JP2001002665A - Thiazoline derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new thiazoline derivative which has an excellent action for inhibiting the adhesion of VCAM-1 on vascular endothelial cells to cells expressing VLA-4 and is useful for preventing and treating arteriosclerosis and so on. SOLUTION: A thiazoline derivative of formula I R1 is a group of formula II [(m) is 0 or 1; Z is N or CH; R10 to R12 are each H, a halogen, a 1 to 5C alkyl or the like], a group of formula III (Y is O, S or the like) or the like; R2 is H or a 1 to 5C alkyl; R3 and R4 are each a 1 to 14C alkyl or the like} or its pharmaceutically acceptable salt, for example, 3-isopentyl-4-methyl-2-[4-(3- trifluoromethylbenzamido) benzoylimino]-3H-thiazoline-5-carboxylic acid. The compound of formula I is obtained by successively reacting a compound of formula IV (R13 is a 1 to 5C alkyl) with sulfuryl chloride and thiourea and further successively with nitrobenzoyl chloride and an alkylating agent, reducing the product, and then reacting the product with a compound of formula V (W is OH or the like) or the like.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to novel thiazoline derivatives as inhibitors of leukocyte adhesion and migration and antagonists of cell adhesion molecules.

[0002]

2. Description of the Related Art The binding between VCAM-1, which is a cell adhesion factor expressed in various inflammatory tissues, and VLA-4, which is expressed on cell surfaces such as lymphocytes, monocytes, and eosinophils,
It is known that the infiltration of these cells into tissues is promoted, causing an exacerbation and chronicity of the inflammatory response (Ca
rdiovasc. Res., 1996; 32: 733). For this reason,
A compound that inhibits the adhesion between VCAM-1 and VLA-4 is
It is considered to be effective in treating various inflammatory diseases (J. Clin. Invest., 1993; 92: 538; Clin. Exp.
Allergy, 1998; 28: 1518, Lab. Invest., 19
91; 64: 313). A compound that inhibits the adhesion between VCAM-1 and VLA-4 is disclosed in WO96-22966, but has insufficient action.

[0003]

SUMMARY OF THE INVENTION An object of the present invention is to have an excellent effect of inhibiting adhesion between VCAM-1 and VLA-4.
An object of the present invention is to provide a novel thiazoline derivative.

[0004]

Means for Solving the Problems As a result of intensive studies, the present inventors have found that a certain thiazoline derivative can achieve the above object, and have completed the present invention. That is, the present invention uses the formula

[0005]

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[Wherein R ¹ is a formula

[0007]

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(Wherein m represents 0 or 1, Z represents a nitrogen atom or CH, and R ¹⁰ , R ¹¹ and R ¹² are the same or different and are a hydrogen atom, a halogen atom, a carbon atom number of 1 to 1).
5 alkyl groups, nitro groups, amino groups, 1 carbon atom
An amino group, a hydroxyl group, a cyano group, an alkyl group having 1 to 3 carbon atoms, which is substituted with 1 to 3 alkyl groups,
Represents an alkoxy group, a phenyl group or a carboxyl group. ), A group represented by the formula

[0009]

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Wherein m is as defined above, and Y represents an oxygen atom, a sulfur atom, —NCH ₃ or NH, and an alkyl group having 1 to 14 carbon atoms. Represents a cycloalkyl group having 3 to 6 carbon atoms or an adamantyl group, R ² represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and R ³ and R ⁴ are the same or different. Alkyl group having 1 to 14 carbon atoms, 3 to 6 carbon atoms
A cycloalkyl group, a cycloalkylmethyl group having 4 to 7 carbon atoms, a cycloalkylethyl group having 5 to 8 carbon atoms, a phenyl group, or a phenylalkyl group having 7 to 10 carbon atoms. Or a pharmaceutically acceptable salt thereof.

In the present invention, the term "alkyl group" used by itself or as a part of a group means a straight-chain or branched-chain alkyl group. And methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl, isopentyl, and the like. ,
Examples include hexyl, isohexyl, heptyl, octyl, decyl, and tetradecyl groups. Further, the cycloalkyl group having 3 to 6 carbon atoms is a cyclopropyl group, a cyclobutyl group, a cyclopentyl group or a cyclohexyl group. The halogen atom is a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.
The “amino group substituted with one or two alkyl groups having 1 to 3 carbon atoms” is preferably an amino group substituted with a methyl group, and more preferably a dimethylamino group. "The alkyl group having 1 to 3 carbon atoms substituted with 1 to 3 halogens" is preferably an alkyl group substituted with a fluorine atom, more preferably a methyl group substituted with a fluorine atom. And most preferably a trifluoromethyl group. The alkoxy group having 1 to 5 carbon atoms is a linear or branched one.
Preferred are a methoxy group and an ethoxy group. The phenylalkyl group having 7 to 10 carbon atoms includes a benzyl group, a phenethyl group, a phenylpropyl group and the like. Formula (I)
The pharmaceutically acceptable salts of the compounds of the above are salts with alkali metals, alkaline earth metals, ammonium, alkylammonium and the like. They are, for example, sodium, potassium, calcium, ammonium, aluminum, triethylammonium salts and the like.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION The compound of the present invention can be synthesized by the following method. That is, the compound of the present invention can be obtained, for example, by the following formula (a) obtained by the method described in Org. Synth. Coll., Vol. 7, page 359.

[0013]

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(Wherein R ⁴ has the same meaning as described above, and R ¹³ represents R ² other than a hydrogen atom). The compound is reacted with sulfuryl chloride at 0 ° C. to 20 ° C. in the absence of a solvent. The following formula (b)

[0015]

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(Wherein R ⁴ and R ¹³ have the same meanings as described above) by reacting the compound with thiourea in a solvent to give the following formula (c):

[0017]

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(Wherein R ⁴ and R ¹³ are as defined above) or a salt thereof.

Next, the compound represented by the following formula (d) obtained by reacting the compound represented by the formula (c) or a salt thereof with nitrobenzoyl chloride in the presence of a base:

[0020]

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(Wherein R ⁴ and R ¹³ have the same meanings as described above) by the method described in JP-A-8-199553, that is, R ³ -X (where X is a halogen) An alkylating agent such as a halide represented by the formula (e), wherein R ³ is the same as defined above, in the presence of a base.

[0022]

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(Wherein R ³ , R ⁴ and R ¹³ have the same meanings as described above).

The compound represented by the formula (e) can be produced as an alternative by the following method. That is, for example, the following formula (f) obtained by the method described in Org. Synth. Coll., Vol. 3, page 735.

[0025]

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(Wherein R ³ has the same meaning as described above) by reacting the compound represented by the formula (b) with the compound represented by the formula (b) by the method described in JP-A-8-199553. The compound represented by (e) can be obtained.

Next, the compound represented by the formula (e) is reduced by a commonly used reduction method by a commonly used method to obtain the following formula (g):

[0028]

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(Wherein R ³ , R ⁴ and R ¹³ have the same meanings as described above). Next, this compound is represented by the following formula (I)

[0030]

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(Wherein, m, R ¹⁰ , R ¹¹ and R ¹² have the same meanings as described above, and W represents a hydroxyl group or a halogen atom.)
Or a compound represented by the following formula (m)

[0032]

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(Wherein m, W and Y have the same meanings as defined above), and R ² is amidated by a general method for forming an amide bond, so that R ² has 1 to 5 carbon atoms. Of the present invention, which is an alkyl group of

The invention compounds wherein R ² is a hydrogen atom, R ²
Is an alkyl group having 1 to 5 carbon atoms, which can be obtained by ester hydrolysis.
For the hydrolysis of the ester, a general method such as an alkali treatment, a mineral acid treatment or an organic acid treatment can be used. When a base is used in the above reaction, examples of the base include alkali metals such as sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, sodium hydroxide, dimesyl sodium, sodium hydride, sodium amide, and tertiary butyl potassium. Salts, amines such as triethylamine, diisopropylethylamine, and pyridine; sodium acetate, potassium acetate, and the like can be used. The acid is, for example, acetic acid, methanesulfonic acid, p-toluenesulfonic acid and the like. Water, methanol, ethanol, isopropyl alcohol, tertiary
Alcohols such as butyl alcohol, ethers such as dioxane and tetrahydrofuran, dimethylformamide, dimethylsulfoxide, pyridine, methylene chloride,
Inert solvents such as chloroform, acetone, and acetic acid can be used for the reaction.

[0035]

The compound of formula (I) thus obtained has an action of inhibiting adhesion between VCAM-1 and VLA-4 expressing cells on vascular endothelial cells. Therefore, the compound of the present invention can be used as an agent for preventing and treating arteriosclerosis and various acute and chronic inflammatory diseases.

For this purpose, the compound of formula (1) can be prepared as an oral drug or an injection by a conventional formulation technique by adding a conventional bulking agent, pH adjuster, solubilizer and the like. . This dose depends on the type of disease, the age of the patient,
It can be increased or decreased as appropriate according to body weight and symptoms.

[0037]

The present invention will be described in more detail with reference to the following examples. Example 1 3-Isopentyl-4-methyl-2- [4- (3-trifluoromethylbenzamido) benzoylimino] -3
Synthesis of H-thiazoline-5-carboxylic acid (1) To a solution of ammonium thiocyanate (45.6 g) in acetone (700 ml) was added 4-nitrobenzoyl chloride (111.3 g), and the mixture was stirred at room temperature for 30 minutes. After filtering off the insoluble matter, the filtrate was concentrated under reduced pressure to obtain 4-nitrobenzoyl isothiocyanate (122.5 g).

(2) Isoamylamine (11.6 ml) was added to a mixture of 4-nitrobenzoyl isothiocyanate (20.8 g) and toluene (300 ml) to obtain a mixture.
Heated to reflux for minutes. Ethyl 2-chloroacetoacetate (27.6 ml) was added to the mixture, and the mixture was heated under reflux for 3 hours while removing water produced by the reaction. After allowing the reaction mixture to cool, the precipitated crystals were filtered, washed with toluene, and yellow needles of 3-isopentyl-4-methyl-2- (4-nitrobenzoylimino) -3H-thiazoline-5-carboxylic acid were obtained. Ethyl (29.6 g) was obtained. 166-167 ° C.

(3) 3-Isopentyl-4-methyl-2
Ethyl-(4-nitrobenzoylimino) -3H-thiazoline-5-carboxylate (2.0 g) was added to ethanol (1
00 ml), ammonium formate (1.8 g) and 10% palladium on carbon (0.2 g) were added, and the mixture was stirred at room temperature for 1 hour and 30 minutes. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in ethyl acetate and washed with water and saturated saline. After drying the organic layer over magnesium sulfate,
The solvent was distilled off to obtain a yellow powder of ethyl 2- (4-aminobenzoylimino) -3-isopentyl-4-methyl-3H-thiazoline-5-carboxylate (1.9 g). Mp 200.0-201.0 ° C (decomposition).

(4) Ethyl 2- (4-aminobenzoylimino) -3-isopentyl-4-methyl-3H-thiazoline-5-carboxylate (1.9 g) was dissolved in methylene chloride (100 ml), and triethylamine ( 0.76
ml) and 3- (trifluoromethyl) benzoyl chloride (1.1 g) were added, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with ethyl acetate and washed with 1 N hydrochloric acid and saturated saline. After drying the organic layer over anhydrous magnesium sulfate,
The solvent was distilled off. The obtained crude crystals were recrystallized from ethyl acetate to give 3-isopentyl-4-methyl-2-colorless needles.
Ethyl [4- (3-trifluoromethylbenzamido) benzoylimino] -3H-thiazoline-5-carboxylate (2.1 g) was obtained. 228.5-230.0 [deg.] C.

(5) 3-Isopentyl-4-methyl-2
Ethyl-[4- (3-trifluoromethylbenzamido) benzoylimino] -3H-thiazoline-5-carboxylate (2.1 g) was added to a mixed solvent of tetrahydrofuran (hereinafter abbreviated as THF) (20 ml) and methanol (20 ml). Dissolve and add 10% aqueous sodium hydroxide solution (7.3m
l) was added and the mixture was heated under reflux for 20 minutes. The solvent was distilled off under reduced pressure, and a 7% aqueous citric acid solution was added, followed by extraction with ethyl acetate. The organic layer was washed with saturated saline, dried over anhydrous magnesium sulfate, and the solvent was distilled off. The obtained crude crystals were recrystallized from ethyl acetate to give the title compound (Compound 1 in Table 1).

Example 2 The same operation as in Example 1 was performed to obtain the compounds shown in Table 1.

[0043]

[Table 1]

Example 3 4-Isobutyl-3-methyl-2- [4- (3-trifluoromethylbenzamido) benzoylimino] -3H
Synthesis of -thiazoline-5-carboxylic acid (1) Sulfuryl chloride (11.3 ml) was added dropwise to methyl isovaleryl acetate (21.1 g) at 0 ° C over 15 minutes. After raising the temperature to room temperature and stirring for 1 hour, the pressure was reduced to remove dissolved gas. Next, ethanol (200 ml)
And thiourea (11.2 g) were added, and the mixture was heated under reflux for 2.5 hours. After ethanol was distilled off under reduced pressure, water and aqueous ammonia were added, and the resulting precipitate was collected by filtration. This was washed with water to obtain a pale yellow powder of methyl 2-amino-4-isobutylthiazole-5-carboxylate (22.4 g). 161.0-164.0 ° C.

(2) Methyl 2-amino-4-isobutylthiazole-5-carboxylate (21.4 g) was dissolved in pyridine (150 ml), 4-nitrobenzoyl chloride (17.7 g) was added, and the mixture was added at room temperature for 1 hour. Stirred. After distilling off pyridine under reduced pressure, 1N hydrochloric acid was added and the resulting precipitate was collected by filtration. This was washed with 1N hydrochloric acid and water, and methyl 4-isobutyl-2- (4-nitrobenzamide) thiazole-5-carboxylate (34.2 g) was obtained as a pale yellow powder.
I got 218.0-219.0 ° C (dec).

(3) 60% oily sodium hydride (4.
0 g) in DMF (300 ml) at 0 ° C. with 4-methyl-2- (4-nitrobenzamido) thiazole-5.
-Methyl carboxylate (33.0 g) was added, and the mixture was stirred for 1 hour while gradually warming to room temperature. Methyl iodide (5.6 ml) was added to the reaction mixture, and the mixture was stirred for 1 hour. Methyl iodide (5.6 ml) was added, and the mixture was further stirred for 1 hour. The reaction mixture was poured into 1N hydrochloric acid containing ice chips, and the resulting precipitate was collected by filtration. This is washed with water, hexane and toluene,
Light brown powder of 4-isobutyl-3-methyl-2- (4-nitrobenzoylimino) -3H-thiazoline-5-carboxylate (29.9 g) was obtained. 235.0-236.0 ° C.

(4) Using the compound obtained in the above (3),
The same operation as in (3) to (5) of Example 1 was performed to obtain the title compound (Compound 9 in Table 2).

Example 4 The same operation as in Example 3 was performed to obtain the compounds shown in Table 2.

[0049]

[Table 2]

Hereinafter, the inhibitory effect of the compound of formula (I) on the binding between VCAM-1 and VLA-4 will be described with reference to Test Examples.

Test Example (1) Human recombinant solubilized VCAM-1 protein (Funakoshi) was dissolved in bovine serum albumin solution (10 μg / ml in 50 mM sodium bicarbonate solution) to 1 μg / ml. This solution was applied to a flat bottom 96 well plate (Linbro T
itertech), dispensing 100 μl per well,
Incubated overnight at 4 ° C. After removing the solution in each well, the well was replaced with 1% BSA / PBS buffer, incubated at 37 ° C. for 1 hour to perform blocking, and then TBS solution (24 mM Tris-HCl,
Washed once with 137 mM NaCl, 2.7 mM KCl, 0.1% BSA, 2 mM glucose).

(2) Literature (J. Exp. Med., 1992; 17)
6:99.).
Amos cells (ATCC) were suspended in a TBS solution containing ₂ mM MnCl2.

(3) A solution obtained by diluting a dimethyl sulfoxide solution of the test drug to a specified concentration with a TBS solution was added to the plate prepared in (1) at 50 μl / well.
l was added. Subsequently, 50 μl (2 × 10 ⁵ cells) of the suspension prepared in (2) was added per well, and 5% C
After incubation at 37 ° C. for 45 minutes in an O ₂ incubator, the plate was washed five times with a TBS solution.

(4) 1% NP
Solubilized with -40, fluorescent plate reader (Millipore)
The number of Ramos cells remaining on the plate was measured by measuring the fluorescence intensity in the above, and the inhibition rate of cell adhesion by the test drug was calculated. From the inhibition rates at various concentrations of the test drug, the concentration (IC ₅₀ value) at which 50% adhesion was inhibited was calculated.
α4 integrin monoclonal antibody HP1 / 2 to VC
It was used as a positive control for AM-1 / VLA-4 binding inhibition.

(5) As a result of calculating the IC ₅₀ value of the compound of the present invention, the compound 1 was 2.3 μM.

Continued on the front page (51) Int.Cl. ⁷ Identification code FI Theme coat II (reference) C07D 417/12 C07D 417/12 F term (reference) 4C033 AD03 AD16 AD17 4C063 AA01 BB09 CC62 CC75 CC92 DD04 DD12 DD62 EE01 4C086 AA02 AA03 BC82 GA08 MA01 NA14 ZA45 ZB02 ZB11 ZB21

Claims (1)

[Claims] (1) Formula (1) Wherein R ¹ is of the formula (In the formula, m represents 0 or 1, Z represents a nitrogen atom or CH, and R ¹⁰ , R ¹¹ and R ¹² are the same or different and each represent a hydrogen atom, a halogen atom, a carbon atom having 1 to 5 carbon atoms. An alkyl group, a nitro group, an amino group, an amino group substituted with one or two alkyl groups having 1 to 3 carbon atoms, a hydroxyl group, a cyano group, and a carbon atom substituted with 1 to 3 halogen atoms; Represents an alkyl group having 1 to 3 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, a phenyl group or a carboxyl group). (Wherein, m has the same meaning as described above, and Y represents an oxygen atom, a sulfur atom, —NCH ₃ or NH.)
Alkyl group having 1 to 14 carbon atoms, 3 to 6 carbon atoms
Cycloalkyl groups or adamantyl groups, R ²
Represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and R ³ and R ⁴ are the same or different and each have 1 carbon atom.
１４14 alkyl groups, cycloalkyl groups having 3-6 carbon atoms, cycloalkylmethyl groups having 4-7 carbon atoms, cycloalkylethyl groups having 5-8 carbon atoms, phenyl groups or carbon atoms Shows several 7 to 10 phenylalkyl groups. Or a pharmaceutically acceptable salt thereof.