JP2000086641A - 2-Substituted benzothiazole derivatives and method for producing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a 2-substituted benzothiazole derivative having a binding inhibitory activity between cell adhesion molecules and useful as an immunosuppressant, an anti-inflammatory agent, an anti-allergic agent and a cancer metastasis inhibitor And methods for their production. SOLUTION: General formula (1) [Wherein, R ¹ represents a hydrogen atom, a lower alkyl group having 1 to 4 carbon atoms, X represents an oxygen atom, an amino group optionally substituted with a lower alkyl group having 1 to 4 carbon atoms, and Y represents a methoxy group. in an optionally substituted benzene ring, a 1,3-thiazole ring, Z is - _(CH 2) n- (n is an integer of 0~3), - HC = CH - , - OCH 2 - and , R ² represents a lower alkyl group having 1 to 4 carbon atoms which may be substituted,
R ³ represents a hydrogen atom, or R ² and R ³ combine to form an optionally substituted piperazine ring or the like], and a pharmacologically characterized 2-substituted benzothiazole derivative; Acceptable salts.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to 2-substituted benzothiazole derivatives which have an activity of inhibiting the binding between cell adhesion molecules and are useful as immunosuppressants, anti-inflammatory agents, anti-allergic agents, and cancer metastasis inhibitors. A method for producing the same.

2. Description of the Related Art Adhesion between vascular endothelial cells and leukocytes is an extremely important step in immune and inflammatory reactions (Mebio, No. 5, Vol. 10, 1993). The main adhesion molecules on the vascular endothelial cell side involved in this adhesion include ICAM-1, VC
AM-1, E-selectin, P-selectin and the like have been reported, and the expression of each adhesion molecule differs depending on the time since the inflammation was induced (Diagnosis and Therapy, 83, 1).
164, (1995), Springer Semin Immunopathol,
Vol. 11, 163, (1989), Cell, Vol. 76, 301, (199)
Four)). That is, P-selectin exhibits a peak of expression 5 to 30 minutes after the inflammation is induced (immediately) and decreases in expression thereafter. ECAM-1 and VCAM-1 show that E-selectin shows a peak of expression 12 to 48 hours later (late stage). Among them, ICAM-1, VC which is expressed in large amounts in the late period
Adhesion to leukocytes via AM-1 is the strongest, and it is said that these two adhesion molecules play important roles even in actual various diseases. Therefore, if adhesion through adhesion molecules such as ICAM-1 and VCAM-1 which play a central role in inflammation can be inhibited, autoimmune diseases such as rheumatoid arthritis, nephritis, osteoarthritis of the knee and chronic inflammatory diseases. It is considered to be effective as a therapeutic drug for allergic diseases such as asthma and dermatitis, and a cancer metastasis inhibitor. Cell adhesion inhibitors reported to date include so-called expression inhibitors that inhibit adhesion by suppressing the expression of adhesion molecules, and so-called binding inhibitors that inhibit adhesion by inhibiting the binding between adhesion molecules. Classified as Most of the cell adhesion inhibitors for ICAM-1 and VCAM-1 are expression inhibitors (JP-A-9-1106).
89, JP-A-8-283156, JP-A-8-19875
2, JP-A-7-304667 and JP-A-7-25816
8), the only binding inhibitor is J. Med. Except for peptide macromolecules such as antibodies and ligands of adhesion molecules.
Chem., Vol. 38, 1057 (1995) only reports non-peptidic low molecular weight compounds. The expression inhibitor often acts on the intracellular signal transduction system, and may suppress functions other than expression of the adhesion molecule. Unlike such an expression inhibitor, the binding inhibitor only inhibits the binding between the adhesion molecules, so it is thought that it can be a drug excellent in safety, but it is still not satisfactory.

DISCLOSURE OF THE INVENTION The present invention relates to ICAM-1 and VCAM- which play a central role in cell adhesion molecules.
By providing a substance that inhibits the binding between 1 and leukocytes, excellent immunosuppressants, anti-inflammatory agents, anti-allergic agents,
An object of the present invention is to provide a cancer metastasis inhibitor.

Means for Solving the Problems The present inventors have developed human monocytes.
-Like cell line (U937) and humans 24 hours after IL-1β stimulation
Binding to umbilical vein-derived vascular endothelial cells (HUVEC)
As a result of intensive studies on compounds that inhibit the substance,
A new cell with a different structure from the cell adhesion inhibitor known in
Regular benzothiazole derivatives are ICAM-1, VCA
Found to inhibit binding between cells via M-1;
The present invention has been completed. That is, the present invention relates to the general formula (1)[Wherein, R¹Is a hydrogen atom, lower alkyl having 1 to 4 carbon atoms
X is an oxygen atom, a lower alkyl group having 1 to 3 carbon atoms.
An amino group which may be substituted, Y is a methoxy group;
Optionally substituted benzene ring, 1,3-thiazole ring
And Z is-(CH ₂) N- (n represents an integer of 0 to 3)
), -HC = CH-, -OCH₂-Is R²Is replaced
A lower alkyl group having 1 to 3 carbon atoms which may be
A phenyl group which may be substituted with a cyclocarbonyl group,
R³Represents a hydrogen atom, or R²And R³Are joined and replaced
Piperazine ring, sulfur atom is oxidized
Thiomorpholine ring, optionally substituted piperi
Which represents a gin ring].
Benzothiazole derivatives and pharmacologically acceptable salts,
And at least one of them as an active ingredient
Cell adhesion inhibitor. Table represented by the general formula (1) in the present invention
The pharmacologically acceptable salts of the compounds that are
Hydrobromide, methanesulfonate, citrate, tartar
Acid addition salts such as acid salts. In addition, one of the present invention
In general formula (1), a lower alkyl group, a lower alkyl group
An optionally substituted amino group, which may be substituted
Lower alkyl groups such as lower alkyl groups having 1 to 4 carbon atoms
"Group" means, for example, methyl, ethyl, propyl, isopropyl
Linear or branched hydrocarbons having 1 to 4 carbon atoms such as pills
And represents "an optionally substituted lower alkyl having 1 to 4 carbon atoms."
A `` alkyl group '' is an ethoxy group at any position of a lower alkyl group.
Xycarbonyl, hydroxy, phenyl, hydro
Xyphenyl group, ethoxyphenyl group, dimethoxyphen
Nyl group, lower alkoxyaminocarboni having 1 to 4 carbon atoms
Group, lower alkylaminocarbonyl having 1 to 4 carbon atoms
Group, 1,3-thiazolidinedione ring, 1,3-oxazolyl
Those having a zindion ring are exemplified. Also, "Replace
"An optionally substituted piperazine ring" refers to one nitrogen atom
A lower aliphatic acyl group having 1 to 4 carbon atoms,
Lower alkoxycarbonyl group, lower alkyl having 1 to 4 carbon atoms
Those having a killsulfonyl group or methylene chains in the ring
And those which have become a rubonyl group.
Or a good piperidine ring "is a lower alcohol having 1 to 4 carbon atoms.
Those having a xycarbonyl group are exemplified. In the present invention
According to the compound represented by the general formula (1),
Compound represented by formula (2) and compound represented by formula (3)
It can be produced by condensing a product.[Wherein, R¹Is a hydrogen atom, lower alkyl having 1 to 4 carbon atoms
X is an oxygen atom, a lower alkyl group having 1 to 4 carbon atoms.
An amino group which may be substituted, Y is a methoxy group;
Optionally substituted benzene ring, 1,3-thiazole ring
And Z is-(CH ₂) N- (n represents an integer of 0 to 3)
), -HC = CH-, -OCH₂-, R⁴Is hydroxy
Represents a halogen group or a halogen atom][R²Is a lower alkyl having 1 to 3 carbon atoms which may be substituted
May be substituted with a kill group or an ethoxycarbonyl group
A phenyl group is represented by R³Represents a hydrogen atom, or R²And R³But
A piperazine ring which may be bonded and substituted, and a sulfur atom
Optionally oxidized thiomorpholine ring, substituted
A piperidine ring which may be represented by the following formula:⁴Is a halogen atom
In the case of chloride, the use of organic bases such as triethylamine and pyridine
Methylene chloride, 1,4-dioxane, etc. in the presence
And can be carried out at 0 ° C. to room temperature. Also, the formula
R of (2)⁴Is a hydroxy group, the usual peptide bond
Mixed acid anhydride method and active ester used in the formation reaction
Process, preferably using a condensing agent.
Is the most appropriate method. The reaction is dicyclohexylcar
Bodiimide (DCC), diisopropylcarbodiimide
(DIPC), diphenylphosphonyl azide (DPP)
A), diethylphosphonyl cyanide (DEPC), 1-
Ethyl-3- (3-dimethylaminopropyl) -carbo
In the presence of a condensing agent such as diimide (WSC)
Uses 4-dimethylaminopyridine (DMAP) as a catalyst
And the reaction solvent is tetrahydrofuran (TH
F), methylene chloride, dimethyl sulfoxide (DMS
O), dimethylformamide (DMF), etc.
The temperature can be from 0 ° C. to room temperature. Also,
In the compound represented by the general formula (1), R²And R³But
If the thiomorpholine ring is an oxidized sulfur atom,
General formula (5)[Wherein, m represents an integer of 1 or 2;¹, X, Y,
Z is as described above], and the compound represented by the general formula (4)
Can also be produced by oxidizing the compound
it can.[Wherein, R¹, X, Y, and Z are as described above.] The reaction is the oxidation of aqueous hydrogen peroxide and m-chloroperbenzoic acid.
Using a solvent such as methylene chloride and DMF in the presence of
C. to room temperature. Furthermore, the above general formula
In the compound represented by (1), R²And R³Is piperazine
When it is a ring, that is, the general formula (7)[Wherein, R¹, X, Y, and Z are as described above]
The compound is obtained by converting the compound represented by the general formula (6) in the presence of an acid.
It can also be produced by hydrolysis.[Wherein, R⁵Represents a lower alkyl group having 1 to 4 carbon atoms;
R¹, X, Y, and Z are as described above.] The reaction was conducted using ethanol containing hydrochloric acid and 1,4-dioxane containing hydrochloric acid.
Organic solvent containing hydrochloric acid such as sun and hydrochloric acid-containing ethyl acetate
In acetic acid-hydrochloric acid mixture, or trifluoroacetic acid
May be used also as a solvent.
Can be. In the compound represented by the general formula (1), R²
And R³Is substituted with a lower alkylsulfonyl group.
In the case of a perazine ring, that is, the general formula (9)[Wherein, R⁶Represents a lower alkyl group having 1 to 4 carbon atoms;
R¹, X, Y, and Z are as described above],
The compound represented by the general formula (7) and the compound represented by the general formula (8)
Can also be produced by condensing the compound
it can.[Wherein, R⁶The reaction is as described above.
In the presence, methylene chloride, DMSO, 1,4-dioxane,
The reaction can be performed in a solvent such as DMF at 0 ° C. to room temperature.
In the compound represented by the general formula (1), R²And R³But lower grade fat
Substituted with an aliphatic acyl group or a lower alkoxycarbonyl group
Where the piperazine ring is represented by the general formula (11)[Wherein, R⁷Is a lower alkyl group having 1 to 4 carbon atoms,
1 to 4 lower alkoxy groups;¹, X, Y, Z
As described above], the compound represented by the general formula (7)
The compound represented by the general formula (10)
It can also be manufactured by combining them.[Wherein, R⁷The reaction is as described above.
In the presence, methylene chloride, DMSO, 1,4-dioxane,
It can be carried out at 0 ° C. to room temperature using a solvent such as DMF.
You.

Next, the present invention will be described with reference to specific examples.
The present invention is not limited by these examples.Reference Example 1  4-[(benzothiazol-2-yl) amino] benzoic acid
Ethyl acid2-chlorobenzothiazole (2.48 g), 4-amino ammonium
Ethyl benzoate (2.00g) mixture at 120 ° C for 3 hours
The mixture was heated and stirred. After cooling, add THF and saturated bicarbonate to the reaction mixture.
Add an aqueous solution of lithium and heat to dissolve the solidified crystals.
Was. Extract with ethyl acetate and wash the organic layer with saturated saline
Then, it was dried over anhydrous sodium sulfate. The solvent is distilled off under reduced pressure
Thereafter, the obtained powder was washed with hot ethyl acetate, and the desired product (0.
96g) was obtained as a colorless powder.Reference example 2  3- [4- (benzothiazol-2-yl) aminophen
Nyl] propionic acid ethyl ester2-chlorobenzothiazole and 3- (4-aminophenyl
G) Same as Reference Example 1 using propionic acid ethyl ester
The desired product was obtained as pale yellow needles. Melting point 8
6.5-87.5 ° C (isopropyl ether) Reference Example 3  4- (benzothiazol-2-yl) phenylacetic acid ethyl
Ruster4-aminophenylacetic acid ethyl ester (2.99 g) and 2
-Chlorobenzothiazole (3.12 g) in DMF (30 ml)
And added with paratoluenesulfonic acid (0.32 g).
Stirred at 10 ° C. for 3 hours. After cooling, the reaction solution was
The mixture was poured into a sodium aqueous solution and extracted with ethyl acetate. Organic
After the layer was dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure.
And the residue is subjected to silica gel column chromatography (development
Solvent: methylene chloride: ether = 30: 1)
Was. The desired product (3.93 g) was obtained as a light brown powder. Isop
Recrystallization from propyl ether gave pale brown needles. Fusion
Point 123-124 ° C Reference example 44-[(benzothiazol-2-yl) amido
No] ethyl phenyloxyacetate2-chlorobenzothiazole (19.2 g) and 4-aminophen
1,3-dimethyl of ethyl phenyloxyacetate (22.0 g)
-2-Imidazolidinone (DMI) (300 ml)
Add ridinium paratoluenesulfonic acid (2.83g),
The mixture was stirred at 140 ° C. for 2 hours. Water and saturated carbonated water in the reaction solution
An aqueous sodium hydrogen chloride solution was added, and the mixture was extracted with ethyl acetate. Yes
The organic layer was washed with water and saturated saline and then anhydrous sodium sulfate.
Dried. After evaporating the solvent under reduced pressure, silica gel column chromatography
Chromatography (developing solvent n-hexane: ethyl acetate =
The crystals obtained by purification in 2: 1) are washed with methanol,
The desired product (21.5 g) was obtained as a colorless powder.Reference Examples 5 to 10  The following compounds were synthesized in the same manner as in Reference Example 4.
1). Reference Example 11  4-[(benzothiazol-2-yl) oxy] phenyl
Ethyl acetate4-hydroxyphenylacetic acid ethyl ester (1.79
g), DMF of 2-chlorobenzothiazole (1.73 g)
(30ml) 60% sodium hydride oily solution (0.70g)
Was added and stirred at 90 ° C. for 1 hour. After cooling, pH 3 phosphoric acid
Adjust the pH to 7 by adding a buffer, dilute with water and extract with ethyl acetate.
did. After washing the organic layer with saturated saline, anhydrous sodium sulfate
Dried. The solvent is distilled off under reduced pressure.
Ram chromatography (Developing solvent n-hexane: vinegar
(Ethyl acetate = 4: 1) to give the desired product (1.65 g) as colorless
Obtained as an oil.Reference Example 12  N-benzoyl-N '-(6-methylbenzothiazole
-2-yl) -thioureaAmmonium thiocyanate (3.80 g) in THF (100 ml)
Under stirring at room temperature, benzoyl chloride (7.00 g) was added to the solution.
The mixture was heated under reflux for 5 minutes (benzoylisothio
Preparation of cyanate). 2-Amino-6-methyl
Add benzothiazole (8.20 g) and heat for an additional hour
Refluxed. The solvent was distilled off under reduced pressure, and water was added to the residue to precipitate.
The crystals were collected by filtration. The crystals are washed with hot ethanol,
(7.71 g) as a pale yellow powder.Reference Example 13  N-benzoyl-N '-(benzothiazol-2-i
Le) ThioureaAs in Reference Example 12, using 2-aminobenzothiazole
The target product was obtained as pale yellow needles.Reference Example 14  2-[(6-methylbenzothiazol-2-yl) amido
[No] -1,3-thiazole-4-acetic acid ethyl esterCompound of Reference Example 12 (7.52 g), lithium hydroxide monohydrate
(5.58 g) dissolved in water (70 ml) and heated under reflux for 20 minutes
did. After cooling, the reaction solution was adjusted to pH 1 by adding dilute hydrochloric acid.
After adjusting the pH to 10 with ammonia water, heat it on a water bath and let it cool.
Was. The precipitated crystals were collected by filtration and subjected to ethyl acetate-THF-DM.
It was recrystallized from F-isopropyl ether. N- (6-
Methylbenzothiazol-2-yl) thiourea (1.37
g) was obtained as colorless needles. The resulting thiourea (1.30
g) was dissolved in THF (50 ml) and 4-chloroacetoacetic acid was dissolved.
Ethyl (5.00 g) and DMAP (0.05 g) were added, and 20 hours
Heated to reflux. The solvent was distilled off under reduced pressure and the obtained powder was saturated.
Add aqueous sodium bicarbonate solution and extract with ethyl acetate.
Was. After washing the organic layer with saturated saline, anhydrous sodium sulfate
And the solvent was distilled off under reduced pressure. The obtained crystals are washed with THF
-From isopropyl ether-isopropyl alcohol
Recrystallize to give the desired product (0.60 g) as milky white needles.
Was.Reference Example 15  2-[(benzothiazol-2-yl) amino] -1,
3-thiazole-4-ethyl acetateUsing the compound of Reference Example 13, hydrolysis was performed in the same manner as in Reference Example 14.
N- (benzothiazol-2-yl) thiourea
Obtained as a colored powder. Further ethyl 4-chloroacetoacetate
And the desired product was obtained as a colorless powder.Reference Example 16  2-[(benzothiazol-2-yl) amino] -1,
Ethyl 3-thiazole-4-carboxylateN- (benzothiazo) obtained in the middle of the synthesis of Reference Example 15
2-chloro) thiourea and 4-chloroacetovinegar
Use ethyl bromopyruvate instead of ethyl acrylate
Reaction was carried out in the same manner as in Example 14 to obtain the desired product as a colorless powder.
Was.Reference Example 17  4-[(benzothiazol-2-yl) amino] phenyl
Luoxyacetic acidIn ethanol (300 ml), the compound of Reference Example 4 (21.5 g), 1
A 0% aqueous sodium hydroxide solution (150 ml) was added, and 90 minutes
The mixture was refluxed while heating. After cooling, pour into ice water and adjust to pH 4 with dilute hydrochloric acid.
The precipitated crystals were collected by filtration. Filter the crystal with ethanol
The mixture was pulverized well in a filter, filtered again, and dried. Object
(16.7 g) was obtained as a colorless powder.Reference Examples 18 to 30  The compounds shown in Table 2 were synthesized in the same manner as in Reference Example 17. Example 1  3- [4- (benzothiazol-2-yl) aminoben
Zoyl] aminopropionic acid ethyl esterCompound of Reference Example 18 (0.79 g), β-alanine ethyl ether
Trihydrochloride (0.80 g) in DMF (40 ml)
Ethylamine (1.50 g), WSC (1.35 g), DMAP
(0.05 g), and the mixture was stirred at room temperature for 18 hours. phosphoric acid
After adding a buffer, the mixture was extracted with ethyl acetate, and the organic layer was saturated.
After washing with brine, the extract was dried over anhydrous sodium sulfate. solvent
Was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography.
Crystals obtained after purification with ethyl acetate (developing solvent: ethyl acetate)
With isopropyl ether-isopropyl alcohol-T
Recrystallize from HF to give the desired product (0.45 g) as colorless needles
I got it. 145-146 ° C Examples 2 to 48  The reaction was carried out using the compound of Reference Example in the same manner as in Example 1 to obtain a table.
Compound 3 was synthesized. Example 49  N- [4- (benzothiazol-2-yl) aminophen
Nylacetyl] piperazine dihydrochlorideThe compound of Example 41 (1.14 g) was treated with acetic acid (30 ml) and concentrated hydrochloric acid.
(30 ml) and stirred at room temperature for 1 hour. Dissolution
The solvent was distilled off under reduced pressure, dilute hydrochloric acid was added to the residue, and the solvent was distilled off again under reduced pressure.
Thereafter, methanol was added and the solvent was further distilled off under reduced pressure. Remaining
After adding ether to crystallize, the ether was decompressed.
After distilling off, the obtained crystals are thoroughly washed with ethanol and collected by filtration.
did. The desired product (0.74 g) was obtained as a colorless powder. Melting point 1
69-172 ℃ Elemental analysis value (%): C₁₉H₂₀N₄OS 2HCl 9 / 5H₂As O, CHN Calculated: 49.85 5.64 12.24 Actual: 49.85 5.61 12.21Example 50  N- [4- (benzothiazol-2-yl) aminophen
Nylacetyl] -N'-isopropoxycarbonylpipe
RazineDMF (3 ml) was added to the compound of Example 49 (0.15 g).
Triethylamine (3 ml), isopropyl chloroformate (0.0
6 ml) and stirred at room temperature for 8 hours. Ice water
And extract with ethyl acetate.
Dried over sodium hydrogen sulfate. After distilling off the solvent under reduced pressure,
The residue is subjected to silica gel column chromatography (developing solvent).
 Purification with n-hexane: ethyl acetate = 1: 2)
(0.11 g) as a colorless powder. Melting point 185-186 ° C Example 51  N- [4- (benzothiazol-2-yl) aminophen
Nylacetyl] -N'-methanesulfonylpiperazineDissolve the compound of Example 49 (0.20 g) in pyridine (2 ml)
And add methanesulfonyl chloride (0.04 ml)
For 6 hours. After evaporating the solvent under reduced pressure, ethyl acetate
And washed with water and saturated saline in this order. Organic layer
After drying over anhydrous sodium sulfate, the residue was distilled off under reduced pressure.
Kagel column chromatography (developing solvent
Purification with ren: methanol = 50: 1) gave the desired product (0.15
g) was obtained as a colorless powder. 203-205 ° C Example 52N- [4- (benzothiazol-2-yl)
Aminophenylacetyl] -1,4-thiomorpholine-
S-oxideCompound of Example 43 (1.19 g) in methylene chloride (30 l)
Dissolve and stir with ice-cooling, m-chloroperbenzoic acid (0.67 g)
Was added little by little, and the mixture was stirred at the same temperature for 8 hours. Salt in the reaction solution
Add methylene chloride, add saturated aqueous sodium bicarbonate solution, water
After washing in this order, the organic layer was dried over anhydrous sodium sulfate.
Was. After evaporating the solvent under reduced pressure, the desired product (1.24 g) was converted to a colorless powder.
I got it. When recrystallized from methanol,
Zum crystals were obtained. 213-215 ° C Example 53  N- [4- (benzothiazol-2-yl) aminophen
Nylacetyl] -1,4-thiomorpholine-S, S-dio
OxideCompound (0.58 g) of the above Example 52 was added to DMF (15 ml).
Dissolve and add m-chloroperbenzoic acid (0.31 g) under ice-cooling and stirring.
added. After stirring at the same temperature for 30 minutes, the mixture was further
Stirred for hours. The reaction solution was poured into water and extracted with ethyl acetate.
After that, the organic layer is washed with a saturated aqueous solution of sodium hydrogen carbonate and water in that order.
And dried over anhydrous sodium sulfate. Decompress the solvent
Evaporation gave the desired product (0.50 g) as a pale yellow powder. D
The crystals were recrystallized from MF-water to obtain light brown prism crystals. Melting point
265-267 ° CNext, results supporting the usefulness of the compound of the present invention were obtained.
Shown by experimental examples. Human vascular endothelial cells and U937 cells
Of test compounds on adhesion to human (monocyte cell line)
Harmful effects Human umbilical vein-derived vascular endothelial cells (HUVECs) were
By stimulating with interleukin-1 (IL-1)
Adhesion of ICAM-1, VCAM-1, ELAM-1, etc.
Expression of the offspring is induced. 24 hours after stimulation, mainly IC
AM-1 and VCAM-1 expression is observed (J. Immun
ol., 144, 2558 (1990), ibid., 149, 698 (1992)). I
Cell adhesion using HUVEC stimulated with L-1 for 24 hours
By conducting the test, ICAM-1 and VCAM-1
The adhesion reaction can be tested. In addition, when the test compound is added
The period between the stimulation of HUVEC with IL-1 and the HUVEC
Test by separating C and U937
The adhesion inhibition effect of the compound is mainly due to the inhibition of the binding of the adhesion molecule.
Whether it is due to suppression of adhesion molecule expression
it can.Experimental example Adhesion molecule binding inhibition test 20% fetal bovine serum and 10 ng / ml vascular endothelial cell expansion
Floating on M199 medium (for culture) containing growth factor (ECGF)
96-well collagen-coated plate
(Flat bottom) 2 × 10⁴/ Well, seeded at 37 ° C, 5
% CO₂Cultured underneath. After culturing for about 24 hours, fetal bovine blood
M199 medium without washing and ECGF (for washing)
The HUVEC was washed twice. Next, human interleukin
M1 for culture containing 10 U / ml of -1β (IL-1β)
The cells were cultured in 99 medium for 24 hours. On the other hand, U937 cell suspension
Liquid (1 × 10⁷/ Ml) 1 mM BCE per ml
10 μl of CF-AM solution (Wako Pure Chemical Industries) was added,
The mixture was incubated for 1 hour under ice-cooling and fluorescently labeled. fluorescence
Label U937 cells with phosphate buffered saline (PBS)
After washing twice with (-)), RP containing 10% fetal bovine serum
Floated in MI-1640 medium (1 × 10⁷/ Ml).
HUVEC were washed three times with M199 medium for washing. test
Dissolve the compound in dimethyl sulfoxide for further culture
80 μl / 1000-fold diluted with M199 medium
Each well was added. Subsequently, fluorescence-labeled U937 cell suspension
The solution was added at 20 μl / well (final of test compound).
Concentration 50 μM). 1000 rpm, room temperature, centrifugation for 1 minute
After that, 37 ° C, 5% CO₂Incubated for 30 minutes under. Each way
Was washed twice with 100 μl of PBS (-), and
Cells were removed. 0.1% sodium dodecyl sulfate aqueous solution
Add 100 μl / well of solution to solubilize cells
Was. Measure the fluorescence intensity of each well (Excitation 490nm,
Emission 530nm), the number of U937 cells adhered from the calibration curve
I asked. The adhesion inhibition rate was calculated according to the following equation.Table 4 shows the results. Note that the suppression rate exceeding 100%
And 100%.As described above, the compounds of the present invention have excellent cell adhesion inhibition.
Production use was approved.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 31/54 ADU A61K 31/54 ADU C07D 277/68 C07D 277/68 417/12 211 417/12 211 241 241 263 263 (72) Inventor Yasuhiko Sakoe 750-3 F-Term, Inabago, Oyama City, Tochigi Pref. ZB11 ZB13 ZB26 ZC02

Claims (7)

[Claims] 1. The general formula (1)[Wherein, R¹Is a hydrogen atom, lower alkyl having 1 to 4 carbon atoms
X is an oxygen atom, a lower alkyl group having 1 to 4 carbon atoms.
An amino group which may be substituted, Y is a methoxy group;
Optionally substituted benzene ring, 1,3-thiazole ring
And Z is-(CH ₂) N- (n represents an integer of 0 to 3)
), -HC = CH-, -OCH₂-Is R²Is replaced
A lower alkyl group having 1 to 4 carbon atoms which may be
A phenyl group which may be substituted with a cyclocarbonyl group,
R³Represents a hydrogen atom, or R²And R³Are joined and replaced
Piperazine ring, sulfur atom is oxidized
Thiomorpholine ring, optionally substituted piperi
Which represents a gin ring].
Nzothiazole derivatives and pharmacologically acceptable salts. 2. The general formula (1)[Wherein, R¹Is a hydrogen atom, lower alkyl having 1 to 4 carbon atoms
X is an oxygen atom, a lower alkyl group having 1 to 4 carbon atoms.
An amino group which may be substituted, Y is a methoxy group;
Optionally substituted benzene ring, 1,3-thiazole ring
And Z is-(CH ₂) N- (n represents an integer of 0 to 3)
), -HC = CH-, -OCH₂-Is R²Is replaced
A lower alkyl group having 1 to 4 carbon atoms which may be
A phenyl group which may be substituted with a cyclocarbonyl group,
R³Represents a hydrogen atom, or R²And R³Are joined and replaced
Piperazine ring, sulfur atom is oxidized
Thiomorpholine ring, optionally substituted piperi
Benzothia]
At least one of a sol derivative and a pharmacologically acceptable salt.
A cell adhesion inhibitor comprising one or more active ingredients. 3. The general formula (2)[Wherein, R¹Is a hydrogen atom, lower alkyl having 1 to 4 carbon atoms
X is an oxygen atom, a lower alkyl group having 1 to 4 carbon atoms.
An amino group which may be substituted, Y is a methoxy group;
Optionally substituted benzene ring, 1,3-thiazole ring
And Z is-(CH ₂) N- (n represents an integer of 0 to 3)
), -HC = CH-, -OCH₂-Is R⁴Is hydro
Represents a xy group or a halogen atom].
General formula (3)[Wherein, R²Is an optionally substituted low carbon number of 1 to 4
Substituted with a lower alkyl group or an ethoxycarbonyl group
A phenyl group represented by R³Represents a hydrogen atom, or R²When
R³May be bonded to an optionally substituted piperazine ring, sulfur
Thiomorpholine ring with optionally oxidized atoms, substituted
Represents an optionally substituted piperidine ring]
General formula (1) characterized by condensing[Wherein, R¹, R², R³, X, Y and Z are as described above]
A method for producing a compound represented by the formula: 4. The formula (4)[Wherein, R¹Is a hydrogen atom, lower alkyl having 1 to 4 carbon atoms
X is an oxygen atom, a lower alkyl group having 1 to 4 carbon atoms.
An amino group which may be substituted, Y is a methoxy group;
Optionally substituted benzene ring, 1,3-thiazole ring
And Z is-(CH ₂) N- (n represents an integer of 0 to 3)
), -HC = CH-, -OCH₂-Indicate]
General formula (5) characterized in that the compound is oxidized[Wherein, m represents an integer of 1 or 2;¹, X, Y,
Z is as described above]. 5. The general formula (6)[Wherein, R¹Is a hydrogen atom, lower alkyl having 1 to 4 carbon atoms
X is an oxygen atom, a lower alkyl group having 1 to 4 carbon atoms.
An amino group which may be substituted, Y is a methoxy group;
Optionally substituted benzene ring, 1,3-thiazole ring
And Z is-(CH ₂) N- (n represents an integer of 0 to 3)
), -HC = CH-, -OCH₂-Is R⁵Is the carbon number
Represents a lower alkyl group of 1 to 4].
Wherein the compound is hydrolyzed in the presence of[Wherein, R¹, X, Y, and Z are as described above]
Manufacturing method of compound. 6. General formula (7)[Wherein, R¹Is a hydrogen atom, lower alkyl having 1 to 4 carbon atoms
X is an oxygen atom, a lower alkyl group having 1 to 4 carbon atoms.
An amino group which may be substituted, Y is a methoxy group;
Optionally substituted benzene ring, 1,3-thiazole ring
And Z is-(CH ₂) N- (n represents an integer of 0 to 3)
), -HC = CH-, -OCH₂-Indicate]
Compound and general formula (8)[Wherein, R⁶Represents a lower alkyl group having 1 to 4 carbon atoms]
General formula characterized by condensing a compound represented by
(9)[Wherein, R¹, R⁶, X, Y, and Z are as described above].
Of producing a compound to be prepared. 7. The general formula (7)[Wherein, R¹Is a hydrogen atom, lower alkyl having 1 to 4 carbon atoms
X is an oxygen atom, a lower alkyl group having 1 to 4 carbon atoms.
An amino group which may be substituted, Y is a methoxy group;
Optionally substituted benzene ring, 1,3-thiazole ring
And Z is-(CH ₂) N- (n represents an integer of 0 to 3)
), -HC = CH-, -OCH₂-Indicate]
Compound and general formula (10)[Wherein, R⁷Is a lower alkyl group having 1 to 4 carbon atoms,
And represents a lower alkoxy group of 1 to 4].
General formula (11) characterized by being condensed[Wherein, R¹, R⁷, X, Y, and Z are as described above].
Of producing a compound to be prepared.