TW201731824A - INTEGRIN [alpha]4[beta]7 INHIBITOR - Google Patents

Abstract

Provided is a compound having an excellent integrin [alpha]4 inhibiting action. A sulfonamide derivative represented by general formula (I) or a pharmaceutically acceptable salt thereof. (In the formula, R1-R5, e-h, D, and B are as defined in the specification.).

Description

44β7 integrin inhibitor

The present invention relates to a sulfonamide derivative or a pharmaceutically acceptable salt thereof and a pharmaceutical composition containing the compound as an active ingredient. In particular, a compound which is useful as a therapeutic or prophylactic agent for an inflammatory disease associated with a lesion, which is an α4 integrin-dependent subsequent process.

Compounds which can be orally administered as a therapeutic or prophylactic agent for an inflammatory disease associated with a lesion and which have an inhibitory action of α4 integrin are known as an α4 integrin-dependent follow-up process. For example, Patent Document 1 discloses a phenylalanine derivative represented by the following formula or a pharmaceutically acceptable salt thereof, and the representative compound has the following chemical structure.

Further, Patent Document 1 discloses the results of VCAM inhibitory activity (VCAM-1/α4β1 binding assay) and (VCAM-1/α4β7 binding assay).

Further, Patent Document 2 discloses a phenylalanine derivative represented by the following formula having a R12(R13)N-X1- group at the terminal or a pharmaceutically acceptable salt thereof.

This compound appeared to have a higher VCAM-1/α4β1 integrin inhibitory activity in the presence of serum than the compound of Example 1 of Patent Document 1. Further, Patent Document 3 also discloses a compound having an α4 integrin inhibitory action.

In Patent Document 4 (WO 2005/077915), a phenylalanine derivative having an α4 integrin inhibitory action represented by the following formula is described, but a 2,6-dichlorobenzylidene group is bonded at the N-terminus of phenylalanine. Or an amino acid residue or the like.

Patent Document 5 (JP-A-2003-321358) discloses a phenylalanine derivative having an inhibitory action of α4 integrin represented by the following formula, but a bond of 2,6-dichlorobenzene at the N-terminus of phenylalanine醯基等.

In Patent Document 6 (WO 01/56994), a phenylalanine derivative having an α4 integrin inhibitory activity represented by the following formula is disclosed, but a guanine acid or the like is bonded to the N-terminus of phenylalanine.

In Patent Document 7 (WO2006/127584), a phenylalanine derivative having an α4 integrin inhibitory action represented by the following formula is described, but a pyrimidine ring or the like is directly bonded to the N-terminus of phenylalanine.

Patent Document 8 (WO01/42215) describes the following formula The phenylalanine derivative having an inhibitory action of α4 integrin is represented, but a 2-chloro-6-methylbenzhydryl group is bonded to the N-terminus of phenylalanine.

Patent Document 9 (WO 2013/161904) discloses a phenylalanine derivative having an α4β7 integrin inhibitory action represented by the following formula. In this document, the results of an evaluation of the VCAM-1/α4β1 integrin binding inhibitory activity of a specific phenylalanine derivative and the evaluation of the MAdCAM-1/α4β7 integrin binding inhibitory activity in the presence of serum are disclosed, and the α4β1 integrin is described. The effect is lower, and the effect is higher than that of α4β7 integrin.

Patent Document 10 (WO2015/064580) discloses a phenylalanine derivative having an α4β7 integrin inhibitory action represented by the following formula. There are also reports in this literature that assess the VCAM-1/α4β1 integrin binding inhibitory activity of specific phenylalanine derivatives and the evaluation of the MAdCAM-1/α4β7 integrin binding inhibitory activity assay in the presence of serum. The results are described as having a lower effect relative to the α4β1 integrin and a higher effect than the α4β7 integrin.

[Previous Technical Literature] [Patent Literature]

[Patent Document 1] WO02/16329

[Patent Document 2] WO05/061466

[Patent Document 3] WO03/070709

[Patent Document 4] WO2005/077915

[Patent Document 5] JP-A-2003-321358

[Patent Document 6] WO01/56994

[Patent Document 7] WO2006/127584

[Patent Document 8] WO01/42215

[Patent Document 9] WO2013/161904

[Patent Document 10] WO2015/064580

[Non-patent literature]

[Non-Patent Document 1] Nat Med. 2014 Dec; 20(12): 1397-400.

SUMMARY OF THE INVENTION An object of the present invention is to provide a novel compound having a chemical structure which is not known in the past and which has an excellent α4 integrin inhibitory action.

In particular, it is an object of the present invention to provide a novel compound having a low effect on α4β1 and a high selectivity to α4β7, which has a high α4 integrin inhibitory effect.

Further, it is an object of the present invention to provide a compound which is capable of oral administration and which has an excellent α4 integrin inhibitory action.

Further, it is an object of the present invention to provide a compound having α11 integrin inhibitory activity which is excellent in safety.

Further, it is an object of the present invention to provide a compound having a sustained long α4 integrin inhibitory activity.

Moreover, it is an object of the present invention to provide a novel compound having excellent α4 integrin inhibition in human whole blood.

Further, it is an object of the present invention to provide a pharmaceutical composition comprising the above novel compound and a pharmaceutically acceptable carrier.

Moreover, it is an object of the present invention to provide a medicament containing the above novel compound.

Further, it is an object of the present invention to provide a therapeutic or prophylactic agent for an α4β7 integrin-dependent subsequent process and a disease-related inflammatory disease.

Moreover, it is an object of the present invention to provide an α4 integrin inhibitor.

The inventors of the present invention investigated the inhibitory activity of α4β7 integrin in human whole blood for compounds having various structures. As a result, it has been found that a sulfonamide derivative having a specific chemical structure bonded to a sulfonyl group having an amine carbonyl group as a substituent or a sulfonyl group of a phenyl group, or a pharmaceutically acceptable salt thereof, is excellent in α4β7 in human whole blood. Integrin inhibitory activity.

That is, the present invention encompasses the following forms [1] to [17].

[1] a sulfonamide derivative represented by the following general formula (I), or a pharmaceutically acceptable salt thereof,

(in the formula, Represents a single bond or a double bond, and R ₁ and R ₂ each independently represent a hydrogen atom, a halogen atom, a lower alkyl group, a lower alkenyl group, a lower alkoxy group, a lower alkoxy lower alkyl group, a hydroxyl group, or a hydroxy lower alkyl group. R ₁ and R ₂ may also be bonded to each other to form a benzene ring which may have a substituent, an alicyclic hydrocarbon having 4 to 7 carbon atoms which may have a substituent, a heteroaromatic ring which may have a substituent, or a heterocyclic ring which may have a substituent, R ₃ represents a lower alkyl group, and e, f, g and h each independently represent CH or a nitrogen atom, B represents a hydroxyl group, an alkoxy group having a carbon number of 1 to 10, and -O- a cyclic group, a oximeoxy group, or an olmesartan esteroxy group, D represents a benzene ring or a heteroaryl ring which may have a substituent, and R ₄ and R ₅ each independently represent a hydrogen atom and a lower alkane which may have a substituent a lower alkyl group which may have a substituent, a phenyl group which may have a substituent, or a heterocyclic group which may have a substituent, and R ₄ and R ₅ may be bonded to each other to form a substituent which may have a substituent. ring).

[2] The sulfonamide derivative of the above [1], or a pharmaceutically acceptable salt thereof, Represents a double bond.

[3] The sulfonamide derivative of the above [1] or [2], or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ are bonded to each other to form a benzene ring which may have a substituent, or a alicyclic hydrocarbon having 4 to 7 carbon atoms having a substituent, a heteroaromatic ring which may have a substituent, or a heterocyclic ring which may have a substituent selected from a lower alkyl group, a lower alkoxy group, and a hydroxyl group. Lower alkyl, amine, lower alkylamino, and lower alkylamino lower alkyl.

[4] The sulfonamide derivative according to the above [1] or [2], wherein R ₁ and R ₂ each independently represent a hydrogen atom, a lower alkyl group or a lower alkoxymethyl group, or a pharmaceutically acceptable salt thereof. R ₁ and R ₂ may be bonded to each other to form an alicyclic hydrocarbon having 4 to 7 carbon atoms which may have a substituent, a heteroaromatic ring which may have a substituent, or a heterocyclic ring which may have a substituent, and the substitution The group is selected from the group consisting of a lower alkyl group and a lower alkoxy group.

[5] The sulfonamide derivative according to the above [1] or [2], or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ each independently represent a hydrogen atom, a lower alkyl group or a lower alkoxymethyl group.

[6] The sulfonamide derivative of the above [1] or [2], or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ are bonded to each other to form a cyclohexene which may have a substituent, or a pyridine having a substituent, a dihydropyran which may have a substituent, or a tetrahydropyran which may also have a substituent selected from the group consisting of a lower alkyl group, a lower alkoxy group, a hydroxy lower alkyl group, and an amine group. a lower alkylamino group and a lower alkylamino lower alkyl group.

[7] The sulfonamide derivative according to any one of the above [1] to [6] or a pharmaceutically acceptable salt thereof, wherein e represents a nitrogen atom, and f, g and h represent C-H.

[8] The sulfonamide derivative according to any one of [1] to [7], wherein the B represents a hydroxyl group or an alkoxy group having 1 to 6 carbon atoms, or a pharmaceutically acceptable salt thereof.

[9] The sulfonamide derivative or a pharmaceutically acceptable salt according to any one of the above [1] to [8] wherein the substituent of D is selected from a halogen atom, a lower alkyl group, a lower alkoxy group, and Hydroxyl.

[10] The sulfonamide derivative according to any one of [1] to [8], wherein the D represents a benzene ring which may have a substituent, and the substituent is selected from a halogen atom, or a pharmaceutically acceptable salt thereof. , lower alkyl, lower alkoxy, and hydroxyl.

[11] The sulfonamide derivative of any one of the above [1] to [10], or a pharmaceutically acceptable salt thereof, wherein R ₄ and R ₅ each independently represent a hydrogen atom, and may have a lower substituent. An alkyl group, a lower alkenyl group which may have a substituent, a phenyl group which may have a substituent, or a heterocyclic group which may have a substituent selected from a halogen atom, a cyano group, a hydroxyl group, a lower alkyl group, a lower alkoxy group, a trifluoromethyl group, and a heterocyclic group, and R ₄ and R ₅ may be bonded to each other to form a heterocyclic group which may have a substituent selected from a lower alkyl group and a lower alkoxy group. Hydroxyl.

[12] The sulfonamide derivative of any one of the above [1] to [10], or a pharmaceutically acceptable salt thereof, wherein R ₄ and R ₅ each independently represent a hydrogen atom and may have a lower substituent An alkyl group or a heterocyclic group which may have a substituent selected from a halogen atom, a cyano group, a hydroxyl group, a lower alkoxy group, a trifluoromethyl group, and a heterocyclic group, and R ₄ and R ₅ may also be bonded. A heterocyclic group which may have a lower alkyl group as a substituent is formed.

[13] The sulfonamide derivative of the above [1], or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ each independently represent a hydrogen atom, a lower alkyl group, a lower alkoxy group, or a lower alkoxy group. The alkyl group, R ₁ and R ₂ may also be bonded to form an alicyclic hydrocarbon having 4 to 7 carbon atoms, a heteroaromatic ring, or a heterocyclic ring which may also be substituted by a lower alkyl group, and D may be selected from a halogen atom. a phenyl ring or a heteroaryl ring substituted with a substituent in a group of a lower alkyl group, a lower alkoxy group and a hydroxyl group, and each of R ₄ and R ₅ independently represents a hydrogen atom, or may have a lower alkoxy group or a hetero group a lower alkyl group having a substituent in a group of a ring group, a hydroxyl group, a cyano group, and a halogen atom, but R ₄ and R ₅ may be bonded to each other to form a heterocyclic group which may be substituted with a lower alkyl group, but, D When a benzene ring substituted with a halogen atom is represented, R ₁ and R ₂ each represent a lower alkyl group, and when D is a benzene ring having a lower alkyl group, a lower alkoxy group or a hydroxyl group as a substituent, the substituent is adjacent to carbon atom bonded to a carbon atom of S and D and bonded, R ₄ or R ₅ represents a substituent of the hydroxyl group by lower alkyl, lower alkyl substituted by hydroxyl groups which By the following formula (a) or (b) represented by,

However, when R ₄ or R ₅ is represented by the formula (b), R ₁ and R ₂ each represent a lower alkyl group, and when R ₄ or R ₅ is a lower alkyl group substituted by a cyano group, D represents a benzene ring.

[14] The sulfonamide derivative of the above [1] or a pharmaceutically acceptable salt thereof, which is represented by the following formula,

(wherein R ₁ , R ₂ , R ₃ , e, f, g, h, and B are as defined in the above [1])

[15] The sulfonamide derivative, or a pharmaceutically acceptable salt thereof, according to the above [1], which is represented by the following formula:

(where B is as defined in [1])

[16] A pharmaceutical composition comprising the sulfonamide derivative according to any one of the above [1] to [15], or a pharmaceutically acceptable salt thereof.

[17] A therapeutic or prophylactic agent for an inflammatory disease associated with a lesion, which is a sulfonamide derivative according to any one of the above [1] to [15], or a pharmaceutical thereof The salt allowed on it.

[18] An α4β7 integrin inhibitor comprising the sulfonamide derivative according to any one of the above [1] to [15], or a pharmaceutically acceptable substance thereof Salt.

According to the present invention, it is possible to provide a novel compound having a chemical structure which is not known in the past and which has an excellent α4 integrin inhibitory action.

In particular, according to the present invention, it is possible to provide a novel compound having a low effect on α4β1 and a high selectivity to α4β7, which has a high α4 integrin inhibitory effect.

Further, according to the present invention, it is possible to provide a compound which is capable of oral administration and which has an excellent α4 integrin inhibitory action.

Further, according to the present invention, it is possible to provide a compound which is safe and has excellent α4 integrin inhibitory activity.

Further, according to the present invention, it is possible to provide a compound having a long-lasting inhibitory activity of α4 integrin.

Moreover, with the present invention, it is possible to provide a novel compound having an excellent α4 integrin inhibitory action in human blood.

Further, according to the present invention, it is possible to provide a pharmaceutical composition comprising the above novel compound and a pharmaceutically acceptable carrier.

Moreover, according to the present invention, it is possible to provide a medicine containing the above novel compound.

Further, according to the present invention, it is possible to provide a therapeutic or prophylactic agent for an inflammatory disease associated with a lesion, which is followed by an α4β7 integrin-dependent process.

Moreover, an α4 integrin inhibitor is provided by the present invention.

In the present specification, "may also have a substituent" means "substituted or unsubstituted". The position and the number of the substituents are not particularly limited as long as they are not particularly limited, and are not particularly limited. When substituted with two or more substituents, the substituents may be the same or different. The substituent may, for example, be a halogen atom, a nitro group, a cyano group, a hydroxyl group, a lower alkyl group, a lower alkenyl group, a lower alkynyl group, a lower alkoxy group, a lower alkylthio group, a hydroxy lower alkyl group or a hydroxy lower alkenyl group. Hydroxy lower alkoxy, lower alkoxyalkyl, halogenated lower alkyl, halogenated lower alkenyl, halogenated lower alkoxy, halogenated lower alkylthio, amine, lower alkylamino, lower alkylamine carbonyl, carboxyl, Lower alkoxycarbonyl, amine mercapto, lower alkylamine, aryl fluorenyl, lower alkyl sulfinyl, lower alkyl sulfonyl, amine sulfonyl, ammonium, aryl, heterocyclic, Aryl lower alkyl, heterocyclic lower alkyl, aryloxy, heterocyclooxy, arylsulfonyl, heterocyclosulfonyl, dihydroxyboroboryl, lower alkylamino lower alkyl, aryl lower An alkoxycarbonyl group, a lower alkenyloxy group, a lower alkoxy group, a lower amidino group, and the like.

The term "lower" in this specification means a group having a carbon number of 1 to 6, and "lower alkyl group" means a straight or branched chain or a cyclic alkyl group having 1 to 6 carbon atoms. For example, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, Tert-pentyl, neopentyl, 2-pentyl, 3-pentyl, n-hexyl, 2-hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclopropyl The ethyl group or the like is preferably a methyl group, an ethyl group or an n-propyl group.

"Lower alkenyl" means the number of carbons containing each isomer A linear or branched chain alkenyl group of 2 to 6. For example, a vinyl group, an allyl group, a propenyl group, a butenyl group, a pentenyl group, a hexenyl group or the like is preferred, and a vinyl group, an allyl group or a propylene group is preferred.

The "halogen atom" may, for example, be a fluorine atom, a chlorine atom, a bromine atom or an iodine atom, and is preferably a fluorine atom or a chlorine atom.

The "lower alkoxy group" means a straight or branched chain having a carbon number of 1 to 6, or an alkoxy group having a cyclic alkyl group. For example, methoxy, ethoxy, n-propoxy, n-butoxy, n-pentyloxy, n-hexyloxy, isopropoxy, isobutoxy, sec-butoxy And tert-butoxy, cyclopropyloxy, cyclobutoxy, cyclopentyloxy, and cyclohexyloxy, preferably methoxy, ethoxy, n-propoxy.

The "lower alkoxymethyl group" means a methyl group substituted with one or more of the above-mentioned "lower alkoxy group". For example, a methoxymethyl group, an ethoxymethyl group, an isopropoxymethyl group, a tert-butoxymethyl group or the like is preferred, and a methoxymethyl group or an ethoxymethyl group is preferred.

The "hydroxyl lower alkyl group" means a lower alkyl group substituted with a hydroxyl group, and examples thereof include a hydroxymethyl group, a hydroxyethyl group and the like, and a hydroxymethyl group is preferred.

The "lower alkylamino lower alkyl group" means a lower alkyl group substituted with an amine group, and the amine group is substituted by one of the aforementioned "lower alkyl groups" or by two. For example, methylaminomethyl, ethylaminomethyl, propylaminomethyl, isopropylaminomethyl, methylaminoethyl, ethylaminoethyl, dimethylamine The methyl group, the methyl ethylaminomethyl group and the like are preferably a methylaminomethyl group, an ethylaminomethyl group or a methylaminoethyl group.

The "alicyclic hydrocarbon" means a cyclic structure composed of a carbon atom and a hydrogen atom, and a cycloalkane formed by a single bond or a cyclic olefin which may further contain a double bond. For example, cyclobutane, cyclopentane, cyclohexene, cycloheptane, cyclohexene or the like is preferred, and cyclohexene and cyclohexene are preferred.

The "heteroaromatic ring" means an aromatic ring containing 4 to 4 members selected from the group consisting of an oxygen atom, a sulfur atom and a nitrogen atom as a ring atom. There are, for example, pyridine rings, hydrazine Ring, pyrimidine ring, pyridyl Ring, furan ring, thiophene ring, pyrrole ring, isoxazole ring, oxazole ring, isothiazole ring, thiazole ring, pyrazole ring, imidazole ring, oxadiazole ring, thiadiazole ring, triazole ring, tetrazole Ring, benzofuran ring, benzothiophene ring, anthracene ring, isoindole ring, benzoxazole ring, benzopyrene Oxazole ring, benzothiazole ring, benzisothiazole ring, benzimidazole ring, indazole ring, anthracene ring, quinoline ring, isoquinoline ring, oxazoline ring, anthracene Ring, quinazoline ring, quin The phenyl ring and the pteridine ring are preferably a pyridine ring or a pyrimidine ring.

The "heterocyclic ring" means a monocyclic to bicyclic heterocyclic ring containing 4 to 4 members selected from the group consisting of an oxygen atom, a sulfur atom and a nitrogen atom as a ring atom. Further, any carbon atom of the ring atom may be substituted by an oxy group, and the sulfur atom or the nitrogen atom may also be oxidized to form an oxide. Moreover, it can also be condensed with a benzene ring. For example, a tetracyclic ether ring, a tetrahydrofuran ring, a dihydropyran ring, a tetrahydropyran ring, two Ring, tetrahydrothiophene ring, tetrahydrothiopyran ring, tetrahydrothiazole ring, trimethylene ring, pyrrolidine ring, piperidine ring, piperazine Ring, same piperidine ring, same pipe Ring, pyrazolidine ring, imidazolium ring, tetrahydropyran ring, tetrahydropyrimidine ring, morpholine ring, thiomorpholine ring, porphyrin ring, isoporphyrin ring, Ring, different Anthracycline, azaporphyrin ring, pyridone ring, imidazole a ring, an imidazole thiazoline ring, a pyrimidinone ring, a carbendazim ring, a quinuclidine ring, etc., preferably a piperidine ring, a piperidine Ring, tetrahydropyran ring, morpholine ring.

The "heterocyclic group" means a monocyclic ?-ring heterocyclic group having 4 to 4 members selected from the group consisting of an oxygen atom, a sulfur atom and a nitrogen atom as a ring atom. Further, any carbon atom of the ring atom may be substituted by an oxy group, and the sulfur atom or the nitrogen atom may also be oxidized to form an oxide. Moreover, it can also be condensed with a benzene ring. There are tetracyclic ether groups, tetrahydrofuranyl groups, dihydropiperidyl groups, tetrahydropyranyl groups, and Base, tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydrothiazolyl, trimethyleneimine, pyrrolidinyl, piperidinyl, piperidine Base, same piperidinyl, same pipe , pyrazolidine, imidazolidinyl, tetrahydropyridyl, tetrahydropyrimidinyl, fluorenyl, thiomorphinyl, porphyrinyl, isoindolyl, 唍基,异 Sulfhydryl, azaindole, pyridinone, imidazolium, imidazolyl, pyrimidinone, beta-ureidourea, quinuclyl, etc., preferably piperidinyl, piperidin Base, tetrahydropyranyl, decyl, pyridone, and endocarbazide.

In the present invention, the sulfonamide derivative represented by the general formula (I) or a pharmaceutically acceptable salt thereof is preferred in the formula.

In the general formula (I), R ₁ and R ₂ each independently represent a hydrogen atom, a lower alkyl group or a lower alkoxymethyl group, and are preferably a hydrogen atom or a lower alkyl group, and particularly preferably a hydrogen atom and a methyl group.

In the general formula (I), the ring formed by the bonding of R ₁ and R _{2 is} preferably pyridine, cyclohexene, dihydropyran, and tetrahydropyran, and is cyclohexene, dihydropyran, and tetra. Hydropyran is further preferred, and dihydropyran and tetrahydropyran are particularly preferred.

In the general formula (I), R ₃ is preferably a lower alkyl group, and particularly preferably a methyl group.

In the general formula (I), e, f, g, and h are preferably C-H and a nitrogen atom, and any one of them is preferably a nitrogen atom, and e or f is particularly preferably a nitrogen atom.

In the general formula (I), B is preferably a hydroxyl group or a lower alkoxy group, and is preferably a hydroxyl group, a methoxy group, an ethoxy group, an isopropoxy group or a cyclohexyl group, and is preferably a hydroxyl group, a methoxy group or an ethoxy group. Isopropyloxy is especially preferred.

In the general formula (I), D is preferably a benzene ring which may have a substituent, and a heteroaryl ring which may have a substituent, and is preferably a benzene ring or a pyridine ring, and particularly preferably a benzene ring.

In the general formula (I), the substituent of D is a halogen atom, a lower alkyl group, a lower alkoxy group, a hydroxyl group, and preferably a fluorine atom, a methyl group, a methoxy group or a hydroxyl group.

In the general formula (I), when D represents a benzene ring or a pyridine ring, the substitution position of the amidoxime group bonded to D and the amine carbon group is preferably a para position.

In the general formula (I), R ₄ and R ₅ are each independently a lower alkyl group which may have a substituent, and a heterocyclic group which may have a substituent, and is preferably a methyl group, an ethyl group, a tert-butyl group or a ring group. Further preferred are propyl, 2-isopentyl, 3,3-dimethyl-2-butyl and tetrahydropyranyl, and ethyl, tert-butyl and cyclopropyl are particularly preferred.

In the general formula (I), the substituents of R ₄ and R _{5 are} preferably a methoxy group, a morpholinyl group, a trifluoromethyl group, a hydroxyl group or a cyano group, and are particularly preferably a morpholinyl group, a trifluoromethyl group or a cyano group. .

In the general formula (I), the ring formed by the bonding of R ₄ and R _{5 is} preferably a heterocyclic ring which may have a substituent, and is a morpholine or a piperazine. Especially good.

Further, it is preferred that the compound is a general formula (I), and R ₁ and R ₂ each independently represent a hydrogen atom, a lower alkyl group, a lower alkoxy group or a lower alkoxy lower alkyl group, and R ₁ and R ₂ may also be bonded to form an alicyclic hydrocarbon having 4 to 7 carbon atoms, a heteroaromatic ring, or a heterocyclic ring which may also be substituted by a lower alkyl group, and D may also be selected from a halogen atom, a lower alkyl group, and a lower stage. a benzene ring or a heteroaryl ring substituted with a substituent in a group of alkoxy groups and a hydroxyl group, and R ₄ and R ₅ each independently represent a hydrogen atom, or may have a group selected from the group consisting of lower alkoxy groups, heterocyclic groups, hydroxyl groups, and cyanogens. a lower alkyl group having a substituent in a group of a halogen atom, but R ₄ and R ₅ may be bonded to each other to form a heterocyclic group which may be substituted by a lower alkyl group, but D represents a halogen atom substituted In the case of a benzene ring, R ₁ and R ₂ each represent a lower alkyl group, and when D is a benzene ring having a lower alkyl group, a lower alkoxy group or a hydroxyl group as a substituent, the substituent may be a carbon bonded adjacent to S. a carbon atom of an atom is bonded to D, and when R ₄ or R ₅ represents a lower alkyl group substituted by a hydroxy group, the lower alkyl group substituted by the hydroxy group is represented by the following formula (a) or (b) Said,

However, when R ₄ or R ₅ is represented by the formula (b), R ₁ and R ₂ each represent a lower alkyl group, and when R ₄ or R ₅ is a lower alkyl group substituted by a cyano group, D represents a benzene ring. Such compounds have a particularly high selectivity.

When the compound represented by the general formula (I) of the present invention can form a salt, the salt thereof can be, for example, a chemical group such as a carboxyl group in the formula, and is an ammonium salt, a sodium salt or a potassium salt. Salts of alkali metals, salts with alkaline earth metals such as calcium and magnesium, aluminum salts, zinc salts, salts with organic amines such as triethylamine, ethanolamine, morpholine, piperidine, dicyclohexylamine, and arginine a salt of a basic amino acid such as lysine. The basic group in the case where a basic group is present may be a salt with an inorganic acid such as hydrochloric acid, sulfuric acid or phosphoric acid, or acetic acid, citric acid, benzoic acid, maleic acid, butenedioic acid or tartaric acid. a salt of an organic carboxylic acid such as succinic acid or a salt of an organic sulfonic acid such as methanesulfonic acid or p-toluenesulfonic acid. As a method of forming a salt, a compound of the general formula (I) and a necessary acid or base may be mixed in a suitable amount in a solvent, a dispersant, or cation exchange or anion exchange in the form of other salts.

The compound of the present invention may also contain a solvent of a compound represented by the general formula (I), such as a hydrate, an alcohol adduct or the like.

The compound of the present invention comprises a form of a prodrug of a compound represented by the general formula (I). The compounding of the compound of the present invention means a compound which is converted into a compound represented by the general formula (I) by a reaction of an enzyme or a gastric acid under physiological conditions in the living body, that is, causing enzyme oxidation, reduction, hydrolysis, etc. The compound which is converted into a compound represented by the general formula (I), or a compound which is converted into a compound represented by the general formula (I) by hydrolysis or the like by gastric acid or the like. The prodrug of the compound represented by the general formula (I) is exemplified in the compounds of the examples, but is not limited thereto. For example, the compound represented by the general formula (I) has When an amine group is used, as a precursor thereof, a compound which is deuterated, alkylated, or phosphorylated by the amine group (for example, an amine group of the compound represented by the general formula (I) is subjected to eicosylation and amiodalation. , pentamine carbonylation, (5-methyl-2-oxo-1,3-dioxolan-4-yl) methoxycarbonylation, tetrahydrofurylation, pyrrolidinylmethylated trimethyl When a compound represented by the formula (I) has a hydroxyl group, the compound represented by the formula (I) has a hydroxyl group, and as a precursor thereof, the hydroxyl group is deuterated, alkylated, and phosphorylated. a borated compound (for example, a hydroxyl group of a compound represented by the general formula (I) is acetylated, softened, propylated, trimethylacetylated, amber thiolated, butylene a compound which is dibasinized, propylamined, dimethylaminomethylcarbonylated, etc. When the compound represented by the formula (I) has a carboxyl group, as a precursor thereof, esterification of the carboxyl group and decylamine are mentioned. a compound (for example, a carboxyl group of a compound represented by the general formula (I) is methyl esterified, ethyl esterified, n-propyl esterified, phenyl esterified, isopropyl esterified, isobutyl esterified, Butyl esterification, cyclopentyl esterification, cyclohexyl esterification, cycloheptyl esterification, cyclobutyl methyl esterification, cyclohexyl methyl esterification, n-hexyl esterification, sec-butyl esterification, tert-butyl Esterification, (4-tetrahydropyranyl)methyl esterification, (4-tetrahydropyranyl) esterification, carboxymethyl esterification, dimethylaminomethyl esterification, trimethyl b Mercaptooxymethyl esterification, ethoxycarbonyloxyethyl esterification, mercaptoesterification, (5-methyl-2-oxo-1,3-dioxolan-4-yl) a compound which is esterified, cyclohexyloxycarbonylethyl esterified, methylammonium, etc. In particular, when the compound represented by the general formula (I) has a carboxyl group, as a precursor thereof, the carboxyl group is represented by carbon. A compound of a straight chain, a branched chain, or a cyclic alkyl group of 1 to 10 is preferred. These compounds can be represented by a general formula (I) by a conventionally known method. To manufacture.

Further, the compound (I) can be converted into the compound (I) by the physiological conditions described in pp. 163 to 198 of the seventh volume of the development of the pharmaceutical product of Hirokawa Shoten, 1990.

The present invention encompasses all isotopes of the compounds represented by the formula (I). The isotope of the compound of the present invention is substituted by at least one atom having the same atomic number (number of protons) but different mass (the sum of protons and the number of neutrons). Examples of the isotope contained in the compound of the present invention include a hydrogen atom, a carbon atom, a nitrogen atom, an oxygen atom, a phosphorus atom, a sulfur atom, a fluorine atom, a chlorine atom and the like, and each contains ² H, ³ H, ¹³ C, ¹⁴ C. , ¹⁵ N, ¹⁷ O, ¹⁸ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ³⁶ Cl, etc. In particular, it is useful when a radioactive isotope that emits radioactivity and emits neutrons such as ³ H or ¹⁴ C is used in a tissue distribution test of a pharmaceutical or a compound. The stable isotope does not cause disintegration, and the amount of the isotope hardly changes, and there is no radioactivity, so it can be used safely. The isotope of the compound of the present invention can be converted by a usual method by replacing the reagent used for the synthesis with a reagent containing a corresponding isotope.

The compound represented by the general formula (I) or a salt thereof is administered directly or as various pharmaceutical compositions. As a dosage form of such a pharmaceutical composition, it can also be used, for example, as a tablet, a powder, a pill, a granule, a capsule, a suppository, a solution, a sugar coating, an injection, or a syrup, using a common preparation auxiliary and as usual Made by law.

For example, a tablet is made by inertizing a phenylalanine derivative of the active ingredient of the present invention with a known auxiliary substance such as lactose, calcium carbonate or calcium phosphate. a releasing agent such as a release agent, gum arabic, white powder or gelatin, a bulking agent such as alginic acid, white powder or pregelatinized starch, a sweetener such as sucrose, lactose or saccharin, peppermint, red bead oil or cherry, etc. a softening gelatin capsule such as a fragrance, a magnesium stearate, a talc or a carboxymethylcellulose lubricant, a fat, a paraffin, a semi-solid and a liquid polyol, a natural oil or a hardened oil, and a medicinal form A solution of a dose, water, ethanol, glycerin, polyol, sucrose, invert sugar, glucose, vegetable oil or the like is obtained by mixing with an excipient.

The inhibitor of the compound represented by the general formula (I) or a salt thereof as an active ingredient can be used as a therapeutic or prophylactic agent for an inflammatory disease associated with a lesion in an α4 integrin-dependent subsequent process. As such an inflammatory disease, there are, for example, rheumatoid arthritis, inflammatory bowel disease, systemic lupus erythematosus, multiple sclerosis, Sjogren's syndrome, wheezing, dryness, allergy, diabetes, palpitary vascular disease. Arteriosclerosis, restenosis, tumor hyperplasia, tumor metastasis, transplant rejection, and/or human immunodeficiency virus infection (see Non-Patent Document 1).

The dosage for administration for the above purpose is determined according to the therapeutic effect of the purpose, the administration method, the treatment period, the age, the body weight, and the like, but is administered as an adult per day by an oral or parenteral route. In the case of oral administration, usually 1 μg to 5 g is used, and when it is administered orally, 0.01 μg to 1 g is usually used.

The amine carbonyl group may be provided as a substituent of D by the sulfonamide derivative represented by the general formula (I). By adopting such a configuration, excellent α4β7 integrin inhibitory activity can be obtained in human whole blood. In addition, this The sulfonamide derivative of the invention has a higher concentration of migration to the portal vein and an increased exposure amount in the circulating blood, so that a stronger effect can be obtained. From this point of view, it is useful as a therapeutic or prophylactic agent for an inflammatory disease associated with a lesion as an α4β7 integrin-dependent follow-up process. Further, in the general sulfonamide derivative of the formula (I), the 2-position and the 5-position of the phenyl group of the phenylalanine moiety are substituted by a fluorine atom. Thereby, it is possible to obtain a lower inhibitory activity against α4β1 integrin and a higher selectivity for α4β7 integrin.

The compound represented by the general formula (I) of the present invention can be, for example, an intermediate having a carboxyl group at the terminal represented by the general formula (MI) and an intermediate having an amine group at the terminal represented by the general formula (M-II). It is produced by a guanidation reaction.

The hydrazylation reaction is conventional, and examples thereof include (1) a method using a condensing agent, (2) a method using an acid halide, and the like.

(1) A method of using a condensing agent, for example, by using a carboxylic acid with an amine or a salt thereof, for example, dichloromethane, tetrahydrofuran (THF), 1,4-dioxane, N,N-dimethyl group a solvent such as guanamine (DMF) or acetonitrile which does not adversely affect the reaction, for example, in the presence or absence of a base such as pyridine, triethylamine or N,N-diisopropylethylamine. For example, in the presence of a condensation aid such as 1-hydroxybenzotriazole (HOBt), 1-hydroxy-7-azabenzotriazole (HOAt) or N-hydroxysuccinimide (HOSu), or In the presence of, for example, 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide (WSC), 1,3-dicyclohexylcarbodiimide (DCC), or (7) A condensing agent such as -azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) is reacted.

(2) The method of using an acid halide can be carried out by using a carboxylic acid in a solvent such as dichloromethane which does not adversely affect the reaction, or in a solvent-free medium, for example, in the presence of a catalyst such as DMF. In the absence or absence, an acid halide obtained by reacting with, for example, sulfinium chloride, oxalic acid chloride or sulfinium bromide, in a solvent such as dichloromethane or THF which does not adversely affect the reaction, For example, in the presence of a base of pyridine or triethylamine or N,N-diisopropylethylamine, it is carried out by reacting with an amine or a salt thereof.

Among them, an intermediate having a carboxyl group at the terminal represented by the general formula (M-I) can be produced by the following method.

In the intermediate of the compound of the present invention represented by the general formula (M-I) having a carboxyl group at the terminal, the production method of the representative compound is as follows. Further, in the following description, the symbols in the formula are the same as those in the above formula (I) unless otherwise specified.

In the general formula (MI), D is an phenyl group which may have a substituent selected from the group consisting of a lower alkyl group, a lower alkoxy group, a hydroxyl group, and a halogen atom, or an intermediate having a carboxyl group at the terminal of a pyridyl group ( S8) can be synthesized using, for example, the methods (manufacturing methods A, B, and C) described below.

<Manufacturing Method A>

In the formula, D ₁ represents a substituent represented by the above D, or a substituent which can be easily converted into D by an operation such as deprotection, wherein R ₂₁ represents a substituent of a general ester such as a lower alkyl group. In the formula, X ₁ represents a halogen atom such as chlorine, bromine or iodine or a leaving group such as trifluoromethanesulfonyloxy group.

The sulfonyl chloride derivative (S1) and the aniline derivative (S2) are in a solvent such as dichloromethane, acetonitrile, THF, or DMF which do not adversely affect the reaction, and are, for example, pyridine or triethyl The sulfonamide derivative (S3) can be synthesized by reacting in the presence of a base such as an amine. The obtained sulfonamide derivative (S3) is, for example, a solvent which does not adversely affect the reaction, such as DMF or N-methyl nitropentafluorenone (NMP), for example, triethylamine, N,N-diisopropyl In the presence of a base such as a methylamine or the like, a metal catalyst such as 1,1'-bis(diphenylphosphinoferrocene)dichloropalladium (II) or the like is used in the oxidation. The aldehyde derivative (S4) can be synthesized by performing a coupling reaction in a carbon atmosphere. The obtained aldehyde derivative (S4) can synthesize a carboxylic acid derivative (S5) by performing an oxidation reaction such as Pinnick oxidation. The obtained carboxylic acid derivative (S5) and the amine derivative (S6) or a salt thereof, for example, dichloromethane, THF, 1,4-dioxane, DMF or acetonitrile, etc., do not adversely affect the reaction. In the solvent, in the presence or absence of a base such as pyridine, triethylamine or N,N-diisopropylethylamine, in the presence of a condensation aid such as HOBt, HOAt or HOSu In the presence or absence, it can be derivatized into a corresponding guanamine derivative (S7) by using a condensing agent such as WSC, DCC or HATU. Next, the guanamine derivative (S7) is used, for example, in a solvent such as THF, 1,4-dioxane, methanol or ethanol which does not adversely affect the reaction, by using, for example, sodium hydroxide or hydrogen. The base carboxylic acid derivative (S8) can be produced by alkaline hydrolysis of a base such as lithium oxide or by acid hydrolysis using, for example, hydrochloric acid or trifluoroacetic acid.

<Manufacturing method B>

In the formula, D ₁ represents a substituent represented by the above D, or a substituent which can be easily converted into D by an operation such as deprotection, wherein R ₂₁ represents a substituent of a general ester such as a lower alkyl group. .

The sulfonyl chloride derivative (S9) and the aniline derivative (S2) are, for example, in a solvent such as dichloromethane, acetonitrile, THF, or DMF which does not adversely affect the reaction, such as pyridine or triethylamine. The sulfonamide derivative (S5) can be synthesized by reacting it in the presence of a base such as the base. The obtained sulfonamide derivative (S5) and the derivative (S6) or a salt thereof, and a solvent such as dichloromethane, THF, 1,4-dioxane, DMF or acetonitrile which do not adversely affect the reaction In the presence or absence of a base such as pyridine, triethylamine or N,N-diisopropylethylamine, in the presence of a condensation aid such as HOBt, HOAt or HOSu, In the absence or presence of a condensing agent such as WSC, DCC or HATU, a corresponding decylamine derivative (S7) can be derivatized by reacting it. Next, the guanamine derivative (S7) is used, for example, in a solvent such as THF, 1,4-dioxane, methanol or ethanol which does not adversely affect the reaction, by using, for example, sodium hydroxide or hydrogen. The base carboxylic acid derivative (S8) can be produced by alkaline hydrolysis of a base such as lithium oxide or by acid hydrolysis using, for example, hydrochloric acid or trifluoroacetic acid.

<Manufacturing Method C>

In the formula, D ₁ represents a substituent represented by the above D, or a substituent which can be easily converted into D by an operation such as deprotection, wherein R ₂₁ represents a substituent of a general ester such as a lower alkyl group. .

The sulfonyl chloride derivative (S10) and the aniline derivative (S2) are, for example, in a solvent such as dichloromethane, acetonitrile, THF, or DMF which does not adversely affect the reaction, such as pyridine or triethylamine. The sulfonamide derivative (S7) can be synthesized by reacting it in the presence of a base such as the base. Next, the sulfonamide derivative (S7) is used, for example, in a solvent such as THF, 1,4-dioxane, methanol or ethanol which does not adversely affect the reaction, by, for example, using sodium hydroxide or hydroxide. The base carboxylic acid derivative (S8) can be produced by alkaline hydrolysis of a base such as lithium or, for example, hydrolysis with hydrochloric acid or acid hydrolysis of trifluoroacetic acid.

In the general formula (MI), D is a phenyl group or a pyridyl group which may have a substituent selected from the group consisting of a lower alkyl group, a lower alkoxy group, a hydroxyl group, and a halogen atom, and R ₅ is an end of a hydrogen atom. The intermediate (S8) having a carboxyl group can be synthesized by, for example, the method (Production Method D) described below.

<Manufacturing method D>

In the formula, D ₁ represents a substituent represented by the above D, or a substituent which can be easily converted into D by an operation such as deprotection, wherein R ₂₁ represents a substituent of a general ester such as a lower alkyl group. In the formula, X ₁ represents a halogen atom such as chlorine, bromine or iodine or a leaving group such as trifluoromethanesulfonyloxy group.

The halogenated aryl derivative (S3) and the isocyanide reagent (S11) in a solvent such as DMSO which does not adversely affect the reaction, in the presence or absence of, for example, water, for example, cesium carbonate The tert-butyl decylamine derivative (S12) can be synthesized by using a metal catalyst such as bisphosphine palladium (II) or the like in the presence of a base such as phenylphosphonium chloride. The obtained tert-butyl decylamine derivative (S12) is used, for example, in a solvent which does not adversely affect the reaction, such as THF, 1,4-dioxane, methanol or ethanol, by using, for example, sodium hydroxide. The base carboxylic acid derivative (S8) can be produced by alkaline hydrolysis of a base such as lithium hydroxide or by acid hydrolysis using hydrochloric acid or trifluoroacetic acid, for example.

In each step, it can be synthesized by using a reaction condition which can be generally substituted, and it should be selected as appropriate according to the kind of the raw material compound or the like. Further, the compound of the present invention obtained by the above method can be purified by a method such as extraction, distillation, crystallization, column chromatography or the like used in general organic synthesis.

The intermediate (S17) having an amine group at the terminal represented by the general formula (M-II) of the compound of the present invention can be synthesized by using, for example, the production methods (manufacturing methods E, F, and G) shown below. Further, in the following description, the symbols in the formula are the same as those in the above formula (I) unless otherwise specified.

<Manufacturing method E>

In the formula, R ₃₁ represents, for example, a tert-butoxycarbonyl group, a benzyloxycarbonyl group or the like, and a substituent of a general amine which can be removed by an operation such as deprotection, wherein X ₃ represents, for example, chlorine, bromine, iodine or the like. A halogen atom or a leaving group such as a trifluoromethanesulfonyloxy group, wherein B ₁ represents a substituent which can be easily converted into B by an operation such as deprotection.

A halogenated aryl derivative (S13) and a borane derivative such as bis(nonyl alcohol ester) diborane in a solvent such as DMF which does not adversely affect the reaction, for example, a base such as potassium acetate In the presence of a base, for example a metal catalyst such as 1,1'-bis(diphenylphosphinoferrocene)dichloropalladium (II), which is derivatized to a corresponding boronic acid ester derivative by a coupling reaction, followed by the obtained boric acid The ester derivative can be deprotected by adding, for example, sodium periodate, ammonium acetate or water to a solvent which does not adversely affect the reaction, such as acetone, to synthesize the corresponding boric acid. Derivative (S14). The obtained boric acid derivative (S14) and the uracil derivative (S15) are, for example, pyridine, in a solvent such as dichloromethane, dimethyl hydrazine (DMSO), or DMF which does not adversely affect the reaction. Or a metal catalyst such as copper (II) acetate or copper (II) trifluoromethanesulfonate in the presence of a base such as triethylamine, and an amino acid derivative (S16) can be synthesized by performing a coupling reaction. . Then, the amino acid derivative (S16) can be produced by, for example, deprotecting by acid hydrolysis using hydrochloric acid or trifluoroacetic acid or hydrodecomposition, to produce the desired carboxylic acid derivative (S17).

<Manufacturing method F>

In the formula, R ₃₁ represents, for example, a tert-butoxycarbonyl group, a benzyloxycarbonyl group or the like, and a substituent of a general amine which can be removed by an operation such as deprotection, wherein R ₃₂ and R ₃₃ independently represent, for example, a lower alkyl group. Or a substituent of a general ester such as a benzene ring having a substituent, wherein B ₁ represents a substituent which can be easily converted into B by an operation such as deprotection.

The nitrate derivative (S18) is contacted with a metal catalyst such as methanol, ethanol, or isopropyl alcohol which does not adversely affect the reaction, for example, a metal catalyst such as palladium carbon, palladium hydroxide, or Reyk nickel. The aniline derivative (S19) can be synthesized by a reduction reaction or under acidic conditions (for example, hydrochloric acid, acetic acid, or an amine chloride) by, for example, reacting with a metal such as zinc. The obtained aniline derivative (S19) and the carbamate derivative (S20) are in a solvent such as dichloromethane, 1,4-dioxane, THF, or DMF which do not adversely affect the reaction. By using a base such as triethylamine, pyridine or diazabicycloundecene (DBU), an amino acid can be synthesized by reacting the same Derivative (S16). Next, the amino acid derivative (S16) can be produced by, for example, deprotecting by acid hydrolysis using hydrochloric acid or trifluoroacetic acid or hydrodecomposition, etc., to produce the desired carboxylic acid derivative (S17).

<Manufacturing method G>

Wherein R ₃₁ represents, for example, a tert-butoxycarbonyl group, a benzyloxycarbonyl group or the like, and a substituent of a general amine which can be removed by an operation such as deprotection, wherein X ₃ , X ₄ and X ₅ are each independently represented. For example, a halogen atom such as chlorine, bromine or iodine or a leaving group such as trifluoromethanesulfonyloxy group, wherein B ₁ represents a substituent which can be easily converted into B by an operation such as deprotection.

The halogenated aryl derivative (S21) and the uracil derivative (S15) are, for example, DMSO, NMP, or DMF, which do not adversely affect the reaction, such as triethylamine, N, N-di In the presence of a base such as propylethylamine or DBU, it can be synthesized by, for example, a coupling reaction using a metal catalyst such as copper (I) iodide, copper (I) bromide or copper (I) chloride. Compound (S22). The obtained compound (S22) and halide (S23) are, for example, DMF In the solvent which does not adversely affect the reaction, for example, in the presence of zinc powder activated by iodine or the like, for example, a metal catalyst such as bis(phenylideneacetone)dipalladium (0) or the like is used. A ligand generally used in organic synthesis of dicyclohexyl-2',6'-dimethoxybisphenyl (SPhos), etc., capable of synthesizing an amino acid derivative by performing a root-shore coupling reaction (S16) ). Then, the amino acid derivative (S16) can be produced by, for example, deprotecting by acid hydrolysis using hydrochloric acid or trifluoroacetic acid or hydrodecomposition, to produce the desired carboxylic acid derivative (S17).

The present invention will be described in more detail based on the following synthesis examples, examples, and test examples. These are preferred embodiments of the present invention, and the present invention is not limited to the synthesis examples, examples, and test examples, and may be varied within a range not exceeding the scope of the present invention. Further, the reagents, devices, and materials used in the present invention can be commercially purchased unless otherwise specified.

The methyl 4-amino-2,5-difluoro-benzoin ester of the common intermediate can be synthesized according to the method described in the patent document (WO 2013/161904).

The synthesis examples of the intermediates used in the synthesis of the compounds of the examples are shown below.

[Synthesis Example 1]

4-[[4-(tert-butylamine-carbamoyl)phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

(step 1)

Synthesis of methyl 2,5-difluoro-4-[(4-iodophenyl)sulfonylamino]benzoin

Methyl 4-amino-2,5-difluoro-benzoin (3.0 g, 16.0 mmol) was dissolved in pyridine (30 ml), and 4-phenylsulfonyl chloride (12.1 g, 40.1 mmol) was added. Stir at 50 ° C for one night. The residue was concentrated under reduced pressure, and the obtained residue was washed with acetonitrile, and dried in vacuo to give methyl 4-[bis[(4-iodophenyl)sulfonyl]amino]-2,5-difluoro. - benzoin ester (11.6 g). THF (24 mL) was added, and the mixture was cooled to 0 ° C, and then a 1M solution of tetrabutylammonium fluoride in THF (24 mL) was added and stirred at room temperature for one and a half hours. A saturated aqueous solution of ammonium chloride was added, and concentrated under reduced pressure to remove THF. The mixture was diluted with ethyl acetate, washed 4 times with 0.5 M hydrochloric acid, washed once with 1M hydrochloric acid, and washed with brine. It was dried over magnesium sulfate and the solvent was distilled off under reduced pressure. The resulting residue was washed with hexane/ethyl acetate = 7:3. Vacuum drying was carried out to obtain the title compound (5.9 g, yield: 82%).

(Step 2)

Synthesis of methyl 2,5-difluoro-4-[(4-methylnonylphenyl)sulfonylamino]benzoin

The compound obtained in (Step 1) (1.0 g, 2.2 mmol) was dissolved in DMF (20 mL), and 1,1'-bis(diphenylphosphinoferrocene)dichloropalladium(II) (0.16 g, 0.22 mmol), and stirred at 70 ° C for 10 minutes under an argon atmosphere. After returning to room temperature, N,N-diisopropylethylamine (1.1 ml, 6.6 mmol) and triethyl decane (0.88 mL, 5.5 mmol) were added, and the mixture was stirred with carbon monoxide and then stirred at 70 ° C overnight. The residue obtained by distilling off the solvent under reduced pressure was charged into a reverse phase HPLC using ODS as a filler, and dissolved in a mixed solution of water and acetonitrile containing 0.1% (v/v) of trifluoroacetic acid, by freezing the fragment of the target. Drying gave the title compound (0.53 g, 68%).

(Step 3)

Synthesis of 4-[(2,5-difluoro-4-methoxycarbonyl-phenyl)amine sulfonyl]benzoic acid

Tert-butylethanol (15.6 ml), water (3.7 mL), and 2-methyl-2-butene were added to the obtained compound (0.53 g, 1.5 mmol). (1.6 ml, 15 mmol), stirred at room temperature. Sodium chlorite (0.41 g, 4.5 mmol) dissolved in water (4.5 mL), dissolved in water (4.5 mL), and then stirred at room temperature overnight. . The reaction solution was adjusted to pH 6 with 2M hydrochloric acid, and ethyl acetate was added to extract. After washing with a saturated aqueous solution of sodium chloride and drying over magnesium sulfate, the residue was washed with acetonitrile to give the title compound (0.56 g, quant.).

(Step 4)

Synthesis of 4-[[4-(tert-butylamine-carbamoyl)phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

The compound (0.56 g, 1.5 mmol), WSC hydrochloride (0.58 g, 3.0 mmol), HOBt (0.20 g, 1.5 mmol), DMF (1 mL), dichloromethane (5 mL) Stir for 2 hours at room temperature. After diluting with dichloromethane, saturated brine was added, and extracted with dichloromethane and a small amount of methanol, dried over magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was dissolved in 1,4-dioxane (4 mL), and a 2M aqueous lithium hydroxide solution (2.3 mL) was added and stirred at room temperature overnight. After confirming the completion of the reaction, it was neutralized with a 1 M aqueous solution of trifluoroacetic acid, and the solvent was distilled off under reduced pressure. The residue was purified in the same manner as (Step 2), and the title compound (0.38 g, 61%).

[Synthesis Example 2] to [Synthesis Example 8], [Synthesis Example 11], [Synthesis Example 13], [Synthesis Example 14], [Synthesis Example 20], and [Synthesis Example 23] to [Synthesis Example 26] By reacting the compound obtained in (Step 3) of [Synthesis Example 1] with the corresponding amine reagent, it can be synthesized in the same manner as the compound of [Synthesis Example 1].

[Synthesis Example 2]

2,5-Difluoro-4-[[4-(morpholinyl-4-carbonyl)phenyl]sulfonylamino]benzoic acid

[Synthesis Example 3]

2,5-Difluoro-4-[[4-(4-dipropylpiperazinyl-1-carbonyl)phenyl]sulfonylamino]benzoic acid

[Synthesis Example 4]

4-[[4-(cyclopropylaminemethanyl)phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

[Synthesis Example 5]

2,5-Difluoro-4-[[4-(methylaminocarbamimidyl)phenyl]sulfonylamino]benzoic acid

[Synthesis Example 6]

2,5-Difluoro-4-[[4-(ethylamine-mercapto)phenyl]sulfonylamino]benzoic acid

[Synthesis Example 7]

2,5-Difluoro-4-[[4-(2-methoxyethylaminecarbamimidyl)phenyl]sulfonylamino]benzoic acid

[Synthesis Example 8]

2,5-Difluoro-4-[[4-(2-morpholinylethylamine-carbenyl)phenyl]sulfonyl Amino acid benzoic acid

[Synthesis Example 9], [Synthesis Example 10], [Synthesis Example 22], and [Synthesis Example 27] In [Synthesis Example 1] (Step 1), methyl 4-amino-2 was used. , 5-difluoro-benzoin ester and 5-iodopyridyl-2-sulfonyl chloride, or 6-iodopyridyl-3-sulfonyl chloride, can be the same as the compound of [Synthesis Example 1] To synthesize.

[Synthesis Example 9]

4-[[5-(tert-butylamine-mercapto)-2-pyridyl]sulfonylamino]-2,5-difluoro-benzoic acid

[Synthesis Example 10]

2,5-Difluoro-4-[[5-(tetrahydropyran-4-ylaminecarbhydryl)-2-pyridyl]sulfonylamino]benzoic acid

[Synthesis Example 11]

2,5-Difluoro-4-[[4-(tetrahydropyran-4-ylaminecarbamimidyl)phenyl]sulfonylamino]benzoic acid

[Synthesis Example 12]

4-[[4-(tert-butylamine-mercapto)-3-methoxy-phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

(step 1)

Synthesis of methyl 4-[(4-bromo-3-methoxy-phenyl)sulfonylamino]-2,5-difluoro-benzoin ester

Methyl 4-amino-2,5-difluoro-benzoin (1.5 g, 18.0 mmol) was dissolved in pyridine (15 ml), and 4-bromo-3-methoxybenzenesulfonyl chloride (4.6 g, After stirring at room temperature for one night, ethyl acetate (20 ml) and water (50 ml) were added. Extracted twice with ethyl acetate, the organic layer The mixture was washed with a saturated aqueous solution of ammonium chloride and brine, and dried over anhydrous sodium sulfate. The residue was concentrated under reduced pressure, and the obtained residue was dissolved in THF (10 mL), and cooled to 0° C., and then butyl butyl ammonium sulfate (3.0 mL) was added and stirred at room temperature for one and a half hours. Water (50 ml) was added, and the mixture was extracted with ethyl acetate three times. The organic layer was washed with 1M hydrochloric acid and brine, and dried over anhydrous sodium sulfate. The residue was concentrated under reduced pressure, and the obtained residue was washed with diethyl ether to give the title compound (2.0 g, yield: 57%).

1H NMR (CDCl ₃ , 300MHz): δ 7.69-7.59 (m, 2H), 7.47-7.41 (m, 1H), 7.34-7.31 (m, 2H), 7.03 (s, 1H), 3.93 (s, 3H) , 3.90 (s, 3H)

(Step 2)

Synthesis of methyl 4-[[4-(tert-butylamine-mercapto)-3-methoxy-phenyl]sulfonylamino]-2,5-difluoro-benzoin

Water (0.10 ml), tert-butyl isocyanide (57 mg, 0.69 mmol), cesium carbonate (75 mg) were added sequentially to the DMSO solution (0.90 ml) of the compound (0.10 g, 0.23 mmol) obtained in (Step 1). , 0.23 mmol), and di-phenylphenylphosphine dichloropalladium (II) (8.0 mg, 0.012 mmol), and the mixture was stirred at 180 ° C for 30 minutes. After cooling to room temperature, a saturated aqueous solution of ammonium chloride (30 ml) was added. Extracted twice with ethyl acetate, and washed the organic layer with saturated brine net. The title compound (24 mg, 23%) was obtained.

_{1H NMR (DMSO-d 6,} 300MHz): δ 11.12 (s, 1H), 7.86 (s, 1H), 7.69-7.64 (m, 2H), 7.51-7.46 (m, 2H), 7.33-7.2 (m, 1H), 3.87 (s, 3H), 3.81 (s, 3H), 1.33 (s, 9H).

(Step 3)

Synthesis of 4-[[4-(tert-butylamine-mercapto)-3-methoxy-phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

A 4 M aqueous lithium hydroxide solution (2.0 ml) was added to a solution (yield: EtOAc, m. Hydrochloric acid was added, the pH was adjusted from 1.0 to 2.0, and the title compound (0.42 g, 90%) was obtained.

1H NMR (DMSO-d6, 300MHz): δ 11.07 (s, 1H), 7.87 (s, 1H), 7.67-7.61 (m, 2H), 7.50-7.46 (m, 2H), 7.28-7.24 (m, 1H) ), 3.87 (s, 3H), 1.33 (s, 9H).

[Synthesis Example 13]

2,5-Difluoro-4-[[4-[[1-(trifluoromethyl)cyclopropyl]amine-carbamoyl]phenyl]sulfonylamino]benzoic acid

[Synthesis Example 14]

2,5-Difluoro-4-[[4-[(2-hydroxy-1,1-dimethyl-ethyl)amine-carbamoyl]phenyl]sulfonylamino]benzoic acid

[Synthesis Example 15] ~ [Compound 18] The compound [Synthesis Example 12] (Step 1), methyl 4-amino-2,5-difluoro-benzoin and the corresponding aryl bromide The reagent action can be synthesized in the same manner as the compound of [Synthesis Example 12].

[Synthesis Example 15]

4-[[4-(tert-butylamine-carbamoyl)-2-methyl-phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

[Synthesis Example 16]

4-[[4-(tert-butylamine-mercapto)-3-methyl-phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

[Synthesis Example 17]

4-[[4-(tert-butylamine-mercapto)-3-fluoro-phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

[Synthesis Example 18]

4-[[4-(tert-butylamine-methyl)-2-methoxy-phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

[Synthesis Example 19]

4-[[4-(tert-butylamine-mercapto)-3-hydroxy-phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

A 1 M solution of tribromoborane/dichloromethane (15 ml) was added to a solution of the compound (0.24 g, 0.54 mmol. Stir for 18 hours. Ice water was added thereto, and the mixture was stirred at room temperature for 15 minutes, and then extracted with ethyl acetate three times. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The residue was washed with ethyl acetate / diethyl ether (3 / 1) to give the title compound (85 mg, 37%).

1H NMR (DMSO-d ₆ , 300MHz): δ 12.27 (s, 1H), 11.08 (s, 1H), 8.24 (s, 1H), 7.98-7.95 (m, 1H), 7.65-7.59 (m, 1H) , 7.32-7.20 (m, 3H), 1.40 (s, 9H)

[Synthesis Example 20]

4-[[4-[(1-cyanocyclopropyl)aminemethanyl]phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

The compound of [Synthesis Example 21] can be synthesized by the same method as the compound of [Synthesis Example 19].

[Synthesis Example 21]

4-[[4-(tert-butylamine-mercapto)-2-hydroxy-phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

[Synthesis Example 22]

4-[[6-(tert-butylamine-mercapto)-3-pyridyl]sulfonylamino]-2,5-difluoro-benzoic acid

[Synthesis Example 23]

4-[[4-[(2-Cyano-1,1-dimethyl-ethyl)amine-carbamoyl]phenyl]sulfonylamino]-2,5-difluoro-benzoic acid

[Synthesis Example 24]

2,5-Difluoro-4-[[4-[[(1S)-1-(hydroxymethyl)-2-methyl-propyl]aminocarbamoyl]phenyl]sulfonylamino] benzoin acid

[Synthesis Example 25]

2,5-Difluoro-4-[[4-[[(1R)-1-(hydroxymethyl)-2-methyl-propyl]aminocarbamoyl]phenyl]sulfonylamino]benzoin acid

[Synthesis Example 26]

2,5-Difluoro-4-[[4-[[(1S)-1-(hydroxymethyl)-2,2-dimethyl-propyl]aminecarboxylidene]phenyl]sulfonylamine Benzoic acid

[Synthesis Example 27]

4-[[5-[(1-Cyanocyclopropyl)aminemethanyl]-2-pyridyl]sulfonylamino]-2,5-difluoro-benzoic acid

[Synthesis Example 28] to [Synthesis Example 30], [Synthesis Example 35] to [Synthesis Example 37], [Synthesis Example 41], and [Synthesis Example 45] can be described in the patent document (WO 2013/161904). Method to synthesize.

[Synthesis Example 28]

Methyl (2S)-2-amino-3-[4-(1-methyl-2,4-dioxo-pyrido[3,4-d]pyrimidin-3-yl)phenyl]propionate

[Synthesis Example 29]

Methyl (2S)-2-amino-3-[6-(1-methyl-2,4-dioxo-pyrido[3,4-d]pyrimidin-3-yl)-3-pyridyl] Propionate

[Synthesis Example 30]

Methyl (2S)-2-amino-3-[4-(3-methyl-2,6-dioxo-pyrimidin-1-yl)phenyl]propionate

[Synthesis Example 31]

Methyl (2S)-2-amino-3-[4-(3,4,5-trimethyl-2,6-dioxo-pyrimidin-1-yl)phenyl]propionate

(step 1)

Methyl (2S)-2-(tert-butoxycarbonylamino)-3-[4-(5,6-dimethyl-2,4-dioxo-1H-pyrimidin-3-yl)phenyl Synthesis of propionate

[4-[(2S)-2-(tert-Butoxycarbonylamino)-3-methoxy-3-oxo-propyl]phenyl] as described in the process of the patent document (WO 2013/161904) Copper acetate (5.2 g, 29 mmol), 5,6-dimethyluracil (4.0 g, 29 mmol), and triethylamine were added sequentially to a solution of boronic acid (9.2 g, 29 mmol) in dichloromethane (100 ml). 10 ml), stirred at room temperature for 18 hours. The reaction solution was subjected to a sepalite filtration, and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography ( petroleum ether: ethyl acetate = 5:1 to 1:1) to give the title compound (0.83 g, 7%).

1H NMR (CDCl ₃ , 400MHz) δ 8.90 (s, 1H), 7.26 (d, J = 8.4 Hz, 2H), 7.17 (d, J = 7.8 Hz, 2H), 5.06 (d, J = 8.7 Hz, 1H) ), 4.64-4.59 (m, 1H), 3.72 (s, 3H), 3.15 (d, J = 6.3 Hz, 2H), 2.16 (s, 3H), 1.95 (s, 3H), 1.44 (s, 9H) .;MS(ESI)m/z 418[M+H]+

(Step 2)

Methyl (2S)-2-(tert-butoxycarbonylamino)-3-[4-(3,4,5-trimethyl-2,6-dioxo-pyrimidin-1-yl)phenyl Synthesis of propionate

Potassium carbonate (0.85 g, 6.0 mmol) and methyl iodide (0.4 ml) were added sequentially at room temperature in a solution of the compound (0.83 g, 2.0 mmol) in DMF (15 ml). Stir for 1 hour and 30 minutes. The reaction solution was concentrated under reduced pressure and water (20 ml) and ethyl acetate (30 ml). The mixture was extracted with EtOAc (3 mL).

1H NMR (CDCl ₃ , 400MHz) δ 8.02 (s, 1H), 7.24 (d, J = 8.4 Hz, 2H), 7.13 (d, J = 8.4 Hz, 1H), 5.05 (d, J = 8.0 Hz, 1H) ), 4.63-4.58 (m, 1H), 3.71 (s, 3H), 3.47 (s, 3H), 3.13 (d, J = 6.0 Hz, 2H), 2.33 (s, 3H), 2.02 (s, 3H) , 1.43(s,9H).;MS(ESI)m/z 432[M+H]+

(Step 3)

Methyl (2S)-2-amino-3-[4-(3,4,5-trimethyl-2,6-dioxo-pyrimidine- Synthesis of 1-yl)phenyl]propionate

The compound obtained in (Step 2) (0.64 g, 1.5 mmol) was dissolved in 4M hydrochloric acid / ethyl acetate (40 ml), and stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure and ethyl acetate (30 ml). After stirring at room temperature for 30 minutes, the title compound was obtained (yield: <RTIgt;

1H NMR (CD ₃ OD, 400MHz) δ 7.31 (d, J = 8.0 Hz, 2H), 7.13 (d, J = 8.4 Hz, 2H), 4.31-4.27 (m, 1H), 3.78 (s, 3H), 3.38(s,3H), 3.37-3.32(m,1H), 3.09-3.03(m,1H), 2.30(s,3H),1.91(s,3H).;MS(ESI)m/z 332[M +H]+

[Synthesis Example 32]

Methyl (2S)-2-amino-3-[6-(3-methyl-2,6-dioxo-pyrimidin-1-yl)-3-pyridyl]propionate

(step 1)

Synthesis of tert-butyl 3-benzhydryl-2,4-dioxy-pyrimidine-1-carboxylate

Di-tert-butyl dicarbonate (10.2 g, 47 mmol) and 4-dimethylaminopyridine (55 mg, 0.45 mmol) were sequentially added to a solution of uracil (5.0 g, 45 mmol) in acetonitrile (50 ml). Stir at room temperature for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was washed with ethyl acetate. The obtained solid was dissolved in dichloromethane (50 ml), and N,N-diisopropylethylamine (7.9 ml, 45 mmol) was added. Benzoyl chloride (5.3 g, 37 mmol) was added under ice cooling, and stirred at room temperature for 12 hr. Water was added to the reaction solution, and the mixture was extracted twice with dichloromethane. The organic layer was washed successively with 0.5 M hydrochloric acid and saturated aqueous sodium chloride, and dried over magnesium sulfate. After concentrating under reduced pressure, the residue was washed with ethyl acetate to give the title compound (9.4 g, 66%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.15 (dd, J = 8.5, 1.6 Hz, 1H), 8.11-8.04 (m, 2H), 7.87-7.76 (m, 1H), 7.68-7.56 (m, 2H), 5.98 (dd, J = 8.4, 1.7 Hz, 1H), 1.54 (d, J = 1.6 Hz, 9H); MS (ESI) m/z 317 [M+H] ⁺

(Step 2)

Synthesis of 3-benzylidenyl-1-methyl-pyrimidine-2,4-dione

Trifluoroacetic acid (8.0 ml) was added to a dichloromethane solution (m. The reaction solution was concentrated under reduced pressure and the residue was dissolved in acetonitrile (50 ml). Potassium carbonate (4.5 g, 33 mmol) and methyl iodide (2.8 ml, 45 mmol) were added sequentially and stirred at room temperature for 12 hours. Water (30 ml) was added to the reaction solution, and the mixture was stirred at room temperature for 30 minutes. The title compound (7.4 g, quant.) was obtained by filtration of solid.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.96 (dd, J = 8.0, 1.3 Hz, 2H), 7.78 (d, J = 7.9 Hz, 1H), 7.83 - 7.72 (m, 1H), 7.60 (dd , J = 8.4, 7.2 Hz, 2H), 5.81 (dd, J = 7.9, 0.6 Hz, 1H), 3.32 (s, 3H); MS (ESI) m/z 231 [M+H] ⁺ .

(Step 3)

Synthesis of 1-methyluracil

An 8 M ammonia/methanol solution (50 ml) was added to the compound (7.4 g, 30 mmol) obtained in (Step 2), and stirred at room temperature for 5 hr. After the solid was filtered off (first crystal), the filtrate was concentrated under reduced pressure. The residue was washed with ethyl acetate (second crystal), and the obtained solid was collected to give the title compound (4.2 g, quant.).

^{1 H NMR (400MHz, DMSO-} d 6) δ 11.21 (s, 1H), 7.61 (d, J = 7.8Hz, 1H), 5.51 (d, J = 7.8Hz, 1H), 3.22 (s, 3H); MS (ESI) m/z 127 [M+H] ⁺ .

(Step 4)

3-(5-bromo-2-pyridyl)-1-methyl-pyrimidine-2,4-dione, and 3-(5-iodo-2-pyridyl)-1-methyl-pyrimidine-2, Synthesis of a mixture of 4-dione

5-Bromo-2-iodopyridine (0.74 g, 2.6 mmol) and copper iodide (0.50 g, 2.6) were sequentially added to a solution of the compound (0.22 g, 1.7 mmol) in DMF (5.0 ml). Methyl) and triethylamine (1.0 ml, 6.9 mmol) were stirred at 140 ° C for 18 h. The reaction solution was cooled to room temperature, and water (25 ml) and dichloromethane (25 ml) were added. The mixture was subjected to sepalite filtration, and after extracting twice with dichloromethane, the organic layer was washed with a saturated aqueous solution of sodium chloride. After drying over magnesium sulfate, it was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give a mixture (1:1) (0.29 g).

(Step 5)

Methyl (2S)-2-(tert-butoxycarbonylamino)-3-[6-(3-methyl-2,6-dioxo-pyrimidin-1-yl)-3-pyridyl]-propyl Acid ester synthesis

The zinc powder (96 mg, 1.5 mmol) was suspended in DMF (2.0 ml), and iodine (26 mg, 0.10 mmol) was added, and the mixture was stirred at room temperature for 15 minutes. Add methyl (2R)-2-(tert-butoxycarbonylamino)-3-iodo-propionate (0.19 g, 0.59 mmol), and iodine (26 mg, 0.10 mmol) at room temperature Stir for 30 minutes.

The resulting mixture (0.29 g) was added to a separate container and dissolved in DMF (1.0 mL). Paragonimide (diphenylideneacetone) dipalladium (0) (22 mg, 0.024 mmol) and SPhos (20 mg, 0.049 mmol) were added sequentially and stirred for 10 minutes. This mixed solution was added to the previously prepared mixed solution, and after degassing and argon substitution operation three times, it was stirred at 60 ° C for 18 hours. The reaction solution was cooled to room temperature, and water (25 ml) and dichloromethane (25 ml) were added. The mixture was subjected to sepalite filtration, and after extracting twice with dichloromethane, the organic layer was washed with a saturated aqueous solution of sodium chloride. After drying over magnesium sulfate, it was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give the title compound (0.21 g).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.42 (d, J = 2.3 Hz, 1H), 7.92 - 7.72 (m, 2H), 7.44 (d, J = 8.1 Hz, 1H), 7.30 (d, J) = 8.0 Hz, 1H), 5.77 (d, J = 7.9 Hz, 1H), 4.35-4.20 (m, 1H), 3.64 (s, 3H), 3.31 (s, 3H), 3.11 (dd, J = 13.9, 4.8 Hz, 1H), 2.94 (dd, J = 14.1, 10.6 Hz, 1H), 1.34 (s, 9H); MS (ESI) m/z 405 [M+H] ⁺ .

(Step 6)

Methyl (2S)-2-amino-3-[6-(3-methyl-2,6-dioxo-pyrimidin-1-yl)- Synthesis of 3-pyridyl]propionate

The compound obtained in (Step 5) (0.21 g, 0.52 mmol) was dissolved in 1,4-dioxane (2.0 ml) and methanol (1.0 ml). A 4 M hydrochloric acid / 1,4-dioxane solution (2.0 ml) was added, and the mixture was stirred at room temperature for 5 hr.

MS (ESI) m/z 305 [M+H] ⁺ .

The compound of [Synthesis Example 33] can be synthesized in the same manner as in [Synthesis Example 32] by using thymine in [Synthesis Example 32] (Step 1).

[Synthesis Example 33]

Isopropyl (2S)-2-amino-3-[6-(3,5-dimethyl-2,6-dioxo-pyrimidin-1-yl)-3-pyridyl]propionate

^{1 H NMR (400MHz, DMSO-} d 6) δ 8.58 (s, 3H), 8.45 (d, J = 2.4Hz, 1H), 7.87 (dd, J = 8.1,2.4Hz, 1H), 7.71 (d, J =1.3 Hz, 1H), 7.36 (d, J = 8.1 Hz, 1H), 4.91 (p, J = 6.2 Hz, 1H), 4.40 (s, 1H), 3.29 (s, 3H), 3.24 (dd, J =14.3, 5.9 Hz, 1H), 3.11 (dd, J = 14.3, 8.6 Hz, 1H), 1.83 (d, J = 1.0 Hz, 3H), 1.16 (d, J = 6.2 Hz, 3H), 1.06 (d) , J = 6.2 Hz, 3H); MS (ESI) m/z 437 [M+H] ⁺ .

The compound of [Synthesis Example 34] can be used in [Synthesis Example 31] (Step 1) by using 6-(ethoxymethyl)-1H-pyrimidine-2,4-dione, and The compound of Example 31] was synthesized in the same manner.

[Synthesis Example 34]

Methyl (2S)-2-amino-3-[4-[4-(ethoxymethyl)-3-methyl-2,6-dioxo-pyrimidin-1-yl]phenyl]propanoic acid ester

MS (ESI) m/z 362 [M+H] ⁺ .

[Synthesis Example 35]

Methyl (2S)-2-amino-3-[5-(3-methyl-2,6-dioxo-pyrimidin-1-yl)-2-pyridyl]propionate

[Synthesis Example 36]

Methyl (2S)-2-amino-3-[4-(3,5-dimethyl-2,6-dioxo-pyrimidin-1-yl)phenyl]propionate

[Synthesis Example 37]

Methyl (2S)-2-amino-3-[5-(3,5-dimethyl-2,6-dioxo-pyrimidin-1-yl)-2-pyridyl]propionate

[Synthesis Example 38]

Isopropyl (2S)-2-amino-3-[4-(1,7-dimethyl-2,4-dioxo-6,8-dihydro-5H-pyrido[3,4-d Pyrimidin-3-yl)phenyl]propionate

(step 1)

Synthesis of ethyl 1-benzyl-5-[(4-nitrophenoxy)carbonylamino]-3,6-dihydro-2H-pyridine-4-carboxylate

Ammonium acetate (5.2 g, 67 mmol) was added to a solution of ethyl 1-benzyl-3-oxo-piperidine-4-carboxylate hydrochloride (2.0 g, 6.7 mmol) in ethanol (20 ml) Stir for 18 hours under temperature. The reaction solution was concentrated under reduced pressure, and dichloromethane and saturated sodium bicarbonate were added. It was extracted twice with dichloromethane, and the organic layer was washed with a saturated aqueous solution of sodium chloride. After drying over magnesium sulfate, the residue was concentrated under reduced pressure and the residue was dissolved in dichloromethane (50 ml). Pyridine (0.6 ml) was added, and 4-nitrophenyl chloroformate (1.4 g, 6.7 mmol) was added and the mixture was stirred at the same temperature for 5 hr. The reaction solution was concentrated under reduced pressure, and the residue was purified to silica gel eluting

(Step 2)

Synthesis of isopropyl (2S)-3-(4-aminophenyl)-2-(tert-butoxycarbonylamino)propionate

Potassium carbonate (3g) was added sequentially to a solution of (2S)-2-(tert-butoxycarbonylamino)-3-(4-nitrophenyl)propanoic acid (2 g, 6.4 mmol) in DMF (15 mL) 22 mmol) and 2-iodopropane (2.0 ml) were stirred at room temperature for 18 hours. Water was added to the reaction solution, and the mixture was extracted three times with a mixed solution of ethyl acetate and hexane (1:1), and the organic layer was washed with a saturated aqueous sodium chloride solution. After drying over magnesium sulfate, the residue was concentrated under reduced pressure and the residue was dissolved in methanol (40 ml) and water (4.0 ml). Zinc powder (3.5 g, 54 mmol) and amine chloride (0.52 g, 9.7 mmol) were added in that order, and the mixture was stirred at 70 ° C for 1 hour and 30 minutes. The reaction solution was subjected to filtration of calcium sulphate, and concentrated under reduced pressure. The residue was subjected to reverse phase HPLC using ODS as a filler, and dissolved in a mixed solution of water and acetonitrile containing 0.1% (v/v) of trifluoroacetic acid. The trifluoroacetate salt of the title compound is obtained by freeze-drying the fragment of the object.

(Step 3)

Isopropyl (2S)-3-[4-(7-benzyl-2,4-dioxo-1,5,6,8-tetrahydropyrido[3,4-d]pyrimidin-3-yl) Synthesis of phenyl]-2-(tert-butoxycarbonylamino)propionate

The compound (0.50 g, 1.1 mmol) and DBU (Step 2) were added sequentially (Step 2) to a 1,4-dioxane solution (20 ml) of Compound (0.49 g, 1.2 mmol). 0.42 ml), stirred at 60 ° C for 18 hours. The reaction solution was concentrated under reduced pressure, and ethyl acetate and water were added. The organic layer was washed with a saturated aqueous solution of sodium chloride and dried over magnesium sulfate. After concentrating under reduced pressure, the residue was purified by ethylamine.

(Step 4)

Isopropyl (2S)-3-[4-(7-benzyl-1,7-dimethyl-2,4-dioxo-6,8-dihydro-5H-pyrido[3,4-d Synthesis of pyrimidine-7-n-n-yl)phenyl]-2-(tert-butoxycarbonylamino)propionate

Potassium carbonate (0.30 g, 2.2 mmol) and methyl iodide (0.22 ml, 3.5 mmol) were added sequentially to a solution of the compound (0.40 g, 0.59 mmol) in DMF (3.5 ml). Stir at room temperature for 3 hours. After the reaction solution was concentrated under reduced pressure, the residue was purified and purified to purified crystals eluted eluted elute ).

(Step 5)

Isopropyl (2S)-2-amino-3-[4-(1,7-dimethyl-2,4-dioxo-6,8-dihydro-5H-pyrido[3,4-d Synthesis of pyrimidin-3-yl)phenyl]propionate

10% palladium carbon (50 mg) was added to a solution of the compound (0.30 g, 0.43 mmol) in isopropyl alcohol (5.0 ml), and the mixture was stirred at room temperature for 18 hours under hydrogen atmosphere. The reaction solution was subjected to sepalite filtration, and concentrated under reduced pressure. The residue was dissolved in 1,4-dioxane (2.0 ml) and isopropylethanol (1.0 ml). A 4 M hydrochloric acid / 1,4-dioxane solution (2.0 ml) was added, and the mixture was stirred at room temperature for 5 hr.

MS (ESI) m/z 401 [M+H] ⁺

[Synthesis Example 39], [Synthesis Example 40], [Synthesis Example 42], and [Synthesis Example 44] The compound (in Synthesis Example 38) (Step 1) can be used by using the corresponding 1,2-ketone The ester reagent was synthesized in the same manner as the compound of [Synthesis Example 38].

[Synthesis Example 39]

Isopropyl (2S)-2-amino-3-[4-(1,6-dimethyl-2,4-dioxo-7,8-dihydro-5H-pyrido[4,3-d Pyrimidin-3-yl)phenyl]propionate

MS (ESI) m/z 401 [M+H] ⁺

[Synthesis Example 40]

Isopropyl (2S)-2-amino-3-[4-(1-methyl-2,4-dioxo-5,6,7,8-tetrahydroquinazolin-3-yl)phenyl Propionate

MS (ESI) m/z 386 [M+H] ⁺

[Synthesis Example 41]

Methyl (2S)-2-amino-3-[4-(3,4-dimethyl-2,6-dioxo-pyrimidin-1-yl)phenyl]propionate

[Synthesis Example 42]

Isopropyl (2S)-2-amino-3-[4-(1-methyl-2,4-dioxo-6,8-dihydro-5H-pyrano[3,4-d]pyrimidine- 3-yl)phenyl]propionate

MS (ESI) m/z 388 [M+H] ⁺

[Synthesis Example 43]

Methyl (2S)-2-amino-3-[6-(1-methyl-2,4-dioxo-7,8-dihydro-5H-pyrano[4,3-d]pyrimidine-3 -yl)-3-pyridyl]propionate

(step 1)

Synthesis of ethyl 4-amino-3,6-dihydro-2H-pyran-5-carboxylate

Ammonium acetate (2.4 g, 31 mmol) was added to a solution of ethyl 4-oxotetrahydropyran-3-carboxylate (0.54 g, 3.1 mmol) in ethanol (10 ml), and stirred at 60 ° C for 18 hours. The reaction solution was concentrated under reduced pressure, and dichloromethane and saturated sodium bicarbonate were added. Extract twice with dichloromethane and saturate the organic layer Wash with sodium chloride solution. After drying over magnesium sulfate, the title compound (0.49 g, quant.) was obtained.

(Step 2)

Synthesis of 1,5,7,8-tetrahydropyrano[4,3-d]pyrimidine-2,4-dione

Trichloroethane oxime isocyanate (0.74 ml, 6.2 mmol) was added to a solution of EtOAc (EtOAc, m. The solid was filtered off and suspended in 8M ammonia/methanol (5.0 mL). After stirring at 70 ° C for 18 hours, the title compound (0.30 g, 57%).

(Step 3)

Synthesis of 3-(5-bromo-2-pyridyl)-1,5,7,8-tetrahydropyrano[4,3-d]pyrimidine-2,4-dione

5-Bromo-2-iodopyridine (1.1 g, 3.7 mmol) and copper iodide (0.30 g, 1.6) were sequentially added to a solution of the compound (0.53 g, 3.2 mmol) in acetonitrile (10.0 ml). Methyl) and DBU (0.93 ml, 6.4 mmol) were stirred at 70 ° C for 18 h. The reaction solution was cooled to room temperature, filtered with calcium sulphate, concentrated under reduced pressure, and the residue was charged with ODS as a filling. The reverse phase HPLC of the solvent was dissolved in a mixture of water and acetonitrile containing 0.1% (v/v) of trifluoroacetic acid, and the title compound (0.26 g, 26%) was obtained.

(Step 4)

Synthesis of 3-(5-bromo-2-pyridyl)-1-methyl-7,8-dihydro-5H-pyrano[4,3-d]pyrimidine-2,4-dione

Potassium carbonate (0.33 g, 2.4 mmol) and methyl iodide (0.25 ml) were added sequentially to a solution of the compound (0.26 g, 0.80 mmol) in DMF (2.0 mL) Stir for 5 hours. The reaction solution was concentrated under reduced pressure, and the residue was subjected to reverse phase HPLC using ODS as a filler, and dissolved in a mixed solution of water and acetonitrile containing 0.1% (v/v) of trifluoroacetic acid, by freezing the fragment of the target. Drying gave the title compound (0.20 g, 74%).

(Step 5)

Methyl (2S)-2-(tert-butoxycarbonylamino)-3-[6-(1-methyl-2,4-dioxo-7,8-dihydro-5H-pyran[4 Synthesis of 3-D]pyrimidin-3-yl)-3-pyridyl]propionate

The zinc powder (0.12 g, 1.8 mmol) was suspended in DMF (2.0 ml), iodine (34 mg, 0.13 mmol) was added, and the mixture was stirred at room temperature for 15 minutes. Add methyl (2R)-2-(tert-butoxycarbonylamino)-3-iodo-propionate (0.23 g, 0.70 mmol), and iodine (34 mg, 0.13 mmol) at room temperature Stir for 30 minutes.

The compound obtained in (Step 4) (0.20 g, 0.59 mmol) was added to the mixture, and then dissolved in DMF (1.0 ml). Paragonimide (diphenylideneacetone) dipalladium (0) (14 mg, 0.015 mmol) and SPhos (24 mg, 0.058 mmol) were added sequentially and stirred for 10 minutes. This mixed solution was added to the previously prepared mixed solution, and after degassing and argon substitution operation three times, it was stirred at 60 ° C for 18 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the residue was poured into a reverse phase HPLC using ODS as a filler, and dissolved in a mixed solution of water and acetonitrile containing 0.1% (v/v) of trifluoroacetic acid. The title fragment was freeze-dried to give the title compound (0.12 g, 45%).

(Step 6)

Methyl (2S)-2-amino-3-[6-(1-methyl-2,4-dioxo-7,8-dihydro-5H-pyrano[4,3-d]pyrimidine-3 Synthesis of -yl)-3-pyridyl]propionate

The compound obtained in (Step 5) (0.12 g, 0.26 mmol) was dissolved in 1,4-dioxane (1.0 ml) and methanol (1.0 ml). A 4 M hydrochloric acid / 1,4-dioxane solution (1.0 ml) was added, and the mixture was stirred at room temperature for 5 hr.

MS (ESI) m/z 361 [M+H] ⁺

[Synthesis Example 44]

Isopropyl (2S)-2-amino-3-[4-(1-methyl-2,4-dioxo-7,8-dihydro-5H-pyrano[4,3-d]pyrimidine- 3-yl)phenyl]propionate

MS (ESI) m/z 388 [M+H] ⁺

[Synthesis Example 45]

Methyl (2S)-2-amino-3-[4-(6-methoxy-1-methyl-2,4-dioxo-pyrido[3,4-d]pyrimidin-3-yl) Phenyl]propionate

[Synthesis Example 46] and the compound of [Synthesis Example 47] In [Synthesis Example 43] (Step 3), by using 6-methyluracil or 5,6-dimethyluracil, The synthesis was carried out in the same manner as in the compound of [Synthesis Example 43].

[Synthesis Example 46]

Methyl (2S)-2-amino-3-[6-(3,4-dimethyl-2,6-dioxo-pyrimidin-1-yl)-3-pyridyl]propionate

MS (ESI) m / z 319 [M + H] +

[Synthesis Example 47]

Methyl (2S)-2-amino-3-[6-(3,4,5-trimethyl-2,6-dioxo-pyrimidin-1-yl)-3-pyridyl]propionate

MS (ESI) m/z 333 [M+H] ⁺

[Synthesis Example 48]

Isopropyl (2S)-2-amino-3-[5-(1-methyl-2,4-dioxo-5,6,7,8-tetrahydroquinazolin-3-yl)-2 -pyridyl]propionate

(step 1)

Synthesis of ethyl 2-[(4-nitrophenoxy)carbonylamino]cyclohexene-1-carboxylate

Ethyl 2-oxocyclohexanecarboxylate (1.5 mL, 8.823 mmol) was dissolved in methanol (90 mL). Ammonium acetate (6.80 g, 88.23 mmol) was added to the solution, and the mixture was stirred at 60 ° C for 14 hours. After the solvent of the reaction solution was removed under reduced pressure, ethyl acetate was added to the residue, and the organic layer was washed with saturated aqueous sodium hydrogen carbonate, water, and brine, and the organic layer was dried over anhydrous sodium sulfate. After the insoluble material was filtered, the residue was dissolved in dichloromethane (45 mL). Pyridine (0.86 mL, 10.65 mmol) was added to the solution, and after cooling to 0 ° C, (4-nitrophenyl) carbamic acid (1.879 g, 9.322 mmol) was added, and the mixture was gradually stirred at room temperature. hour. After the solvent of the reaction solution was removed under reduced pressure, the obtained residue was purified using silica gel column chromatography (SiO ₂ , hexane / ethyl acetate) to give the title compound (2.72 g, 94.0 %).

MS (ESI) m/z 335 [M+H]+

(Step 2)

Synthesis of 3-(6-bromo-3-pyridyl)-5,6,7,8-tetrahydro-1H-quinazoline-2,4-dione

The compound obtained in (Step 1) (1.47 g, 4.401 mmol) was dissolved in 1,4-dioxane (45 mL), and 6-bromopyridin-3-amine (0.795 g, 4.621 mmol) and 1,8- Diazabicyclo[5.4.0]undec-7-ene (1.31 mL, 8.802 mmol) was stirred at room temperature for 17 h. After the solvent of the reaction solution was removed under reduced pressure, the obtained residue was subjected to reverse phase HPLC using ODS as a filler, and purified by the same method as in [Synthesis Example 1] (Step 2) to obtain a title compound. (719 mg, 30.2%).

MS (ESI) m/z 322 [M+H]+

(Step 3)

Synthesis of 3-(6-bromo-3-pyridyl)-1-methyl-5,6,7,8-tetrahydroquinazoline-2,4-dione

The compound obtained in (Step 2) (719 mg, 2.232 mmol) was dissolved in dimethylformamide (8 mL), and potassium carbonate (770 mg, 5.580 mmol) and methyl iodide (207 uL, 3.348 mmol) were added at room temperature. Stir under 3 hours. Ethyl acetate was added to the reaction solution, and the organic layer was washed with water and brine and dried over anhydrous sodium sulfate. After the insoluble matter was filtered, the solvent was removed under reduced pressure to obtain a residue. The residue was purified by reverse phase HPLC using ODS as a solvent to afford the title compound (269 mg, 35.9%).

MS (ESI) m/z 336 [M+H]+

(Step 4)

Isopropyl (2S)-2-(tert-butoxycarbonylamino)-3-[5-(1-methyl-2,4-dioxo-5,6,7,8-tetrahydroquinazoline Synthesis of oxa-3-yl)-2-pyridyl]propionate

The zinc powder (157 mg, 2.40 mmol) was suspended in DMF (2.0 ml), and iodine (46.3 mg, 0.18 mmol) was added, and the mixture was stirred at room temperature for 15 minutes. Add methyl (2R)-2-(tert-butoxycarbonylamino)-3-iodo-propionate (269 mg, 0.801 mmol), and iodine (46.3 mg, 0.18 mmol) at room temperature. Stir for 30 minutes.

The obtained compound (256 mg, 0.801 mmol) was added to a separate container and dissolved in DMF (2.0 ml). Ginseng (diphenylideneacetone) dipalladium (0) (18.3 mg, 0.020 mmol) and SPhos (32.9 mg, 0.0801 mmol) were added in that order and stirred for 10 minutes. This mixed solution was added to the previously prepared mixed solution, and after degassing and argon substitution operation three times, it was stirred at 60 ° C for 15 hours. The reaction solution was cooled to room temperature, and water (25 ml) and dichloromethane (25 ml) were added. The mixture was subjected to sepalite filtration, and after extracting twice with dichloromethane, the organic layer was washed with a saturated aqueous solution of sodium chloride. After drying over anhydrous sodium sulfate, it was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give the title compound (223 mg, 57.3%).

MS (ESI) m/z 487 [M+H]+

(Step 5)

Isopropyl (2S)-2-amino-3-[5-(1-methyl-2,4-dioxo-5,6,7,8-tetrahydroquinazolin-3-yl)-2 Synthesis of pyridyl]propionate hydrochloride

The compound obtained in (Step 4) (223 mg, 0.459 mmol) was dissolved in ethyl acetate (3mL). 4N Hydrochloric acid/ethyl acetate solution (1.2 mL) was added to the solution, and the mixture was stirred at room temperature for 7 hours. After the solvent was removed under reduced pressure, the title compound (219 mg, quant.) was obtained by freeze-drying.

MS (ESI) m/z 387 [M+H]+

[Example 1]

(2S)-2-[[4-[[4-(tert-butylamine-carbamoyl)phenyl]sulfonylamino]-2,5-difluoro-benzylidenyl]amino]- 3-[6-(1-methyl-2,4-dioxo-pyrido[3,4-d]pyrimidin-3-yl)-3-pyridyl]propionic acid

(step 1)

Methyl (2S)-2-[[4-[[4-(tert-butylamine-carbamoyl)phenyl]sulfonylamino]-2,5-difluoro-benzylidene]amino Synthesis of -3-[6-(1-methyl-2,4-dioxo-pyrido[3,4-d]pyrimidin-3-yl)-3-pyridyl]propionate

HATU (44 mg, 0.12) was added to the DMF solution (1.5 ml) of the compound (40 mg, 0.097 mmol) of [Comp. Methyl), and N,N-diisopropylamine (58 μl, 0.34 mmol) were stirred at room temperature for 5 hours.

The residue was concentrated under reduced pressure, and the residue was subjected to reverse phase HPLC using ODS as a filler, and dissolved in a mixed solution of water and acetonitrile containing 0.1% (v/v) of trifluoroacetic acid, and the fraction of the object was freeze-dried. The title compound (47 mg, 67%) was obtained.

(Step 2)

(2S)-2-[[4-[[4-(tert-butylamine-carbamoyl)phenyl]sulfonylamino]-2,5-difluoro-benzylidenyl]amino]- Synthesis of 3-[6-(1-methyl-2,4-dioxo-pyrido[3,4-d]pyrimidin-3-yl)-3-pyridyl]propionic acid

To the 1,4-dioxane solution (1.0 ml) of the compound (47 mg, 0.063 mmol) obtained in (Step 1), water (1.0 ml), and 1M aqueous sodium acid aqueous solution (0.19 ml), Stir for 5 hours at room temperature. After adding 1 M hydrochloric acid and neutralizing, and concentrating under reduced pressure, the residue was charged to a reverse phase HPLC using ODS as a filler, and a mixture containing water and acetonitrile containing 0.1% (v/v) of trifluoroacetic acid. The solution was dissolved and the title compound (40 mg, 87%) was obtained.

1H NMR (400MHz, DMSO-d6) δ 10.92 (brs, 1H), 8.99 (s, 1H), 8.69 (dd, J = 8.1, 2.2 Hz, 1H), 8.57 (d, J = 5.0 Hz, 1H), 8.46 (d, J = 2.3 Hz, 1H), 8.02 (s, 1H), 7.97-7.81 (m, 6H), 7.40 (d, J = 8.0 Hz, 1H), 7.32 - 7.17 (m, 2H), 4.72 -4.63(m,1H), 3.60(s,3H),3.33-3.26(m,1H),3.14-3.04(m,1H), 1.35(s,9H).;MS(ESI)m/z 736[ M+H]+

The compound shown in Table 1 can be used by using any one of the carboxylic acid intermediates selected from [Synthesis Example 1] to [Synthesis Example 27] and from any of the amines of [Synthesis Example 28] to [Synthesis Example 48]. The compound or a salt thereof was synthesized in the same manner as the compound of [Example 1].

[Embodiment 2]

Isopropyl (2S)-2-[[4-[[4-(tert-butylamine-carbamoyl)phenyl]sulfonylamino]-2,5-difluoro-benzylidene]amine 3-[4-(1-methyl-2,4-dioxy-pyrido[3,4-d]pyrimidin-3-yl)phenyl]propionate

A 4 M hydrochloric acid / 1,4-dioxane solution (1.0 ml) was added to a solution of A4 (35 mg, 0.048 mmol) in isopropyl alcohol (2.0 ml), and stirred at 60 ° C for 5 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the residue was poured into a reverse phase HPLC using ODS as a filler, and dissolved in a mixed solution of water and acetonitrile containing 0.1% (v/v) of trifluoroacetic acid. The title compound (34 g, 91%) was obtained by freeze-drying the title compound.

The compound shown in Table 2 can be the same as the compound of [Example 2] by using any one of sulfonamide derivatives selected from A1 to A58, and the corresponding ethanol (isopropyl alcohol or cyclohexylethanol). To synthesize.

Test example 1

(1) Evaluation test of VCAM-1/α4β1 integrin binding inhibitory activity

The ability of the test substance which inhibits the binding of the human T cell line cell line Jurkat known to express α4β1 integrin to VCAM-1 was determined.

Recombinant human VCAM-1/Fc (R&D systems) solution (1 μg/mL) diluted in buffer A (carbonate buffer, pH 9.6) at 50 μL/well in a 96-well microtiter plate at 4 °C Cultivate one night. After washing once with PBS, Block Ace (Snow Print Dairy) was added at 150 μL/well, and cultured at room temperature for 2 hours. After removal, it was washed once with PBS.

100 μL of various concentrations of test substance diluted in binding buffer (DMEM containing 40 mM HEPES, 0.2% BSA, and 4 mM MnCl ₂ ) and Jurkat cells (2×10 ⁶ cells/mL) were added to each VCAM-1/Fc coating. In a disc (5x10 ⁵ cells/well), incubate at 30 ° C for 15 minutes to 30 minutes. After the cells were allowed to bind to the wells, the unbound cells were removed by washing with PBS. Buffer C (containing 1.5% Triton X-100 in PBS) was added to the disc at 50 μL/well to dissolve the bound Jurkat cells. 30 μL of Substrate Buffer (Promega, CytoTox 96 Non-Radioactive Cytotoxicity Assay) was added to 30 μL of the cell lysate, and the mixture was allowed to react at room temperature in the dark for 30 minutes. 30 μL of Stop Solution (Promega, CytoTox 96 Non-Radioactive Cytotoxicity Assay) was added, and the absorbance at 490 nm was measured using a disk reader. The absorbance obtained here is a value for detecting the activity of lactic dehydrogenase (LDH) eluted from the supernatant, i.e., proportional to the number of Jurkat cells bound to VCAM-1 and remaining on the disk. The test was carried out by repeating. When the absorbance of the well containing no test substance was set to 100%, the binding ratio of the cells in various concentrations was determined, and the concentration IC _{50 which} brings about a 50% binding inhibitory effect was calculated. The results obtained are collectively shown in Table 3.

Test example 2

(2) Evaluation test of MAdCAM-1/α4β7 integrin binding inhibitory activity

The ability to inhibit the test substance which is known to exhibit the binding of the human B cell line cell line RPMI-8866 of α4β7 integrin to MAdCAM-1 was determined.

A recombinant mouse MAdCAM-1/Fc (R&D systems) solution (1 μg/mL) diluted in buffer A (carbonate buffer, pH 9.6) was added to a 96-well microplate at 50 μL/well, and cultured at 4 °C. one night. After washing once with PBS, Block Ace (Snow Print Dairy) was added at 150 μL/well, and cultured at room temperature for 2 hours. After removal, it was washed once with PBS.

100 μL of various concentrations of test substance diluted in binding buffer (DMEM containing 40 mM HEPES, 0.2% BSA, and 4 mM MnCl ₂ ) and RPMI-8866 cells (2×10 ⁶ cells/mL) were added to each MAdCAM-1/Fc. coating the disk (5x10 ⁵ cells / well) and cultured at 30 deg.] C for about 60 minutes. After the cells were allowed to bind to the wells, the unbound cells were removed by washing with PBS. Buffer C (PBS containing 1.5% Triton X-100) was added to the disc at 50 μL/well to dissolve the bound RPMI-8866 cells. 30 μL of Substrate Buffer (Promega, CytoTox 96 Non-Radioactive Cytotoxicity Assay) was added to 30 μL of the cell lysate, and the mixture was allowed to react at room temperature in the dark for 30 minutes. Each of 30 μL of Stop Solution (Promega, CytoTox 96 Non-Radioactive Cytotoxicity Assay) was added, and the absorbance at 490 nm was measured using a disk reader. The absorbance obtained here is the value of detecting the lactic acid dehydrogenase (LDH) activity eluted from the supernatant, that is, proportional to the number of RPMI-8866 cells bound to MAdCAM-1 and remaining on the disc. . The test was carried out by repeating. When the absorbance of the well containing no test substance was set to 100%, the binding ratio of the cells in various concentrations was determined, and the concentration IC _{50 which} brings about a 50% binding inhibitory effect was calculated. The results obtained are collectively shown in Table 3.

Test Example 3

(3) Evaluation of MAdCAM-1/α4β7 integrin binding inhibitory activity in the presence of serum (1)

The ability to inhibit the test substance which is known to exhibit the binding of the human B cell line cell line RPMI-8866 of α4β7 integrin to MAdCAM-1 was determined.

A recombinant mouse MAdCAM-1/Fc (R&D systems) solution (1 μg/mL) diluted in buffer A (carbonate buffer, pH 9.6) was added to a 96-well microplate at 50 μL/well, and cultured at 4 °C. one night. After washing once with PBS, Block Ace (Snow Print Dairy) was added at 150 μL/well, and cultured at room temperature for 2 hours. After removal, it was washed once with PBS.

100 μL of various concentrations of test substance diluted in binding buffer (DMEM containing 40 mM HEPES, 0.2% BSA, and 4 mM MnCl ₂ ) and RPMI-8866 cells (2×10 ⁶ cells/mL) were added to each MAdCAM-1/Fc. the coated discs in (7.5x10 ⁵ cells / well), containing a final concentration of 50% human serum, cultured at 30 deg.] C for about 60 minutes. After the cells were allowed to bind to the wells, the unbound cells were removed by washing with PBS. Buffer C (PBS containing 1.5% Triton X-100) was added to the disc at 50 μL/well to dissolve the bound RPMI-8866 cells. 30 μL of Substrate Buffer (Promega, CytoTox 96 Non-Radioactive Cytotoxicity Assay) was added to 30 μL of the cell lysate, and the mixture was allowed to react at room temperature in the dark for 30 minutes. Each of 30 μL of Stop Solution (Promega, CytoTox 96 Non-Radioactive Cytotoxicity Assay) was added, and the absorbance at 490 nm was measured using a disk reader. The absorbance obtained here is the value of detecting the lactic acid dehydrogenase (LDH) activity eluted from the supernatant, that is, proportional to the number of RPMI-8866 cells bound to MAdCAM-1 and remaining on the disc. . The test was carried out by repeating. When the absorbance of the well containing no test substance was set to 100%, the binding ratio of the cells in various concentrations was determined, and the concentration IC _{50 which} brings about a 50% binding inhibitory effect was calculated. The results obtained are collectively shown in Table 3.

[Table 3-1]

Comparing the results of Test Example (1) with Test Example (2), it was judged that the compound of the present invention had a lower effect on α4β1, a higher effect on α4β7, and a higher selectivity. Thus, the effect on α4β1 is lower, and the effect on α4β7 is higher. If it has higher selectivity, it will reduce the effect on α4β1 which inhibits the circulation of lymphocytes in the whole body, and will greatly inhibit the effect on α4β7 which is specifically expressed in the intestinal tract. Therefore, it has the advantage of being able to treat the disease more efficiently.

Test Example 4

(4) Evaluation of MAdCAM-1/α4β7 integrin binding inhibitory activity in human whole blood

The binding inhibitory activity of T cell α4β7 integrin to MAdCAM-1 in human whole blood of the test substance was measured. Blood samples are voluntary by healthy people The blood of the person is provided.

A 4 mM MnCl ₂ solution was added to human whole blood and incubated with various test substance dilutions for 10 minutes. 10 μg/mL of recombinant mouse MAdCAM-1/Fc (R&D Systems) was added and cultured in a total amount of 50 μL for 30 minutes. 950 μL of Lyse/Fix (BD Biosciences) was added, hemolyzed at 37 ° C, and fixed for 10 minutes. After centrifugation for 5 minutes, the supernatant was removed, and 600 μL of RPMI-1640 culture solution (hereinafter referred to as a culture solution) supplemented with 10% of unactivated bovine embryo serum was added, and after centrifugation for 5 minutes, the supernatant was removed and washed. After washing with a culture solution, 0.625 μg/mL of Rat Anti-Mouse MAdCAM-1 antibody (Southern Biotech) was added and cultured for 30 minutes or longer. After washing with the culture solution, 50 μg/mL of Goat Anti-Rat IgG (H+L) Antibody and FITC (Life Technologies) were added and cultured for 30 minutes or longer. After washing with the culture solution, 10 μg/mL of PE Rat Anti-Mouse CD4 (BD Pharmigen) was added and cultured for 30 minutes or longer. After washing with the culture solution, the ratio of the MAdCAM-1 positive cell rate in the CD4-positive cells was measured using a flow cytometer.

In the test, the results of independent tests using blood of two to three people were used. The concentration of the ligand in the well containing no test substance was 100%, and the ligand was set to 0%. The inhibitory rate of MAdCAM-1 binding of the test substance was calculated, and the IC _{50 which} caused 50% inhibition of binding was calculated.

Test Example 5

The portal vein migration concentration of the test substance was measured to evaluate the oral absorption.

The test substance was dissolved or uniformly suspended in a 0.5% (w/v) methylcellulose aqueous solution, and a female mouse (BALB/cAnNCrlCrlj, 7 to 9 weeks old) was orally administered with 3 compounds using a gastric probe ( 3mg/10mL/kg).

After 30 minutes of administration, the laparotomy was performed under fluoroether anesthesia, and about 0.2 mL of blood was collected from the portal vein using a syringe treated with DDVP (esterase inhibitor) and heparin sodium, and stored on ice.

The blood was taken by centrifugation at 18,000 g x 3 minutes using a cooling centrifuge to obtain a plasma sample, and after extracting the test substance with acetonitrile, the plasma concentration was quantified by LC/MS/MS.

Moreover, the plasma concentration is the concentration of the test substance and its active metabolite. The calculated plasma concentration is shown in Table 5.

Claims (18)

a sulfonamide derivative represented by the following general formula (I), or a pharmaceutically acceptable salt thereof, (in the formula, Represents a single bond or a double bond, and R ₁ and R ₂ each independently represent a hydrogen atom, a halogen atom, a lower alkyl group, a lower alkenyl group, a lower alkoxy group, a lower alkoxy lower alkyl group, a hydroxyl group, or a hydroxy lower alkyl group. R ₁ and R ₂ may also be bonded to each other to form a benzene ring which may have a substituent, an alicyclic hydrocarbon having 4 to 7 carbon atoms which may have a substituent, a heteroaromatic ring which may have a substituent, or a heterocyclic ring which may have a substituent, R ₃ represents a lower alkyl group, and e, f, g and h each independently represent CH or a nitrogen atom, B represents a hydroxyl group, an alkoxy group having a carbon number of 1 to 10, and -O- a ring group, a cilexetiloxy group, or a melamine alcohol (Medoxomiloxy), D represents a benzene ring or a heteroaryl ring which may have a substituent, and R ₄ and R ₅ each independently represent a hydrogen atom. a lower alkyl group having a substituent, a lower alkenyl group which may have a substituent, a phenyl group which may have a substituent, or a heterocyclic group which may have a substituent, and R ₄ and R ₅ may also bond and form a heterocyclic ring which may have a substituent). A sulfonamide derivative according to claim 1, or a pharmaceutically acceptable salt thereof, wherein Represents a double bond. The sulfonamide derivative of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ are bonded to each other, and a benzene ring which may have a substituent, or a carbon number which may have a substituent a 4 to 7 alicyclic hydrocarbon, a heteroaromatic ring which may have a substituent, or a heterocyclic ring which may further have a substituent selected from the group consisting of a lower alkyl group, a lower alkoxy group, a hydroxy lower alkyl group, and an amine group. a lower alkylamino group and a lower alkylamino lower alkyl group. The sulfonamide derivative of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ each independently represent a hydrogen atom, a lower alkyl group or a lower alkoxymethyl group, and R ₁ and R _{2 are} also Bonding to form an alicyclic hydrocarbon having 4 to 7 carbon atoms which may have a substituent, a heteroaromatic ring which may have a substituent, or a heterocyclic ring which may also have a substituent selected from a lower alkyl group And a lower alkoxy group. The sulfonamide derivative of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ each independently represent a hydrogen atom, a lower alkyl group or a lower alkoxymethyl group. The sulfonamide derivative of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ are bonded to each other to form a cyclohexene which may have a substituent, a pyridine which may have a substituent, a dihydropyran having a substituent or a tetrahydropyran which may also have a substituent selected from the group consisting of a lower alkyl group, a lower alkoxy group, a hydroxy lower alkyl group, an amine group, a lower alkylamino group, And a lower alkylamino lower alkyl group. The sulfonamide derivative according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, wherein e represents a nitrogen atom, and f, g and h represent C-H. The sulfonamide derivative according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, wherein B represents a hydroxyl group or an alkoxy group having 1 to 6 carbon atoms. The sulfonamide derivative according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein the substituent of D is selected from the group consisting of a halogen atom, a lower alkyl group, a lower alkoxy group, and a hydroxyl group. The sulfonamide derivative according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein D represents a benzene ring which may have a substituent selected from a halogen atom, a lower alkyl group, and a lower level Alkoxy group, and hydroxyl group. The sulfonamide derivative according to any one of claims 1 to 10, wherein R ₄ and R ₅ each independently represent a hydrogen atom or a lower alkyl group which may have a substituent, or may have a lower alkenyl group of a substituent, a phenyl group which may have a substituent, or a heterocyclic group which may have a substituent selected from a halogen atom, a cyano group, a hydroxyl group, a lower alkyl group, a lower alkoxy group, and the like. A fluoromethyl group and a heterocyclic group, and R ₄ and R ₅ may be bonded to each other to form a heterocyclic group which may have a substituent selected from the group consisting of a lower alkyl group, a lower alkoxy group and a hydroxyl group. The sulfonamide derivative, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 10, wherein R ₄ and R ₅ each independently represent a hydrogen atom, a lower alkyl group which may have a substituent, or a heterocyclic group having a substituent selected from a halogen atom, a cyano group, a hydroxyl group, a lower alkoxy group, a trifluoromethyl group, and a heterocyclic group, and R ₄ and R ₅ may be bonded to each other to form A heterocyclic group in which a lower alkyl group is a substituent. The sulfonamide derivative of claim 1, or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ each independently represent a hydrogen atom, a lower alkyl group, a lower alkoxy group, or a lower alkoxy lower alkyl group, R ₁ and R ₂ may also be bonded to form an alicyclic hydrocarbon having 4 to 7 carbon atoms, a heteroaromatic ring, or a heterocyclic ring which may also be substituted by a lower alkyl group, and D may be selected from a halogen atom or a lower alkyl group. a benzene ring or a heteroaryl ring substituted with a substituent in a group of a lower alkoxy group and a hydroxyl group, and R ₄ and R ₅ each independently represent a hydrogen atom or may have a lower alkoxy group, a heterocyclic group or a hydroxyl group. a lower alkyl group having a substituent in a group of a cyano group and a halogen atom, but R ₄ and R ₅ may be bonded to each other to form a heterocyclic group which may be substituted by a lower alkyl group, but D is a halogen atom. When a benzene ring is substituted, R ₁ and R ₂ each represent a lower alkyl group, and when D is a benzene ring having a lower alkyl group, a lower alkoxy group or a hydroxyl group as a substituent, the substituent is bonded adjacent to S. The carbon atom of the carbon atom is bonded to D, and when R ₄ or R ₅ represents a lower alkyl group substituted by a hydroxyl group, the lower alkyl group substituted by the hydroxyl group is represented by the following formula ( As indicated by a) or (b), However, when R ₄ or R ₅ is represented by the formula (b), R ₁ and R ₂ each represent a lower alkyl group, and when R ₄ or R ₅ is a lower alkyl group substituted by a cyano group, D represents a benzene ring. The sulfonamide derivative of claim 1, or a pharmaceutically acceptable salt thereof, which is represented by the following formula, (wherein R ₁ , R ₂ , R ₃ , e, f, g, h, and B are as defined in claim 1). The sulfonamide derivative of claim 1, or a pharmaceutically acceptable salt thereof, which is represented by the following formula, (where B is as defined in claim 1). A pharmaceutical composition comprising the sulfonamide derivative according to any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof. A therapeutic or prophylactic agent for an inflammatory disease associated with a lesion, which is a sulfonamide derivative according to any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof. An α4β7 integrin inhibitor comprising the sulfonamide derivative according to any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof.