JP2002212070A - Sphingosine-1-phosphate antagonist containing aspartic acid derivative as active ingredient - Google Patents

Abstract

(57) Abstract: A sphingosine-1-phosphate antagonist comprising, as an active ingredient, an aspartic acid derivative represented by the general formula (I) or a non-toxic salt thereof. [In the formula, ^{R 1} is halogen, trihalomethyl, OH, alkyl, alkoxy,
R represents alkylthio, alkylsulfinyl or alkylsulfonyl; n represents 0 or 1-3;
² represents H or an alkyl group; R ³ represents H, -COR
It represents ^{9, etc.; R 13} represents H, halogen, trihalomethyl, alkyl or alkoxy groups. ]. Embedded image The compound of the general formula (I) is sphingosine-1
-Having phosphate antagonism, inflammatory diseases, diseases accompanied by abnormal angiogenesis, cancer-related diseases, cerebral infarction, nephritis, immune diseases,
It is useful for treating and / or preventing Crohn's disease, colitis, chronic diarrhea and the like.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a sphingosine-1-phosphate antagonist containing an aspartic acid derivative or a non-toxic salt thereof as an active ingredient. More specifically, the present invention relates to a compound of the general formula (I)

Embedded image (Wherein all symbols have the same meanings as described below). The present invention relates to a sphingosine-1-phosphate antagonist containing, as an active ingredient, an aspartic acid derivative or a non-toxic salt thereof.

[0002]

2. Description of the Related Art Sphingosine-1-phosphate (hereinafter referred to as S1P
Abbreviated as sphingolipids) is a lipid produced by the turnover of sphingolipids, and has been proposed to act as an intercellular or intracellular messenger. First, results suggesting the possibility of S1P as an intracellular second messenger were reported, and subsequently, S1P intracellular injection experiments revealed that S1P expresses physiological functions in cells. However, to date, no intracellular molecule on which S1P acts directly has yet been found. Recently, G protein-coupled seven-transmembrane receptor Endothelial Differentiation
Gene-1 (EDG-1) and its related molecules EDG-3 and EDG-5
It has been shown one after another to function as an S1P receptor, suggesting that various physiological actions of S1P may occur via the receptor. This is because EDG-1 converts human umbilical vein endothelial cells (HUVEC) into Phorbol 12-myristate 13-acetate (PM
A) It originated from the first report by Hla et al. In 1990 as an orphan receptor induced by stimulation. Thereafter, a plurality of EDG genes were cloned, and it was found that the homology between the receptors at the amino acid level was about 44 to 50%, indicating that these genes form a family. EDG-1 was first reported by the group to be an S1P receptor in 1998, indicating that other receptors also react with S1P. In addition, it has been revealed that these receptors also have high homology (about 30%) with the lysophosphatidic acid (LPA) receptor, and are positioned as an S1P / LPA receptor family. Recently, it has been shown that EDG-1 interacts with Gi protein and causes activation of p42MAPK / ERK2 via Gi protein. Many cell responses are due to pertussis toxin sensitivity, and platelets with high sphingosine kinase activity do not work similarly to S1P even if sphingosine is added to the culture medium.
It has been suggested that this may be a receptor-mediated effect. In addition, since S1P is stored in platelets and is released into the blood upon activation of platelets, it is highly likely that the physiological action of S1P is expressed via a receptor on the cell membrane.

[0003] S1P is known to act as an inhibitor of smooth muscle cell and cancer cell motility, regulation of blood pressure, aggregation of platelets, etc. Recently, S1P plays an important role also in angiogenesis. I've found it to work. Menq-Jer Lee et al. HU
Using VEC, S1P has been reported to promote the survival, elongation of adhesion points, and microangiogenesis of these cells via EDG-1 and EDG-3 receptors. Also they are S1P
Have a synergistic effect with fibroblast growth factor (FGF) and vascular endothelial cell growth factor (VEGF) on angiogenesis in vivo. OK-Hee Lee et al. Also clarified the effect of S1P on DNA synthesis promotion and migration on HUVEC using HUVEC, and reported that S1P alone promotes angiogenesis for angiogenesis in vivo as well. are doing. From these facts, it is considered that S1P has an angiogenesis promoting action as one of the physiological actions in vivo.

[0004] Accordingly, S1P antagonists are useful for inflammatory diseases (eg, immune and non-immune diseases, rheumatoid arthritis, psoriasis), diseases accompanied by abnormal angiogenesis (eg, restenosis, diabetic retinopathy, angiogenic diseases). Glaucoma, post lens fibroplasia, atherosclerotic plaque capillary proliferation, thyroid hyperplasia (including Graves' disease), pulmonary inflammation, nephrotic syndrome,
And osteoporosis), cancer-related diseases (eg, solid tumor, solid tumor metastasis, hemangiofibromas, myeloma, multiple myeloma, Kaposi's sarcoma), cerebral infarction, nephritis, immune disease, Crohn's disease, colitis, chronic It is useful as a therapeutic and / or prophylactic agent for diarrhea and the like. Among the solid tumors described above, there are breast cancer, lung cancer, gastric cancer, esophageal cancer, colorectal cancer, liver cancer, ovarian cancer, follicular cell tumor, male germinoma, cervical cancer, endometrial cancer, prostate cancer, kidney cancer , Skin cancer, osteosarcoma, pancreatic cancer, urinary tract cancer, thyroid cancer, brain tumor and the like.

[0005]

DISCLOSURE OF THE INVENTION The present inventors have conducted research to find a compound having excellent antagonism to S1P and high safety, and an aspartic acid derivative represented by the general formula (I) has been developed. The inventors have found that the present invention has been achieved and completed the present invention.
Further, the aspartic acid derivative represented by the general formula (I) or a non-toxic salt thereof of the present invention is a known compound described in JP-A No. 2-209861.

[0006]

DISCLOSURE OF THE INVENTION The present invention relates to 1) the general formula (I)

Embedded image [Wherein, R ¹ represents (1) a halogen atom, (2) a trihalomethyl group, (3) a hydroxyl group, (4) a C1-15 alkyl group,
(5) C1-4 alkoxy group, (6) C1-4 alkylthio group, (7) C1-4 alkylsulfinyl group,
(8) C1-4 alkylsulfonyl group, (9)

Embedded image(In the group, R⁴Is hydrogen atom, halogen atom, trihalomethyl
Group, hydroxyl group, C1-4 alkyl group or C1-4 alkyl group
Represents a coxy group. ), (10) -NR⁵R⁶Group (group
Medium, R⁵And R⁶Are each independently a hydrogen atom or
Represents a C1-4 alkyl group. ), (11) -COR⁷
Group (in the group, R⁷Is a C1-4 alkyl group or R⁴(Base
Medium, R ⁴Represents the same meaning as described above. )
Represents a phenyl group. ), (12) -COOR⁸Group (group
Medium, R⁸Represents a hydrogen atom or a C1-4 alkyl group
You. ), (13) -CONR⁵R⁶Group (in the group, R⁵And
And R⁶Represents the same meaning as described above. ), (14) -SO
₂NR⁵R⁶Group (in the group, R⁵And R⁶Is the same as above
Represents taste. ), (15) cyano group, (16) nitro group
Or (17) -NHCO-R⁷Group (in the group, R⁷Is
Has the same meaning as ) Wherein n is 0 or 1-3
R represents an integer²Is a hydrogen atom or C1-4 alkyl
And represents R³Is (1) hydrogen atom, (2) -COR
⁹Group (in the group, R⁹Is a C1-4 alkyl group or R¹⁰Base
(In the group, R¹⁰Is hydrogen atom, halogen atom, trihalo
Cyl group, C1-4 alkyl group or C1-4 alkoxy
Represents a group. ) Represents a substituted phenyl group. ),
(3)

Embedded image (In the group, R ¹⁰ has the same meaning as described above.), (4)

Embedded image (In the group, R ¹¹ and R ¹² are each independently a hydrogen atom, a C1-4 alkyl group or an R ¹⁰ group (in the group, R ¹⁰
Represents the same meaning as described above. Or R ¹¹ and R ^{12 taken} together represent a C4-5 alkylene group, and p represents an integer of 1 or 2. ), (5)

Embedded image (In the group, two R ¹⁰ each independently represent the same meaning as described above.), (6)

Embedded image (In the group, R ¹⁰ has the same meaning as described above, and q is 1 to 4
Represents an integer. ) Or (7)

Embedded image Wherein R ¹⁰ represents the same meaning as described above, and R ¹³ represents a hydrogen atom, a halogen atom, a trihalomethyl group, a C1-4 alkyl group or a C1-4 alkoxy group. ] Or a non-toxic salt thereof as an active ingredient.

In the general formula (I), the halogen atom represented by R ¹ , R ⁴ , R ¹⁰ or R ¹³ is fluorine, chlorine, bromine and iodine.

In the general formula (I), the trihalomethyl group represented by R ¹ , R ⁴ , R ¹⁰ or R ¹³ is a methyl group tri-substituted by a halogen atom. For example, a trifluoromethyl group, a trichloromethyl group, a tribromomethyl group, a triiodomethyl group and the like can be mentioned.

[0009] In the formula (I), a C1~15 alkyl group represented by ^{R 1,} methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl Pentadecyl group and isomer groups thereof.

In the general formula (I), the C1-4 alkoxy group represented by R ¹ , R ⁴ , R ¹⁰ or R ¹³ is methoxy, ethoxy, propoxy, butoxy and isomers thereof.

[0011] In the formula (I), a C1~4 alkylthio group represented by ^{R 1,} it is methylthio, ethylthio, propylthio, butylthio and the isomers thereof.

[0012] In the formula (I), a C1~4 alkylsulfinyl group represented by R ^1, a methylsulfinyl, ethylsulfinyl, propyl sulfinyl, butylsulfinyl and isomeric groups thereof.

[0013] In the formula (I), a C1~4 alkylsulfonyl group represented by R ^1, methylsulfonyl, ethylsulfonyl, propylsulfonyl, butylsulfonyl groups and the isomers thereof.

In the general formula (I), R ² , R ⁴ , R ⁵ ,
^{R 6, R 7, R 8} , R 9, R 10, R 1 1, R 12, R
^The C1-4 alkyl group represented by ¹³ is a methyl, ethyl, propyl, butyl group and isomers thereof.

In the general formula (I), the C4-5 alkylene group represented by R ¹¹ and R ¹² together is a tetramethylene and pentamethylene group.

In the present invention, all isomers are included unless otherwise indicated. For example, the alkyl group, the alkoxy group and the alkylene group include straight-chain and branched ones. Further, isomers (E, Z, cis, trans) in double bonds, rings and condensed rings, isomers due to the presence of asymmetric carbon (R, S, α, β, enantiomers, diastereomers) , An optically active substance having optical activity (D, L, d, l-form), polar forms (high-polarity forms, low-polarity forms) by chromatographic separation, equilibrium compounds, mixtures of any ratio thereof, and racemic mixtures All are included in the present invention.

In the present invention, unless otherwise specified:
Symbol as obvious to the person skilled in the art

Embedded image Indicates that it is connected to the other side of the paper (ie, α-configuration),

Embedded image Indicates that it is connected to the near side of the paper (that is, β-configuration),

Embedded image Represents α-, β- or a mixture thereof.

The compound represented by the formula (I) can be converted into a non-toxic salt by a known method. In the present specification, the non-toxic salt includes an alkali metal salt, an alkaline earth metal salt, an ammonium salt, an amine salt, an acid addition salt, a hydrate and the like.

The salt is preferably a non-toxic, water-soluble salt. Suitable salts include salts of alkali metals (potassium, sodium, etc.), salts of alkaline earth metals (calcium, magnesium, etc.), ammonium salts, and pharmaceutically acceptable organic amines (tetramethylammonium, triethylamine, methylamine) Dimethylamine, cyclopentylamine, benzylamine, phenethylamine, piperidine, monoethanolamine, diethanolamine, tris (hydroxymethyl) aminomethane, lysine, arginine, N-methyl-D-glucamine and the like.

The acid addition salts are preferably non-toxic and water-soluble. Suitable acid addition salts include, for example, inorganic salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate, nitrate, or acetate, lactate, tartrate, benzoate Organic acid salts such as acid, citrate, methanesulphonate, ethanesulphonate, benzenesulphonate, toluenesulphonate, isethionate, glucuronate, gluconate. The hydrate salt is preferably non-toxic and water-soluble.

In the general formula (I), R ¹ is preferably a halogen atom, a trihalomethyl group or a C1-4 alkoxy group, particularly preferably a chlorine atom, an iodine atom, a trifluoromethyl group or a methoxy group. General formula (I)
In the formula, R ² is preferably a hydrogen atom.

In the general formula (I), R ³ is a hydrogen atom or

Embedded image Is preferred. In the general formula (I), R ¹³ is preferably a hydrogen atom. In the general formula (I), n is preferably 0 or 1.

Among the compounds of the present invention represented by formula (I), preferred compounds are those represented by formula (I-1)

Embedded image (Wherein, R ¹ has the same meaning as described above),

Formula (I-2)

Embedded image (Wherein, R ¹ has the same meaning as described above),

Formula (I-3)

Embedded image (Wherein, R ¹ has the same meaning as described above),

General formula (I-4)

Embedded image (Wherein, R ¹ has the same meaning as described above),

Formula (I-5)

Embedded image (Wherein, R ¹ has the same meaning as described above),

General formula (I-6)

Embedded image (Wherein, R ¹ has the same meaning as described above) or

Formula (I-7)

Embedded image (Wherein, R ¹ has the same meaning as described above), and non-toxic salts thereof.

Specific preferred compounds of the present invention include the compounds represented by Compounds 1 to 7 or their non-toxic salts.

Compound 1: N- (3-mercapto-2RS
-Benzylpropionyl) -β-alinino-L-aspartic acid (the compound described in Example 12 of JP-A-2-209861)

Embedded image

Compound 2: N- (3-mercapto-2RS
-Benzylpropionyl) -β- (4-fluoroalinino) -L-aspartic acid (compound described in Example 12 (3) of JP-A No. 2-209861)

Embedded image

Compound 3: N- (3-mercapto-2RS
-Benzylpropionyl) -β- (4-chloroalinino) -L-aspartic acid (the compound described in Example 12 (4) of JP-A-2-209861)

Embedded image

Compound 4: N- (3-mercapto-2RS
-Benzylpropionyl) -β- (4-iodoarinino) -L-aspartic acid (compound described in Example 12 (5) of JP-A-2-209861)

Embedded image

Compound 5: N- (3-mercapto-2RS
-Benzylpropionyl) -β- (4-methoxyalinino) -L-aspartic acid (compound described in Example 12 (6) of JP-A No. 2-209861)

Embedded image

Compound 6: N- (3-mercapto-2RS
-Benzylpropionyl) -β- (4- (N, N-dimethylamino) alinino) -L-aspartic acid
Compound described in Example 12 (7) of 2-209861)

Embedded image

Compound 7: N- (3-mercapto-2RS
-Benzylpropionyl) -β- (4-trifluoromethylalinino) -L-aspartic acid (Japanese Patent Laid-Open No. 2-209861)
(Compound described in Example 12 (8))

Embedded image

[0038]

[Production method of the compound of the present invention] The compound of the present invention represented by the general formula (I) is a known compound described in JP-A No. 2-209861, and its production method is also described in JP-A No. 2-209861. ing.

[0039]

As described above, S1P antagonists are useful for inflammatory diseases (eg, immune and non-immune diseases, rheumatoid arthritis, psoriasis) and diseases accompanied by abnormal angiogenesis (eg, restenosis, diabetic disease). Retinopathy, angiogenesis glaucoma, post lens fibroplasia, atherosclerotic plaque capillary proliferation, thyroid hyperplasia (including Graves' disease), lung inflammation, nephrotic syndrome, and osteoporosis), cancer-related diseases (Eg, solid tumor, solid tumor metastasis, hemangiofibromas, myeloma, multiple myeloma, Kaposi's sarcoma), cerebral infarction, nephritis, immune disease,
It is considered to be useful as an agent for treating and / or preventing Crohn's disease, colitis, chronic diarrhea and the like. Therefore, S1P
The compound of the present invention represented by the general formula (I) having an antagonistic activity can inhibit inflammatory diseases (eg, immune and non-immune diseases, rheumatoid arthritis, psoriasis) and abnormal angiogenesis in animals including humans, particularly humans. Accompanying diseases such as restenosis, diabetic retinopathy, neovascular glaucoma, post lens fibroplasia, atherosclerotic plaque capillary proliferation,
Thyroid hyperplasia (including Graves' disease), pulmonary inflammation, nephrotic syndrome, and osteoporosis), cancer-related diseases (eg, solid tumors, solid tumor metastases, angiofibromas, myeloma, multiple myeloma, Kaposi's sarcoma), It can be used as a therapeutic and / or prophylactic agent for cerebral infarction, nephritis, immune disease, Crohn's disease, colitis, chronic diarrhea, and the like.

[0040]

[Toxicity] The toxicity of each active ingredient and their non-toxic salts contained in the present invention was sufficiently low, and it was confirmed that they can be used safely as pharmaceuticals.

[0041]

[Application to Pharmaceuticals] In order to use the compound represented by the general formula (I), a non-toxic salt thereof or a hydrate thereof for use in the present invention for the above-mentioned purpose, it is usually required to use systemically or locally Or it is administered in parenteral form. The dosage varies depending on the age, body weight, symptoms, therapeutic effect, administration method, treatment time, etc., but is generally from 1 mg per adult per dose.
It is orally administered once to several times a day in the range of 1000 mg, or 1 mg to 1 dose per adult per dose.
Parenteral administration (preferably, intravenous administration) is performed once to several times a day in the range of 00 mg, or continuous intravenous administration is performed in the range of 1 hour to 24 hours a day. Of course, as described above, since the dose varies depending on various conditions, there are cases where a smaller amount than the above dose is sufficient,
In some cases, it may be necessary to go beyond the range.

When administering the compound represented by the general formula (I), solid compositions, liquid compositions and other compositions for oral administration and injections, external preparations and suppositories for parenteral administration are administered. Used as an agent and the like. Solid compositions for oral administration include tablets, pills, capsules, powders, granules and the like. Capsules include hard capsules and soft capsules. In such a solid composition, the one or more active substances comprise at least one inert diluent such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone, aluminum metasilicate. It is mixed with magnesium acid. The composition may contain, in a conventional manner, additives other than an inert diluent, for example, a lubricant such as magnesium stearate, a disintegrant such as calcium cellulose glycolate, a stabilizer such as lactose, glutamic acid or asparagine. A solubilizing agent such as an acid may be contained. The tablet or pill may be coated with a film of a gastric or enteric substance such as sucrose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose phthalate, or may be coated with two or more layers, if necessary. . Also included are capsules of absorbable materials such as gelatin.

Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, syrups, elixirs and the like. In such liquid compositions, one or more active substances are contained in commonly used inert diluents (eg, purified water, ethanol). The composition may contain, in addition to the inert diluent, adjuvants such as wetting agents and suspending agents, sweetening agents, flavoring agents, fragrances, and preservatives. Other compositions for oral administration include sprays which contain one or more active substances and are formulated in a manner known per se. The composition may further comprise a buffering agent which provides isotonicity with a stabilizer other than the inert diluent, such as sodium bisulfite,
For example, they may contain isotonic agents such as sodium chloride, sodium citrate or citric acid. Methods for producing sprays are described in detail, for example, in US Pat. Nos. 2,868,691 and 3,095,355.

The injections for parenteral administration according to the present invention include sterile aqueous and / or non-aqueous solutions, suspensions and emulsions. Aqueous solutions and suspensions include, for example, distilled water for injection and physiological saline.
Examples of the water-insoluble solutions and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, and Polysorbate 80 (registered trademark). In addition, sterile aqueous and non-aqueous solutions, suspensions, and emulsions may be mixed and used. Such compositions may also contain adjuvants such as preserving, wetting, emulsifying, dispersing, stabilizing (eg, lactose) and solubilizing agents (eg, glutamic acid, aspartic acid). . These are sterilized by filtration through a bacteria retaining filter, blending of a bactericide or irradiation. They can also be used to produce sterile solid compositions, for example, sterilized or dissolved in sterile distilled water for injection or other solvents before use of the lyophilized product. Other compositions for parenteral administration include one or more active substances,
It includes external solutions, ointments, liniments, suppositories for rectal administration, and pessaries for vaginal administration, etc., which are formulated in a conventional manner.

[0045]

Hereinafter, the present invention will be described in detail with reference to Examples.
The present invention is not limited to these.

Example 1 [Experimental method] (1)Cell preparation  Human umbilical vein vascular endothelial cells (HUVEC) for vascular endothelial cells
Serum-containing liquid medium (MCDB104 liquid medium containing 5% fetal bovine serum)
Subculture in Tokyo, Nissui Pharmaceutical Co., Ltd., Tokyo)
Using. The medium for vascular endothelial cells is a growth factor solution (endothelial cells).
Vesicle growth supplement 0.5 mg / ml, heparin 1 mg /
ml in Dulbecco's PBS (-) solution)
It was prepared by adding one hundredth of the volume. Once a week
And exchange the medium once between passages.
Was. All cultures are performed in a culture tank at 37 ° C, 5% CO₂In the presence
Done.

(2)HUVEC BrdU incorporation assay  HUVEC in vascular endothelial cell medium (Nissui Pharmaceutical) 1 × 10⁵
Cells / ml, 96-well
1 × 10 for the rate (IWAKI)⁴Pieces / well (100μ
Seed at 1). 48 hours after this time, remove the medium
After washing twice with 150 μl of PBS (-), 1% fetal bovine
90 μl of M199 medium containing serum (GIBCO) was added to each well.
Was. At this time, an experiment to examine only the action of S1P or VEGF
In the case of, the content of S1P (BIOMOL)
It was added so as to be 0 to 10 μM per degree. Also drug S1P
Alternatively, in experiments to examine the inhibitory effect of the effect on VEGF
In this case, the concentration of S1P or VEGF was 1
μM or 10 ng / ml.
Drug solution 10 times the final concentration (0.3 μM to 100 μM)
M) was added to each well in an amount of 10 μl (final concentration 0.03).
μM to 10 μM). Culture for 24 hours from this point
Modeoxyuridine (BrdU) 100 μM solution (Cell pro
liferation ELISA kit, included with Amersham)
10 μl was added (final concentration 10 μM). Another 24 hours
After culturing, Cell proliferation ELISA kit (Amersham)
Measured BrdU incorporation by cells.

(3)Cell ELISA assay  Using Cell proliferation ELISA kit (Amersham),
The experiment was performed according to the package insert. After treating cells with BrdU,
Discard the culture supernatant and add 1 cell fixation / DNA denaturing solution to each well.
50 μl was added and left for 30 minutes. Cell fixation / DNA denaturation
Remove the solution and add 150 μl blocking buffer.
For 30 minutes. Next, remove the Blocking buffer
Then, add the anti-BrdU antibody solution labeled with peroxidase to 1
00 µl was added and left for 30 minutes. Next, remove the antibody solution
And wash three times with 150 μl of Wash buffer (PBS (-))
After removing water sufficiently by tapping, the reaction substrate solution
(3,3 ', 5,5'-tetramethylbenzidine, TMB in 15% DMS
O solution) was added. After a certain time, 2N sulfuric acid solution
Add 25 μl of the solution, and use the microplate reader SPECTRAM.
AXPRO (Startup software SOFTMAXPRO ver.2.0, Nippon molecu
Device, Ashiya City) to measure absorbance at 450 nm
did. Also, ED₅₀Or IC₅₀Was calculated.

[Results] (1)S1P enhances BrdU uptake in HUVEC
About  Figure 1 shows the enhancing effect of S1P on HUVEC uptake of BrdU.
did. ED₅₀The value was about 0.1 μM.

(2)S1P-induced HUVEC enhances BrdU incorporation
On the inhibitory effect of the compound of the present invention in  Fig. 2 shows the incorporation of BrdU into HUVEC by S1P (1 μM).
Showed the inhibitory effect of the compound of the present invention. In addition, IC₅₀value
Are shown in Table 1.

[Table 1]

(3)HUVE induced by VEGF of the compound of the present invention
Examination of the effect of C on BrdU incorporation  Figure 3 BrdU removal of HUVEC by VEGF (10 ng / ml)
The effect of the compound of the present invention on the inflammation was shown.

[Consideration] (1)S1P enhances BrdU uptake in HUVEC
About  As shown in FIG. 1, S1P was dose-dependent on HUVEC.
BrdU uptake was enhanced. In addition, BrdU acquisition of HUVEC
Integration peaked near 1 μM S1P concentration. this thing
Thus, S1P depends on its dose to regulate HUVEC DNA synthesis.
It was found to promote.

(2)S1P-induced HUVEC enhances BrdU incorporation
On the inhibitory effect of the compound of the present invention in  As shown in FIG. 2, the compound of the present invention is BrdU of S1P-induced HUVEC.
Uptake activity was inhibited in a dose-dependent manner. From this,
The compound of the present invention is dose-dependently dependent on DN of S1P-induced HUVEC.
A was found to inhibit the synthesis promoting action.

(3)HUVE induced by VEGF of the compound of the present invention
Examination of the effect of C on BrdU incorporation  As shown in FIG. 3, all the compounds of the present invention
BrdU incorporation of VEGF-induced HUVECs significantly even at high concentrations
Did not inhibit.

[0055]

Formulation Example 1 Formulation Example 1 The following components were mixed in a conventional manner and tabletted to obtain 100 tablets each containing 50 mg of the active ingredient.・ N- (3-mercapto-2RS-benzylpropionyl) -β- (4-trifluoromethylalinino) -L-aspartic acid 5.0 g ・ Carboxymethylcellulose calcium (disintegrant)・ 0.2g ・ Magnesium stearate (lubricant) ・ ・ ・ ・ ・ ・ ・ 0.1g ・ Microcrystalline cellulose ・ ・ ・ ・ ・ 4.7g

Formulation Example 2 After mixing the following components by a conventional method, the solution is sterilized by a conventional method, filled into ampoules in 5 ml portions, freeze-dried by a conventional method, and contains 20 mg of active ingredient per ampule. 100 ampoules were obtained. N- (3-mercapto-2RS-benzylpropionyl) -β- (4-trifluoromethylalinino) -L-aspartic acid 2.0 g mannitol 20 g distilled water・ ・ ・ ・ 1000ml

[Brief description of the drawings]

FIG. 1 shows the dose-dependent enhancing action of S1P on HUVEC BrdU incorporation.

FIG. 2 shows the suppressive effect of the compound of the present invention on the HUVEC BrdU uptake-enhancing effect of S1P (1 μM).

FIG. 3. BrdU of HUVEC by VEGF (10 ng / ml)
It is an effect of the compound of the present invention on an uptake enhancing effect.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61P 5/14 A61P 5/14 9/10 101 9/10 101 9/14 9/14 11/00 11 / 00 13/12 13/12 17/06 17/06 19/10 19/10 29/00 29/00 35/00 35/00 35/04 35/04 37/00 37/00 43/00 111 43/00 111 C07C 323/54 C07C 323/54 327/32 327/32. MA04 NA14 ZA02 ZA33 ZA45 ZA59 ZA66 ZA73 ZA81 ZA89 ZA97 ZB07 ZB11 ZB15 ZB26 ZC06 ZC35 ZC41 4H006 AA01 AB22 AB27 AB28 TA04 TN60

Claims (1)

[Claims] 1. A compound of the general formula (I)[Wherein, R¹Are (1) halogen atom, (2) trihalome
A tyl group, (3) a hydroxyl group, (4) a C1-15 alkyl group,
(5) C1-4 alkoxy group, (6) C1-4 alkyl
A thio group, (7) a C1-4 alkylsulfinyl group,
(8) C1-4 alkylsulfonyl group, (9)(In the group, R⁴Is hydrogen atom, halogen atom, trihalomethyl
Group, hydroxyl group, C1-4 alkyl group or C1-4 alkyl group
Represents a coxy group. ), (10) -NR⁵R⁶Group (group
Medium, R⁵And R⁶Are each independently a hydrogen atom or
Represents a C1-4 alkyl group. ), (11) -COR⁷
Group (in the group, R⁷Is a C1-4 alkyl group or R⁴(Base
Medium, R ⁴Represents the same meaning as described above. )
Represents a phenyl group. ), (12) -COOR⁸Group (group
Medium, R⁸Represents a hydrogen atom or a C1-4 alkyl group
You. ), (13) -CONR⁵R⁶Group (in the group, R⁵And
And R⁶Represents the same meaning as described above. ), (14) -SO
₂NR⁵R⁶Group (in the group, R⁵And R⁶Is the same as above
Represents taste. ), (15) cyano group, (16) nitro group
Or (17) -NHCO-R⁷Group (in the group, R⁷Is
Has the same meaning as ) Wherein n is 0 or 1-3
R represents an integer²Is a hydrogen atom or C1-4 alkyl
And represents R³Is (1) hydrogen atom, (2) -COR
⁹Group (in the group, R⁹Is a C1-4 alkyl group or R¹⁰Base
(In the group, R¹⁰Is hydrogen atom, halogen atom, trihalo
Cyl group, C1-4 alkyl group or C1-4 alkoxy
Represents a group. ) Represents a substituted phenyl group. ),
(3)(In the group, R¹⁰Represents the same meaning as described above. ), (4)(In the group, R¹¹And R¹²Are independently hydrogen sources
, A C1-4 alkyl group or R¹⁰Group (in the group, R¹⁰
Represents the same meaning as described above. ) Replaced by
Represents a nyl group, or R¹¹And R¹²Together
Represents a C4-5 alkylene group, and p is 1 or 2
Represents an integer. ), (5)(In the group, two R¹⁰Is independently the same as above
Indicates meaning. ), (6)(In the group, R¹⁰Represents the same meaning as described above, and q represents 1 to 4.
Represents an integer. ) Or (7)(In the group, R¹⁰Represents the same meaning as described above. )
Then R¹³Is hydrogen atom, halogen atom, trihalomethyl
Group, C1-4 alkyl group or C1-4 alkoxy group
Express. Or an aspartic acid derivative represented by the formula:
Sphingosine containing these non-toxic salts as active ingredients
-1-phosphate antagonists.