WO2001001981A1 - Vegf receptor antagonists - Google Patents

Abstract

VEGF receptor antagonists containing as the active ingredient comopunds of general formula (1) or medicinally acceptable salts thereof: wherein R1 is hydrogen, CO¿2?R?5, or OR6¿ (wherein R?5 and R6¿ are each hydrogen or C¿1?-C6 alkyl); R?2¿ is hydrogen or CO¿2R?7 (wherein R7 is hydrogen or C¿1?-C6 alkyl); R?3¿ is hydrogen or C¿1?-C6 alkyl; R?4 is C¿8-C25 alkyl; X is O, S or NH; and Y is O or NHCO.

Description

 Specification

 VEGF receptor antagonist Technical field

 The present invention relates to a VEGF receptor antagonist that inhibits binding of VEGF, a specific growth factor of vascular endothelial cells, to a receptor.

 This application is based on patent applications to Japan (Japanese Patent Application No. 11-188269 and Japanese Patent Application No. 11-188270), and the description of the Japanese application is incorporated as a part of this specification. And Background art

VEGF (vascular endothelial growth factor) is a highly specific growth factor for vascular endothelial cells, and VEGF and its receptor play a central role in physiological angiogenesis such as development and development, and placental formation. F1t-1 (fms-like tyrosine kinase) and KDR (kinase insert do main containing receptor) have been reported as VEGF receptors (Advances in Cancer Research _N 67: 281: 281). 316, 1995).

 VEGF and its receptors are not only physiological angiogenesis, but also diabetic retinopathy, rheumatoid arthritis, solid tumors (Advances in Cancer Research, Vol. 67, pp. 281-316, 1995), etc. It also plays a central role in the pathological angiogenesis seen in these diseases, suggesting that it is deeply involved in the development of such diseases. VEGF and its receptor are known to be involved not only in angiogenesis but also in vascular permeability enhancement. VEGF-induced vascular hyperpermeability is associated with pathological changes such as cancerous ascites retention and cerebral edema during ischemia-reperfusion injury (J. Clin. Invest., Vol. 104, pp. 1613-1620, 1999). It has been suggested to be involved in the symptoms.

Therefore, substances that inhibit the binding between VEGF and its receptor are useful for treating various diseases in which pathological angiogenesis by VEGF is involved and in pathological disorders in which vascular permeability enhancement by VEGF is involved. It is considered useful for improving symptoms. Disclosure of the invention

 An object of the present invention is to provide a VEGF receptor antagonist for treating a disease associated with VEGF-induced angiogenesis and ameliorating a pathological condition involving VEGF-induced hypervascularization. It is.

 The VEGF receptor antagonist of the present invention has the following formula (1)

It is characterized by containing the compound represented by (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

In the formula (1), R ¹ is a hydrogen atom, CO ₂ R ⁵ or OR ⁶ (where ¹⁵ and ¹⁶ are each a hydrogen atom or a trialkyl group). , R ² is a hydrogen atom or a group represented by CO ₂ R ⁷ (where R ⁷ is a hydrogen atom or a C ₆ alkyl group), and R ³ is a hydrogen atom or a d-6 alkyl group. There, R ⁴ is C ₈ - a ₂₅ § alkyl group, X is 0, S or NH, Y is 0 or NHC0.

It is known that a part of the compound represented by the above formula (1) is used for a color power blur for photographic film (Reinhold Groeger et al. Veroeffentl ichungen Wissenschaftlichen Photo-Laborator ι en Wolfen, (Vol. 10, 309 to 315, 1965) shows that among the compounds represented by the formula (1), R ¹ is a hydrogen atom or C0 ₂ R ⁵ (where R ⁵ is a hydrogen atom or a C physician ₆ alkyl group.) in a group represented, R ² is hydrogen atom, R ³ is a hydrogen atom or a d-6 alkyl group, R ⁴ is C _{8 25} alkyl group, The compound in which X is NH and Y is 0 or a pharmaceutically acceptable salt thereof is a novel compound.

Furthermore, the present invention provides a VEG comprising the VEGF receptor antagonist. Provide a therapeutic agent for a disease involving F.

 Further, the present invention provides a method for treating a disease associated with VEGF, which comprises administering the above VEGF receptor antagonist.

 Further, the present invention provides a use of the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof for the treatment of a therapeutic agent for a disease associated with VEGF, and a therapeutic agent for a disease associated with VEGF. To provide a use of the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof for the production of BEST MODE FOR CARRYING OUT THE INVENTION

 In the present invention, the d alkyl group means a linear or branched alkyl group having 1 to 6 carbon atoms, such as a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, and a t-alkyl group. —Butyl group, pentyl group, isopentyl group, 1-ethylpropyl group, hexyl group, isohexyl group, 1-ethylbutyl group and the like.

The C ₅ alkyl group means a linear or branched alkyl group having a carbon number of 8-25, for example Okuchiru group, 7-Mechiruokuchiru group, 7, 7-Jimechiruoku ethyl group, O click evening decyl , 17-methyloxydecyl group, 17,17-dimethyloxydecyl group, penosylcosyl group, 23-methyltetracosyl group, and 22,22-dimethyltricosyl group.

 The pharmaceutically acceptable salts in the present invention include, for example, salts with mineral acids such as sulfuric acid, hydrochloric acid and phosphoric acid, acetic acid, oxalic acid, lactic acid, tartaric acid, fumaric acid, maleic acid, methanesulfonic acid, benzenesulfonic acid And the like, salts with organic acids such as trimethylamine and methylamine, and salts with metal ions such as sodium ion, potassium ion and calcium ion.

 In addition, some of the compounds according to the present invention have a polymorphism, and the present invention includes any of the crystal forms.

In the present invention, R ¹ is preferably C 0 ₂ R ⁵ (R ⁵ is as defined above), and R ² is preferably a hydrogen atom. Further R ¹ is more preferably a C_〇 ₂ H. R ³ is preferably a hydrogen atom. R ⁴ is preferably an alkyl group having 14 to 22 carbon atoms, and more preferably R ⁴ is an alkyl group having 18 carbon atoms. Examples of the compounds according to the present invention include the following.

 5-[3-(4-octyldecyloxyphenyl) -1-3-oxopropionylamino] -2, 4 '-oxinibenzoic acid

 5-[3-(4-Oxyl decyloxyphenyl) 1-3-oxopropionylamino] -2, 4 '-Oxinibenzoic acid dimethyl ester

 5— [3— (4-octyldecyloxyphenyl) -1-3-oxopropionylamino] —2,3′—oxinibenzoic acid

 5- [3- (4-octyldecyloxyphenyl) -1-3-oxopropionylamino] _2,3'-oxinibenzoic acid dimethyl ester

 5- [3- (4-octyldecyloxyphenyl) -1-3-oxopropionylamino] —2, 2'-oxinibenzoic acid

 5-[3- (4-octyldecyloxyphenyl) -1-3-oxopropionylamino] — 2,2'-oxinibenzoic acid dimethyl ester

 3— [3— (4-octyldecyloxyphenyl) -1-3-oxopropionylamino] —3 ′, 4-oxodibenzoic acid

 3-[3-(4-octyl decyloxyphenyl) 1-3-oxopropionylamino]-3 ', 4- oxodibenzoic acid dimethyl ester

 5— [3— (4-octyldecyloxyphenyl) -1-3-oxopropionylamino] —2,4′—thiodibenzoic acid

 5-[3- (4-octyldecyloxyphenyl) -1-3-oxopropionylamino] —2, 4'-thiodibenzoic acid dimethyl ester

 5— [3— (4-octyldecyloxyphenyl) -1-3-oxopropionylamino] —2,3′—thiodibenzoic acid

 5- [3- (4-octadecyloxyphenyl) -1-3-oxopropionylamino] —2,3'-thiodibenzoic acid dimethyl ester

5— [3— (4-octyldecyloxyphenyl) -1-3-oxopropionylamino] —2,4′—Iminodibenzoic acid _c

 Five

 5- [3- (4-Ocdecyldecyloxyphenyl) -1-3-oxopropionylamino] —2,4'-Iminodibenzoic acid dimethyl ester

 5— [3— (4-octadecyloxyphenyl) -1-3-oxopropionylamino] —2,3′—iminodibenzoic acid

 5- [3- (4-octyldecyloxyphenyl) -1-3-oxopropionylamino] — 2,3'-imino dibenzoic acid dimethyl ester

 5-[3- (4-octyldecyloxyphenyl) -1-3-oxopropionylamino] — 2, 2'-Iminodibenzoic acid

5- [3- (4-octadecyloxyphenyl) -l-oxopropionylamino}-2,2'-iminodibenzoic acid dimethyl ester The compound represented by the above formula (1) is as follows. Wherein X, Y, RR ² , R ³ and R ⁴ are as defined above, halo is a halogen atom, and R is a lower alkyl group. ).

R² [縮合]

R ² [condensation]

C〇 ₂ R

式 （3) の化合物と式 （4) のニトロ化合物を塩基存在下、 及び触媒量の銅粉末 の存在下又は非存在下、 適当な溶媒中、 0°Cから 150°Cの間の温度にて攪拌し、 式 （5) の化合物を得る。 塩基としては、 炭酸カリウム、 炭酸ナトリウム、 炭酸 水素ナトリウム、 炭酸水素カリウム.、 炭酸セシウム、 水素化ナトリウム、 水素化 カリウムなどの無機塩基、 トリェチルァミン、 ジイソプロピルェチルァミン、 ピ リジンなどの有機塩基などが用いられる。 溶媒としては、 N, N—ジメチルホル ムアミ ドなどの反応に不活性な溶媒などが用いられる。 (6) P-R '

The compound of formula (3) and the nitro compound of formula (4) are brought to a temperature between 0 ° C and 150 ° C in a suitable solvent in the presence of a base and in the presence or absence of a catalytic amount of copper powder. To give the compound of formula (5). Examples of the base include inorganic bases such as potassium carbonate, sodium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, cesium carbonate, sodium hydride, potassium hydride, and the like, and organic bases such as triethylamine, diisopropylethylamine, and pyridine. Used. As the solvent, a solvent inert to the reaction such as N, N-dimethylformamide is used.

If necessary, the compound of formula (5) is stirred in a suitable solvent at a temperature between 0 ° C and 100 ° C in the presence of a lower alkyl halide and a base, and R ¹ and R ² are hydrogen atoms. Or a compound of the formula (5) in which R ³ is an alkyl group or an alkoxycarbonyl group is obtained. As the base, potassium carbonate, sodium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, cesium carbonate and the like are used. As the solvent, a solvent inert to the reaction such as N, N-dimethylformamide is used.

Next, the nitro group of the compound of the formula (5) in which R ¹ and R ² are not a carboxyl group and R ³ is an alkyl group is reduced to an amino group to obtain a compound of the formula (6). The reduction methods include reduction using a metal or metal salt such as iron or tin in the presence of an acid such as ammonium chloride, hydrochloric acid or acetic acid, catalytic reduction using a catalyst such as palladium-carbon, Raney nickel, platinum oxide, palladium- Reduction with ammonium formate in the presence of a carbon catalyst. Examples of the solvent include solvents inert to the reaction, such as methanol, ethanol, and isopropyl alcohol.

 The compound of formula (6) obtained here can be treated at a temperature between 100 ° C and 200 ° C in the presence or absence of a solvent inert to the reaction of the ketoester of formula (7) with toluene or xylene. To give the compound of formula (1) according to the present invention.

 The alkoxycarbonyl group in the formula (1) can be converted into a carboxyl group by hydrolyzing the ester by a usual method for hydrolyzing an ester.

The compound represented by the formula (1) or a pharmaceutically acceptable salt thereof is used as the VEGF receptor antagonist, in particular, in the treatment of VEGF-related diseases and in the production thereof. The VEGF receptor antagonist of the present invention inhibits VEGF-dependent vascular endothelial cell proliferation by inhibiting the binding of a ligand (VEGF) to VEGF receptor. Five

 7

In addition, it inhibits angiogenesis and inhibits vascular permeability enhancement by VEGF.

 Here, the diseases and pathological symptoms related to VEGF include, for example, diabetic retinopathy and other retinopathies, rheumatoid arthritis, solid tumors, cerebral edema and injury related to ischemia-reperfusion injury, psoriasis, atheroma Sclerosis, post-lens fibroplasia, neovascular glaucoma, age-related macular degeneration, thyroid hyperplasia (including Graves' disease), chronic inflammation, pneumonia, Neff's syndrome, reduced tumor immune function, ascites retention, endocardial fluid Effusions (such as those associated with pericarditis) and pleural effusion.

 Among the above diseases, in particular, the following diseases have been reported to be improved by VEGF inhibition.

① Diabetic retinopathy and other retinopathies

 Diabetic retinopathy is a disease in which various abnormalities in the retinal blood vessels caused by prolonged exposure to hyperglycemic conditions cause a variety of lesions in the retina and vitreous body, and abnormalities in the eyeball as the disease progresses It is known that angiogenesis and bleeding lead to blindness. On the other hand, it has been reported that there is a positive correlation between elevated intraocular VEGF levels and abnormal intraocular neovascularization in diabetic patients (New Engl. J. Med., Vol. 331, 1480--1487, 1994). Also, in monkey retinopathy model, angiogenesis is suppressed by suppressing intravenous administration of anti-VEGF neutralizing monoclonal antibody by inhibiting VEGF activity (Arch. 0pthalmol., Vol. 114, p. 66—71. P. 1996), Inhibition of retinal neovascularization by administration of a VEGF signaling inhibitor in a mouse reticulomegaly model (Am. J. Pathol., 156, 697-707). , 2000). Based on the above, the VEGF receptor antagonist is considered to be effective for diabetic retinopathy and other ischemic retinopathies.

② Rheumatoid arthritis

Serum VEGF levels in patients with rheumatoid arthritis are significantly higher than in healthy individuals, and it has been reported that VEGF production is increased in localized lesions (JI Adjunct uno l., Vol. 152, pp. 4149-p. 4156, 1994), suggesting that VEGF is deeply involved in the formation of pathological conditions. In a mouse collagen arthritis model, an anti-VEGF antiserum administration has been reported to improve the disease state (J. Immunol., Vol. 164, pp. 5922- 5927, 2000).

VEGF is also thought to play an important role in the neovascularization of malignant tumors (Biochem. Biophys. Res. Commun., Vol. 161, pp. 851-858, 1989).

VEGF is used for brain tumors such as gliomas, malignant lymphomas, pituitary adenomas, meningiomas, melanomas, colorectal cancer, ovarian cancer, Teng cancer, esophageal cancer, rhabdomyosarcoma, leiomyosarcoma, lipopositive sarcoma and lung gland It is known that the production of many solid malignant tumors such as cancer is enhanced (Nature, Vol. 362, pp. 841-844, 1993, Biochem. Biophys. Res. 183, p. 1167-p. 1174, p. 1992) _c VEGF secreted from tumor cells proliferates vascular endothelial cells by binding to tyrosine kinase-type receptors present specifically on vascular endothelial cells, It is thought to be involved in tumor growth or metastasis by inducing tumor angiogenesis (0ncogene, vol. 5, p. 519-p. 524, 1990; Science, vol. 255, p. 899- Pp. 991, 1992).

 It has been reported that administration of an anti-VEGF monoclonal antibody suppresses tumor growth in a nude mouse transplant model of glioblastoma, rhabdomyosarcoma, and leiomyosarcoma (Nature, Vol. 362, No. 841). P. 844, 1993), it has been suggested that VEGF receptor antagonists have antitumor effects on various solid tumors.

脳 Ischemic reperfusion injury-related cerebral edema and injury

VEGF is thought to be involved in the development of edema due to its vascular hyperpermeability, and in a mouse cerebral ischemia model, administration of a fusion protein of mouse VEGF receptor protein [mFlt (1-3)] and IgG Suppresses brain edema and damage (J. Clin. Invest., Vol. 104, pp. 1613-1 pp. 1620, 1999). The VEGF receptor antagonist according to the present invention and the therapeutic agent for a disease associated with VEGF according to the present invention can be administered orally or parenterally.

 The dosage forms are tablets, capsules, granules, powders, powders, troches, ointments, creams, emulsions, suspensions, suppositories, injections, etc. It can be manufactured by the method specified in the 12th revised Japanese Pharmacopoeia). These dosage forms can be appropriately selected depending on the condition, age, and purpose of treatment of the patient. In the preparation of various dosage forms, conventional excipients (eg, crystalline cellulose, starch, lactose, mannitol, etc.), binders (eg, hydroxypropylcellulose, polyvinylpyrrolidone, etc.), lubricants (eg, For example, magnesium stearate, talc, and the like, and disintegrants (eg, carboxymethyl cellulose calcium) can be used.

 The dose of the compound of the present invention is 1 to 2000 mg per day when treating an adult, and the compound is administered once or several times a day. This dosage may be adjusted as appropriate according to the patient's age, weight and condition. Manufacturing example

 [Production Example 1]

48.4 g of anhydrous potassium carbonate was dissolved in a solution of 35.5 g of 4-hydroxybenzoic acid methyl ester and 50.2 g of 2-chloro-5-nitrobenzoic acid methyl ester in 500 ml of N, N-dimethylformamide (DMF). The mixture was stirred at 80 ° C for 3 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated saline and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the obtained crude product was recrystallized from methanol to give 2- (4-methoxycarbonylphenoxy) -15-nittobenzoic acid methyl ester (melting point: 10) 67.7 g (3 to 105 ° C) was obtained (reaction formula (8) below).

(8)

To a mixture of 11.6 g of the compound obtained by the above reaction formula (8) suspended in 300 ml of methanol was added 1.00 g of 10% palladium-carbon, and the mixture was stirred under a hydrogen atmosphere at room temperature for 2 hours. The reaction mixture was filtered to remove the catalyst, this _c the filtrate to give the crude product was distilled off under reduced pressure by silica gel column chromatography (hexane: acetic acid Echiru = 2: Hand eluted 3) purified by Thereafter, the product was recrystallized from methanol to obtain 8.53 g of 2- (4-methoxycarbonylphenoxy) -5-aminobenzoic acid methyl ester (melting point: 144 to 146 ° C.) (the following reaction formula (9)).

(9)

化合物 1 ( 10) A mixture of 2.84 g of the compound obtained by the above reaction formula (9) and 4.34 g of 3- (4-oxodecyloxyphenyl) -3-ethyl oxopropionate was heated at 140 ° C. for 7 hours. The obtained crude product was purified by silica gel column chromatography (eluted with hexane: cloth form: ethyl acetate = 2: 3: 1), and recrystallized from ethyl acetate to give Compound 1 (melting point: 3.15 g (115 ° -117 ° C.) was obtained (the following reaction formula (10)). .

Compound 1 (10)

 [Production Example 2]

By the same operation as in Production Example 1, Compounds 2 to 8 represented by the general formula ( ¹¹ ) and having R ¹¹ and R ¹² shown in Table 1 were obtained. Their melting points are also shown in Table 1.

(1 1)

table 1

R ¹¹ R ¹² Melting point (° c) Compound 2 H 90-92

Me0 ₂ C ^^ O compound 3 H 50-52

C0 ₂ e Compound 4 H 85-87 Compound 5

O. H 42-43 CH e ₂ Compound 6 H chi 138-142 Compound 7 H 113-115 e0 ₂ C ^^ O compound 8 H 83-93

[Production Example 3]

 A mixture of 1.09 g of 2- (4-methoxycarbonylphenylthio) -5-nitrobenzoic acid methyl ester and 1.75 g of iron powder obtained by the same operation as in the reaction formula (8) of Production Example 1 was mixed with isopropyl alcohol 3 ml and aqueous ammonium chloride solution (0.05 g of ammonium chloride, 0.95 ml of water) were added, and the mixture was stirred at 85 ° C for 10 minutes. To the reaction mixture was added chloroform and the mixture was filtered through celite, and then washed with chloroform. The filtrate and the washing solution were combined, washed with a saturated saline solution, and dried with anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain 0.996 g of methyl 5-amino-2- (4-methoxycarbonylphenylthio) benzoate (yellow viscous substance) (the following reaction formula (12)).

(12) Compound 9 (melting point: 121 to 124 ° C) was obtained from the compound obtained in the above reaction formula (12) by the same operation as in the reaction formula (10) in Production Example 1 (the following reaction formula ( 13) ) .

化合物 9 3) [製造例 4]

Compound 9 3) [Production Example 4]

By the same operation as in Production Example 2, Compounds 10 to 18 represented by the general formula (14) and having R ¹³ , R ^14, and R ¹⁵ shown in Table 2 were obtained. Their melting points are also shown in Table 2.

(14)

W 1

 15 Table 2

Compound R ¹³ R ¹⁴ R ¹⁵ Melting point (° c) Compound 10 H OC ₁₀ H ₂₁ 126-128

Compound 11 H OC ₁₈ H ₃₇ 124-128

Compound 12 H OC ₁₈ H ₃₇ 58-60

C0 ₂ Me compound 13 H NHCOC ₁₇ H ₃₅ 116-121

C0 ₂ Me compound 14 H OC ₁₈ H ₃₇ 100-101

 s

 c

Compound 15 H OC ₁₈ H ₃₇ 111-112

化合物 17 H OC₁₈H₃₇ 86-89 Compound 16 H OC ₁₈ H ₃₇ 97-99

Compound 17 H OC ₁₈ H ₃₇ 86-89

Compound 18 H OC ₁₈ H ₃₇ 111-115 [Production Example 5]

 To a mixture of 3.63 g of Compound 1 suspended in 50 ml of ethanol was added an aqueous sodium hydroxide solution (2.03 g of sodium hydroxide, 50 ml of water), and the mixture was stirred at 0 ° C for 90 minutes. The reaction solution was acidified by adding 5% hydrochloric acid, and the precipitated solid was filtered, washed with water, and dried under reduced pressure to obtain 3.31 g of compound 19 (melting point: 219 to 221 ° C) (the following reaction formula). (15)).

Compound 1

 Compound 19

(15)

[Production Example 6]

The same operation as in Production Example 5 was carried out using Compounds 2 to 18, and represented by the general formula (16), wherein R ¹⁶ , R ¹⁷ and R ¹⁸ had the structures shown in Tables 3 and 4 and Compounds 20 to 36 I got Their melting points are also shown in Tables 3 and 4.

(16) Table 3

R ¹⁶ R ¹⁷ R ¹⁸ Melting point (° c) Compound 20 H OCigH ₃₇ 165-169 Compound 21 H OC ₁₈ H ₃₇ 180-182

Compound 22H OC18H37 124-127

 k ^. Compound 23 H 3 lOM 3 /, 106-109

匕 ^ ¾ ク 0 4 4 HJ. H. , 214-217 Compound 25 H OC ₁₈ H ₃₇ 198-201

H0 ₂ C ^^ O compound 26 HC OC ₁₈ H ₃₇ 186-189.

Table 4

R ¹⁶ R ¹⁷ R ¹⁸ Melting point (° c) Compound 27 H OC ₁₈ H ₃₇ 190-193 Compound 28 H OC ₁₀ H ₂₁ 216-219 Compound 29 H OC ₁₈ H ₃₇ 168-171

化合物 30 H OG₁₈H₃₇ 228-231

Compound 30 H OG ₁₈ H ₃₇ 228-231

C0 ₂ H compound 31 H NHCOC ₁₇ H ₃₅ 261-264

C0 ₂ H compound 32 H OC ₁₈ H ₃₇ 54-56 compound 33 H OC ₁₈ H ₃₇ 217-222 compound 34 H OC ₁₈ H ₃₇ 204-209 compound 35 H OC ₁₈ H ₃₇ 166-170 compound 36 H OC ₁₈ H ₃₇ 165-169

[製造例 Ί ]

[Production example Ί]

 To a solution of 11.4 g of 4-aminobenzoic acid and 15.4 g of 2-fluoro-5-nitrobenzoic acid in 500 ml of N, N-dimethylformamide, add 22.9 g of anhydrous potassium carbonate and 0.462 g of powder, and add 100 ° C. The mixture was stirred at C for 1 hour, at 12 CTC for 3 hours, and at 140 ° C for 8 hours. The reaction solution was acidified by adding water and hydrochloric acid, and the precipitated solid was collected by filtration to obtain 20.6 g of a crude product.

 Anhydrous potassium carbonate 14.1 and methyl iodide 19.3 were added to a solution of 20.6 g of the above crude product in 500 ml of N, N-dimethylformamide, and the mixture was stirred at room temperature for 2 hours. Water was added to the reaction solution, extracted with ethyl acetate, and the organic layer was washed with saturated saline and dried over anhydrous magnesium sulfate. The crude product obtained by evaporating the solvent under reduced pressure is suspended in ethyl acetate, filtered and filtered to give 5-dimethyl-2,4'-iminodibenzoic acid dimethyl ester (melting point: 205-206 ° C) 13.5 g was obtained (the following reaction formula (17)).

(17) To a solution of 12.3 g of the compound obtained in the above reaction formula (17) in 300 ml of methanol was added 1.23 g of 10% palladium-carbon, and the mixture was stirred under a hydrogen atmosphere at room temperature for 4 hours. The reaction solution was filtered to remove the catalyst, and the crude product obtained by evaporating the solvent under reduced pressure was purified by silica gel column chromatography (eluted with hexane: ethyl acetate = 1: 1). And recrystallized to give 8.87 of 5-amino-2,4'-iminodibenzoic acid dimethyl ester (melting point: 113 to 115 ° C) (the following reaction formula ( 18))

18) A mixture of the compound 5.14 obtained by the above reaction formula (18) and 8.66 g of 3- (4-octyldecyloxyphenyl) -3-ethyloxoethyl ester was heated and stirred at 140 ° C. for 13 hours. The obtained crude product was purified by silica gel column chromatography (eluted with hexane: chloroform: ethyl acetate = 3: 1: 1) to give Compound 37 (melting point: 112 12: L at 14 ° C) 7.77 g (reaction formula (19) below)

化合物 37 ( 19)

Compound 37 (19)

[Production Example 8]

To a solution of 3.92 g of 3-aminobenzoic acid and 5.30 g of 2-fluoro-5-nitrobenzoic acid in 180 ml of N, N-dimethylformamide was added 9.88 g of anhydrous potassium carbonate and 0.16 g of copper powder. The mixture was stirred at ° C for 10 hours. The reaction solution was acidified by adding water and hydrochloric acid, and the precipitated solid was collected by filtration to obtain 11.7 g of a crude product. To a solution of the above crude product (11.7 g) in N, N-dimethylformamide (200 ml) was added anhydrous potassium carbonate (8.02 g) and methyl iodide (8.23 g), and the mixture was stirred at room temperature for 4 hours. Water was added to the reaction solution, extracted with ethyl acetate, and the organic layer was washed with saturated saline and dried over anhydrous magnesium sulfate. The crude product obtained by evaporating the solvent under reduced pressure was purified by silica gel column chromatography (eluted with hexane: ethyl acetate = 2: 1) to give 5-nitro-1,2,3'-iminodibenzo. 6.23 g of dimethyl ester (melting point: 149 to 151 ° C.) was obtained (the following reaction formula (20)).

 (20) To a solution of 6.23 g of the compound obtained in the above reaction formula (20) in 150 ml of methanol was added 0.623 g of 10% palladium-carbon, and the mixture was stirred at room temperature under a hydrogen atmosphere for 17 hours. The reaction solution was filtered to remove the catalyst, the solvent was distilled off under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (eluted with hexane: ethyl acetate = 2: 1) to give 5-amino. 5.37 g of 1,2′-imino dibenzoic acid dimethyl ester (yellow oily substance) was obtained (the following reaction formula (21)).

(twenty one) A mixture of 3.40 g of the compound obtained by the above reaction formula (21) and 5.22 g of 3- (4-oxodecyloxyphenyl) ethyl 3-oxopropionate was heated and stirred at 140 ° C. for 10 hours. The obtained crude product was purified by silica gel column chromatography (eluted with hexane: chloroform: ethyl acetate = 3: 2: 1), and recrystallized from methanol to give Compound 38 (melting point: 100 4.75 g was obtained (the following reaction formula (22)).

 Compound 38 (22)

[Production Example 9]

By the same operation as in Production Example 7 and Production Example 8, Compound 39 and Compound 40 represented by General Formula (23) and having R ¹⁹ having the structure shown in Table 5 were obtained. Their melting points are also shown in Table 5.

表 5

Table 5

[Production Example 10]

 An aqueous sodium hydroxide solution (2.80 g of sodium hydroxide, 50 ml of water) was added to a mixture of the compound 37 of 5.00 suspended in 50 ml of ethanol, and the mixture was stirred at 80 ° C for 3 hours. The reaction solution was acidified by adding 5% hydrochloric acid, and the precipitate was collected by filtration, washed with water, and dried to obtain 4.28 g of compound 41 (melting point: 214 to 219 ° C) (the following reaction formula (24) )).

Compound 37

 Compound 41 (24)

[Production Example 1 1]

To a mixture of 4.73 g of compound 38 suspended in 140 ml of ethanol was added an aqueous sodium hydroxide solution (2.65 g of sodium hydroxide, 70 ml of water), and the mixture was stirred at 70 ° C for 1 hour. The reaction mixture was acidified by adding 5% hydrochloric acid, and the precipitate was collected by filtration, washed with a trowel and dried to obtain 4.20 g of compound 42 (melting point: 211 to 215 ° C) (the following reaction). (Equation (25))

Compound 38

[Production Example 12]

The same operations as in Production Example 10 and Production Example 11 were carried out using Compound 39 and Compound 40 to give Compound 43 and Compound 44 represented by General Formula (26), wherein R ^2Q had the structure shown in Table 6. Obtained. Their melting points are also shown in Table 6.

(26)

Table 6

R ²⁰ Melting point (° c) Compound 43 C0 ₂ H 231-235 Compound 44 H 147-150 [Test Example 1]

 The following tests were performed according to the method described in the literature (Cell Growth & Differentiation, Volume 7, pages 213 to 221, 1996).

NIH3T3 cells (7 × 10 ⁴ cells / well) in which Flt-1 was forcibly expressed were seeded on a 24-well collagen-coated plate, and Dulbecco's modified Eagle's medium (DMEM) containing 10% calf serum and 200 zg / ml Geneticin G418 was added. ), And cultured at 37 ° C for 24 hours in a 5% carbon dioxide atmosphere. Place the cells in buffer A [containing 10 mM HEPES (Ν-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) and 0.1% BSA (bovine serum a lbumin) in DMEM] at 4 ° C for 30 minutes Planned paste. After that, the medium was exchanged for buffer B (containing 10 mM HEPES and 0.5% BSA in DMEM), and each of the test compounds shown in Table 7 was dissolved in dimethyl sulfoxide and diluted to the specified concentration with buffer B to prepare. The prepared test solution and [ ¹²⁵ I] -VEGF (final concentration: 25 pM) were added, and the binding reaction was performed at 4 ° C for 90 minutes. After completion of the reaction, the cells were washed three times with ice-cold buffer A. Subsequently, 0.5 ml of 0.5 M NaOH was added to each well, and the cells were thawed at room temperature for 30 minutes. The radioactivity of the cell lysate for each Ueru measured at Ganmakaun evening one to calculate the total amount of binding of ^[125 1] -VE GF. ^[125 1] - non-specific binding of VE GF, was determined by competitive Atsusi (competition assay) of unlabeled VE GF presence of ΙΟηΜ, ^[125 I] - from the difference between ¥ £ total binding of 0 The specific binding of [ ¹²⁵¹ ]-£ 0 was calculated.

The binding inhibition rate of the test compound was calculated by the following equation. Binding inhibition rate (%) = [ ¹²⁵ 1] of test compound added group — VEGF specific binding amount

 [1] X 100

[ ¹²⁵ I] -VEGF specific binding amount of control group From this value, 50% binding inhibitory concentration (IC ₅ ) of the test compound was calculated. Table 7 shows the results. Table 7

[Test Example 2]

Using NIH3T3 cells in which KDR was forcibly expressed, each of the test compounds shown in Table 8 was tested in the same manner as in Test Example 1 above. Table 8 shows the results.

Table 8 1 T ¹ 50 (it Μ IU,

J 1 Q7 ft ft 1 1 1 VJ Ω Ω Ω fi

、 し 口 1YJ \

 V ^ H

, Mouth 1¾l <

 I ΰ

Π A

1.4U

ic mouth U.O 1

U.C30 Distilled Mouthpiece 'yyj 1 fi Compound 36 0.63 Compound 41 0.48 Compound 42 0.53 Compound 43 0.75 [Test Example 3]

 In vivo tumor growth inhibition test

The human colon cancer cell line Co 1-1 tumor passaged subcutaneously in a nude mouse was excised, cut into small pieces, and a 2 mm x 2 mm x 2 mm tumor mass was implanted subcutaneously into the right flank of nude mice with a needle. dayO). A test solution prepared by dissolving a test compound (compound 19) in isotonic phosphate buffer (PH9) was administered to tumor-implanted nude mice for 28 days (from day 1 to day 28) daily, as shown in Table 9. The dose was administered intraperitoneally. The tumor volume was calculated from the values of the major axis and minor axis of the tumor measured using calipers on day 30 according to the following formula. Tumor volume (mm ³ ) = 0.5 X [major axis (mm)] x [minor axis (mm)] ² The tumor growth inhibitory effect of the test compound was expressed as T / C ( %) By the following equation. Table 9 shows the results. Tumor volume of test compound administration group

 T / C (%) = X I 00

 Tumor volume of control group Table 9

Dose TZC (%)

(mgZkgZaay) Industrial applicability

 The VEGF receptor antagonist of the present invention is considered to inhibit angiogenesis by inhibiting VEGF-dependent vascular endothelial cell proliferation, and to suppress VEGF-induced increase in vascular permeability.

 Therefore, the VEGF receptor antagonist of the present invention is expected to be used as a therapeutic agent for diseases involving angiogenesis induced by VEGF, such as diabetic retinopathy, rheumatoid arthritis, and solid tumors. In addition, the VEGF receptor antagonist of the present invention is expected to have an effect of suppressing pathological symptoms associated with VEGF-induced vascular hyperpermeability such as cerebral edema during ischemia-reperfusion injury.

Claims

The scope of the claims 1. The following equation (1)

(1) (In the formula (1), R ¹ is a hydrogen atom, C ₂ R ⁵ or OR ⁶ (where R ⁵ and R ⁶ are each a hydrogen atom or a C ₆ alkyl group.) R ² is a hydrogen atom or a group represented by C 0 ₂ R ⁷ (where R ⁷ is a hydrogen atom or d- ₆ alkyl group), and R ³ is a hydrogen atom or C ₆ alkyl R ⁴ is a C ₅ alkyl group, X is 0, S or NH, and Y is 0 or NHCO.) Or a pharmaceutically acceptable salt thereof as an active ingredient A VEGF receptor antagonist. 2. In formula (1), R ⁴ is C ₁₄ - VE GF receptor antagonist of claim 1 which is ₂₂ alkyl group. 3. The VEGF receptor antagonist according to claim 1, wherein in the formula (1), R ⁴ is a C ₁₈ alkyl group. 4. In the formula (1), R ¹ is a group represented by C 0 ₂ R ⁵ (where R ⁵ is a hydrogen atom or a d ₆ alkyl group), and R ² is a hydrogen atom. Item 4. The VEGF receptor antagonist according to Item 3. 5. The following formula (2)

 (2) (In the formula (2), R ³ and R ⁵ are each a hydrogen atom or a CH alkyl group, and is 0, S or NH.) Or a pharmaceutically acceptable salt thereof as an active ingredient 2. The VEGF receptor antagonist according to claim 1, which is contained as 6. The VEGF receptor antagonist according to claim 5, wherein in the formula (2), R ³ and R ⁵ are both hydrogen atoms. 7. A therapeutic agent for a disease associated with VEGF, comprising the VEGF receptor antagonist according to any one of claims 1 to 6. 8. The therapeutic agent according to claim 7, wherein the disease associated with VEGF is selected from the group consisting of diabetic retinopathy, rheumatoid arthritis, solid tumor, and cerebral edema at ischemia-reperfusion injury. 9. A method for treating a disease associated with VEGF, which comprises administering the VEGF receptor antagonist according to any one of claims 1 to 6. 10. The method according to claim 9, wherein the disease associated with VEGF is selected from the group consisting of diabetic retinopathy, rheumatoid arthritis, solid tumor, and cerebral edema at ischemia-reperfusion injury. 1 1. Use of a compound represented by the formula (1) or a pharmaceutically acceptable salt thereof for the treatment of a therapeutic agent for a disease associated with VEGF. 12. The use according to claim 11, wherein the disease associated with VEGF is selected from the group consisting of diabetic retinopathy, rheumatoid arthritis, solid tumors, and cerebral edema during ischemia-reperfusion injury. 13. Use of a compound represented by the formula (1) or a pharmaceutically acceptable salt thereof for producing a VEGF receptor antagonist. 14. Use of a compound represented by the formula (1) or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating a disease associated with VEGF. 15. The use according to claim 14, wherein the disease associated with VEGF is selected from the group consisting of diabetic retinopathy, rheumatoid arthritis, solid tumor, and cerebral edema at ischemia-reperfusion injury.