2-yl-thiazol-4-ylmethyl-
amino)phenyl]pent-4-enoic acid methyl ester
[0363]


In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (78.0 mg) was reacted with N-(4-bromophenyl)-N-((thiazol-4-
yl)methyl)pyrimidin-2-amine (90.0 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-
[4-(pyrimidin-2-yl-thiazol-4-ylmethyl-amino)phenyl]pent-4-enoic acid methyl ester
(87.7 mg). Column chromatography (silica gel, eluent: chloroform—*
chloroform/ethyl acetate = 3/1) and thin layer chromatography (cyclohexane/ethyl
acetate = 1/2) were used for purification.
Example 41
[0365]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyrimidin-2-yl-thiazol-4-ylmethyl-
amino)phenyl]pent-4-enoic acid sodium salt

In the same manner as in Example 8, (E)-2-(2,6-dichlorobenzamido)-5-|4-

(pyrimidin-2-yl-thiazol-4-ylmethyl-amino)phenyl]pent-4-enoic acid methyl ester
(87.7 mg) was hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(pyrimidin-
2-yl-thiazol-4-ylmethyl-amino)phenyl]pent-4-enoic acid sodium salt (63.6 mg).
Example 42
[0368]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-((4-methoxy-pyrimidin-2-yl)-methyI-
amino)phenyl]pent-4-enoic acid methyl ester

In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (43 mg) was reacted with N-(4-iodophenyl)-4-methoxy-N-
methylpyrimidin-2-aminc (51 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-((4-
methoxy-pyrimidin-2-yl)-methyl-amino)phenyl]pent-4-enoic acid methyl ester (32
mg). Column chromatography (silica gel, eluent: hexane/ethyl acetate = 2/1) was
used for purification.
Example 43
[0371]
(E)-2-(2.6-Dichlorobenzamido)-5-[4-((4-methoxy-pyrimidin-2-yl)-methyl-
amino)phenyl]pen1-4-enoic acid
[0372]


In the same manner as in Example 2, (E)-2-(2,6-dichlorobenzamido)-5-[4-((4-
methoxy-pyrimidin-2-yl)-methyl-amino)phenyl]pent-4-enoic acid methyl ester (31
mg) was hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-((4-methoxy-
pyrimidin-2-yl)-methyl-amino)phenyl]pent-4-enoic acid (29 mg).
Example 44
[0374]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-((4,6-dimethoxy-pyrimidin-2-yl)-methyl-
amino)phenyl]pen1-4-enoic acid methyl ester

In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pcnt-4-enoic
acid methyl ester (80 mg) was reacted with N-(4-iodophenyl)-4,6-dimethoxy-N-
mcthylpyrimidin-2-amine (118 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-|4-
((4,6-dimethoxy-pyrimidin-2-yl)-methyI-arnino)phenyl]pent-4-enoic acid methyl
ester (92 mg). Column chromatography (silica gel, eluent: hexane/ethyl acetate =
2/1) was used for purification.

Example 45
[0377]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-((4,6-dimethoxy-pyrimidin-2-yl)-methyl-
amino)phenyl]pent-4-enoic acid

In the same manner as in Example 2, (E)-2-(2,6-dichlorobenzamido)-5-[4-
((4,6-dimethoxy-pyrimidin-2-yl)-methyl-amino)phenyl]pent-4-enoic acid methyl
ester (91 mg) was hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-((4,6-
dimethoxy-pyrimidin-2-yl)-methyl-amino)phenyl]pent-4-enoic acid (76 mg).
Example 46
[0380]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(phenyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid methyl ester


In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (52.4 mg) was reacted with N-(4-bromophenyl)-N-
phenylpyrimidin-2-amine (56.6 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-
(phenyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (40.0 mg).
Column chromatography (silica gel, eluent: chloroform/cyclohexane = 1/1→
chloroform) and thin layer chromatography (silica gel, developing solvent:
cyclohexane/ethyl acetate = 1/2) were used for purification.
Example 47
[0383]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(phenyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid sodium salt

In the same manner as in Example 8, (E)-2-(2,6-dichlorobenzamido)-5-(4-
(phenyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (40.0 mg) was
hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(phenyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid sodium salt (25.3 mg).
Example 48
[0386]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyrimidin-2-yl-(tetrahydro-pyran-4-yl)-
amino)phenyl]pent-4-enoic acid methyl ester


In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (164.0 mg) was reacted with trifluoromethanesulfonic acid 4-
[pyrimidin-2-yl-(tetrahydropyran-4-yl)amino]phenyl ester (220.0 mg) to obtain (Fv)-2-
(2,6-dichlorobenzamido)-5-[4-(pyrimidin-2-yl-(tetrahydro-pyran-4-yl)-
amino)phenyl]pent-4-enoic acid methyl ester (178.5 mg). Column chromatography
(silica gel, eluent: chloroform/cyclohexane = 1/2→chloroform) and thin layer
chromatography (silica gel, developing solvent: cyclohexane/ethyl acetate = 1/1)
were used for purification.
Example 49
[0389]
(E)-2-(2t6-Dichlorobenzamido)-5-[4-(pyrimidin-2-yl-(tetrahydro-pyran-4-yl)-
amino)phenyl]pent-4-enoic acid sodium salt


[0391]
In the same manner as in Example 8, (E)-2-(2,6-dichlorobenzamido)-5-[4-
(pyrimidin-2-yl-(tetrahydro-pyran-4-yl)-amino)phenyl]pent-4-enoic acid methyl ester
(178.5 mg) was hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(pyrimidin-
2-yl-(tetrahydro-pyran-4-yl)-amino)phenyl]pent-4-enoic acid sodium salt (103 mg).
Example 50
[0392]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-((3-methyl-2-butenyl)-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester

In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoie
acid methyl ester (145 mg) was reacted with N-(4-iodophenyl)-N-(3-methyl-2-
butenyl)pyrimidin-2-amine (175 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-
((3-methyl-2-butenyl)-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester
(133 mg). Column chromatography (silica gel, eluent: hexane/ethyl acetate = 1/1)
was used for purification.
Example 51
|0395)
(E)-2-(2,6-Dichlorobenzamido)-5-[4-((3-methyl-2-butenyl)-pyrimidin-2-
ylamino)phcnyl]pent-4-enoic acid sodium salt


In the same manner as in Example 8, (E)-2-(2,6-dichlorobenzamido)-5-[4-((3-
methyl-2-butenyl)-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (133
mg) was hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-((3-methyl-2-
butenyl)-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid sodium sail (95 mg).
Example 52
[0398]
(E)-5-[4-(Cyclopropylmethyl-pyrimidin-2-ylamino)phenyl]-2-(2,6-
dichlorobenzamido)pent-4-enoic acid methyl ester

In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-cnoic
acid methyl ester (1 39 mg) was reacted with N-(cyclopropylmethyl)-N-(4-
iodophenyl)pyrimidin-2-amine (162 mg) to obtain (E)-5-[4-(cyclopropylmethyl-
pyrimidin-2-ylamino)phenyl]-2-(2,6-dichlorobenzamido)pent-4-enoic acid methyl

ester (167 mg). Column chromatography (silica gel, eluent: hexane/ethyl acetate =
1 /1) was used for purification.
Example 53
[0401]
(E)-5-[4-(Cyclopropylmethyl-pyrimidin-2-ylamino)phenyl]-2-(2,6-
dichlorobenzamido)pent-4-enoic acid sodium salt

In the same manner as in Example 8, (E)-5-[4-(cyclopropylrnethyl-pyrimidin-
2-ylamino)phenyl]-2-(2,6-dichlorobenzarnido)pent-4-enoic acid methyl ester (153
mg) was hydrolyzed to obtain (E)-5-[4-(cyclopropylmethyl-pyrimidin-2-
ylamino)phenyl]-2-(2,6-dichlorobenzamido)pent-4-enoic acid sodium salt (101 mg).
Example 54
[0404]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(isobutyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid methyl ester


[0406]
In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (143 mg) was reacted with N-(4-iodophenyl)-N-isobutylpyrimidin-
2-amine (167 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(isobutyl-pyrimidin-
2-ylamino)phenyl]pent-4-enoic acid methyl ester (159 mg). Column
chromatography (silica gel, eluent: hexane/ethyl acetate = 1/1) was used for
purification.
Example 55
[0407]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(isobutyl~pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid sodium salt

In the same manner as in Example 8. (E)-2-(2,6-dichlorobenzamido)-5-[4-
(isobutyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (157 mg) was
hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(isobutyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid sodium salt (128 mg).
Example 56
[0410]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(propyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid methyl ester
[0411]


In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (171 mg) was reacted with N-(4-iodophenyl)-N-propylpyrimidin-2-
amine (192 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(propyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester (209 mg). Column chromatography
(silica gel, eluent: cyclohexane/chloroform = 4/1→3/l) was used for purification.
Example 57
[0413]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(propyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid

In the same manner as in Example 2, (E)-2-(2,6-dichlorobenzamido)-5-[4-
(propyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (127 mg) was
hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(propyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid (95 mg).
Example 58

[0416]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyridin-4-ylmethyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester

In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (72 mg) was reacted with N-(4-iodophenyl)-N-((pyridin-4-
yl)methyl)pyrimidin-2-amine (93 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-
(pyridin-4-ylmethyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (99
mg). Column chromatography (silica gel, eluent: cyclohexane/ethyl acetate = 1/2—>
1/3) was used for purification.
Example 59
[0419]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyridin-4-ylmethyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid
[0420]


In the same manner as in Example 2, (E)-2-(2,6-dichlorobenzamido)-5-[4-
(pyridin-4-ylmethyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (99
mg) was hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(pyridin-4-
ylmethyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid (48 mg).
Example 60
[0422]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyridin-2-ylmethyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester

In the same manner as in Example 1, 2-(2,6-dichlorobenzamido)pent-4-enoic
acid methyl ester (124 mg) was reacted with N-(4-iodophenyl)-N-((pyridin-2-
yl)methyl)pyrimidin-2-amine (159 mg) to obtain (E)-2-(2,6-diehlorobenzamido)-5-
|4-(pyridin-2-ylmethyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester

(208 mg). Column chromatography (silica gel, eluent: cyclohexane/ethyl acetate =
1 /l → 1 /2) was used for purification.
Example 61
[0425]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyridin-2-ylmethyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid

In the same manner as in Example 2, (E)-2-(2,6-dichlorobenzamido)-5-|4-
(pyridin-2-ylmethyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (208
mg) was hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(pyridin-2-
ylmethyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid (123 mg).
Example 62
[0428]
(E)-2-(2,6-Difluorobenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid methyl ester
[0429]


Under an argon atmosphere, triethylamine (0.028 ml) and 2,6-difluorobenzoyl
chloride (0.015 ml) were added to a solution of 2-amino-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester (31.7 mg) in dichloromethane (2 ml),
and the resulting mixture was stirred at room temperature for 2 hours. Saturated
aqueous sodium hydrogen carbonate solution was added to the reaction solution and
the resulting mixture was extracted with ethyl acetate. Organic layer was washed
twice with water and once with saturated brine, and dried over anhydrous sodium
sulfate. After removing anhydrous sodium sulfate by filtration, the filtrate was
concentrated. The residue was purified by thin layer chromatography (silica gel.
developing solvent: cyclohexane/ethyl acetate = 3/2) to obtain (E)-2-(2,6-
difluorobenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid
methyl ester (42 mg).
Example 63
[0431]
(E)-2-(2,6-Difluorobenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid
[0432]


In the same manner as in Example 2, (E)-2-(2,6-difluorobenzamido)-5-[4-
(methyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (42 mg) was
hydrolyzed to obtain (E)-2-(2,6-difluorobenzamido)-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoie acid (29 mg).
Example 64
[0434]
(E)-2-(2-Chloro-6-fluorobenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-
4-enoic acid methyl ester

Under an argon atmosphere, EDC hydrochloride (39 mg) and HOBT (3 mg)
were added to a solution of 2-amino-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-
4-enoic acid methyl ester (51 mg) and 2-chloro-6-fluorobenzoic acid in
dichloromethane (1.5 ml), and the resulting mixture was stirred at room temperature
for 28 hours. To the reaction solution, 1N hydrochloric acid was added, and the
resulting mixture was extracted with ethyl acetate. Organic layer was washed with

saturated aqueous sodium hydrogen carbonate solution and dried over anhydrous
sodium sulfate. After removing anhydrous sodium sulfate by filtration, the filtrate
was concentrated. The residue was purified by thin layer chromatography (silica gel.
developing solvent: hexane/ethyl acetate = 1/2) to obtain (E)-2-(2-chloro-6-
fluorobenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid
methyl ester (39 mg).
Example 65
[0437]
(E)-2-(2-Chloro-6-fluorobenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-
4-enoic acid

In the same manner as in Example 2, (E)-2-(2-chloro-6-fluorobenzamido)-5-
[4-(methyl-pyrimidin-2-ylamino)phenyl]pcnt-4-enoic acid methyl ester (39 mg) was
hydrolyzed to obtain (E)-2-(2-chloro-6-nuorobenzamido)-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid (34 mg).
Example 66
[0440]
(E)-2-(2-Chloro-6-methylbenzamido)-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester
|0441]


Under an argon atmosphere, oxalyl dichloride (0.022 ml) and DMF (0.002
ml) were added to a solution of 2-chloro-6-methylbenzoic acid (29 mg) in
dichloromethane (1.0 ml), and the resulting mixture was stirred at room temperature
for 1 hour. The reaction solution was concentrated and the residue was dissolved in
dichloromethane (1.0 ml). To the solution, 2-amino-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester (50 mg) and triethylamine (0.05 ml)
were added, and the resulting mixture was stirred at room temperature for 2.5 hours.
To the reaction solution, IN hydrochloric acid was added, and the resulting solution
was extracted with dichloromethane. Organic layer was washed with saturated
aqueous sodium hydrogen carbonate solution and dried over anhydrous sodium
sulfate. After removing anhydrous sodium sulfate by filtration, the filtrate was
concentrated. The residue was purified by thin layer chromatography (silica gel,
developing solvent: hexane/ethyl acetate = 1/1) to obtain (E)-2-(2-chloro-6-
methylbenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid
methyl ester (63 mg).
Example 67
[0443]
(E)-2-(2-Chloro-6-methylbenzamido)-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid
[0444]


In the same manner as in Example 2, (E)-2-(2-chloro-6-methylbenzamido)-5-
[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (63 mg) was
hydrolyzed to obtain (E)-2-(2-chloro-6-methylbenzamido)-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid (57 mg).
Example 68
[0446]
(E)-2-(2,6-Dimethylbenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid methyl ester

Under an argon atmosphere, oxalyl dichloride (0.025 ml) and DMF (0.002
ml) were added to a solution of 2,6-dimethylbenzoic acid (24 mg) in
dichloromethane (1.0 ml), and the resulting mixture was stirred at room temperature
for 2 hours. The reaction solution was concentrated and the residue was dissolved
in dichloromethane (1.0 ml). To the solution. 2-amino-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester (50 mg) and triethylaminc (0.05 ml)
were added, and the resulting mixture was stirred at room temperature for 3 hours.
To the reaction solution, 1N hydrochloric acid was added, and the resulting solution

was extracted with dichloromethane, followed by drying the organic layer over
anhydrous sodium sulfate. After removing anhydrous sodium sulfate by filtration,
the filtrate was concentrated. The residue was purified by thin layer
chromatography (silica gel, developing solvent: hexane/ethyl acetate = 3/2) to obtain
(E)-2-(2,6-dimethylbenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid methyl ester (62 mg).
Example 69
[0449]
(E)-2-(2,6-Dimethylbenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid

In the same manner as in Example 2, (E)-2-(2,6-dimethylbenzamido)-5-[4-
(methyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (62 mg) was
hydrolyzed to obtain (E)-2-(2,6-dimethylbenzamido)-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid (47 mg).
Example 70
[0452]
(E)-5-[4-((2-Cyano-ethyl)-pyrimidin-2-yl-amino)-phenyl]-2-(2,6-
difluorobenzamido)-pent-4-enoic acid methyl ester
[0453]


In the same manner as in Example 1, 2-(2,6-difluorobenzamido)pent-4-enoic
acid methyl ester (17 mg) was reacted with 3-(N-(4-iodophenyl)-N-(pyrimidin-2-
yl)amino)propanenitrile (24 mg) to obtain ((E)-5-[4-((2-cyano-ethyl)-pyrimidin-2-yl-
amino)-phenyl]-2-(2,6-difluorobenzamido)-pent-4-enoic acid methyl ester (18 mg).
Column chromatography (silica gel, eluent: hexane/ethyl acetate = 2/1) was used for
purification.
Example 71
[0455]
(E)-5-[4-((2-Cyano-ethyl)-pyrimidin-2-yl-amino)-phenyl]-2-(2,6-
difluorobenzamido)-pent-4-enoic acid

In the same manner as in Example 2, (E)-5-[4-((2-cyano-ethyl)-pyrimidin-2-
yl-amino)-phenyl]-2-(2,6-difluorobenzamido)-pent-4-enoic acid methyl ester (18 mg)

was hydrolyzed to obtain (E)-5-[4-((2-cyano-ethyl)-pyrimidin-2-yl-amino)-phenyl]-2-
(2,6-difluorobenzamido)-pent-4-enoic acid (10 mg).
Example 72
[0458]
2-(2,6-Dichlorobenzamido)-5-[4-(pyriraidin-2-yloxy)phenyl]pent-4-ynoic acid
methyl ester

Under an argon atmosphere, copper iodide (2.8 mg) and
dichlorobis(triphenylphosphine)palladium (5.2 mg) were added to a solution of 2-
(2,6-dichlorobenzamido)pent-4-ynoic acid methyl ester (74 mg) and 2-(4-
iodophenoxy)pyrimidine (81 mg) in THF/diisopropylamine (3/1) mixture (5 ml), and
the resulting mixture was stirred at room temperature for 2 hours. The reaction
solution was concentrated and ethyl acetate was added to the residue. The insoluble
matter was removed by filtration, and the filtrate was washed once with water and
once with saturated brine and dried over anhydrous sodium sulfate. After removing
anhydrous sodium sulfate by filtration, the filtrate was concentrated. The residue
was purified by column chromatography (silica gel, eluent: cyclohexane/chlorofonn
= 2/1→cyclohexane/chloroform = 1/3) to obtain 2-(2,6-dichlorobenzamido)-5-[4-
(pyrimidin-2-yloxy)phenyl]pent-4-ynoic acid methyl ester (100 mg).
Example 73
[0461]
2-(2,6-Dichlorobenzamido)-5-[4-(pyrimidin-2-yloxy)phenyl]pent-4-ynoic acid


In the same maimer as in Example 8, 2-(2,6-dichlorobenzamido)-5-[4-
(pyrimidin-2-yloxy)phenyl]pent-4-ynoic acid methyl ester (100 mg) was hydrolyzed
to obtain 2-(2,6-dichlorobenzamido)-5-[4-(pyrimidin-2-yloxy)phenyl]pent-4-ynoic
acid sodium salt (85 mg).
Example 74
[0464]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyrimidin-2-ylamino)phenyl]pent-4-ynoic acid
methyl ester

In the same manner as in Example 72, 2-(2,6-dichlorobenzamido)pent-4-
ynoic acid methyl ester (50.0 mg) was reacted with N-(4-iodophenyl)pyrimidin-2-
amine (49.5 mg) to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(pyrimidin-2-
ylamino)phenyl]pent-4-ynoic acid methyl ester (80.0 mg). Column chromatography
(silica gel, eluent: chloroform/cyclohexane = 3/1→chloroform/ethyl acetate = 5/1)
and thin layer chromatography (silica gel, developing solvent: chloroform/ethyl

acetate = 3/1) were used for purification.
Example 75
[0467]
(E)-2-(2,6-Dichlorobenzamido)-5-[4-(pyrimidin-2-ylamino)phenyl]pent-4-ynoic acid
sodium salt

In the same manner as in Example 8, (E)-2-(2,6-dichlorobenzamido)-5-[4-
(pyrimidin-2-ylamino)phenyl]pent-4-ynoic acid methyl ester (80.0 mg) was
hydrolyzed to obtain (E)-2-(2,6-dichlorobenzamido)-5-[4-(pyrimidin-2-
ylamino)phenyl]pent-4-ynoic acid sodium salt (62.3 mg).
Example 76
[0470]
(S.E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-methoxytetrahydropyran-4-yl)-
phenyl]pent-4-enoic acid methyl ester

Under an argon atmosphere, palladium acetate (295 mg) and tris(2-

methylphenyl)phosphine (384 mg) were added to a suspension of (S)-2-(2,6-
dichlorobenzamido)pent-4-enoic acid methyl ester (7.60 g), tetrahydro-4-(4-
iodophenyl)-4-methoxy-2H-pyran (8.00 g) and potassium carbonate (5.21 g) in DMF
(90 ml), and the resulting mixture was stirred at 80°C for 2 hours. After cooling the
reaction solution to room temperature, ethyl acetate was added thereto, and the
resulting mixture was washed 3 times with water and once with saturated brine,
followed by drying the organic layer over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was purified by column chromatography (silica gel, eluent:
cyclohexane/chloroform = 2/1→ 1 /4). The obtained crudely purified product was
further purified by column chromatography (silica gel, eluent: cyclohexane/ethyl
acetate = 4/1) to obtain (S,E)-2-(2,6-dichlorobenzamido)-5-[4-(4-
methoxytetrahydropyran-4-yl)-phenyl]pent-4-enoic acid methyl ester (9.80 g).
Example 77
[0473]
(S,E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-methoxytetrahydropyran-4-yl)-
phenyl]pent-4-enoic acid

In a mixed solvent of THF (250 ml) and water (125 ml), (S,E)-2-(2,6-
dichlorobenzamido)-5-f4-(4-methoxytetrahydropyran-4-yl)-phenyl]pent-4-enoicacid
methyl ester (9.80 g) was dissolved, and the resulting mixture was cooled to 0°C.

Barium hydroxide octahydrate (3.14 g) was added thereto and the resulting mixture
was stirred at 0°C for 8 hours. The reaction solution was concentrated to remove
THF, and water (150 ml) was added thereto, followed by washing the resulting
mixture with ether. Aqueous layer was acidified by adding IN hydrochloric acid in
small portions thereto and extracted 3 times with ethyl acetate. Organic layers were
washed with saturated brine and dried over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was purified by column chromatography (silica gel, eluent:
chloroformtertethanol = 10/1). The obtained crudely purified product was further
purified by column chromatography (silica gel, eluent: cyclohexane/ethyl acetate -
1/1→ethyl acetate) to obtain (S,E)-2-(2,6-dichlorobenzamido)-5-[4-(4-
methoxytetrahydropyran-4-yl)-phenyl]pent-4-enoic acid (6.15 g).
Example 78
[0476]
(S,E)-2-(2,6-Dichlorobenzamido)-5-[4-(4-methoxytetrahydropyran-4-yl)-
phenyl]pent-4-enoic acid sodium salt

To(S,E)-2-(2,6-dichlorobenzamido)-5-[4-(4-methoxytetrahydropyran-4-yl)-
phenyl]pent-4-enoic acid (4.24 g), THF (8.86 ml) and IN aqueous sodium hydroxide
solution (8.86 ml) were added, and the resulting mixture was stirred at room
temperature for 5 minutes. The reaction solution was concentrated to dryness to

obtain (S,E)-2-(2,6-dichlorobenzamido)-5-[4-(4-methoxytetrahydropyran-4-yl)-
phenyl]pent-4-enoic acid sodium salt (4.30 g).
Example 79
[0479]
(S,E)-2-(2,6-Dichlorobenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid methyl ester

Under an argon atmosphere, palladium acetate (93.2 mg) and tris(2-
methylphenyl)phosphine (121.2 mg) were added to a suspension of (S)-2-(2,6-
dichlorobenzamido)pent-4-enoic acid methyl ester (1.20 g), N-(4-iodophenyl)-N-
methylpyrimidin-2-arnine (1.24 g) and potassium carbonate (824 mg) in DMF (20
ml), and the resulting mixture was stirred at 80°C for 3 hours. After cooling the
reaction solution to room temperature, ethyl acetate was added thereto, and the
resulting mixture was washed twice with water and once with saturated brine,
followed by drying the organic layer over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was purified by column chromatography (silica gel, eluent:
cyclohexane/chloroform = 1/1 →chloroform). The obtained crudely purified
product was purified again by column chromatography (silica gel, eluent:
cyclohexane/ethyl acetate = 4/1→2/l) to obtain (S,E)-2-(2,6-dichlorobenzamido)-5-
[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (1.28 g).

Example 80
[0482]
(S,E)-2-(2,6-Dichlorobenzamido)-5-[4-(methyl-pyrimidin-2-yiamino)phenyljpent-4-
enoic acid

A solution of (S,E)-2-(2,6-dichlorobenzamido)-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester (1.28 g) in THF (45 ml) was cooled to
0°C. To the solution, 0.1N aqueous lithium hydroxide solution (40 ml) was added,
and the resulting mixture was stirred at 0°C for 40 minutes. Water (50 ml) was
added to the reaction solution, and the resulting mixture was washed with ether.
Aqueous layer was acidified by adding 1N hydrochloric acid in small portions thereto,
and extracted twice with ethyl acetate. Organic layers were washed with saturated
brine and dried over anhydrous sodium sulfate. After removing anhydrous sodium
sulfate by filtration, the filtrate was concentrated to dryness to obtain (S,E)-2-(2,6-
dichlorobenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid
(1.01 g).
Example 81
[0485]
(S,E)-2-(2,6-Dichlorobenzamido)-5-[4-(methyl-pyrimidin-2-ylamino)phenyl]pent-4-
enoic acid sodium salt
[0486]


To a solution of (S,E)-2-(2,6-dichlorobenzamido)-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid (10.60 g) in methanol (200 ml), IN aqueous
sodium hydroxide solution (22.5 ml) was added, and the resulting mixture was stirred
at room temperature for 5 minutes. The reaction solution was concentrated to
dryness to obtain (S,E)-2-(2,6-dichlorobenzamido)-5-[4-(methyl-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid sodium salt (11.08 g). lR(KBr)cm"': 3385, 1584,
1552, 1486, 1431, 1397,1315,1195,1112,968,799.
Example 82
[0488]
(S,E)-2-(2,6-Dichlorobenzamido)-5-[4-(isopropyi-pyrimidin-2-ylamino)phenyI]pent-
4-enoic acid methyl ester

Under an argon atmosphere, palladium acetate (19.9 mg) and tris(2-
methylphenyl)phosphine (25.9 mg) were added to a suspension of (S)-2-(2,6-
dichlorobenzamido)pent-4-enoic acid methyl ester (514.4 mg), N-(4-iodophenyl)-N-

isopropylpyrimidin-2-amine (577.4 mg) and potassium carbonate (352.9 mg) in DMF
(6 ml), and the resulting mixture was stirred at 80°C for 7 hours. After cooling the
reaction solution to room temperature, ethyl acetate was added thereto, and the
resulting mixture was washed twice with water and once with saturated brine,
followed by drying the organic layer over anhydrous sodium sulfate. After
removing anhydrous sodium sulfate by filtration, the filtrate was concentrated. The
residue was purified by column chromatography (silica gel, eluent:
cyclohexane/chloroform = 2/1→1/1→ 1/3). The obtained crudely purified product
was purified again by column chromatography (silica gel, eluent: cyclohexane/ethyl
acetate = 6/1→4/l→2/l) to obtain (S,E)-2-(2,6-dichlorobenzamido)-5-[4-(isopropyl-
pyrimidin-2-ylamino)phenyl]pent-4-enoic acid methyl ester (553.6 mg).
Example 83
[0491]
(S,E)-2-(2,6-Dichlorobenzamido)-5-[4-(isopropyl-pyrimidin-2-ylamino)phcnyl]pent-
4-enoic acid

A solution of (S,E)-2-(2,6-dichlorobenzamido)-5-[4-(isopropy]-pyrimidin-2-
ylamino)phenyl]pent-4-enoic acid methyl ester (526.2 g) in THF (15 ml) was cooled
to 0°C. To the solution, 0. IN aqueous lithium hydroxide solution (15.4 ml) was
added, and the resulting mixture was stirred at 0°C for 40 minutes. Water (20 ml)
was added to the reaction solution and the resulting mixture was washed with ether.

Aqueous layer was acidified by adding 1N hydrochloric acid in small portions thereto,
and extracted twice with ethyl acetate. Organic layers were washed with saturated
brine and dried over anhydrous sodium sulfate. After removing anhydrous sodium
sulfate by filtration, the filtrate was concentrated to dryness to obtain (S,E)-2-(2,6-
dichlorobenzamido)-5-[4-(isopropyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid
(420.7 mg).
Example 84
[0494]
(S,E)-2-(2,6-Dichlorobenzamido)-5-[4-(isopropyl-pyrimidin-2-ylamino)phenyl]pent-
4-enoic acid sodium salt

To a suspension of (S,E)-2-(2,6-dichlorobenzamido)-5-[4-(isopropyl-
pyrimidin-2-ylamino)phenyl]pent-4-enoic acid (390.4 mg) in methanol (15 ml), IN
aqueous sodium hydroxide solution (0.782 ml) was added, and the resulting mixture
was stirred at room temperature for 5 minutes. The reaction solution was
concentrated to dryness to obtain (S,E)-2-(2,6-dichlorobenzamido)-5-[4-(isopropyl-
pyrimidin-2-ylamino)phenyl]pent-4-enoic acid sodium salt (388.8 mg). [R(KBr)cm"
': 3386, 2974, 1585, 1549,1509. 1455, 1292, 1122,968,798,780.
[0497]
The spectral data of the compounds of Examples 1 to 84 are shown in Tables
8-16.

Example 85
[0507]
Pharmacokinetic Evaluation in Rats
Each compound was dissolved in PBS or PBS containing ] 0% PEG, thereby
preparing 0.5 mgtertL dosing solution. The compound was administered orally or
intravenously to male SD rats (7 to 9 weeks old). Blood was collected with time
from the tail vein at time points up to 8 hours after the administration, and obtained
blood was centrifuged to collect blood plasma. The blood plasma was pretreated by
using the solid-phase extraction method, and the concentration of the compound was
analyzed with LCtertStertS (ESI negative mode).
[0508]
The obtained pharmacokinetic parameters are summarized in Table 17. As
the Comparative Compound, 2-[4-((3,5-
dichlorobenzensulfonylamino)methyl)benzoylamino]-5-(4-(methyl-pyrimidin-2-
ylamino)phenyl)pent-4-enoic acid (XXI) described in WO 99/26923 was used.


As is apparent from Table 17. the therapeutic or prophylactic agents for
multiple sclerosis according to the present invention showed excellent
bioavailabilities and low total clearance values compared to the Comparative
Compound. Therefore, the agents have the excellent effects when they are
administered orally, and the effects are sustained because of their excellent in vivo
stability.
Example 86
[0513]

Measurement of Inhibitory Effect on Leukocyte Functions
Jurkat cells, which are the cell line originated from human acute T cell
lymphoma, were allowed to react with BCECF-AM at 37°C for 20 minutes to
fluorescently label the cells. The fluorescently labelled Jurkat cells were allowed to
react with ligand-expressing cells or with a ligand-immobilized plate at 37"C for 30
minutes. After removing non-adherent cells by washing. 1% NP40 was added
thereto to lyse the adherent cells, and fluorescence intensity was measured with
Cytofluor 2300 (Millipore). From the obtained fluorescence intensity, the number
of adherent cells was calculated. Each test compound was reacted with the Jurkat
cells before the beginning of the adhesion reaction. In Table 18, the IC50 of each
compound (i.e. the concentration at which the number of adherent cells is reduced by
50%) is shown.


As is apparent from Table 18, the therapeutic or prophylactic agents of the
present invention clearly inhibit functions of leukocytes involved in the development
of multiple sclerosis, so that they exhibit therapeutic and/or prophylactic actions
against multiple sclerosis.
Example 87

[0516]
Measurement of Inhibitory Effect on Inflammatory Mediator Production by
Leukocytes
A mixture of human peripheral blood and physiological saline containing 3%
dextran was left to stand for 30 minutes and then the upper layer was recovered.
The upper layer was overlaid on Histopaquc 1077 (SIGMA), and the resultant was
centrifuged at 1400 rpm for 30 minutes. The supernatant was removed by
aspiration and a buffer was added to the precipitate to prepare a neutrophil
suspension (4 x 105 cells/mL). To the neutrophil suspension, ionomycin was added
and the mixture was incubated at 37°C for 30 minutes, followed by quantification of
leukotriene B4 by EIA (Amersham, Biotrak EIA system). Each test compound was
added to the neutrophil suspension before adding ionomycin. In Table 19, (he IC50
of each compound (i.e. the concentration at which the production of Leukotriene B4
is inhibited by 50%) is shown.

As is apparent from Table 19, the therapeutic or prophylactic agents of the
present invention inhibit production of inflammatory mediators by leukocytes
involved in the development of multiple sclerosis, so that they exhibit therapeutic
and/or prophylactic actions against multiple sclerosis.
Example 88
[0519]

Inhibitory Effect of Compounds against Mouse Experimental Autoimmune
Encephalomyelitis Model
This method was carried out in accordance with the literature "Int. Immunol.,
9,1243-1251 (1997)".
[0520]
A PBS solution containing 2 mgtertL partial synthetic peptide (PLP139-151)
of proteolipid protein was mixed with an equal amount of Freund's complete
adjuvant, and the resulting mixture was inoculated intracutaneously into both lateral
regions of 7-week-old female SJL mouse (Charles River Laboratories Japan) in a
total amount of 0.1 mL (0.05 mL per one side). The compounds described in
Example 77. Example 78, Example 80, Example 81, Example 83 and Example 84
were orally administered at a dose of 50 mg/kg every day from one day before the
inoculation, and neurological symptoms was scored (0: normal, 1: limp tail or hind
limb weakness, 2: limp tail and hind limb weakness, 3: partial hind limb paralysis, 4:
complete hind limb paralysis, 5: moribund state) at Days 11, 12 or 14. The
scoring was carried out by using the method described in Current Protocols in
Immunology (John Wiley & Sonsjnc).
[0521]
The score of neurological symptoms in mouse rose to 1.0-2.1 by inoculation
of PLP13 9-151. On the other hand, increase in the score of neurological symptoms
was remarkably reduced when the test compounds were orally administered. The
rate of reduction by each compound was as follows: the compound described in
Example 77, 40%; the compound described in Example 78, 60%; the compound
described in Example 80, 60%: the compound described in Example 81, 68%; the
compound described in Example 83, 70%; and the compound described in Example
84, 67%.
[0522]

As is apparent from the results, the therapeutic or prophylactic agents of the
present invention have a remarkable inhibitory effect against neurological symptoms
of multiple sclerosis when orally administered.
Industrial Availability
[0523]
The glycine derivatives or pharmaceutically acceptable salts thereof can be
used as a therapeutic or prophylactic agent for multiple sclerosis.

CLAIMS
1. A therapeutic or prophylactic agent for multiple sclerosis comprising as an
effective ingredient a compound of the Formula (1):

[wherein
R1 represents hydrogen or C1-C5 alkyl;
Xs independently represent fluoro, chloro, bromo, iodo or C1-C3 alkyl;
V represents -CH=CH- or -C=C-;
Y represents Formula (II) or Formula (III):

(wherein
R2 represents C1-C5 alkyl or C1-C3 alkoxy;
R3 represents hydrogen or C1-C5 alkyl;
m represents an integer of 0 to 3;
n represents 0 or 1;
p represents an integer of 0 to 2;
W represents -O- or -N(R4)-
(wherein
R4 represents hydrogen, C1-C5 alkyl, C3-C6, alkenyl, phenyl, benzyl, benzyl
substituted with one or two R5s, tetrahydropyranyl, -(CH2)q-O-CH3, pyridylmethyl,
-(CH2)q-CN, C4-C7 cycloalkylmethyl orthiazol-4-ylmethyl:

R5 represents hydroxy or C1-C3 alkoxy; and
q represents an integer of 1 to 3))]
or a pharmaceutically acceptable salt thereof.
2. The therapeutic or prophylactic agent for multiple sclerosis comprising as an
effective ingredient the compound or the pharmaceutically acceptable salt thereof
recited in claim 1, wherein in Formula (I),
V is -CH=CH-;
when Y is represented by the Formula (II), m is 0; and
when Y is represented by the Formula (III), p is 1.
3. The therapeutic or prophylactic agent for multiple sclerosis comprising as an
effective ingredient the compound or the pharmaceutically acceptable sail thereof
recited in claim 2, wherein in Formula (I),
R! is hydrogen;
when Y is represented by the Formula (II), W is -N(R4)- and R4 is C1-C3 alkyl,
cyanoethyl, tetrahydropyranyl or phenyl; and
when Y is represented by the Formula (III), n is 0 and R3 is C1-C3 alkyl.
4. The therapeutic or prophylactic agent for multiple sclerosis comprising as an
effective ingredient the compound or the pharmaceutically acceptable salt thereof
recited in claim 3, wherein in Formula (I),
Xs are independently chloro or methyl;
V is trans -CH=CH-;
when Y is represented by the Formula (II), W is -N(R4)- and R4 is methyl or
isopropyl; and
when Y is represented by the Formula (III), R3 is methyl.
5. A therapeutic or prophylactic method for multiple sclerosis, comprising
administering to a patient an effective amount of the compound of the Formula (I) or
the pharmaceutically acceptable salt thereof recited in any one of claims 1 to 4.

A therapeutic or prophylactic agent for multiple sclerosis is disclosed. The
therapeutic or prophylactic agent comprises as an effective ingredient a glycine
derivative having a specific structure or a pharmaceutically acceptable salt thereof,
for example, the below-described compound [(E)-2-(2,6-dichlorobenzamido)-5-[4-
(isopropyl-pyrimidin-2-ylamino)phenyl]pent-4-enoic acid]. The therapeutic or
prophylactic agent for multiple sclerosis according to the present invention shows the
excellent absorbability and in vivo stability when orally administered, and exhibits
high therapeutic or prophylactic effects.